tRNA-related sequences trigger systemic mRNA … 1 tRNA-related sequences trigger systemic mRNA transport in plants 2 3 Authors: 4 Wenna Zhanga, Christoph J. Thiemea, Gregor Kollwigb,

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tRNA-related sequences trigger systemic mRNA transport in plants 1

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Authors: 3

Wenna Zhanga, Christoph J. Thiemea, Gregor Kollwigb, Federico Apelta, Lei Yanga, 4

Nikola Wintera, Nadine Andresenc, Dirk Walthera, and Friedrich Kraglera,b,d 5

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Author Affiliations: 7 a Max Planck Institut für Molekulare Pflanzenphysiologie, Wissenschaftspark Golm, Am 8

Mühlenberg 1, Golm, Germany. 9 10 b Department of Biochemistry, Centre of Molecular Biology, Max F. Perutz Laboratories, 11

University of Vienna, Dr. Bohrgasse 9/5, A1030 Vienna, Austria. 12

13 c Institut für Biochemie, CCM, Charité Universitätsmedizin Berlin, Charitéplatz 1, 10117 14

Berlin 15

16 d Corresponding Author 17

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Corresponding Author Email Address: 20

[email protected] 21

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Estimated length of the Manuscript: 23

Words including Introduction, Results, and Discussion: approx. 2800, 24

Figures 4, 25

Supplemental Figures 5, Supplemental Data Sets 3, Supplemental Tables 2. 26

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The author responsible for distribution of materials integral to the findings presented in 28

this article in accordance with the policy described in the Instructions for Authors is: 29

Friedrich Kragler ([email protected]) 30

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Abbreviations: GUS, Glucuronidase; DNDMC1, Dominant-Negative DISRUPTED 31

MEIOTIC cDNA 1; CK1, CHOLINE KINASE 1; tRNA, transfer RNA; mRNA, messenger 32

RNA 33

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Keywords: cell-to-cell mobile mRNA; non-cell-autonomous RNA; mRNA motif, di-35

cistronic RNA; phloem; grafting; flowers; tRNA; mRNA transfer, anthers, pollen, 36

meiosis. 37 38

Synopsis 39

Specific tRNA-derived motifs present in an endogenous di-cistronic transcript mediate 40 mRNA transport along the phloem. 41 42

Abstract 43

In plants, protein-coding messenger RNAs (mRNAs) can move via the phloem 44

vasculature to distant tissues, where they may act as non-cell-autonomous signals. 45

Emerging work has identified many phloem-mobile mRNAs, but little is known 46

regarding RNA motifs triggering mobility, the extent of mRNA transport, and the 47

potential of transported mRNAs to be translated into functional proteins after transport. 48

To address these aspects, we produced reporter transcripts harboring transfer RNA 49

(tRNA) - like structures (TLS) that were found to be enriched in the phloem stream and 50

in mRNAs moving over chimeric graft junctions. Phenotypic and enzymatic assays on 51

grafted plants indicated that mRNAs harboring a distinctive TLS can move from 52

transgenic roots into wild-type leaves and from transgenic leaves into wild-type flowers 53

or roots; these mRNAs can also be translated into proteins after transport. In addition, we 54

provide evidence that di-cistronic mRNA:tRNA transcripts are frequently produced in 55

Arabidopsis thaliana and are enriched in the population of graft-mobile mRNAs. Our 56

results suggest that tRNA-derived sequences with predicted stem-bulge-stem-loop 57

structures are sufficient to mediate mRNA transport and seem to be necessary for the 58

mobility of a large number of endogenous transcripts that can move through graft 59

junctions. 60 61

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Introduction 63

In plants, small interfering RNAs (siRNAs), microRNAs (miRNAs), and messenger 64

RNAs (mRNAs) can move locally from cell to cell via plasmodesmata and can also move 65

over long distances by entering the phloem vasculature. The mobile siRNAs and 66

miRNAs regulate gene expression, affect target mRNAs, and mediate antiviral defense 67

(Ruiz-Medrano et al., 2004; Lough and Lucas, 2006; Kalantidis et al., 2008; Molnar et 68

al., 2010; Melnyk et al., 2011). Distinct mRNAs such as the homeodomain protein-69

encoding transcripts of potato (Solanum tuberosum) BEL5 and maize (Zea mays) 70

knotted1 (KN1) also move to other tissues and trigger developmental decisions in targeted 71

cells (Kim et al., 2001; Banerjee et al., 2006). 72

The molecular mechanisms enabling intercellular mRNA transport and the fate of 73

transported mRNAs in target tissues remain poorly understood. On the one hand, 74

conserved and, thus, predictive mRNA motifs have not been described for known graft-75

mobile mRNA populations (Guo et al., 2013; Thieme et al., 2015; Yang et al., 2015). On 76

the other hand, recent work in potato showed that the 3′ UTR of the phloem-mobile 77

transcript BEL5 supports mRNA stability and trafficking into roots, where BEL5 protein 78

initiates tuber formation (Banerjee et al., 2009; Cho et al., 2015). 79

Viral RNAs can move via the phloem stream in the absence of viral proteins, 80

suggesting that endogenous cellular factors recognize a structural RNA motif and mediate 81

long-distance transport through the phloem (Gopinath and Kao, 2007). Non-conserved 82

viral 3′ UTR sequences, which interact with 5′ UTRs, seem to play a role in facilitating 83

viral RNA cell-to-cell transfer (Lough et al., 2006). Similarly, viroids (infectious, non-84

protein-coding small RNAs) form specific stem-loop structures not yet identified in other 85

mobile RNAs allowing them to enter the plant phloem long-distance transport system 86

(Ding, 2009; Takeda et al., 2011). 87

Many positive-strand RNA viruses harbor conserved stem-loop structures in the 3′ 88

UTR resembling those of canonical tRNAs. Such viral tRNA-like structures (TLS) seem 89

to play a crucial role in viral replication and infectivity (Dreher et al., 1999; Fechter et al., 90

2001; Barends et al., 2004). The TLSs are aminoacylated, and therefore, the viral clover-91

like tRNA structures are likely recognized by plant tRNA-binding and modifying proteins 92

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(Dreher, 2010). Also, viral TLS recognized by the ribosomal elongation factor 93

eEF1A_GTP form a stable RNA-protein complex repressing viral RNA minus-strand 94

synthesis (Matsuda et al., 2004). 95

Viral TLS-mediated intercellular or long-distance transport of viral RNAs remains 96

to be shown, but support for the notion that tRNA-related structures might be bona fide 97

RNA mobility motifs for endogenous transcripts was found in the non-coding RNA 98

population of phloem exudate from pumpkin (Cucurbita pepo) (Zhang et al., 2009). In 99

pumpkin phloem exudate, specific subsets of tRNAs are enriched suggesting selective 100

tRNA import from surrounding tissues into conducting phloem vessels. Taken together 101

the presence of specific tRNAs in the phloem stream, the role of viral TLS, and frequent 102

occurrences of di-cistronic poly(A)-mRNA:tRNA transcripts point towards a potential 103

function of tRNA-related sequences in triggering mobility of transcripts. Here, we 104

present evidence that the TLS can trigger mobility of otherwise non-mobile mRNAs and 105

that TLS are significantly enriched in the mobile mRNA populations found in A. 106

thaliana. 107

108

Results 109

tRNAMet fusion transcripts move into flowers 110

To establish a simple phenotypic scoring system for mRNAs harboring predicted 111

mobility motifs such as TLSs, we used a dominant negative variant of A. thaliana 112

DISRUPTION OF MEIOTIC CONTROL 1 (DNDMC1) that lacks the N-terminal 92 amino 113

acid residues (Figure 1 A) and, therefore, interferes with the progression of meiosis 114

(Habu et al., 1996). DMC1 is a specific meiotic cell-cycle factor and a member of the 115

highly conserved RecA-type recombinase family of DNA-dependent ATPases active 116

during meiosis in sporogenic cells (Doutriaux et al., 1998). Lack of a functional 117

DMC1/RAD51 complex induces achiasmatic meiosis resulting in the formation of 118

anomalously shaped pollen containing an aberrant number of chromosomes and, 119

consequently, is necessary for proper pollen development (Bishop et al., 1992; Zhang et 120

al., 2014). Thus, production of misshaped pollen in anthers and decreased fertility 121

indicate the presence of either DMC1 siRNA (Zhang et al., 2014) (Figure 1B, C) or the 122

product of translation of the dominant negative DNDMC1 mRNA. To implement a 123

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reporter system for mRNA mobility, we produced transgenic Nicotiana tabacum lines 124

expressing YFP-DNDMC1 fusion proteins as a fluorescent reporter (Figure 1B, C) to test 125

for DMC1 silencing (Zhang et al. 2014) potentially induced by the transgenic DNDMC1 126

constructs. We also generated lines expressing DNDMC1 mRNA fused to the full-length 127

potato BEL5 transcript, which is known to be mobile (Cho et al., 2015) (DNDMC1:BEL5) 128

as a positive control (Figure 1A). As a negative control, we made lines expressing 129

DNDMC1 mRNA 5' fused to the vegetative Nicotiana tabacum growth regulator 130

CENTRORADIALIS-like 2 (Amaya et al. 1999) (CET2:DNDMC1) (Supplemental Figure 131

1). Finally, since tRNAMet was detected in the phloem sap of pumpkin (Zhang et al., 132

2009), we made lines expressing DNDMC1 mRNA fused to full-length tRNAMet 133

(AT5G57885; DNDMC1:tRNAMet; tRNAMet:DNDMC1) (Figure 1A). 134

Independent transgenic plants expressing DNDMC1 mRNA fusion constructs were 135

verified to show a pollen sterility phenotype (Figure 1D, E) and used in grafting 136

experiments (Figure 1F, G) to evaluate transcript mobility from transgenic source tissue 137

to wild-type flowers. Transgenic lines expressing the YFP-DNDMC1 fusion did not 138

exhibit a dominant-negative effect on endogenous tobacco DMC1 (Figure 1B); also, the 139

fusion transcript was not graft-mobile (Supplemental Figure 1). Thus, these plants could 140

be used to evaluate grafted DNDMC1 transgenic plants for their potential production of 141

mobile siRNAs targeting endogenous DMC1 in wild-type tobacco flowers, triggering 142

sterility (Zhang et al., 2009). 143

We first confirmed by RT-PCR that DNDMC1 mRNA does not contain the 144

sequences triggering mobility (Supplemental Figure 1) and, thus, is suitable as a reporter 145

for transcript mobility, producing a pollen phenotype (Figure 2 A, Supplemental Figure 146

1). We next addressed the mobility of the fusion transcripts by grafting DNDMC1:BEL5, 147

DNDMC1:tRNAMet, tRNAMet:DNDMC1, or tobacco CET2:DNDMC1 transgenic plants with 148

wild-type plants and examined pollen sterility and presence of the fusion transcript in the 149

wild-type flowers (Figure 2B-G; Supplemental Figure 1). As expected, after induction 150

with estradiol, the DNDMC1:BEL5 and CET2:DNDMC1 scions grafted onto wild-type 151

stocks showed a significantly higher percentage of aberrant pollen formation in their 152

flowers (30.9 ±7.6% and 51.9 ±7.8%, respectively) than grafted wild-type plants (4.0 153

±3.0%) (Figure 2C; Supplemental Figure 1; Supplemental Table 1). Confirming previous 154

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reports that BEL5 fusion transcripts are mobile (Banerjee et al., 2006), wild-type plants 155

grafted onto DNDMC1:BEL5 stocks produced a significantly higher number of misshapen 156

pollen (19.7 ±14.3 %) than wild-type controls and the presence of fusion transcript was 157

confirmed by RT-PCR in closed wild-type flowers (Figure 2E). Confirming that the 158

DNDMC1 RNA itself does not trigger mobility and that DNDMC1 protein itself is not 159

mobile grafted CET2:DNDMC1 stock plants did not significantly induce aberrant pollen 160

formation in wild-type flowers (7.8 ±4.9) (Supplemental Figure 1). Thus, DNDMC1 - 161

RNA fusion constructs can be employed as a RNA mobility reporter system by producing 162

a quantifiable pollen phenotype. 163

Next, to learn whether a phloem-allocated tRNA contains the necessary structural 164

information mediating mRNA movement over long-distances, we grafted transgenic 165

plants expressing the 3′ UTR DNDMC1:tRNAMet (Figure 2B) or the 5′ UTR 166

tRNAMet:DNDMC1 (Supplemental Figure 1) fusion construct. Expression was induced by 167

applying estradiol to the transgenic source leaves (stock) or transgenic stem (scion) ~ 1.5 168

weeks after grafting and prior to flower induction. Estradiol-treated grafted plants formed 169

a significantly higher number of misshapen pollen (14.2 ±7.6 %; Supplemental Table 1) 170

compared to control grafts and wild-type plants (Figure 2A, B), and RT-PCR assays 171

confirmed the presence of DNDMC1:tRNAMet and tRNAMet:DNDMC1 poly(A) transcripts in 172

wild-type flowers formed on transgenic stock plants (Figure 2E, Supplemental Figure 1). 173

To exclude the possibility that the grafted chimeric plants produce a mobile 174

DMC1 siRNA that moves into wild-type flower tissues and silences the endogenous 175

DMC1, triggering a pollen sterility phenotype (Zhang et al., 2014), we grafted the 176

DNDMC1:tRNAMet plants with the YFP-DNDMC1 fusion line, which expresses a reporter 177

protein that can be easily detected by fluorescence microscopy. In contrast to the DMC1 178

siRNA control lines, no systemic siRNA-mediated silencing of the YFP-DNDMC1 179

reporter construct could be detected in sepals (Figure 2F). Thus, the DNDMC1:tRNAMet 180

fusion transcript does not induce systemic silencing and the observed defects in pollen 181

formation in grafted plants (Figure 2G) can be attributed to the systemic delivery of the 182

DNDMC1 fusion transcripts. In summary, the presence of the full-length tRNAMet sequence 183

in the 5′ or 3′ UTR triggers transport of the DNDMC1 poly(A) transcript from stock to 184

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source leaves and into sporogenic tissues, where it is apparently translated, as it interferes 185

with meiosis in male tissues. 186

187

tRNAs harbor a signal for systemic mRNA movement 188

To evaluate whether particular tRNA sequences related to the viral TLS mediate systemic 189

mRNA movement, we used the core sequences of the two phloem-imported tRNAs, 190

tRNAMet (anticodon CAT; 72 bases; TAIR# AT5G57885) and tRNAGly (anticodon TAT; 74 191

bases; TAIR# AT1G71700), and the non-phloem imported tRNAIle (73 bases; TAIR# 192

AT3G05835) (Zhang et al., 2009). tRNAGly is also present in the 3' UTR of the graft-193

mobile A. thaliana CHOLINE KINASE 1 (CK1) transcript (TAIR# AT1G71697; Zhang et 194

al., 2015). We fused these three tRNA sequences to the 3' UTR of the cell-autonomous β-195

