Indian River Research and Education Center, Florida University, USA.
School of Agriculture, Ningxia University, China
I
Contents
Identification, Expression, and Functional Analysis of the Galactose metabolism gene
LBGALA and Gene Family in goji berry (Lycium barbarum L.) .................................. 1
Transcriptomic analysis of periwinkle (Catharanthus roseus) in response to Clas
(Candidatus Liberibacter asiaticus) infection and characterization of
infection-promoting effectors ....................................................................................... 3
Low Concentration of Sodium Chloride by Foliar Spraying Promote the Growth and
Photosynthesis of Non-heading Chinese Cabbage ....................................................... 5
Comprehensive Evaluation of Different Fertilizer Schemes on the Growth of
Watermelon Seedlings................................................................................................... 7
Responses of Ziziphus jujuba Mill.cv.Lingwuchangzao Fruit Coloration and
Anthocyanin Synthesis Related Gene Expression to Elevated Temperature and
Drought Stress ............................................................................................................... 8
Screening , Identification and Optimization of Fermentation Conditions of Biocontrol
Strain WQ-6 to Melon Fusarium Wilt Disease ........................................................... 10
Morphology and Glucosinolate Metabolism of Chinese Kale in Response to
Supplementary Far-red Light ...................................................................................... 13
Effects of Zeolitic Urea on Growth and Yield of Wheat ............................................ 16
Changes in soil organic carbon pools and enhanced crop productivity in sugarcane
bagasse-biochar amended Soil under wheat-maize cropping system of Pakistan ...... 17
Nanotechnology for Reduction and Remediation of Nonpoint Source of Pollution .. 19
Compost Utilization in Horticultural Cropping Systems ............................................ 20
Evaluation of nitrate leaching and ammonia volatilization from carbon based
designers’ fertilizers in alkaline calcareous soils ........................................................ 21
Analysis in Vitro Culture and Light Quality Effect of Lewisia cotyledon ................. 22
Nutrient Use Efficiency in Crops: A Comprehensive Approach................................. 24
Proliferation and Flowering in vitro of Tissue Culture Seedlings of Lewisia cotyledon
based on Orthogonal Design ....................................................................................... 26
II
Plant growth regulators and EDTA improve phytoremediation potential and
antioxidant response of Chenopodium ambrosioides L. in a Cd-spiked soil .............. 28
Plants buffer against high light stress-induced transcript of CsGA2ox8 via alternative
splicing to fine-tune gibberellins level and maintain hypocotyl elongation ............... 30
Seeds priming technology as a key strategy to increase crops production under
adverse environmental conditions ............................................................................... 32
Effects of greenhouse new/waste rockwool and nutrient on growth and quality of
pakchoi ........................................................................................................................ 33
Purification ability from constructed wetlands treating sewage treatment plants
effluent and microbial community succession in epiphytic biofilms ......................... 34
Impacts of water flow and suspended sediment on microbial community in epiphytic
biofilm on two submerged macrophytes ..................................................................... 36
Aeration enhanced nutrient removal and increased microbial diversity during
Hydrilla verticillata decomposition in wetlands in winter ......................................... 38
Detection of chlorophyll content in tomato leaves based on portable hyperspectral
imaging technique ....................................................................................................... 40
Research of Volatile Components on Champagne and Japanese Peppermint Stem and
Leaf Tissues ................................................................................................................ 42
Study on Visualization Distribution of Chlorophyll in Tomato Leaves under brackish
irrigation Based on Hyperspectral Imaging Technology ............................................ 44
Comparison and Screening of Total RNA Extraction Methods from Fennel Leaves 46
Analysis of Flavor Compounds in Grapefruit Mint Leaf Stem Based on GC-IMS.... 48
Comparative Analysis of Volatile Components in Leave and Stem of the Mentha
spicala by GC-IMS ..................................................................................................... 50
The Identity of Pathongen Causing Tomato Powdery Mildew in Yinchuan .............. 52
Applications of Environmental Economic Policies in the Prevention and Control of
Agricultural Non-point Source Pollution in China ..................................................... 54
1
Identification, Expression, and Functional Analysis of the Galactose
metabolism gene LBGALA and Gene Family in goji berry
(Lycium barbarum L.)
Yaping Ma1,2, Yun Xie2, Naigong Wang3, Bing Cao2,
Lihua Song2, Handong Gao1
1 College of Forestry, Nanjing Forestry University, Nanjing, China 2 School of Agriculture, Ningxia University, Yinchuan, China 3 Zhongke Redflag Information Technology Co., Ltd., Beijing, China
Objectives Exposing goji berry to elevated CO2 for long periods has been shown to
reduce sugar and secondary metabolite contents, and gene LBGALA (Lycium
barbarum alpha-galactosidase) has been found to play a key regulatory role in sugar
metabolism pathways. The objective of this study was to identify gene LBGALA
family members based on the transcriptome, analyze their character and expression
patterns, further understand the initial regulatory functions to sugar metabolism.
Methods The experiment was conducted in open-top chambers (OTC) with two
treatments: ambient (400 μmol mol−1 CO2), elevated (700 μmol mol−1 CO2), and after
90 and 120 days the root, stem, leaf, and fruit tissue samples were collected for
qRT-PCR expression analysis and rapid amplification of cDNA ends (RACE).
Potential members of the LBGALA gene family were identified based on the de novo
transcriptome database with the Hidden Markov model (HMM) and BLAST
homology searches. Green fluorescent protein (GFP) was used for subcellular
localization assay.
Results Six genes (named LBGALA1~LBGALA6) were identified as members of the
LBGALA gene family, one of which exhibited complete open reading frames (ORFs).
The full-length cDNA fragments of LBGALA (GenBank accession No. MH025913)
obtained by RACE-PCR was 1233 bp encoding 410 amino acids. Bioinformatics
analysis showed that the length of LBGAL family members proteins ranged from 101
to 634 aa, with the predicted molecular weight varied from 11.50 to 71.46 KD and
the isoelectric point in the range to 4.86 to 8.74. Phylogenetic analysis revealed that
LBGALA family members are grouped. All LBGALAs proteins contain the typical
Melibiase domain and conservative motif analysis shows that there are 10 motifs in
LBGALA family proteins. The expression of LBGALAs was expressed in different
2
tissues of goji berry under different CO2 treatments with 90 and 120 days.
Sub-cellular localization showed that LBGALA and LBGALA1 were located in the
chloroplast.
Conclusions Six LBGALA genes from the Lycium barbarum transcriptome were
identified and contain a typical Melibiase domain. The expression patterns of the
LBGALAs under different CO2 concentrations and tissues suggested that significantly
expressed in root, leaf, stem, and fruit. This study provides a basis for further
research on the regulation function of LBGALA to sugar metabolism.
Acknowledgments This work was supported by the National Natural Science
Foundation of China (Grants No. 31660199).
3
Transcriptomic analysis of periwinkle (Catharanthus roseus) in
response to Clas (Candidatus Liberibacter asiaticus) infection
and characterization of infection-promoting effectors
Qiaolin Zheng1, Marco Pitino1, YongPing Duan2
and Liliana M. Cano1
1 University of Florida, Institute of Food and Agricultural Sciences IFAS, Department of Plant
Pathology, Indian River Research and Education Center, Fort Pierce, FL 34945, USA 2 U.S. Horticultural Research Laboratory, USDA-ARS, Fort Pierce, FL 34945, FL, USA
Background Huanglongbing (HLB), the most destructive disease of citrus, causes
quick decline of citrus production all over the world and has rendered millions of
others useless for production. HLB-associated Candidatus Liberibacter asiaticus
(Clas) is the most widespread pathogen in citrus and transmitted by insect vector
Asian citrus psyllid (ACP, Diaphorina citri). HLB was first discovered in the United
States in Florida in 2005, which reduced the value of Florida citrus output by $7
billion during the years from 2005 to 2014 and caused 72.2% reduction in the
production of oranges for processing and 20.5% reduction in the fresh fruit market in
the United States from 2007-08 to 2017-18. In order to develop novel strategies to
completely eliminate HLB in citrus, one of the most important aspects is to clarify
the mechanism of citrus-Clas interactions. Periwinkle (Catharanthus roseus) is
HLB-susceptible host plant and presents similar infected-symptoms observed in
citrus. Moreover, it is easily grown and quickly infected by Clas through grafting.