GUS mRNA sequence (Figure 3 A, Supplemental Figure 2). To evaluate the mobility of 196

the fusion transcripts, A. thaliana Col-0 lines expressing 35Spro:GUS or 35Spro:GUS:tRNA 197

fusion constructs were produced and hypocotyl-grafted with Col-0 wild type (shoot or 198

root). Two weeks after grafting, GUS enzyme activity was visualized in situ (Figure 3B, 199

C; Supplemental Figure 3). Control grafts with transgenic 35Spro:GUS plants lacking the 200

tRNA sequences in the 3′ UTR showed no GUS activity and GUS mRNA presence in 201

distal wild-type root (n=0/55 grafts) or leaf (n=0/43 grafts) tissues indicating that neither 202

the GUS mRNA nor the GUS protein moves over graft junctions (Figure 3C, D). 203

However, GUS activity was detected in phloem-associated cells in wild-type roots after 204

hypocotyl grafting with transgenic scion plants expressing GUS:tRNAMet (n=9/44 grafts) 205

or GUS:tRNAGly (n=6/25 grafts). No GUS activity was observed in wild-type roots grafted 206

with plants expressing GUS:tRNAIle (n=0/57 grafts). RT-PCR assays confirmed the 207

presence of GUS:tRNAMet and GUS:tRNAGly and the absence of GUS:tRNAIle transcripts in 208

wild-type roots after grafting (Figure 3D). Notably, the reverse grafts with transgenic 209

roots and wild-type scions indicate that the shoot-to-root mobile GUS:tRNAMet fusion 210

transcript does not move from root to shoot (n=0/36 grafts) and that GUS:tRNAGly barely 211

moves from root to shoot (n=3/26 grafts). 212

To learn whether the whole tRNA sequence or a subsequence is sufficient to mediate 213

mRNA mobility, we used tRNAMet deletion constructs lacking the assigned dihydrouridine 214

(D)-, anticodon (A)-, or TψC (T) –arm/loop structures and combinations thereof (Figure 215

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3C). Again, plants expressing these GUS mRNA fusion constructs were grafted with wild 216

type and then tested for GUS activity and presence of the fusion transcript. As indicated 217

by GUS and RT-PCR assays, the ΔD, ΔDT, and ΔDA, but not the ΔAT tRNAMet deletion 218

construct, were sufficient to mediate GUS transport into wild-type roots and, with a very 219

low frequency, to scion leaves (Figure 3C, D). Presence of the GUS:ΔDtRNAMet transcript 220

and translation in phloem-associated cells of wild-type roots and leaves suggests that only 221

part of the tRNAMet sequence is required to trigger mobility. This also indicates that A-, 222

and TψC hairpin-loop sequences have redundant roles in triggering mRNA transport as 223

only deletions of both the Α and TψC hairpin-loop sequences eliminated mobility of the 224

GUS fusion transcript. 225

To elucidate whether tRNA sequences or sequences related to viral TLS motifs 226

confer mRNA mobility, we first evaluated whether the endogenous mobile mRNA 227

population found in A. thaliana is enriched for TLS motifs. We screened the A. thaliana 228

graft-mobile transcriptome database (n=3606) (Thieme et al., 2015) for the presence of 229

TLS motifs in the mRNA UTRs and coding sequences (CDS) (Figure 4 A). We 230

performed scans for sequence-independent structure motifs using the established 231

consensus tRNA descriptor (Macke et al., 2001) recognizing the stem-loop arrangements 232

found in most tRNAs (Supplemental Figure 4). The analysis revealed that a significant 233

number of mobile A. thaliana transcripts (Thieme et al., 2015) (11.4%; n= 411 of 3606) 234

or grapevine (Vitis spp.) transcripts (Yang et al., 2015) (7.5%; n= 249 of 3333) harbor a 235

TLS motif in the CDS or 3′ UTR (Figure 4A; Supplemental Data Set 1). Furthermore, 236

annotated tRNA genes were found in closer proximity to genes encoding mobile 237

transcripts than to genes encoding non-mobile transcripts (p<0.0003; Cohen's D d=0.313; 238

see Methods). Independent of DNA-strand assignment, of all 1,125 genes flanked by a 239

tRNA gene, 158 produced mobile RNAs and of these, 113 are located within 1,000 bp of 240

the tRNA gene (34.7% enrichment for mobile transcripts, p=0.005, Fisher’s exact test, 241

Figure 4B). 242

To determine whether these neighboring tRNAs are actually co-transcribed to 243

form di-cistronic mRNA-tRNA molecules, we analyzed paired-end RNA-Seq data from 244

A. thaliana (Thieme et al. 2015, Ito et al. 2015) for the presence of poly(A) RNA:tRNA 245

matching sequences and performed RT-PCR assays on selected transcripts (Figure 4C, 246

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Supplemental Figure 5, Supplemental Data Set 2). Both analyses revealed that di-247

cistronic mRNA-tRNA transcripts are produced relatively frequently. In total, 132 di-248

cistronic transcripts spanning protein-coding genes (120 unique loci) and tRNAs (118 249

unique loci) were supported by RNA-Seq data (Supplemental Data Set 2). Of the 120 250

genes, 27 genes (22.5%) are annotated to produce mobile mRNAs and of the 118 tRNA-251

genes, 24 tRNA genes were found as di-cistronic transcripts in conjunction with a mobile 252

transcript (Figure 4C). Of all mRNA-tRNA tandem sequences within 1000 bp of the 253

respective gene loci, evidence for a di-cistronic nature was found 1.6 times more often 254

when the mRNA was annotated as mobile than for non-mobile mRNAs, albeit statistical 255

significance could not be established as the numbers were low (p=0.1, Fisher’s exact test, 256

Supplemental Data Set 2). 257

To confirm these findings and to substantiate the notion that tRNAs play a role in 258

transcript mobility, we analyzed insertion mutants of the CK1 (TAIR# AT1G71697) gene, 259

which produces a mobile transcript (Thieme et al. 2015) and expresses an enzyme 260

catalyzing the reaction of choline to phosphatidylcholine (Tasseva et al., 2004). 261

According to the paired-end sequencing data the tRNAGly core sequence (TAIR# 262

AT1G71700) is present in the CK1 3′ UTR region forming a di-cistronic CK1:tRNAGly 263

transcript (Supplemental Figure 5, Supplemental Data Set 2). tRNAGly fused to GUS 264

mRNA mediated mobility of the otherwise non-mobile GUS sequence (Figure 3). 265

To test whether CK1 mRNA mobility depends on the presence of the tRNAGly in 266

the 3′ UTR, we first confirmed and then used two SALK T-DNA insertion lines for 267

grafting experiments: ck1.1 (SALK_070759) and ck1.2 (SALK_023420) (Figure 4D). In 268

ck1.1 mutants the T-DNA is located within the first intron and the ck1.2 mutants have an 269

insertion between the CK1 stop codon and the annotated tRNAGly sequence. We performed 270

Arabidopsis stem grafting experiments with ck1.2 and wild type (Col-0) and assayed the 271

presence of wild-type CK1:tRNAGly and truncated ck1.2 mRNA in stock and scion 272

samples via RT-PCR (Figure 4E). Although ck1.2 mutants produce a full-length CK1 273

poly(A) transcript containing all protein-coding sequences, the truncated transcript 274

lacking the tRNAGly sequence could not be detected in wild-type samples. By contrast, 275

wild-type CK1:tRNAGly transcript was present in both ck1.2 scion and ck1.2 stock tissue 276

samples. This suggests that the CK1:tRNAGly transcript was bi-directionally mobile from 277

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stock to scion (Figure 4E) whereas the mutant ck1.2 transcript lacking the tRNAGly 278

sequence was not transported over graft junctions. Thus, the endogenously produced di-279

cistronic CK1:tRNAGly transcript seems to be graft-mobile due to the presence of the 3′ 280

UTR tRNAGly sequence. 281

As lack of detectable ck1.2 transcript mobility could be a result of low expression 282

levels, we performed quantitative RT-PCR assays to evaluate CK1 expression levels in 283

the two ck1.1 and ck.2 mutants and wild-type plants. Here, only marginal expression 284

could be detected in the ck1.1 mutant whereas CK1 poly(A)-RNA transcript levels in the 285

ck1.2 mutant were similar to that found in the wild type (Figure 4F). Despite comparably 286

high CK1 transcript levels in wild-type and ck1.2 mutant plants, both the ck1.2 line and 287

the ck1.1 line showed a significant decrease in rosette leaf size compared to wild type 288

(Figure 4F). This implies that either ck1.2 plants produce less functional CK1 enzyme 289

due to the lack of the 3' tRNA sequence or that CK1 mRNA presence in expressing cells 290

as well as CK1 mRNA mobility is equivalently essential for normal growth behavior of 291

Arabidopsis. 292

293

Discussion 294

RNAs are arguably the most functionally diverse biological macromolecules found in 295

cells. Their diverse roles are determined by both their complex three-dimensional 296

structure and by their primary sequence. Our study reveals an additional biological role of 297

tRNA sequences in plants. They harbor a motif mediating mRNA transport to distant 298

plant cells. Interestingly, transcript mobility was induced by tRNAMet and tRNAGly, but not 299

by tRNAIle, consistent with the absence of tRNAIle in the pumpkin phloem sap (Zhang et 300

al., 2009). As our results indicate that transcript mobility is mediated by a particular RNA 301

structure, a tRNA motif-scanning algorithm did indeed reveal a significantly high number 302

of identified mobile mRNAs that harbor a TLS motif or are transcribed from genes in 303

close proximity to annotated tRNA genes (Figure 4A, Supplemental Data Set 1) which 304

seem to frequently produce di-cistronic poly(A)-RNA:tRNAs (Figure 4C, Supplemental 305

Figure 5, Supplemental Data Set 2). While the functional role of many mobile mRNAs in 306

distant tissues remains to be elucidated, evidence supports the notion that trafficking of 307

small si/miRNAs and large mRNAs via the phloem plays an important role in regulating 308

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plant development (Lucas et al., 2001). A surprisingly high number of mRNAs can be 309

found in phloem exudates (Guo et al., 2013) and move across graft junctions (Thieme et 310

al., 2015; Yang et al., 2015), but no general and easily predictable RNA motif or 311

conserved sequence mediating mobility could be identified in the graft-mobile transcript 312

populations (Calderwood et al. 2016). However, our data suggest that a significant 313

fraction of mobile mRNAs carries a TLS motif potentially mediating mobility across 314

graft junctions. 315

Highlighting the complexity of the translocation system, mRNA transfer does not 316

strictly follow the source to sink phloem flow as GUS:tRNA fusions not only moved from 317

shoot (source) to root (sink), but also vice versa (Figure 3B, C; Supplemental Figure 3). 318

Hence, it is likely that two transport pathways exist for delivering mRNA molecules. One 319

could be based on passive, non-selective delivery from source to sink via the phloem 320

vessels. Obviously, passive delivery such as diffusion depends on transcript stability and 321

abundance. However, also another pathway in the form of a targeted and active transport 322

system seems to be in place mediating the delivery of mRNAs from root to shoot. This 323

notion finds support in the observed directional and tissue-specific distribution of graft-324

mobile mRNAs in A. thaliana (Thieme et al., 2015). Here two aspects suggest that a 325

number of mRNAs move in an active and regulated fashion: i) transport against the 326

phloem flow from root to shoot (sink to source) followed by ii) transfer of distinct mobile 327

mRNAs to specific aboveground tissues such as leaves or flowers. Furthermore, the 328

presence of an active mRNA delivery mechanism is supported by two additional findings 329

presented here: i) specific sequences derived from TLS are sufficient to confer mobility 330

to heterologous mRNAs, and ii) deletion of a TLS in the plant endogenous CK1:tRNAGly 331

di-cistronic transcript makes it immobile. This suggests that TLS or closely related RNA 332

structures mediate transport of a number of graft-mobile transcripts. Here, TLS motifs 333

could provide an evolutionary link between RNA virus transport and mRNA transport. In 334

both systems, TLS appear to support RNA transport along the phloem vasculature to 335

distant cells. Also, as numerous tRNA genes are dispersed over the genome, it is 336

conceivable that di-cistronic gene products are relatively often created by genomic 337

rearrangements. Such randomly created mobile transcripts could complement distant 338

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mutant cells harboring non-sense mutations, which seem to occur frequently in higher 339

eukaryotes (McConnell et al. 2013). 340

In any case, specific RNA transport motifs should interact with plant endogenous 341

RNA-binding proteins. This finds support in studies on viroid RNA structures that are 342

specifically necessary for entering specific vascular tissues (Takeda et al., 2011) and in 343

the observed interaction of phloem-delivered mRNAs such as GAI and BEL5 with a 344

phloem-expressed polypyrimidine-tract binding protein (PTB) recognizing poly-cysteine 345

(C) - uracil (U) nucleotide stretches present in the 3′ UTRs of some phloem mobile 346

transcripts (Ham et al., 2009; Cho et al., 2015). However, a predicted RNA sequence 347

related to PTB binding motifs was not found in the tRNA fusion constructs used in this 348

study and, thus, is unlikely to play the primary role in triggering their movement. A 349

structural analysis revealed that the level of overall base pairings in the 3′ end of A. 350

thaliana transcripts assigned to be mobile is similar to non-mobile transcripts (two-351

sample Kolmogorov-Smirnov [KS] test, p=0.011), while the 3′-terminal sequence regions 352

of mobile transcripts form energetically less stable folds (two-sample KS test, p<2.2E-353

16). This is caused by a significantly lower GC content in the respective region compared 354

to transcripts not found to be mobile (Wilcoxon rank sum test, p<2.518e-10) 355

(Supplemental Figure 4). Hence, the mobile mRNA population seems to be generally less 356

stable and less likely to bind to PTB proteins. 357

In general, predicted folds of mobile tRNA variants (Figure 4G) point towards an 358

RNA hairpin motif triggering transport. All tRNA sequences and variants thereof eliciting 359

bi-directional GUS mRNA transfer over graft junctions seem to form a predicted long 360

hairpin motif with a stem (8-12 nucleotides) - variable bulge(s) - stem (4-7 nucleotides) - 361

and variable loop (Figure 4G), which is surprisingly similar to structures formed by 362

precursor miRNAs having predicted stem - short bulge – stem – extensive loop core 363

folds. Interaction of a D stem-loop with a T stem-loop present in mature L-shaped tRNAs 364

seems not to be essential as the D loop deletion actually enhanced the transport activity of 365

GUS fusions. Requirements for TLS-mediated transport seem to be either within the A- 366

or the T- loop and the acceptor stem sequences present in all mobile GUS-tRNA fusion 367

transcripts. Here it is important to note that not all tRNAs seem to confer mobility to 368

transcripts. The tRNAIle(TAT) core sequence fused to GUS did not trigger GUS transcript 369

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mobility over graft junctions. This observation is in line with previous studies in which 370

tRNAIle (TAT) is not or rarely detected in the phloem exudate of pumpkin (Zhang et al. 371

2009). Also we did not identify tRNAIle(TAT) di-cistronic transcripts in RNA-seq data 372

(Figure 4C, Supplemental Data Set 2) and found evidence that tRNAIle(TAT) has a 373

predicted A- and T-loop folding structure distinct from tRNAGly, tRNAMet , and variants 374

thereof, triggering mobility (Figure 4G). 375

Another important aspect is that GUS enzyme activity assays and sterility 376

phenotypes detected in distant wild-type tissues corroborate the notion that mobile 377

mRNAs are translated into functional proteins after transport. Interestingly, the observed 378

plant size phenotype in mutants producing a non-mobile CK1 transcript (ck1.2) lacking 379

the di-cistronic tRNAGly sequence is equivalent to that observed with the ck1.1 null 380

mutant (Figure 4F). Here, although it seems implausible, CK1 transcript mobility appears 381

to have a similar crucial function as the gene product itself. In any case, we could show 382

that specific tRNA sequences such as tRNAGly, tRNAMet, and tRNAMet -derived sequences 383

trigger transport of otherwise non-mobile transcripts and that a significant number of 384

mobile mRNAs harbor a TLS motif. 385

386

Methods 387

Plant Material and Growth Conditions 388

Tobacco (Nicotiana tabacum cv. Petite Havana) plants were grown under aseptic 389

conditions on agar-solidified medium containing 30 gL-1 sucrose. Rooted tobacco plants 390

were transferred to soil and grown to maturity under standard greenhouse conditions 391

(relative humidity: 55%; day temperature: 25°C; night temperature: 20°C; diurnal cycle: 392

16 h light / 8 h darkness; light intensity: 190-600 μE·m−2·s−1; mixed light (ratio 1:1) 393

from Metal-halide light (HPIT) and Sodium-vapor light (AgroSonT) sources plus 394

sunlight). Arabidopsis thaliana seeds of wild type (Col-0), and transgenic 35Spro:GUS, 395

ck1.1 (SALK_070759), ck1.2 (SALK_023420) plants of ecotype Col-0 were used and 396

grown in controlled environmental chambers (light source: mixed fluorescent tubes 50% 397

cold white, 50% warm white) for growth assays or on soil in the greenhouse (relative 398

humidity: 60%; day temperature: 22°C; night temperature: 19°C; diurnal cycle: 16 h light 399

14

/ 8 h darkness; light intensity: 170-200 μE·m−2·s−1; mixed light (ratio 1:1) from Metal-400

halide light (HPIT) and Sodium-vapor light (AgroSonT) sources plus sunlight). The 401

SALK- lines were obtained from the Salk Institute Genomic Analysis Laboratory, 402

California (Alonso et al., 2003). 403

Grafting and Estradiol Treatment 404

Tobacco plants used for grafting experiments were grown two to three months on soil in 405

the greenhouse. A standard splice grafting procedure was used as previously described 406

(Zhang et al., 2014). In short, plants with the same stem diameter carrying five fully 407

expanded leaves were used as stock and scion material; rootstocks were prepared by 408

removing the apical leaves from the top of the plant and keeping two to three source 409

leaves. Scions were prepared by cutting the stem 3 to 4 cm below the apex and removing 410

the source leaves. A long slanting cut was made on the rootstock stem (about 30 degrees 411

from vertical) with a matching cut at the scion base. The surfaces of both cuts were 412

immediately pressed together and the junction was tightly wrapped with parafilm. The 413

first week after grafting the scion was covered with a plastic bag and kept under high 414

humidity. After the graft junction was established, axillary branches and leaves emerging 415

at the stock were removed to enforce apical dominance of the scion. Before flower 416

induction 5 µM 17-β-estradiol mixed with Lanolin (Sigma-Aldrich) (1000x stock 417

solution: 5 mM 17-β-estradiol in DMSO, stored at -20°C) was applied with soaked tissue 418

paper onto the adaxial side of stock plant leaf surfaces to induce gene expression. The 419

tissue paper was left on the surface to mark the side of induction. After flowers appeared 420

on the scion, part of the induced leaf and emerging first closed flowers were sampled for 421

fusion transcript presence by RT-PCR. 422

A. thaliana hypocotyl grafting was performed as described (Thieme et al., 2015). In short, 423

plants were grown vertically on solid 0.5 MS medium (1% sucrose) at 22°C with a 424

photoperiod of 8 h light (fluence rate of 100 µmol m-2 s-1). The temperature was increased 425

to 26°C 4 days after germination to reduce adventitious root formation. 6 to 7 days after 426

germination seedlings were used for grafting under sterile conditions as described 427

(Thieme et al., 2015). In short, seedlings were cut transversely in the middle of the 428

15

hypocotyl with a razor blade (Dumont; No.5), and a silicon collar (NeoTecha; Ø 0.30x 429

0.60 mm) was slid over the stock in which the scion was inserted. Grafted plantlets were 430

placed on solid 0.5 MS medium (supplemented with 1% agar and 1% sucrose) and grown 431

at 22°C (8 h light). Appearing adventitious roots were cut every two days and after two 432

weeks successfully grafted plants were submitted to histochemical GUS stain assays, or 433

root and shoot material were harvested separately for RT-PCR detection of GUS 434

transcripts. The detailed procedure of Arabidopsis inflorescence stem grafting used for 435

CK1 mobility assays was performed as described (Nisar et al., 2012) and samples were 436

harvested for RNA extraction and RT-PCR detection one week post grafting. 437

Expression Constructs 438

To produce a dominant-negative Arabidopsis DMC1 with a N-terminal 92 amino acid 439

deletion in A. thaliana DMC1 (DNDMC1; provided by Rijk Zwaan) transcripts with 3′ 440

UTR and 5' UTR fusions an expression binary constructs named pRD1 and pRD4 were 441

created based on a pMDC7 (Curtis and Grossniklaus, 2003) backbone. The DNDMC1 442

fragment was introduced 5′ or 3′ of the pMDC7 gateway cloning cassette, which resulted 443

in a template binary vector used to clone via a gateway reaction the RNA sequences of 444

BEL5, or tRNAMet between the DNDMC1 ORF and promoter or terminator (Figure 1A). 445

Synthetic oligonucleotides were used to produce gateway ENTRY clones with the 446

according sequence for gateway recombination with the binary vector (Supplemental 447

Table 2). The binary vector constructs based on pMDC7 allow estradiol-induced 448

DNDMC1:RNA or RNA:DNDMC1 expression. pEarleyGate104 used for 35Spro:YFP-449

DNDMC1 expression and the DMC1 siRNA N. tabacum line (35Spro:BcDMC1 hpRNAi) 450

and its function was previously described (Zhang et al., 2014). 451

GUS fusion constructs harboring tRNAMet (AUG), tRNAGly

(GGC), or tRNAIle (AUA) and 452

tRNAMet (AUG) variants in the 3′ UTR were created by PCR amplification using an NcoI 453

GUS forward primer covering the GUS start codon and by a BstEII GUS reverse primer 454

covering the GUS stop codon and the tRNA sequence. The resulting PCR fragment was 455

amplified again with an unspecific XbaI reverse primer harboring an XbaI site for 456

identification of the cloned fragment. The resulting NcoI-BstEII digested fragments were 457

16

cloned into the accordingly digested pCambia1305.1 (Chen et al., 1998) allowing 458

expression of the GUS:tRNA constructs driven by a 35S promoter. All synthetic 459

oligonucleotides used in the PCR reactions are listed in Supplemental Table 2. 460

RNA Isolation and Reverse Transcription Reactions 461

Samples were prepared in 1 mL Trizol reagent (Invitrogen) (0.5 mL / 100 mg tissue) as 462

described previously (Zhang et al., 2009). After centrifugation (10,000 xg, 10 minutes at 463

4°C), the supernatant (~1 mL) was transferred to a new RNase-free tube and extracted 464

once with 200 µL and once with 50 µL chloroform. To precipitate the RNA the 465

supernatant was supplemented with two volumes of 99% isopropanol, 0.1 volumes of 3 466

M sodium acetate (pH 5.2), 1 µg of linear acrylamide (Invitrogen), and incubated >1 h at 467

-20°C. After centrifugation (16,000 xg, 30 minutes at 4°C), the resulting pellet was 468

washed twice with 80% ethanol, once with 99% ethanol, air dried, and resuspended in 20 469

µL RNase-free water. To determine RNA quality and concentration, 1 µL of each RNA 470

sample was submitted to agarose gel electrophoresis (2%, agarose, 1x TBE) and 471

quantified using a NanoDrop ND–1000 (Thermo Scientific). 472

Reverse transcription reaction was performed with 1 U/µL AMV reverse transcriptase 473

(Promega) with following modifications: total RNA (~4 µg) was denatured at 70°C for 10 474

minutes in the presence of oligo(dT) primer followed by a 5 minutes annealing 475

incubation at 37°C prior to the RT-reaction, then incubated at 42°C for one hour, and 476

72°C for 10 minutes for deactivation. RT-PCR was conducted under standard PCR 477

conditions with 40-45 cycles (Zhang et al 2015). Oligonucleotides used for RT-PCR are 478

listed in Supplemental Table 2. 479

Quantitative RT-PCR 480

Quantitative Real-time PCR was performed according to the SYBR Green method in a 5 481

μl volume using 4 μg total RNA, 2.5 μl SYBR Green Master Mix (Applied Biosystems), 482

0.2 μM forward and reverse primers. For each genomic confirmed ck1 mutant RNA from 483

3-5 individual plants was isolated and used. At least three technical replicas were 484

performed. An ABI System Sequence Detector (Applied Biosystems 7900HT fast Real 485

17

time PCR) was used with the following regimen of thermal cycling: Stage 1: 1 cycle, 2 486

minutes at 50°C; Stage 2: 1 cycle, 10 minutes at 95°C; Stage 3: 40 cycles, 15 seconds at 487

95°C, 1 minute 60°C. Dissociation stage: 15 seconds at 95°C, 15 seconds at 60°C, 15 488

seconds at 95°C. Oligonucleotides used for RT-PCR are listed in Supplemental Table 2. 489

Microscopy and Pollen Shape Analysis 490

The statistical pollen shape analysis indicating sterility was performed as described 491

previously (Zhang et al., 2014). Tobacco pollen was collected and stained with propidium 492

iodide (0.01 mg/ml, Molecular Probes, USA). To image the shape and size of the pollen a 493

Confocal Laser-Scanning Microscope (CLSM; TCS SP5; Leica Microsystems) was used. 494

The system had following settings: Detection Channel 2 (red): 570-650 nm. The Channel 495

2 gain (PMT) was set between 500-600 V, Pinhole: 1.0 Airy Units, 5 Z-stacks with 5-6 496

μm were merged and used for the shape recognition algorithm as described (Zhang et al., 497

2014). 498

YFP fluorescence was detected as described (Zhang et al., 2014) with following settings: 499

Sequential channel scan mode with a maximum aerial pinhole of 1.5 Airy Units. To 500

compare the YFP fluorescence intensity between plants the same settings such as laser 501

power, gain voltage, pinhole, objective, magnification, and channel/filter wavelengths 502

were used. Z-stack images were assembled and processed using the Image J software 503

package (NIH). Detection Channel 1 (green): 535 to 617 nm; Detection Channel 2 (red): 504

not used; Detection Channel 3 (blue; Chloroplast/plastid auto-fluorescence): 695 to 765 505

nm. Channel 1 gain (PMT) was set between 500 - 600 V. 506

β-Glucuronidase (GUS) Detection 507

Histochemical reactions with substrate X-Gluc were performed with plant material 508

incubated in 80% acetone for 20 minutes at -20°C, washed 2x with 50 mM NaPO4 buffer 509

pH 7.0. The staining solution (1 mM X-Gluc diluted to 25 mg/mL, in 50 mM NaPO4 pH 510

7.0 buffer, supplemented with 2 mM potassium ferricyanide, 2 mM potassium 511

ferrocyanide, 0.1 % Triton X-100) was vacuum infiltrated for 15 minutes. The staining 512

reaction was carried out at 37°C overnight and stopped by rinsing the tissues three times 513

18

in 70% ethanol for 1 h. The stained plant material was examined by stereo-microscopy 514

(Leica, DFC300, FX). For thin sections GUS stained samples were dehydrated in an 515

ethanol series including a fixation step 20% ethanol, 35% ethanol, 50% ethanol, FAA 516

prepared fresh (50% ethanol, 3.7% formaldehyde, 5% acetic acid), 70% ethanol for 30 517

minutes each at room temperature. Then the samples were embedded in paraffin using the 518

enclosed tissue processor Leica ASK300S and the embedding center Leica EG1160. 10 519

µm and 20 µm longitudinal and traverse sections were placed on poly L-Lysine-coated 520

slides. After drying the samples overnight at 42°C the slides were de-waxed twice in 521

histoclear for 10 minutes and then incubated twice in 99.8% ethanol for 10 minutes under 522

constant movement at room temperature. After drying overnight the cover slips were 523

mounted with Entellan new (Merck Millipore) and examined by an epi-fluorescence 524

microscope (BX61, Olympus). 525

Bioinformatic Analysis 526

tRNA motif scans. Reference sequences of all protein-encoding genes [available cDNA 527

sequence data associated with all protein-coding Arabidopsis genes, TAIR10 (Lamesch et 528

al., 2012), excluding organellar genomes] were partitioned into distinct sets based on 529

their annotation as mobile or non-mobile as detected in heterografted Arabidopsis 530

accessions or Cuscuta-parasite Arabidopsis-host interactions (Thieme et al., 2015). 531

Subsets were generated for genes common to both mobile sets (n=486), present in at least 532

one of them (n=3606), as well as according to the observed movement direction (root-to-533

shoot, shoot-to-root, and bidirectional). All genes and associated transcripts assigned as 534

non-mobile were used as controls. All sets were filtered for duplicate sequences, and 535

annotated tRNA genes were removed. tRNA sequence data were obtained from the 536

tRNAdb (Juhling et al., 2009). Prior to structure motif scans, each sequence was padded 537

with 50 "N" leading and trailing characters to facilitate the detection of terminally located 538

tRNA structures without asymmetric ends at the tRNA acceptor arm which are required 539

by the default tRNA descriptor. All sets were analyzed by RNAMotif version 3.1.1 540

(Macke et al., 2001) using the provided tRNA structure descriptor and default parameter 541

settings. Motif enrichment associated with genes encoding mobile transcripts compared 542

to background data was assessed by Fisher’s exact test. Specificity of the searched tRNA-543

19

like structure was assessed by permutation scans of the default tRNA descriptor. 20,000 544

different tRNA descriptors were produced by randomly altering the accepted minimum 545

and maximum lengths limits for the stems and the single-stranded loops in the model 546

(normal distribution using µ=0, sigma=5; minimum stem length set to 3 nt). Each 547

descriptor was evaluated against the mobile/non-mobile data by RNAMotif with default 548

settings. Structuredness; i.e. the percentage of base-paired nucleotides and associated 549

energetics, within the 3′ UTR was addressed by excising the 150 nt 3′ -terminal sequence 550

portion and subsequent analysis of its predicted secondary structure (RNAfold, default 551

settings). 552

553

tRNA-mRNA tandem scans. Genes adjacent to tRNA loci were identified according to 554