Clas bacteria present in periwinkle plants can grow faster and reach higher titer level
than in citrus.
Methods Two-month old healthy periwinkle plants were grafted inoculation with
Clas infected branches. The leaves of the grafted branches were collected at 2 and 4
weeks after grafting (wag) for RNA extraction. The expression of a total of 27 Clas
effectors was assayed with RT-qPCR methods in Clas-infected periwinkle leaves.The
transcriptome analysis of periwinkle infected with Clas was performed as well.
Results A total of 14,062 genes were considered differentially expressed using a fold
change cutoff of Gfold (0.01) less than -1 and greater than 1 in Clas-infected
periwinkle at 2 wag, and 9,509 DEGs (Differentially expressed genes) were
identified at 4 wag. Eleven DEGs were validated via qRT-PCR, and all of them were
up-regulated in response to Clas infection in periwinkle, including DEGs involved in
4
transcription factors, secondary metabolism pathways, signal pathways, stress, cell
wall modification, transporters, and starch synthesis. Gene expression of 19 putative
effectors was detectable in Clas-infected periwinkle leaves at early infection stage, and
effector ‘0470’showed the highest expression level. In total, 27 putative effectors were
highly expressed at 4 wag in Clas-infected periwinkle, and the expression level of
each effector was thousands of times higher than that expressed at 2 wag.
Conclusions Transcriptional profiling of the genes involved in Clas infection was
investigated in Clas-infected periwinkle at 2 and 4 wag. Our study indicated larger
numbers of genes with much higher expression level were induced at 2 wag than 4 wag
in response to Clas infection, as well as the up-regulated genes much more than the
down-regulated ones. The numbers of detectable effectors and the transcriptional level
of effector were related to the titers of bacteria in infected periwinkle leaves. The
expression of 19 and 27 effectors were observed at 2 and 4 wag, respectively. The
expression level of effectors at 4 wag was thousands of times higher than that at 2 wag.
5
Low Concentration of Sodium Chloride by Foliar Spraying Promote
the Growth and Photosynthesis of Non-heading Chinese Cabbage
Wei Zhang1, Shuang Gao2, Junlin Wang3, Mei Wang4,
Zhikui Gao1*, Zhanjun Xue1*
1 Collage of Horticulture,Hebei Agricultural University, Baoding , China 2 College of Food Science and Technology, Agricultural University of Hebei, Baoding,China 3 College of Life Sciences, Hebei Agricultural University, Baoding , China 4 Center of Science and Education Service,Hebei Agricultural University, Baoding, China
Objectives Many researches have been paied more attentions for the stress effects of
higher concentration (50-300 mmol·L-1) sodium chloride (NaCl), which were directly
used into soil or nutrient solution. Recently, studies have shown that there are
beneficial effects from the addition of low NaCl as a nutrient solution to the
rhizosphere and this practice promotes the growth of various plants. Although low
concentration NaCl is applied repeatedly in soil cultivation, it is likely to also create
hidden dangers of secondary soil salinization. So the foliar spraying method was used
to study the regulatory effect of low concentration of NaCl on the growth and
photosynthetic activity of non-heading Chinese cabbage and provide a basis for
broadening the application of NaCl.
Methods The non-heading Chinese Cabbage variety 'Hanyue F1' in the three–leaf
stage was used as the test material growing in nutritive bowl under the greenhouse.
Low concentrations of NaCl (6 mmol·L-1) were applied once a day as a foliar spray at
every morning during the growth period with distilled water (0 mmol·L-1) as the
control. After the leaves were sprayed with the low concentrations of NaCl lasting for
18 days, the growth, photosynthesis, water holding and nutrient patameters were
determined.
Results The fresh biomass, dry biomass and water content of the whole plant
increased significantly by 49.5%, 28.7%, and 51.5%, respectively. The water holding
capacity of the WHCplant, leaf area, leaf succulence, the photosynthetic carbon
absorption rate(Pn) and the stomatal conductance Gs also increased significantly.