TAIR10 gene models including protein-coding, non-coding genes, and pseudogenes. 555

Statistical significance of the difference of gene proximity distributions (distances 556

between tRNA-genes and mobile vs. non-mobile gene neighbors) was estimated by the 557

non-parametric Kolmogorov-Smirnov test; relevance was assessed by the effect size 558

(Cohen's D) based on the mean observed differences and associated standard deviations. 559

560

Di-cistronic tRNA analysis. A. thaliana reference genome (TAIR10) sequence 561

information was obtained from The Arabidopsis Information Resource (TAIR) associated 562

gene model descriptions (gtf version 10.30) were taken from http://plants.ensembl.org. 563

Paired-end RNA-Seq data (100nt reads from both ends) was retrieved from the Sequence 564

Read Archive (SRA) (http://www.ncbi.nlm.nih.gov/sra), accessions SRX853394 (14.1G 565

bases, root sample) and SRX853395 (15.3G bases, shoot sample) (Thieme et al., 2015) as 566

well as DRX014481 (19G bases, root sample) and DRX014482 (32.7G bases, root 567

sample) (Ito et al. 2015). Read data were quality trimmed and Illumina adapter sequences 568

were clipped by using Trimmomatic (Lohse et al. 2012) standard settings 569

(ILLUMINACLIP:<adapterfile>:2:40:15, LEADING:3, TRAILING:3, 570

SLIDINGWINDOW:4:15, and MINLEN:36). 571

Mapping of sequences mate pairs to the A. thaliana reference genome (TAIR10) was 572

done by STAR v2.5.1 (Dobin et al. 2012) based on Ensembl gene model descriptions. 573

Considering the high number of tRNA genes in the Arabidopsis genome and their similar 574

20

sequences, reads with multiple alignments were excluded, minimum overhang for gene 575

junctions was set to 10 nt for annotated junctions and 20 nt for unannotated junctions, 576

maximum number of allowed mismatches per pair was 10 nt (outFilterMultimapNmax 1, 577

alignSJDBoverhangMin 10, alignSJoverhangMin 20, outFilterMismatchNmax 10). 578

Subsequently, all read pairs mapping to chromosomes 1 to 5 with a minimum alignment 579

quality Q≥10 were checked to be intersecting with both, tRNA and mRNA gene 580

annotations. Finally, identified 132 di-cistronic poly(A)-RNA::tRNA transcripts were 581

grouped by their tRNA gene identity (118 unique tRNA genes, Figure 4C) as well as by 582

the protein-coding gene (120 unique genes) and the assigned transcript mobility. Results 583

were compared to the list of annotated tRNA-mRNA tandems and statistical significance 584

for the observed overlap to the di-cistronic transcripts was assessed by Fisher’s exact test. 585

586

Accession Numbers 587

Sequence data from this article can be found in the Arabidopsis Genome Initiative or 588

GenBank/EMBL databases under the following accession numbers: DMC1 TAIR 589

AT3G22880; BEL5 GenBank AF406697; CEN2/CET2 GenBank AF145260; tRNAMet 590

TAIR AT5G57885; tRNAGly TAIR AT1G71700; tRNAIle TAIR AT3G05835; ACTIN2 591

(ACT2) TAIR At3g18780; CK1 TAIR AT1G71697; GUS GenBank AKK29426. 592

593

Supplemental Data 594

Supplemental Figure 1. tRNAMet:DNDMC1 movement into flowers and pollen phenotype. 595

Supplemental Figure 2. tRNA sequences fused to the 3'UTR of GUS. 596

Supplemental Figure 3. Images of hypocotyl-grafted wild-type / GUS:tRNA 597

Supplemental Figure 4. Computational analysis of tRNA-like sequences (TLS) present 598

in mobile mRNAs. 599

Supplemental Figure 5. RT-PCR assays confirming the presence of di-cistronic poly(A)-600

RNA:tRNA transcripts in wild-type A. thaliana flowers and leaves. 601

Supplemental Table 1. Pollen shape analysis of wild-type, transgenic, and grafted plants. 602

21

Supplemental Table 2. Oligonucleotides used in the study. 603

Supplemental Data Set 1. Distribution of tRNA-like sequence (TLS) motifs in graft-604

mobile A. thaliana and grapevine transcripts. 605

Supplemental Data Set 2. tRNA proximity to genes, and occurrences of di-cistronic 606

poly(A) RNA:tRNA transcripts. 607

Supplemental Data Set 3. Number of independent transgenic lines used to perform graft 608

experiments. 609

610

Acknowledgments 611

We would like to thank Dana Schindelasch and Marina Stratmann (MPI-MPP) for their 612

outstanding technical support. This work was partially funded by MPI-MPP and Rijk 613

Zwaan to FK. 614

615

Author Contributions 616

WZ performed grafting experiments, evaluated pollen phenotypes, constructed GUS 617

fusions, analyzed GUS transgenic and CK1 mutants, and performed RT-PCR 618

experiments. GK constructed DMC1 fusions and made transgenic N. tabacum lines. FA 619

with WZ conducted the pollen shape analysis. NA supervised and supported by WZ 620

analyzed ck1 mutant plants and performed ck1 and wild-type grafting experiments. LY 621

performed some A. thaliana grafts and harvested pollen from grafted N. tabacum plants. 622

NW embedded and analyzed GUS stained tissue from grafted plants. CJT and DW 623

performed the bioinformatic analysis of graft-mobile mRNA sequences data. FK outlined 624

the project, suggested experiments, analyzed data, and wrote the manuscript with WZ 625

supported by CJT, DW, and NW. 626 627

22

628

References 629

Amaya. I., Ratcliff O.J., and Brafley D.J. (1999). Expression of CENTRORADIALIS 630 (CEN) and CEN-like genes in tobacco reveals a conserved mechanism controlling phase 631 change in diverse species. Plant Cell 11: 1405–1417. 632

Alonso, J.M., Stepanova, A.N., Leisse, T.J., Kim, C.J., Chen, H., Shinn, P., 633 Stevenson, D.K., Zimmerman, J., Barajas, P., Cheuk, R., Gadrinab, C., Heller, C., 634 Jeske, A., Koesema, E., Meyers, C.C., Parker, H., Prednis, L., Ansari, Y., Choy, N., 635 Deen, H., Geralt, M., Hazari, N., Hom, E., Karnes, M., Mulholland, C., Ndubaku, 636 R., Schmidt, I., Guzman, P., Aguilar-Henonin, L., Schmid, M., Weigel, D., Carter, 637 D.E., Marchand, T., Risseeuw, E., Brogden, D., Zeko, A., Crosby, W.L., Berry, C.C., 638 and Ecker, J.R. (2003). Genome-wide insertional mutagenesis of Arabidopsis thaliana. 639 Science 301: 653-657. 640

Banerjee, A.K., Lin, T., and Hannapel, D.J. (2009). Untranslated regions of a mobile 641 transcript mediate RNA metabolism. Plant Physiol. 151: 1831-1843. 642

Banerjee, A.K., Chatterjee, M., Yu, Y., Suh, S.G., Miller, W.A., and Hannapel, D.J. 643 (2006). Dynamics of a mobile RNA of potato involved in a long-distance signaling 644 pathway. Plant Cell 18: 3443-3457. 645

Barends, S., Rudinger-Thirion, J., Florentz, C., Giege, R., Pleij, C.W., and Kraal, B. 646 (2004). tRNA-like structure regulates translation of Brome mosaic virus RNA. J. Virol. 647 78: 4003-4010. 648

Bishop, D.K., Park, D., Xu, L., and Kleckner, N. (1992). DMC1: a meiosis-specific 649 yeast homolog of E. coli recA required for recombination, synaptonemal complex 650 formation, and cell cycle progression. Cell 69: 439-456. 651

Calderwood A., Kopriva S., and Morris R.J. (2016). Transcript abundance explains 652 mRNA mobility data in Arabidopsis thaliana. Plant Cell 28: 610-615. 653

Cho, S.K., Sharma, P., Butler, N.M., Kang, I.H., Shah, S., Rao, A.G., and Hannapel, 654 D.J. (2015). Polypyrimidine tract-binding proteins of potato mediate tuberization through 655 an interaction with StBEL5 RNA. J. Exp. Bot. 66: 6835-6847. 656

Curtis, M.D., and Grossniklaus, U. (2003). A gateway cloning vector set for high-657 throughput functional analysis of genes in planta. Plant Physiol. 133: 462-469. 658

Ding, B. (2009). The biology of viroid-host interactions. Annual Review of 659 Phytopathology 47, 105-131. 660

Dobin A., Davis C.A., Schlesinger F., Drenkow J., Zaleski C., Jha S., Batut P., 661 Chaisson M., and Gingeras T.R. (2012) STAR: ultrafast universal RNA-seq aligner, 662 bioinformatics. Bioinformatics 1:15-21 663

23

Doutriaux, M.P., Couteau, F., Bergounioux, C., and White, C. (1998). Isolation and 664 characterisation of the RAD51 and DMC1 homologs from Arabidopsis thaliana. Mol. 665 Gen. Genet. 257: 283-291. 666

Dreher, T.W. (2010). Viral tRNAs and tRNA-like structures. RNA 1: 402-414. 667

Fechter, P., Rudinger-Thirion, J., Florentz, C., and Giege, R. (2001). Novel features 668 in the tRNA-like world of plant viral RNAs. Cell Mol. Life. Sci. 58: 1547-1561. 669

Gopinath, K., and Kao, C.C. (2007). Replication-independent long-distance trafficking 670 by viral RNAs in Nicotiana benthamiana. Plant Cell 19: 1179-1191. 671

Guo, S., Zhang, J., Sun, H., Salse, J., Lucas, W.J., Zhang, H., Zheng, Y., Mao, L., 672 Ren, Y., Wang, Z., Min, J., Guo, X., Murat, F., Ham, B.K., Zhang, Z., Gao, S., 673 Huang, M., Xu, Y., Zhong, S., Bombarely, A., Mueller, L.A., Zhao, H., He, H., 674 Zhang, Y., Zhang, Z., Huang, S., Tan, T., Pang, E., Lin, K., Hu, Q., Kuang, H., Ni, P., 675 Wang, B., Liu, J., Kou, Q., Hou, W., Zou, X., Jiang, J., Gong, G., Klee, K., Schoof, 676 H., Huang, Y., Hu, X., Dong, S., Liang, D., Wang, J., Wu, K., Xia, Y., Zhao, X., 677 Zheng, Z., Xing, M., Liang, X., Huang, B., Lv, T., Wang, J., Yin, Y., Yi, H., Li, R., 678 Wu, M., Levi, A., Zhang, X., Giovannoni, J.J., Wang, J., Li, Y., Fei, Z., and Xu, Y. 679 (2013). The draft genome of watermelon (Citrullus lanatus) and resequencing of 20 680 diverse accessions. Nat. Genet. 45: 51-58. 681

Habu, T., Taki, T., West, A., Nishimune, Y., and Morita, T. (1996). The mouse and 682 human homologs of DMC1, the yeast meiosis-specific homologous recombination gene, 683 have a common unique form of exon-skipped transcript in meiosis. Nucleic Acids Res 684 24: 470-477. 685

Ham, B.K., Brandom, J.L., Xoconostle-Cazares, B., Ringgold, V., Lough, T.J., and 686 Lucas, W.J. (2009). A polypyrimidine tract binding protein, pumpkin RBP50, forms the 687 basis of a phloem-mobile ribonucleoprotein complex. Plant Cell 21: 197-215. 688

Ito S., Nozoye T., Sasaki E., Shiwa Y., Shibata-Hatta M., Ishige T., Fukui K., Ito K., 689 Nakanishi H., Nishizawa N.K., Yajima S., and Asami T. (2015). Strigolactone 690 regulates anthocyanin accumulation, acid phosphatases production and plant growth 691 under low phosphate condition in Arabidopsis. PLoS ONE 10: e0119724. 692

Juhling, F., Morl, M., Hartmann, R.K., Sprinzl, M., Stadler, P.F., and Putz, J. (2009). 693 tRNAdb 2009: compilation of tRNA sequences and tRNA genes. Nucleic Acids Res. 37: 694 D159-162. 695

Kalantidis, K., Schumacher, H.T., Alexiadis, T., and Helm, J.M. (2008). RNA 696 silencing movement in plants. Biol. Cell 100: 13-26. 697

Kim, M., Canio, W., Kessler, S., and Sinha, N. (2001). Developmental changes due to 698 long-distance movement of a homeobox fusion transcript in tomato. Science 293: 287-699 289. 700

24

Lamesch, P., Berardini, T.Z., Li, D., Swarbreck, D., Wilks, C., Sasidharan, R., 701 Muller, R., Dreher, K., Alexander, D.L., Garcia-Hernandez, M., Karthikeyan, A.S., 702 Lee, C.H., Nelson, W.D., Ploetz, L., Singh, S., Wensel, A., and Huala, E. (2012). The 703 Arabidopsis Information Resource (TAIR): improved gene annotation and new tools. 704 Nucleic Acids Res. 40: D1202-1210. 705

Lohse, M., Bolger, A.M., Nagel, A., Fernie, A.R., Lunn, J.E., Stitt, M., and Usadel, B. 706 (2012). RobiNA: a user-friendly, integrated software solution for RNA-Seq-based 707 transcriptomics. Nucleic Acids Res. 40: W622-627. 708

Lough, T.J., and Lucas, W.J. (2006). Integrative plant biology: role of phloem long-709 distance macromolecular trafficking. Annu. Rev. Plant Biol. 57: 203-232. 710

Lough, T.J., Lee, R.H., Emerson, S.J., Forster, R.L., and Lucas, W.J. (2006). 711 Functional analysis of the 5' untranslated region of potexvirus RNA reveals a role in viral 712 replication and cell-to-cell movement. Virology 351: 455-465. 713

Lucas, W.J., Yoo, B.C., and Kragler, F. (2001). RNA as a long-distance information 714 macromolecule in plants. Nat. Rev. Mol. Cell Biol. 2: 849-857. 715

McConnell, M.J., Lindberg, M.R., Brennand, K.J., Piper, J.C., Voet, T., Cowing-716 Zitron, C., Shumilina, S., Lasken, R.S., Vermeesch, J.R., Hall, I.M., and Gage, F.H. 717 (2013). Mosaic copy number variation in human neurons. Science 342: 632-637. 718

Macke, T.J., Ecker, D.J., Gutell, R.R., Gautheret, D., Case, D.A., and Sampath, R. 719 (2001). RNAMotif, an RNA secondary structure definition and search algorithm. Nucleic 720 Acids Res. 29: 4724-4735. 721

Matsuda, D., Yoshinari, S., and Dreher, T.W. (2004). eEF1A binding to aminoacylated 722 viral RNA represses minus strand synthesis by TYMV RNA-dependent RNA polymerase. 723 Virology 321: 47-56. 724

Melnyk, C.W., Molnar, A., and Baulcombe, D.C. (2011). Intercellular and systemic 725 movement of RNA silencing signals. EMBO J. 30: 3553-3563. 726

Molnar, A., Melnyk, C.W., Bassett, A., Hardcastle, T.J., Dunn, R., and Baulcombe, 727 D.C. (2010). Small silencing RNAs in plants are mobile and direct epigenetic 728 modification in recipient cells. Science 328: 872-875. 729

Nisar, N., Verma, S., Pogson, B.J., and Cazzonelli, C.I. (2012). Inflorescence stem 730 grafting made easy in Arabidopsis. Plant Methods 8: 50. 731

Ruiz-Medrano, R., Xoconostle-Cazares, B., and Kragler, F. (2004). The 732 plasmodesmatal transport pathway for homeotic proteins, silencing signals and viruses. 733 Curr. Opin. Plant Biol. 7: 641-650. 734

25

Takeda, R., Petrov, A.I., Leontis, N.B., and Ding, B. (2011). A three-dimensional RNA 735 motif in Potato spindle tuber viroid mediates trafficking from palisade mesophyll to 736 spongy mesophyll in Nicotiana benthamiana. Plant Cell 23: 258-272. 737

Tasseva, G., Richard, L., and Zachowski, A. (2004). Regulation of phosphatidylcholine 738 biosynthesis under salt stress involves choline kinases in Arabidopsis thaliana. FEBS 739 Lett. 566: 115-120. 740

Thieme, C.J., Rojas-Triana, M., Stecyk, E., Schudoma, C., Zhang, W., Yang, L., 741 Miambres, M., Walther, D., Schulze, W.X., Paz-Ares, J., Scheible, W.-R.D., and 742 Kragler, F. (2015). Endogenous Arabidopsis messenger RNAs transported to distant 743 tissues. Nature Plants 1: 15025. 744

Yang, Y., Mao, L., Jittayasothorn, Y., Kang, Y., Jiao, C., Fei, Z., and Zhong, G.Y. 745 (2015). Messenger RNA exchange between scions and rootstocks in grafted grapevines. 746 BMC Plant Biol. 15: 251. 747

Zhang, S., Sun, L., and Kragler, F. (2009). The phloem-delivered RNA pool contains 748 small noncoding RNAs and interferes with translation. Plant Physiol. 150: 378-387. 749

Zhang, W., Kollwig, G., Stecyk, E., Apelt, F., Dirks, R., and Kragler, F. (2014). Graft-750 transmissible movement of inverted-repeat-induced siRNA signals into flowers. Plant J. 751 80: 106-121. 752

753

Figure 1. Dominant negative DMC1 as a reporter construct causes male sterile

flowers.

(A) Schematic drawing of the DNDMC1 RNA fusion constructs used. A. thaliana

DNDMC1 codes for a truncated protein lacking the N-terminal 92 amino acids and

dominantly interferes with meiosis resulting in misshaped pollen and partial male

sterility. The DNDMC1 coding sequence was fused to graft-mobile S. tuberosum BEL5

sequences or phloem tRNAMet at the 3′ UTR to evaluate their potential to trigger

DNDMC mRNA transport over graft junctions. (B) to (E) Fertile anthers of wild-type

Nicotiana tabacum plants show regular pollen production with minimal abnormally

shaped pollen (2-3%), whereas hpDMC1 siRNA transgenic tobacco plants produce high

numbers of abnormally shaped pollen and are sterile as previously described (Zhang et

al., 2014). YFP-DNDMC1 transgenic plants have normal pollen production similar to wild

type because the N-terminal YFP fusion abolishes the dominant negative effect of

truncated DMC1. Transgenic plants expressing DNDMC1 fused with tRNAMet or BEL5 at

the 3′UTR exhibit increased male sterility. (C) and (E) Propidium iodide-stained

pollen harvested from transgenic plants were imaged by confocal laser scanning

microscopy and evaluated by an automatic imaging analysis algorithm to count

abnormally shaped pollen (Zhang et al., 2014) indicated by % numbers. Arrows indicate

normal pollen; arrowheads indicate abnormally shaped pollen. Scale bars: 30 µm. (F)

and (G) Scheme of performed stem-grafts to evaluate transport of mRNA to wild-type

flowers.

Figure 2. DNDMC1 fusion transcript transport induces aberrantpollenformation.

(A) Flowers of grafted wild-type stock plants supported by 35Spro:YFP-DNDMC1

transgenic scions and reciprocal grafts are fertile. (B) Upper panel: Grafted wild-type /

wild-type or wild-type / DNDMC1:tRNAMet plants showed normal pollen production when

mock treated. Lower panel: Estradiol-induced expression of DNDMC1:tRNAMet in scion or

stock plant parts resulted in partially sterile anthers in both transgenic and wild-type

flowers. The latter suggests DNDMC1:tRNAMet mRNA transport into and expression of the

truncated DMC1 protein in wild-type male meiocytes. (C) Flowers of grafted

DNDMC1:BEL5 transgenic plants. Upper panel: Mock-treated wild-type /

estradiol>>DNDMC1:BEL5 grafts showed weak male sterility. Lower panel: Flowers of

grafted plants treated with estradiol exhibit partial male sterility. (D) RT-PCR assays on

RNAs samples from grafted wild-type tissues revealed that the YFP-DNDMC1 control

transcript is not allocated over graft junctions into wild-type stock leaves (n=6) or scion

flowers (n=8). ACTIN2 (ACT2) specific RT-PCR was used as a positive control. (E) RT-

PCR assays on RNA samples from grafted plants. DNDMC1:tRNAMet and DNDMC1:BEL5

is detected in transgenic and in wild-type scion flowers. Appearance of a specific PCR

product in samples from grafted wild-type stock leaves and wild-type flowers (red

asterisks) suggests mobility of the DNDMC1:tRNAMet fusion transcript. Number of tested

grafted plants is shown on the right. (F) CLSM images of sepals formed on YFP-

DNDMC1 producing scions. Upper panel: YFP-DNDMC1 / YFP-DNDMC1 control graft

with expected high green fluorescence emitted by YFP-DNDMC1. Middle panel: Control

graft with siRNA-producing stock plants (hpDMC1) with expected low YFP fluorescence

and distribution in YFP-DNDMC1 flowers (Zhang et al., 2014). Lower panel: YFP-

DNDMC1 scion grafted onto DNDMC1:tRNAMet transgenic stock shows similar YFP

fluorescence levels as YFP-DNDMC1 / YFP-DNDMC1 control grafts. Note that YFP-

DNDMC1 fusion protein is detected in all epidermal leaf cells except when grafted with

hpDMC1 producing DMC1 siRNA lines. Green indicates presence of YFP-DNDMC1;

Blue: plastid auto-fluorescence. Scale bar: 300 µm. (G) Statistical analysis of data from

automated detection of misshaped pollen appearing on grafted plants. Misshapen pollen

formation was significantly higher on wild-type scions supported by DNDMC1:tRNAMet

and DNDMC1:BEL5 stock plants than in control grafts. Asterisks indicate highly

significant differences against controls using Chi-square test for independence of

variables in a contingency table. Biological replicates: n>8. Error bars indicate S. D. For

details see Supplemental Table 1.

Figure 3.GUS:tRNAfusiontranscriptsandmobilityingraftedArabidopsis

thaliana.

(A)Schematicdrawingofused35Spro:GUS:tRNAfusionconstructs(for sequences of

tRNAMet, tRNAGly, tRNAIle, and tRNAMet deletions see Supplemental Figure 2). (B)

Example of a hypocotyl grafted GUS:tRNAMet/wild-type (Col-0) plant. Blue color

indicates presence of GUS activity in the hypocotyl above the graft junction (arrow) and

in the wild-type root tip. (C) GUS activity in leaves and primary root tips detected in

GUS:tRNA / wild-type grafts. The numbers indicate the fraction of GUS staining

detected in the wild-type root tips or wild-type leaf vasculature (arrows) of plants grafted

with the indicated transgenic line. At least 3 independent transgenic lines were used for

each graft combination (for details see Supplemental Data Set 3 and for additional images

of grafted plants see Supplemental Figure 3. (D) RT-PCR on poly(A)-RNA samples

harvested from grafted plants. Three samples from 3-5 grafted plants were pooled and

tested for presence of GUS transcripts in wild-type tissue (asterisks). Numbers indicate

occurrence of GUS poly(A) RNA in the tested wild-type root or wild-type leaves RNA

samples. ACTIN2 (ACT2) specific RT-PCR was used as a positive control confirming

mRNA presence in the samples.

Figure 4. Mobile A.thalianamRNAsandoccurrenceoftRNA-likemotifs

(A) Number of all identified mobile transcripts (n=3606) with predicted tRNA-like

structures found by the default RNAMotif tRNA descriptor, which does not capture the

tRNAIle (TAT) (Supplemental Figure 4). Absolute counts as well as enrichment in relation

to transcripts not found in the mobile database are shown. Asterisks indicate significant

counts (p<0.05) according to Fisher’s exact test. (B) Normalized frequency (estimated

density) and cumulative relative frequency (ecdf) of inter-gene distances of tRNA-mRNA

tandem gene pairs with the tRNA being located within 1000 nucleotides up- or

downstream of genes coding for mobile transcripts (blue) or non-mobile predicted

transcripts (grey). Vertical dashed lines indicate medians of shown distributions. Mobile

transcript encoding loci in comparison to loci not producing mobile transcript show a

significantly closer proximity to tRNA genes (two-sample, two-sided Kolmogorov-

Smirnov test, p<0.0003; Cohen's D d=0.313). (C) Number of tRNA genes according to

their anticodon, which were detected as poly(A)-RNA:tRNA di-cistronic transcripts in

RNA-Seq data. Orange: distribution of the 94 tRNA genes observed di-cistronically;

blue: tRNA genes (n=24) associated with mobile transcripts; gray: TAIR10 annotated

tRNA genes; asterisks: tRNA genes with di-cistronic transcripts confirmed by RT-PCR;

arrows: experimentally tested tRNA fusions. (D) Schematic diagram of A. thaliana

CHOLINE KINASE 1 (CK1, AT1G71697) gene and analyzed insertion mutants. CK1

mRNA exists as a di-cistronic poly(A) - tRNA transcript. The ck1.2 mutant harbors a T-

DNA insertion between the CK1 stop codon and the annotated tRNAGly (AT1G71700) in

the 3′ UTR resulting in a truncated poly(A) transcript lacking the tRNAGly sequences.

RT-PCR primers are indicated as follows: P1 binding to exon 7 of CK1 CDS and P3

binding to tRNAGly sequences were used for wild-type CK1:tRNAGly identification. P1

together with P2 which is stretching the T-DNA left border and CK1 3′ UTR were used

for specific ck1.2 detection. (E) RT-PCR with the indicated primers revealed that the

CK1 poly(A) transcript is present in ck1.2 mutant samples (asterisks) originating from

stem-grafted wild-type Col-0 tissue. In the reciprocal wild-type samples a mutant CK1

poly(A) transcript produced in ck1.2 and lacking the 3′ UTR tRNAGly sequence was not

detected. (F) Phenotype of ck1.1 and ck1.2 and reverse transcription quantitative PCR of

transcripts. Wild-type and ck1.2 plants show similarly high levels of CK1 transcript

whereas ck1.1 mutants show very low levels. Rosette area size measurements on adult

plants revealed that both ck1.1 and ck1.2 mutants are significantly smaller than wild type

(Students T-test, mutant vs. wild type; p-value ck1.1 = 0.006; p-value ck1.2 = 0.002;

n=16 plants/line). Error bars: S. D. (G) Schematic folding structure of the GUS TLS 3′

UTR motifs predicted according to their minimal free energy (MFE).

Parsed CitationsAmaya. I., Ratcliff O.J., and Brafley D.J. (1999). Expression of CENTRORADIALIS (CEN) and CEN-like genes in tobacco reveals aconserved mechanism controlling phase change in diverse species. Plant Cell 11: 1405-1417.

Pubmed: Author and TitleCrossRef: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Alonso, J.M., Stepanova, A.N., Leisse, T.J., Kim, C.J., Chen, H., Shinn, P., Stevenson, D.K., Zimmerman, J., Barajas, P., Cheuk, R.,Gadrinab, C., Heller, C., Jeske, A., Koesema, E., Meyers, C.C., Parker, H., Prednis, L., Ansari, Y., Choy, N., Deen, H., Geralt, M.,Hazari, N., Hom, E., Karnes, M., Mulholland, C., Ndubaku, R., Schmidt, I., Guzman, P., Aguilar-Henonin, L., Schmid, M., Weigel, D.,Carter, D.E., Marchand, T., Risseeuw, E., Brogden, D., Zeko, A., Crosby, W.L., Berry, C.C., and Ecker, J.R. (2003). Genome-wideinsertional mutagenesis of Arabidopsis thaliana. Science 301: 653-657.


Banerjee, A.K., Lin, T., and Hannapel, D.J. (2009). Untranslated regions of a mobile transcript mediate RNA metabolism. PlantPhysiol. 151: 1831-1843.


Banerjee, A.K., Chatterjee, M., Yu, Y., Suh, S.G., Miller, W.A., and Hannapel, D.J. (2006). Dynamics of a mobile RNA of potatoinvolved in a long-distance signaling pathway. Plant Cell 18: 3443-3457.


Barends, S., Rudinger-Thirion, J., Florentz, C., Giege, R., Pleij, C.W., and Kraal, B. (2004). tRNA-like structure regulates translationof Brome mosaic virus RNA. J. Virol. 78: 4003-4010.


Bishop, D.K., Park, D., Xu, L., and Kleckner, N. (1992). DMC1: a meiosis-specific yeast homolog of E. coli recA required forrecombination, synaptonemal complex formation, and cell cycle progression. Cell 69: 439-456.


Calderwood A., Kopriva S., and Morris R.J. (2016). Transcript abundance explains mRNA mobility data in Arabidopsis thaliana. PlantCell 28: 610-615.


Cho, S.K., Sharma, P., Butler, N.M., Kang, I.H., Shah, S., Rao, A.G., and Hannapel, D.J. (2015). Polypyrimidine tract-binding proteinsof potato mediate tuberization through an interaction with StBEL5 RNA. J. Exp. Bot. 66: 6835-6847.


Curtis, M.D., and Grossniklaus, U. (2003). A gateway cloning vector set for high-throughput functional analysis of genes in planta.Plant Physiol. 133: 462-469.


Ding, B. (2009). The biology of viroid-host interactions. Annual Review of Phytopathology 47, 105-131.Pubmed: Author and TitleCrossRef: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Dobin A., Davis C.A., Schlesinger F., Drenkow J., Zaleski C., Jha S., Batut P., Chaisson M., and Gingeras T.R. (2012) STAR: ultrafastuniversal RNA-seq aligner, bioinformatics. Bioinformatics 1:15-21


Doutriaux, M.P., Couteau, F., Bergounioux, C., and White, C. (1998). Isolation and characterisation of the RAD51 and DMC1homologs from Arabidopsis thaliana. Mol. Gen. Genet. 257: 283-291.