Simultaneously, treatment with NaCl not only increased the dry biomass of leaves,
particularly the petioles, but also increased the water content of all the organs,
particularly the stem. In addition, the treatment increased the dry biomass water
holding capacity of the leaves and stems of non-heading Chinese cabbage. The
6
content of free amino acids in the leaves and petioles of Chinese cabbage increased
significantly by 73.4% and 58.8%, respectively, following the application of NaCl to
the leaves. In addition, the NaCl treatment in three–leaf stage stimulated the
expanding leaves (the second and third euphyllas), particularly for the leaves that
were about to unfold (the fourth to the eighth euphyllas).
Conclusions Spraying treatment of a low concentration of NaCl not only increased
the fresh content of non-heading Chinese cabbage through the accumulation of water
and dry biomass, but also increased the expansion of leaf area, the light interception
area and the stomatal conductance (GS), so as to improve the Pn and dry biomass
accumulation.
Acknowledgments This work was supported by the Key R & D Projects of Hebei
Province (18226928D).
E-mail to corresponding author [email protected]; [email protected]
7
Comprehensive Evaluation of Different Fertilizer Schemes
on the Growth of Watermelon Seedlings
LIANG Huan, ZHU Juhong, GE Mihong, WANG Dehuan,
ZHOU Mobing, SHI Xianfeng *
Wuhan Academy of Agricultural Sciences,Wuhan 430345
Abstract 【Objective】The effects of different fertilization amount and concentration
on the growth and development of watermelon seedlings were studied in order to
identification the fertilization scheme of watermelon seedling.【Method】Taking the
‘zaojia 84-24’ watermelon seedlings as the research objects, the morphological
parameters, such as the plant height, stem diameter and so on, were measured. A
comprehensive evaluation method containing principal component, membership
functions and system clustering was applied to analyze indices in different fertilizer
treatment. 【Result】There were significant differences in the 11 indexes after
different fertilizer treatment, such as plant height, stem thickness, leaf area, fresh
weight and so on. By principal component analysis, 11 indexes were transformed into
3 independent comprehensive indexes. Cluster analysis showed that D5、E5 and E6
treated seedlings had the best quality. 【Conclusion】The fertilization scheme of
watermelon seedling was as follows: the fertilization rate respectively were 250
mg∙L-1, 250~300 mg∙L-1and 250~350 mg∙L-1 at three developmental stage, including
cotyledon extension, the 1st leave stage and 2nd leave stage.
Key words Watermelon, Plug seedling, Fertilization scheme; Comprehensive
evaluation
8
Responses of Ziziphus jujuba Mill.cv.Lingwuchangzao Fruit
Coloration and Anthocyanin Synthesis Related Gene
Expression to Elevated Temperature
and Drought Stress
Wenqian Jiang, Lihua Song
School of Agriculture, Ningxia University, Yinchuan,China
Background Increased temperature and drought are among the most important
factors challenging sustainable agricultural production worldwide. The world average
temperature is likely to increase by 1.5 °C between 2030 and 2052 and 2.3 ± 0.3 °C
by 2076. Along with these increased temperatures, it is also predicted that there will
be changes to precipitation patterns in various regions. Ningxia, located in
northwestern China, is an arid and semi-arid region. In the past 60 years, the
temperature has been rising and the amount of evaporation has also been increasing.
Therefore, the elevated temperature and drought in Ningxia will become more and
more obvious, which will impact the fruit coloration of ‘Lingwuchangzao’ (Ziziphus
jujuba Mill. cv. ‘Lingwuchangzao’), which is one of featured economic forest tree
species.
Methods We applied the air temperature and drought interaction design. The
treatment of drought stress included three levels (D1, soil moisture is 70 - 75 % of
field capacity; D2, soil moisture is 50 - 55% of field capacity and D3, soil moisture is
30 - 35 % of field capacity), whereas the treatment of air temperature included two
levels (T1, natural air temperature; T2, elevated air temperature=T1+(2 (±0.5) °C).
With 5-year-old grafted seedlings of jujube cultivar ‘Lingwuchangzao’ as the the
experimental objects, the Open-top chambers (OTC) were used to simulate the
elevated temperature environment, solar automatic irrigation control system to
control the soil moisture. By measuring the fruit sugar, acid content, color index, peel
pigment content, anthocyanin synthase activity and gene expression under different
treatments at different stages, to analyze the effects of elevated temperature and
drought stress on fruit coloration and anthocyanin synthesis of ‘Lingwuchangzao’.