Dreher, T.W. (2010). Viral tRNAs and tRNA-like structures. RNA 1: 402-414.Pubmed: Author and TitleCrossRef: Author and Title

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Plant Cell%5BTitle%5D%29 AND 11%5BVolume%5D AND 1405%5BPagination%5D AND Amaya%2E I%2E%2C Ratcliff O%2EJ%2E%2C and Brafley D%2EJ%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Amaya. I., Ratcliff O.J., and Brafley D.J. (1999). Expression of CENTRORADIALIS (CEN) and CEN-like genes in tobacco reveals a conserved mechanism controlling phase change in diverse species. Plant Cell 11: 1405-1417.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Amaya.&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Expression of CENTRORADIALIS (CEN) and CEN-like genes in tobacco reveals a conserved mechanism controlling phase change in diverse species&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Expression of CENTRORADIALIS (CEN) and CEN-like genes in tobacco reveals a conserved mechanism controlling phase change in diverse species&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Amaya.&as_ylo=1999&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Science%5BTitle%5D%29 AND 301%5BVolume%5D AND 653%5BPagination%5D AND Alonso%2C J%2EM%2E%2C Stepanova%2C A%2EN%2E%2C Leisse%2C T%2EJ%2E%2C Kim%2C C%2EJ%2E%2C Chen%2C H%2E%2C Shinn%2C P%2E%2C Stevenson%2C D%2EK%2E%2C Zimmerman%2C J%2E%2C Barajas%2C P%2E%2C Cheuk%2C R%2E%2C Gadrinab%2C C%2E%2C Heller%2C C%2E%2C Jeske%2C A%2E%2C Koesema%2C E%2E%2C Meyers%2C C%2EC%2E%2C Parker%2C H%2E%2C Prednis%2C L%2E%2C Ansari%2C Y%2E%2C Choy%2C N%2E%2C Deen%2C H%2E%2C Geralt%2C M%2E%2C Hazari%2C N%2E%2C Hom%2C E%2E%2C Karnes%2C M%2E%2C Mulholland%2C C%2E%2C Ndubaku%2C R%2E%2C Schmidt%2C I%2E%2C Guzman%2C P%2E%2C Aguilar%2DHenonin%2C L%2E%2C Schmid%2C M%2E%2C Weigel%2C D%2E%2C Carter%2C D%2EE%2E%2C Marchand%2C T%2E%2C Risseeuw%2C E%2E%2C Brogden%2C D%2E%2C Zeko%2C A%2E%2C Crosby%2C W%2EL%2E%2C Berry%2C C%2EC%2E%2C and Ecker%2C J%2ER%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Alonso, J.M., Stepanova, A.N., Leisse, T.J., Kim, C.J., Chen, H., Shinn, P., Stevenson, D.K., Zimmerman, J., Barajas, P., Cheuk, R., Gadrinab, C., Heller, C., Jeske, A., Koesema, E., Meyers, C.C., Parker, H., Prednis, L., Ansari, Y., Choy, N., Deen, H., Geralt, M., Hazari, N., Hom, E., Karnes, M., Mulholland, C., Ndubaku, R., Schmidt, I., Guzman, P., Aguilar-Henonin, L., Schmid, M., Weigel, D., Carter, D.E., Marchand, T., Risseeuw, E., Brogden, D., Zeko, A., Crosby, W.L., Berry, C.C., and Ecker, J.R. (2003). Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301: 653-657.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Alonso,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Genome-wide insertional mutagenesis of Arabidopsis thaliana&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

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http://www.ncbi.nlm.nih.gov/pubmed?term=%28Plant Physiol%5BTitle%5D%29 AND 151%5BVolume%5D AND 1831%5BPagination%5D AND Banerjee%2C A%2EK%2E%2C Lin%2C T%2E%2C and Hannapel%2C D%2EJ%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Banerjee, A.K., Lin, T., and Hannapel, D.J. (2009). Untranslated regions of a mobile transcript mediate RNA metabolism. Plant Physiol. 151: 1831-1843.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Banerjee,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Untranslated regions of a mobile transcript mediate RNA metabolism&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Untranslated regions of a mobile transcript mediate RNA metabolism&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Banerjee,&as_ylo=2009&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Plant Cell%5BTitle%5D%29 AND 18%5BVolume%5D AND 3443%5BPagination%5D AND Banerjee%2C A%2EK%2E%2C Chatterjee%2C M%2E%2C Yu%2C Y%2E%2C Suh%2C S%2EG%2E%2C Miller%2C W%2EA%2E%2C and Hannapel%2C D%2EJ%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Banerjee, A.K., Chatterjee, M., Yu, Y., Suh, S.G., Miller, W.A., and Hannapel, D.J. (2006). Dynamics of a mobile RNA of potato involved in a long-distance signaling pathway. Plant Cell 18: 3443-3457.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Banerjee,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Dynamics of a mobile RNA of potato involved in a long-distance signaling pathway&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

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http://www.ncbi.nlm.nih.gov/pubmed?term=%28J%2E Virol%5BTitle%5D%29 AND 78%5BVolume%5D AND 4003%5BPagination%5D AND Barends%2C S%2E%2C Rudinger%2DThirion%2C J%2E%2C Florentz%2C C%2E%2C Giege%2C R%2E%2C Pleij%2C C%2EW%2E%2C and Kraal%2C B%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Barends, S., Rudinger-Thirion, J., Florentz, C., Giege, R., Pleij, C.W., and Kraal, B. (2004). tRNA-like structure regulates translation of Brome mosaic virus RNA. J. Virol. 78: 4003-4010.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Barends,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=tRNA-like structure regulates translation of Brome mosaic virus RNA&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=tRNA-like structure regulates translation of Brome mosaic virus RNA&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Barends,&as_ylo=2004&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28DMC%5BTitle%5D%29 AND 1%5BVolume%5D AND Bishop%2C D%2EK%2E%2C Park%2C D%2E%2C Xu%2C L%2E%2C and Kleckner%2C N%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Bishop, D.K., Park, D., Xu, L., and Kleckner, N. (1992). DMC1: a meiosis-specific yeast homolog of E. coli recA required for recombination, synaptonemal complex formation, and cell cycle progression. Cell 69: 439-456.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Bishop,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Bishop,&as_ylo=1992&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Plant Cell%5BTitle%5D%29 AND 28%5BVolume%5D AND 610%5BPagination%5D AND Calderwood A%2E%2C Kopriva S%2E%2C and Morris R%2EJ%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Calderwood A., Kopriva S., and Morris R.J. (2016). Transcript abundance explains mRNA mobility data in Arabidopsis thaliana. Plant Cell 28: 610-615.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Calderwood&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Transcript abundance explains mRNA mobility data in Arabidopsis thaliana&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Transcript abundance explains mRNA mobility data in Arabidopsis thaliana&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Calderwood&as_ylo=2016&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28J%2E Exp%2E Bot%5BTitle%5D%29 AND 66%5BVolume%5D AND 6835%5BPagination%5D AND Cho%2C S%2EK%2E%2C Sharma%2C P%2E%2C Butler%2C N%2EM%2E%2C Kang%2C I%2EH%2E%2C Shah%2C S%2E%2C Rao%2C A%2EG%2E%2C and Hannapel%2C D%2EJ%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Cho, S.K., Sharma, P., Butler, N.M., Kang, I.H., Shah, S., Rao, A.G., and Hannapel, D.J. (2015). Polypyrimidine tract-binding proteins of potato mediate tuberization through an interaction with StBEL5 RNA. J. Exp. Bot. 66: 6835-6847.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Cho,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Polypyrimidine tract-binding proteins of potato mediate tuberization through an interaction with StBEL5 RNA&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Polypyrimidine tract-binding proteins of potato mediate tuberization through an interaction with StBEL5 RNA&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Cho,&as_ylo=2015&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Plant Physiol%5BTitle%5D%29 AND 133%5BVolume%5D AND 462%5BPagination%5D AND Curtis%2C M%2ED%2E%2C and Grossniklaus%2C U%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Curtis, M.D., and Grossniklaus, U. (2003). A gateway cloning vector set for high-throughput functional analysis of genes in planta. Plant Physiol. 133: 462-469.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Curtis,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=A gateway cloning vector set for high-throughput functional analysis of genes in planta&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=A gateway cloning vector set for high-throughput functional analysis of genes in planta&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Curtis,&as_ylo=2003&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Annual Review of Phytopathology%5BTitle%5D%29 AND 47%5BVolume%5D AND 105%5BPagination%5D AND Ding%2C B%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Ding, B. (2009). The biology of viroid-host interactions. Annual Review of Phytopathology 47, 105-131.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Ding,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=The biology of viroid-host interactions&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=The biology of viroid-host interactions&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Ding,&as_ylo=2009&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Bioinformatics%5BTitle%5D%29 AND 1%5BVolume%5D AND 15%5BPagination%5D AND Dobin A%2E%2C Davis C%2EA%2E%2C Schlesinger F%2E%2C Drenkow J%2E%2C Zaleski C%2E%2C Jha S%2E%2C Batut P%2E%2C Chaisson M%2E%2C and Gingeras T%2ER%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Dobin A., Davis C.A., Schlesinger F., Drenkow J., Zaleski C., Jha S., Batut P., Chaisson M., and Gingeras T.R. (2012) STAR: ultrafast universal RNA-seq aligner, bioinformatics. Bioinformatics 1:15-21

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Dobin&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=STAR: ultrafast universal RNA-seq aligner, bioinformatics&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=STAR: ultrafast universal RNA-seq aligner, bioinformatics&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Dobin&as_ylo=2012&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Mol%2E Gen%2E Genet%5BTitle%5D%29 AND 257%5BVolume%5D AND 283%5BPagination%5D AND Doutriaux%2C M%2EP%2E%2C Couteau%2C F%2E%2C Bergounioux%2C C%2E%2C and White%2C C%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Doutriaux, M.P., Couteau, F., Bergounioux, C., and White, C. (1998). Isolation and characterisation of the RAD51 and DMC1 homologs from Arabidopsis thaliana. Mol. Gen. Genet. 257: 283-291.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Doutriaux,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Isolation and characterisation of the RAD51 and DMC1 homologs from Arabidopsis thaliana&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Isolation and characterisation of the RAD51 and DMC1 homologs from Arabidopsis thaliana&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Doutriaux,&as_ylo=1998&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28RNA%5BTitle%5D%29 AND 1%5BVolume%5D AND 402%5BPagination%5D AND Dreher%2C T%2EW%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Dreher, T.W. (2010). Viral tRNAs and tRNA-like structures. RNA 1: 402-414.

Google Scholar: Author Only Title Only Author and Title

Fechter, P., Rudinger-Thirion, J., Florentz, C., and Giege, R. (2001). Novel features in the tRNA-like world of plant viral RNAs. CellMol. Life. Sci. 58: 1547-1561.


Gopinath, K., and Kao, C.C. (2007). Replication-independent long-distance trafficking by viral RNAs in Nicotiana benthamiana.Plant Cell 19: 1179-1191.


Guo, S., Zhang, J., Sun, H., Salse, J., Lucas, W.J., Zhang, H., Zheng, Y., Mao, L., Ren, Y., Wang, Z., Min, J., Guo, X., Murat, F., Ham,B.K., Zhang, Z., Gao, S., Huang, M., Xu, Y., Zhong, S., Bombarely, A., Mueller, L.A., Zhao, H., He, H., Zhang, Y., Zhang, Z., Huang, S.,Tan, T., Pang, E., Lin, K., Hu, Q., Kuang, H., Ni, P., Wang, B., Liu, J., Kou, Q., Hou, W., Zou, X., Jiang, J., Gong, G., Klee, K., Schoof,H., Huang, Y., Hu, X., Dong, S., Liang, D., Wang, J., Wu, K., Xia, Y., Zhao, X., Zheng, Z., Xing, M., Liang, X., Huang, B., Lv, T., Wang, J.,Yin, Y., Yi, H., Li, R., Wu, M., Levi, A., Zhang, X., Giovannoni, J.J., Wang, J., Li, Y., Fei, Z., and Xu, Y. (2013). The draft genome ofwatermelon (Citrullus lanatus) and resequencing of 20 diverse accessions. Nat. Genet. 45: 51-58.


Habu, T., Taki, T., West, A., Nishimune, Y., and Morita, T. (1996). The mouse and human homologs of DMC1, the yeast meiosis-specific homologous recombination gene, have a common unique form of exon-skipped transcript in meiosis. Nucleic Acids Res24: 470-477.


Ham, B.K., Brandom, J.L., Xoconostle-Cazares, B., Ringgold, V., Lough, T.J., and Lucas, W.J. (2009). A polypyrimidine tract bindingprotein, pumpkin RBP50, forms the basis of a phloem-mobile ribonucleoprotein complex. Plant Cell 21: 197-215.


Ito S., Nozoye T., Sasaki E., Shiwa Y., Shibata-Hatta M., Ishige T., Fukui K., Ito K., Nakanishi H., Nishizawa N.K., Yajima S., andAsami T. (2015). Strigolactone regulates anthocyanin accumulation, acid phosphatases production and plant growth under lowphosphate condition in Arabidopsis. PLoS ONE 10: e0119724.


Juhling, F., Morl, M., Hartmann, R.K., Sprinzl, M., Stadler, P.F., and Putz, J. (2009). tRNAdb 2009: compilation of tRNA sequencesand tRNA genes. Nucleic Acids Res. 37: D159-162.


Kalantidis, K., Schumacher, H.T., Alexiadis, T., and Helm, J.M. (2008). RNA silencing movement in plants. Biol. Cell 100: 13-26.Pubmed: Author and TitleCrossRef: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Kim, M., Canio, W., Kessler, S., and Sinha, N. (2001). Developmental changes due to long-distance movement of a homeoboxfusion transcript in tomato. Science 293: 287-289.


Lamesch, P., Berardini, T.Z., Li, D., Swarbreck, D., Wilks, C., Sasidharan, R., Muller, R., Dreher, K., Alexander, D.L., Garcia-Hernandez, M., Karthikeyan, A.S., Lee, C.H., Nelson, W.D., Ploetz, L., Singh, S., Wensel, A., and Huala, E. (2012). The ArabidopsisInformation Resource (TAIR): improved gene annotation and new tools. Nucleic Acids Res. 40: D1202-1210.


Lohse, M., Bolger, A.M., Nagel, A., Fernie, A.R., Lunn, J.E., Stitt, M., and Usadel, B. (2012). RobiNA: a user-friendly, integratedsoftware solution for RNA-Seq-based transcriptomics. Nucleic Acids Res. 40: W622-627.


Lough, T.J., and Lucas, W.J. (2006). Integrative plant biology: role of phloem long-distance macromolecular trafficking. Annu. Rev.Plant Biol. 57: 203-232.


http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Dreher,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Viral tRNAs and tRNA-like structures&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Viral tRNAs and tRNA-like structures&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Dreher,&as_ylo=2010&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Cell Mol%2E Life%2E Sci%5BTitle%5D%29 AND 58%5BVolume%5D AND 1547%5BPagination%5D AND Fechter%2C P%2E%2C Rudinger%2DThirion%2C J%2E%2C Florentz%2C C%2E%2C and Giege%2C R%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Fechter, P., Rudinger-Thirion, J., Florentz, C., and Giege, R. (2001). Novel features in the tRNA-like world of plant viral RNAs. Cell Mol. Life. Sci. 58: 1547-1561.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Fechter,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Novel features in the tRNA-like world of plant viral RNAs&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Novel features in the tRNA-like world of plant viral RNAs&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Fechter,&as_ylo=2001&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Plant Cell%5BTitle%5D%29 AND 19%5BVolume%5D AND 1179%5BPagination%5D AND Gopinath%2C K%2E%2C and Kao%2C C%2EC%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Gopinath, K., and Kao, C.C. (2007). Replication-independent long-distance trafficking by viral RNAs in Nicotiana benthamiana. Plant Cell 19: 1179-1191.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Gopinath,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Replication-independent long-distance trafficking by viral RNAs in Nicotiana benthamiana&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Replication-independent long-distance trafficking by viral RNAs in Nicotiana benthamiana&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Gopinath,&as_ylo=2007&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Nat%2E Genet%5BTitle%5D%29 AND 45%5BVolume%5D AND 51%5BPagination%5D AND Guo%2C S%2E%2C Zhang%2C J%2E%2C Sun%2C H%2E%2C Salse%2C J%2E%2C Lucas%2C W%2EJ%2E%2C Zhang%2C H%2E%2C Zheng%2C Y%2E%2C Mao%2C L%2E%2C Ren%2C Y%2E%2C Wang%2C Z%2E%2C Min%2C J%2E%2C Guo%2C X%2E%2C Murat%2C F%2E%2C Ham%2C B%2EK%2E%2C Zhang%2C Z%2E%2C Gao%2C S%2E%2C Huang%2C M%2E%2C Xu%2C Y%2E%2C Zhong%2C S%2E%2C Bombarely%2C A%2E%2C Mueller%2C L%2EA%2E%2C Zhao%2C H%2E%2C He%2C H%2E%2C Zhang%2C Y%2E%2C Zhang%2C Z%2E%2C Huang%2C S%2E%2C Tan%2C T%2E%2C Pang%2C E%2E%2C Lin%2C K%2E%2C Hu%2C Q%2E%2C Kuang%2C H%2E%2C Ni%2C P%2E%2C Wang%2C B%2E%2C Liu%2C J%2E%2C Kou%2C Q%2E%2C Hou%2C W%2E%2C Zou%2C X%2E%2C Jiang%2C J%2E%2C Gong%2C G%2E%2C Klee%2C K%2E%2C Schoof%2C H%2E%2C Huang%2C Y%2E%2C Hu%2C X%2E%2C Dong%2C S%2E%2C Liang%2C D%2E%2C Wang%2C J%2E%2C Wu%2C K%2E%2C Xia%2C Y%2E%2C Zhao%2C X%2E%2C Zheng%2C Z%2E%2C Xing%2C M%2E%2C Liang%2C X%2E%2C Huang%2C B%2E%2C Lv%2C T%2E%2C Wang%2C J%2E%2C Yin%2C Y%2E%2C Yi%2C H%2E%2C Li%2C R%2E%2C Wu%2C M%2E%2C Levi%2C A%2E%2C Zhang%2C X%2E%2C Giovannoni%2C J%2EJ%2E%2C Wang%2C J%2E%2C Li%2C Y%2E%2C Fei%2C Z%2E%2C and Xu%2C Y%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Guo, S., Zhang, J., Sun, H., Salse, J., Lucas, W.J., Zhang, H., Zheng, Y., Mao, L., Ren, Y., Wang, Z., Min, J., Guo, X., Murat, F., Ham, B.K., Zhang, Z., Gao, S., Huang, M., Xu, Y., Zhong, S., Bombarely, A., Mueller, L.A., Zhao, H., He, H., Zhang, Y., Zhang, Z., Huang, S., Tan, T., Pang, E., Lin, K., Hu, Q., Kuang, H., Ni, P., Wang, B., Liu, J., Kou, Q., Hou, W., Zou, X., Jiang, J., Gong, G., Klee, K., Schoof, H., Huang, Y., Hu, X., Dong, S., Liang, D., Wang, J., Wu, K., Xia, Y., Zhao, X., Zheng, Z., Xing, M., Liang, X., Huang, B., Lv, T., Wang, J., Yin, Y., Yi, H., Li, R., Wu, M., Levi, A., Zhang, X., Giovannoni, J.J., Wang, J., Li, Y., Fei, Z., and Xu, Y. (2013). The draft genome of watermelon (Citrullus lanatus) and resequencing of 20 diverse accessions. Nat. Genet. 45: 51-58.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Guo,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=The draft genome of watermelon (Citrullus lanatus) and resequencing of 20 diverse accessions&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=The draft genome of watermelon (Citrullus lanatus) and resequencing of 20 diverse accessions&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Guo,&as_ylo=2013&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28The%5BTitle%5D%29 AND 1%5BVolume%5D AND Habu%2C T%2E%2C Taki%2C T%2E%2C West%2C A%2E%2C Nishimune%2C Y%2E%2C and Morita%2C T%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Habu, T., Taki, T., West, A., Nishimune, Y., and Morita, T. (1996). The mouse and human homologs of DMC1, the yeast meiosis-specific homologous recombination gene, have a common unique form of exon-skipped transcript in meiosis. Nucleic Acids Res 24: 470-477.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Habu,&hl=en&c2coff=1


http://scholar.google.com/scholar?as_q=&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Habu,&as_ylo=1996&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28A%5BTitle%5D%29 AND 50%5BVolume%5D AND Ham%2C B%2EK%2E%2C Brandom%2C J%2EL%2E%2C Xoconostle%2DCazares%2C B%2E%2C Ringgold%2C V%2E%2C Lough%2C T%2EJ%2E%2C and Lucas%2C W%2EJ%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Ham, B.K., Brandom, J.L., Xoconostle-Cazares, B., Ringgold, V., Lough, T.J., and Lucas, W.J. (2009). A polypyrimidine tract binding protein, pumpkin RBP50, forms the basis of a phloem-mobile ribonucleoprotein complex. Plant Cell 21: 197-215.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Ham,&hl=en&c2coff=1


http://scholar.google.com/scholar?as_q=&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Ham,&as_ylo=2009&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28PLoS ONE%5BTitle%5D%29 AND 10%5BVolume%5D AND 0119724%5BPagination%5D AND Ito S%2E%2C Nozoye T%2E%2C Sasaki E%2E%2C Shiwa Y%2E%2C Shibata%2DHatta M%2E%2C Ishige T%2E%2C Fukui K%2E%2C Ito K%2E%2C Nakanishi H%2E%2C Nishizawa N%2EK%2E%2C Yajima S%2E%2C and Asami T%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Ito S., Nozoye T., Sasaki E., Shiwa Y., Shibata-Hatta M., Ishige T., Fukui K., Ito K., Nakanishi H., Nishizawa N.K., Yajima S., and Asami T. (2015). Strigolactone regulates anthocyanin accumulation, acid phosphatases production and plant growth under low phosphate condition in Arabidopsis. PLoS ONE 10: e0119724.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Ito&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Strigolactone regulates anthocyanin accumulation, acid phosphatases production and plant growth under low phosphate condition in Arabidopsis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Strigolactone regulates anthocyanin accumulation, acid phosphatases production and plant growth under low phosphate condition in Arabidopsis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Ito&as_ylo=2015&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28tRNAdb%5BTitle%5D%29 AND 2009%5BVolume%5D AND compilation of tRNA sequences and tRNA genes%2E Nucleic Acids Res%2E 37%3A D159%5BPagination%5D AND Juhling%2C F%2E%2C Morl%2C M%2E%2C Hartmann%2C R%2EK%2E%2C Sprinzl%2C M%2E%2C Stadler%2C P%2EF%2E%2C and Putz%2C J%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Juhling, F., Morl, M., Hartmann, R.K., Sprinzl, M., Stadler, P.F., and Putz, J. (2009). tRNAdb 2009: compilation of tRNA sequences and tRNA genes. Nucleic Acids Res. 37: D159-162.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Juhling,&hl=en&c2coff=1


http://scholar.google.com/scholar?as_q=&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Juhling,&as_ylo=2009&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Biol%2E Cell%5BTitle%5D%29 AND 100%5BVolume%5D AND 13%5BPagination%5D AND Kalantidis%2C K%2E%2C Schumacher%2C H%2ET%2E%2C Alexiadis%2C T%2E%2C and Helm%2C J%2EM%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Kalantidis, K., Schumacher, H.T., Alexiadis, T., and Helm, J.M. (2008). RNA silencing movement in plants. Biol. Cell 100: 13-26.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Kalantidis,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=RNA silencing movement in plants&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=RNA silencing movement in plants&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Kalantidis,&as_ylo=2008&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Science%5BTitle%5D%29 AND 293%5BVolume%5D AND 287%5BPagination%5D AND Kim%2C M%2E%2C Canio%2C W%2E%2C Kessler%2C S%2E%2C and Sinha%2C N%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Kim, M., Canio, W., Kessler, S., and Sinha, N. (2001). Developmental changes due to long-distance movement of a homeobox fusion transcript in tomato. Science 293: 287-289.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Kim,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Developmental changes due to long-distance movement of a homeobox fusion transcript in tomato&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Developmental changes due to long-distance movement of a homeobox fusion transcript in tomato&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Kim,&as_ylo=2001&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Nucleic Acids Res%5BTitle%5D%29 AND 40%5BVolume%5D AND D1202%5BPagination%5D AND Lamesch%2C P%2E%2C Berardini%2C T%2EZ%2E%2C Li%2C D%2E%2C Swarbreck%2C D%2E%2C Wilks%2C C%2E%2C Sasidharan%2C R%2E%2C Muller%2C R%2E%2C Dreher%2C K%2E%2C Alexander%2C D%2EL%2E%2C Garcia%2DHernandez%2C M%2E%2C Karthikeyan%2C A%2ES%2E%2C Lee%2C C%2EH%2E%2C Nelson%2C W%2ED%2E%2C Ploetz%2C L%2E%2C Singh%2C S%2E%2C Wensel%2C A%2E%2C and Huala%2C E%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Lamesch, P., Berardini, T.Z., Li, D., Swarbreck, D., Wilks, C., Sasidharan, R., Muller, R., Dreher, K., Alexander, D.L., Garcia-Hernandez, M., Karthikeyan, A.S., Lee, C.H., Nelson, W.D., Ploetz, L., Singh, S., Wensel, A., and Huala, E. (2012). The Arabidopsis Information Resource (TAIR): improved gene annotation and new tools. Nucleic Acids Res. 40: D1202-1210.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Lamesch,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=The Arabidopsis Information Resource (TAIR): improved gene annotation and new tools&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=The Arabidopsis Information Resource (TAIR): improved gene annotation and new tools&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Lamesch,&as_ylo=2012&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Nucleic Acids Res%5BTitle%5D%29 AND 40%5BVolume%5D AND W622%5BPagination%5D AND Lohse%2C M%2E%2C Bolger%2C A%2EM%2E%2C Nagel%2C A%2E%2C Fernie%2C A%2ER%2E%2C Lunn%2C J%2EE%2E%2C Stitt%2C M%2E%2C and Usadel%2C B%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Lohse, M., Bolger, A.M., Nagel, A., Fernie, A.R., Lunn, J.E., Stitt, M., and Usadel, B. (2012). RobiNA: a user-friendly, integrated software solution for RNA-Seq-based transcriptomics. Nucleic Acids Res. 40: W622-627.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Lohse,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=RobiNA: a user-friendly, integrated software solution for RNA-Seq-based transcriptomics&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=RobiNA: a user-friendly, integrated software solution for RNA-Seq-based transcriptomics&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Lohse,&as_ylo=2012&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Annu%2E Rev%2E Plant Biol%5BTitle%5D%29 AND 57%5BVolume%5D AND 203%5BPagination%5D AND Lough%2C T%2EJ%2E%2C and Lucas%2C W%2EJ%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Lough, T.J., and Lucas, W.J. (2006). Integrative plant biology: role of phloem long-distance macromolecular trafficking. Annu. Rev. Plant Biol. 57: 203-232.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Lough,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Integrative plant biology: role of phloem long-distance macromolecular trafficking&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Integrative plant biology: role of phloem long-distance macromolecular trafficking&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Lough,&as_ylo=2006&as_allsubj=all&hl=en&c2coff=1

Lough, T.J., Lee, R.H., Emerson, S.J., Forster, R.L., and Lucas, W.J. (2006). Functional analysis of the 5' untranslated region ofpotexvirus RNA reveals a role in viral replication and cell-to-cell movement. Virology 351: 455-465.


Lucas, W.J., Yoo, B.C., and Kragler, F. (2001). RNA as a long-distance information macromolecule in plants. Nat. Rev. Mol. Cell Biol.2: 849-857.


McConnell, M.J., Lindberg, M.R., Brennand, K.J., Piper, J.C., Voet, T., Cowing-Zitron, C., Shumilina, S., Lasken, R.S., Vermeesch,J.R., Hall, I.M., and Gage, F.H. (2013). Mosaic copy number variation in human neurons. Science 342: 632-637.


Macke, T.J., Ecker, D.J., Gutell, R.R., Gautheret, D., Case, D.A., and Sampath, R. (2001). RNAMotif, an RNA secondary structuredefinition and search algorithm. Nucleic Acids Res. 29: 4724-4735.


Matsuda, D., Yoshinari, S., and Dreher, T.W. (2004). eEF1A binding to aminoacylated viral RNA represses minus strand synthesis byTYMV RNA-dependent RNA polymerase. Virology 321: 47-56.


Melnyk, C.W., Molnar, A., and Baulcombe, D.C. (2011). Intercellular and systemic movement of RNA silencing signals. EMBO J. 30:3553-3563.


Molnar, A., Melnyk, C.W., Bassett, A., Hardcastle, T.J., Dunn, R., and Baulcombe, D.C. (2010). Small silencing RNAs in plants aremobile and direct epigenetic modification in recipient cells. Science 328: 872-875.


Nisar, N., Verma, S., Pogson, B.J., and Cazzonelli, C.I. (2012). Inflorescence stem grafting made easy in Arabidopsis. Plant Methods8: 50.


Ruiz-Medrano, R., Xoconostle-Cazares, B., and Kragler, F. (2004). The plasmodesmatal transport pathway for homeotic proteins,silencing signals and viruses. Curr. Opin. Plant Biol. 7: 641-650.