Results 1.The elevated temperature significantly increased a* (red-green difference)
(13.68%) and a*/b* (red-green difference/yellow-blue difference) (10.63%), soluble
sugar content (39.92%), the content of flavonoid and carotenoid, the expression of
9
ZjCHS, ZjF3H, ZjDFR, ZjUFGT and ZjLDOX genes. Drought treatments
significantly decreased a*, a*/b*, soluble sugar content, the content of flavonoid and
carotenoid and this genes expression. Elevated temperature significantly increased
the activity of UFGT (6.90%), PAL (6.20%), and DFR (5.47%) during the coloration
stage, while drought treatments significantly reduced the activity of UFGT, PAL, and
DFR, especially the severe drought treatment.
2.The L* and h* of the D3T1 treatment combination were the highest, which
were significantly higher than other treatment combinations, indicating that the fruits
under D3T1 treatment combination had poor coloration. In the full-ripening stage, the
peel anthocyanin content, the activity of UFGT, PAL and CHI, the ZjF3H gene
expression of the D1T2 treatment combination was significantly higher than D1T1
treatment combination.
3. Soluble sugar content was always significantly or highly significantly
positively correlated with anthocyanin content during the coloration and full-ripening
stage (r=0.526*, 0.677**), and the organic acid content is highly significantly
negatively correlated with anthocyanin content during the coloration stage (r=
-0.894**).
4. During the fruit coloration stage and the full-ripening stage, the activity of
UFGT, PAL, and DFR were significantly or highly significantly positively correlated
with the synthesis and accumulation of anthocyanin. And ZjLDOX, ZjUFGT and
ZjDFR are the key genes for the anthocyanin synthesis pathway of ‘Lingwuchangzao’
peel under elevated temperature and drought in the fruit coloration process.
Conclusions The elevated temperature by about 2.0℃ promoted the coloration of
‘Lingwuchangzao’ by increasing the soluble sugar content of fruits, and promoted the
activity of UFGT, PAL, DFR and the expression of ZjCHS, ZjDFR, ZjUFGT, and
ZjLDOX genes. Drought stress is not conducive to fruit coloration. ZjLDOX, ZjUFGT
and ZjDFR are the key genes in the anthocyanin synthesis pathway of the pericarp of
‘Lingwucahngzao’. Elevated temperature with normal soil moisture is the best
environment of ‘Lingwuchangzao’ fruit coloration by promoting UFGT, PAL, CHI
activity and ZjCHS, ZjF3H gene expression.
Acknowledgements This work was supported by the National Nature Science
Foundation of China (Grants No. 31760203).
10
Screening , Identification and Optimization of Fermentation
Conditions of Biocontrol Strain WQ-6 to
Melon Fusarium Wilt Disease
Wenwen Zhang, Qing WU, Cui Yin, and Yune Cao
School of Agriculture,Ningxia University,Yinchuan,China
Background A lot of studies have been done on the screening and application of
antagonistic bacteria against wilt of cucurbit crops. But most of the studies. The
biocontrol bacteria in the plant is only targeted at a single type of fusarium oxysporum,
which has a single effect in disease control and has great limitations in production and
application. It is necessary to screen the biocontrol bacteria that have broad spectrum
resistance to various fusarium oxysporum. Metabolic active substances are the key to
the biocontrol effect of bacterial strains. Appropriate fermentation technology is the
basis for obtaining a large number of metabolically active substances from
microorganisms. Based on this, in this study different cucumber, watermelon and The
fusarium wilt bacteria of muskmelon was selected as the target bacteria, and the
antibacterial properties of the strains and their metabolites were selected as the
screening conditions to be screened from the earthworm compost. The birth control
strain was selected to study its broad-spectrum control effect and application
effect.The antagonistic strains were optimized by single factor test and response
surface analysis. In this paper, the effect of antagonistic strain on melon fusarium wilt
was verified by seed germination test and pot experiment, and the antagonistic strain
was studied. In order to provide new species resources for the biological control of
cucurbit fusarium wilt, the biocontrol effect of plant compound and earthworm
composting extract on fusarium wilt was studied. Field application lays the foundation.