Takeda, R., Petrov, A.I., Leontis, N.B., and Ding, B. (2011). A three-dimensional RNA motif in Potato spindle tuber viroid mediatestrafficking from palisade mesophyll to spongy mesophyll in Nicotiana benthamiana. Plant Cell 23: 258-272.


Tasseva, G., Richard, L., and Zachowski, A. (2004). Regulation of phosphatidylcholine biosynthesis under salt stress involvescholine kinases in Arabidopsis thaliana. FEBS Lett. 566: 115-120.


Thieme, C.J., Rojas-Triana, M., Stecyk, E., Schudoma, C., Zhang, W., Yang, L., Miambres, M., Walther, D., Schulze, W.X., Paz-Ares,J., Scheible, W.-R.D., and Kragler, F. (2015). Endogenous Arabidopsis messenger RNAs transported to distant tissues. NaturePlants 1: 15025.


Yang, Y., Mao, L., Jittayasothorn, Y., Kang, Y., Jiao, C., Fei, Z., and Zhong, G.Y. (2015). Messenger RNA exchange between scionsand rootstocks in grafted grapevines. BMC Plant Biol. 15: 251.


Zhang, S., Sun, L., and Kragler, F. (2009). The phloem-delivered RNA pool contains small noncoding RNAs and interferes with

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Virology%5BTitle%5D%29 AND 351%5BVolume%5D AND 455%5BPagination%5D AND Lough%2C T%2EJ%2E%2C Lee%2C R%2EH%2E%2C Emerson%2C S%2EJ%2E%2C Forster%2C R%2EL%2E%2C and Lucas%2C W%2EJ%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Lough, T.J., Lee, R.H., Emerson, S.J., Forster, R.L., and Lucas, W.J. (2006). Functional analysis of the 5' untranslated region of potexvirus RNA reveals a role in viral replication and cell-to-cell movement. Virology 351: 455-465.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Lough,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Functional analysis of the 5' untranslated region of potexvirus RNA reveals a role in viral replication and cell-to-cell movement&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Functional analysis of the 5' untranslated region of potexvirus RNA reveals a role in viral replication and cell-to-cell movement&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Lough,&as_ylo=2006&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Nat%2E Rev%2E Mol%2E Cell Biol%5BTitle%5D%29 AND 2%5BVolume%5D AND 849%5BPagination%5D AND Lucas%2C W%2EJ%2E%2C Yoo%2C B%2EC%2E%2C and Kragler%2C F%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Lucas, W.J., Yoo, B.C., and Kragler, F. (2001). RNA as a long-distance information macromolecule in plants. Nat. Rev. Mol. Cell Biol. 2: 849-857.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Lucas,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=RNA as a long-distance information macromolecule in plants&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=RNA as a long-distance information macromolecule in plants&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Lucas,&as_ylo=2001&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Science%5BTitle%5D%29 AND 342%5BVolume%5D AND 632%5BPagination%5D AND McConnell%2C M%2EJ%2E%2C Lindberg%2C M%2ER%2E%2C Brennand%2C K%2EJ%2E%2C Piper%2C J%2EC%2E%2C Voet%2C T%2E%2C Cowing%2DZitron%2C C%2E%2C Shumilina%2C S%2E%2C Lasken%2C R%2ES%2E%2C Vermeesch%2C J%2ER%2E%2C Hall%2C I%2EM%2E%2C and Gage%2C F%2EH%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=McConnell, M.J., Lindberg, M.R., Brennand, K.J., Piper, J.C., Voet, T., Cowing-Zitron, C., Shumilina, S., Lasken, R.S., Vermeesch, J.R., Hall, I.M., and Gage, F.H. (2013). Mosaic copy number variation in human neurons. Science 342: 632-637.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=McConnell,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Mosaic copy number variation in human neurons&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Mosaic copy number variation in human neurons&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=McConnell,&as_ylo=2013&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Nucleic Acids Res%5BTitle%5D%29 AND 29%5BVolume%5D AND 4724%5BPagination%5D AND Macke%2C T%2EJ%2E%2C Ecker%2C D%2EJ%2E%2C Gutell%2C R%2ER%2E%2C Gautheret%2C D%2E%2C Case%2C D%2EA%2E%2C and Sampath%2C R%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Macke, T.J., Ecker, D.J., Gutell, R.R., Gautheret, D., Case, D.A., and Sampath, R. (2001). RNAMotif, an RNA secondary structure definition and search algorithm. Nucleic Acids Res. 29: 4724-4735.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Macke,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=RNAMotif, an RNA secondary structure definition and search algorithm&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=RNAMotif, an RNA secondary structure definition and search algorithm&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Macke,&as_ylo=2001&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Virology%5BTitle%5D%29 AND 321%5BVolume%5D AND 47%5BPagination%5D AND Matsuda%2C D%2E%2C Yoshinari%2C S%2E%2C and Dreher%2C T%2EW%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Matsuda, D., Yoshinari, S., and Dreher, T.W. (2004). eEF1A binding to aminoacylated viral RNA represses minus strand synthesis by TYMV RNA-dependent RNA polymerase. Virology 321: 47-56.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Matsuda,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=eEF1A binding to aminoacylated viral RNA represses minus strand synthesis by TYMV RNA-dependent RNA polymerase&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=eEF1A binding to aminoacylated viral RNA represses minus strand synthesis by TYMV RNA-dependent RNA polymerase&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Matsuda,&as_ylo=2004&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28EMBO J%5BTitle%5D%29 AND 30%5BVolume%5D AND 3553%5BPagination%5D AND Melnyk%2C C%2EW%2E%2C Molnar%2C A%2E%2C and Baulcombe%2C D%2EC%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Melnyk, C.W., Molnar, A., and Baulcombe, D.C. (2011). Intercellular and systemic movement of RNA silencing signals. EMBO J. 30: 3553-3563.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Melnyk,&hl=en&c2coff=1

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http://scholar.google.com/scholar?as_q=Intercellular and systemic movement of RNA silencing signals&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Melnyk,&as_ylo=2011&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Science%5BTitle%5D%29 AND 328%5BVolume%5D AND 872%5BPagination%5D AND Molnar%2C A%2E%2C Melnyk%2C C%2EW%2E%2C Bassett%2C A%2E%2C Hardcastle%2C T%2EJ%2E%2C Dunn%2C R%2E%2C and Baulcombe%2C D%2EC%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Molnar, A., Melnyk, C.W., Bassett, A., Hardcastle, T.J., Dunn, R., and Baulcombe, D.C. (2010). Small silencing RNAs in plants are mobile and direct epigenetic modification in recipient cells. Science 328: 872-875.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Molnar,&hl=en&c2coff=1

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http://scholar.google.com/scholar?as_q=Small silencing RNAs in plants are mobile and direct epigenetic modification in recipient cells&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Molnar,&as_ylo=2010&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Plant Methods%5BTitle%5D%29 AND 8%5BVolume%5D AND 50%5BPagination%5D AND Nisar%2C N%2E%2C Verma%2C S%2E%2C Pogson%2C B%2EJ%2E%2C and Cazzonelli%2C C%2EI%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Nisar, N., Verma, S., Pogson, B.J., and Cazzonelli, C.I. (2012). Inflorescence stem grafting made easy in Arabidopsis. Plant Methods 8: 50.

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http://scholar.google.com/scholar?as_q=Inflorescence stem grafting made easy in Arabidopsis&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Inflorescence stem grafting made easy in Arabidopsis&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Nisar,&as_ylo=2012&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Curr%2E Opin%2E Plant Biol%5BTitle%5D%29 AND 7%5BVolume%5D AND 641%5BPagination%5D AND Ruiz%2DMedrano%2C R%2E%2C Xoconostle%2DCazares%2C B%2E%2C and Kragler%2C F%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Ruiz-Medrano, R., Xoconostle-Cazares, B., and Kragler, F. (2004). The plasmodesmatal transport pathway for homeotic proteins, silencing signals and viruses. Curr. Opin. Plant Biol. 7: 641-650.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Ruiz-Medrano,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=The plasmodesmatal transport pathway for homeotic proteins, silencing signals and viruses&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=The plasmodesmatal transport pathway for homeotic proteins, silencing signals and viruses&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Ruiz-Medrano,&as_ylo=2004&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Plant Cell%5BTitle%5D%29 AND 23%5BVolume%5D AND 258%5BPagination%5D AND Takeda%2C R%2E%2C Petrov%2C A%2EI%2E%2C Leontis%2C N%2EB%2E%2C and Ding%2C B%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Takeda, R., Petrov, A.I., Leontis, N.B., and Ding, B. (2011). A three-dimensional RNA motif in Potato spindle tuber viroid mediates trafficking from palisade mesophyll to spongy mesophyll in Nicotiana benthamiana. Plant Cell 23: 258-272.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Takeda,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=A three-dimensional RNA motif in Potato spindle tuber viroid mediates trafficking from palisade mesophyll to spongy mesophyll in Nicotiana benthamiana&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=A three-dimensional RNA motif in Potato spindle tuber viroid mediates trafficking from palisade mesophyll to spongy mesophyll in Nicotiana benthamiana&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Takeda,&as_ylo=2011&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28FEBS Lett%5BTitle%5D%29 AND 566%5BVolume%5D AND 115%5BPagination%5D AND Tasseva%2C G%2E%2C Richard%2C L%2E%2C and Zachowski%2C A%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Tasseva, G., Richard, L., and Zachowski, A. (2004). Regulation of phosphatidylcholine biosynthesis under salt stress involves choline kinases in Arabidopsis thaliana. FEBS Lett. 566: 115-120.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Tasseva,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Regulation of phosphatidylcholine biosynthesis under salt stress involves choline kinases in Arabidopsis thaliana&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Regulation of phosphatidylcholine biosynthesis under salt stress involves choline kinases in Arabidopsis thaliana&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Tasseva,&as_ylo=2004&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Nature Plants%5BTitle%5D%29 AND 1%5BVolume%5D AND 15025%5BPagination%5D AND Thieme%2C C%2EJ%2E%2C Rojas%2DTriana%2C M%2E%2C Stecyk%2C E%2E%2C Schudoma%2C C%2E%2C Zhang%2C W%2E%2C Yang%2C L%2E%2C Miambres%2C M%2E%2C Walther%2C D%2E%2C Schulze%2C W%2EX%2E%2C Paz%2DAres%2C J%2E%2C Scheible%2C W%2E%2DR%2ED%2E%2C and Kragler%2C F%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Thieme, C.J., Rojas-Triana, M., Stecyk, E., Schudoma, C., Zhang, W., Yang, L., Miambres, M., Walther, D., Schulze, W.X., Paz-Ares, J., Scheible, W.-R.D., and Kragler, F. (2015). Endogenous Arabidopsis messenger RNAs transported to distant tissues. Nature Plants 1: 15025.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Thieme,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Endogenous Arabidopsis messenger RNAs transported to distant tissues&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Endogenous Arabidopsis messenger RNAs transported to distant tissues&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Thieme,&as_ylo=2015&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28BMC Plant Biol%5BTitle%5D%29 AND 15%5BVolume%5D AND 251%5BPagination%5D AND Yang%2C Y%2E%2C Mao%2C L%2E%2C Jittayasothorn%2C Y%2E%2C Kang%2C Y%2E%2C Jiao%2C C%2E%2C Fei%2C Z%2E%2C and Zhong%2C G%2EY%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Yang, Y., Mao, L., Jittayasothorn, Y., Kang, Y., Jiao, C., Fei, Z., and Zhong, G.Y. (2015). Messenger RNA exchange between scions and rootstocks in grafted grapevines. BMC Plant Biol. 15: 251.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Yang,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Messenger RNA exchange between scions and rootstocks in grafted grapevines&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Messenger RNA exchange between scions and rootstocks in grafted grapevines&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Yang,&as_ylo=2015&as_allsubj=all&hl=en&c2coff=1

translation. Plant Physiol. 150: 378-387.Pubmed: Author and TitleCrossRef: Author and TitleGoogle Scholar: Author Only Title Only Author and Title

Zhang, W., Kollwig, G., Stecyk, E., Apelt, F., Dirks, R., and Kragler, F. (2014). Graft-transmissible movement of inverted-repeat-induced siRNA signals into flowers. Plant J. 80: 106-121.


http://www.ncbi.nlm.nih.gov/pubmed?term=%28Plant Physiol%5BTitle%5D%29 AND 150%5BVolume%5D AND 378%5BPagination%5D AND Zhang%2C S%2E%2C Sun%2C L%2E%2C and Kragler%2C F%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Zhang, S., Sun, L., and Kragler, F. (2009). The phloem-delivered RNA pool contains small noncoding RNAs and interferes with translation. Plant Physiol. 150: 378-387.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Zhang,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=The phloem-delivered RNA pool contains small noncoding RNAs and interferes with translation&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=The phloem-delivered RNA pool contains small noncoding RNAs and interferes with translation&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Zhang,&as_ylo=2009&as_allsubj=all&hl=en&c2coff=1

http://www.ncbi.nlm.nih.gov/pubmed?term=%28Plant J%5BTitle%5D%29 AND 80%5BVolume%5D AND 106%5BPagination%5D AND Zhang%2C W%2E%2C Kollwig%2C G%2E%2C Stecyk%2C E%2E%2C Apelt%2C F%2E%2C Dirks%2C R%2E%2C and Kragler%2C F%2E%5BAuthor%5D&dopt=abstract

http://search.crossref.org/?page=1&rows=2&q=Zhang, W., Kollwig, G., Stecyk, E., Apelt, F., Dirks, R., and Kragler, F. (2014). Graft-transmissible movement of inverted-repeat-induced siRNA signals into flowers. Plant J. 80: 106-121.

http://scholar.google.com/scholar?as_q=&num=10&as_occt=any&as_sauthors=Zhang,&hl=en&c2coff=1

http://scholar.google.com/scholar?as_q=Graft-transmissible movement of inverted-repeat-induced siRNA signals into flowers&num=10&btnG=Search+Scholar&as_epq=&as_oq=&as_eq=&as_occt=any&as_publication=&as_yhi=&as_allsubj=all&hl=en&lr=&c2coff=1

http://scholar.google.com/scholar?as_q=Graft-transmissible movement of inverted-repeat-induced siRNA signals into flowers&num=10&btnG=Search+Scholar&as_occt=any&as_sauthors=Zhang,&as_ylo=2014&as_allsubj=all&hl=en&c2coff=1

DOI 10.1105/tpc.15.01056; originally published online June 7, 2016;Plant Cell

Andresen, Dirk Walther and Friedrich KraglerWenna Zhang, Christoph Thieme, Gregor Kollwig, Federico Apelt, Lei Yang, Winter Winter, Nadine

tRNA-related sequences trigger systemic mRNA transport in plants

This information is current as of June 17, 2018

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tRNA-related sequences trigger systemic mRNA … 1 tRNA-related sequences trigger systemic mRNA transport in plants 2 3 Authors: 4 Wenna Zhanga, Christoph J. Thiemea, Gregor Kollwigb,

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