Methods Four kinds of fusarium wilt pathogens (2 kinds of cucumber, 1 kind of
watermelon and 1 kind of melon) were selected as the pathogens, and the flat plate
confrontation method combined with the method of inhibiting mycelium growth rate
was adopted to select 1 strain of antagonistic bacteria from earthworm compost that
had significant inhibitory effect on all four kinds of pathogenic bacteria. Physiological,
biochemical and molecular identification were carried out for the screened strains with
inhibitory effect to determine the species. The optimum fermentation technology was
obtained by using single factor test and response surface analysis method, which
11
increased the antibacterial diameter and antibacterial rate of bacterial strain
fermentation liquid against fusarium wilt. Finally, the antagonistic bacteria were tested
to meet the environmental requirements of biocontrol agents to ensure that they could
colonize the root system of crops.
Results A new strain WQ-6 exhibiting antagonism towards all four pathogens tested,
was screened out using the methods of plate-confrontation and inhibition mycelial
growth and was identified as Bacillus siamensis. Based on single factor test and
response surface analysis,an response surface analysis,an optimal fermentation
condition for strain WQ-6(i.e. glucose 30 g·L -1,peptone 30 g·L-1 ,magnesium sulfate
1g·L-1,glycerol 1g·L-1,dipotassium hydrogen phosphate 1.5g·L-1,temperature 32.2 ℃,
liquid-loading rate 19.2%,rotating speed 236.2 r·min-1,pH 6.0,inoculation rate 4%
and cultural time 24 hours)was obtained. Under the optinal condition,the inhibition
zone diameter increased by 3.52–10.45 mm and the inhibition rate of pathogens
increased by 2.28%–32.16%,as compared to the unoptimized control. The seed
germination test (cucumber,watermelon and muskmelon)and pot experiment showed
that the strain WQ-6 reduced the disease incidence of seeds by 32%–68%,increased
germination rate by 100%–259.6%,and decreased the disease index of plants by 20%–
34%. In addition, the WQ-6 fermentation broth(WFB)mixed with water extract of
vermicompost significantly enhanced the root length,stem length and vigor index of
seeds,and reduced the disease index of plants by 6.78%–36.44%,as compared with
WFB alone.
Conclusions Among the existing biocontrol bacteria against cucurbit fusarium wilt,
most of them are bacteria, especially bacillus. Bacillus has the ability to inhibit a
variety of plant pathogens and is a widely used biocontrol strain. In this study, B.
Siamensis had a good antagonistic effect on cucumber Fusarium wilt, melon Fusarium
wilt and watermelon fusarium wilt (the inhibitory diameter was 28.32 ~ 31.32 mm, and
the inhibitory rate was 51.65% ~ 59.15%).The selection of medium components
suitable for the growth of the strain and the optimum fermentation conditions were
beneficial to increase the fermentation yield of the strain and give full play to its
biocontrol potential. In this study, based on the single factor experiment, response
surface analysis method designed by Plackett Burman experiment Design and Central
Composite Design was adopted to optimize the fermentation process of strain WQ-6 as
a whole, which simplifies the test steps, is intuitive and has high reliability. Compared
12
with that before optimization, the biocontrol ability of the strain was significantly
increased, and the inhibition diameter and inhibition rate of fusarium wilt increased by
14.1% ~ 50.1% and 4.3% ~119.2%, respectively. At the same time, the control effect
test results showed that the incidence of seed decreased by 32% ~ 68%, the
germination rate increased by 100% ~ 259.58%, and the disease index of plant
decreased by 20% ~ 34%.The results showed that wQ-6 had a good effect of
antagonizing cucurbit wilt and promoting growth. The root length, stem length and
vigor index of cucurbit seed germination were significantly higher than that of
pathogen treatment with wQ-6 fermentation broth alone, and the prevention and
control effect on plants was improved by 6.78% ~ 36.44%, indicating that wQ-6
combined with earthworm compost extract could improve its ability to prevent
diseases and promote growth. At the same time, the fermentation conditions of wQ-6
strain were optimized as a whole to determine a set of fermentation mode with the
highest yield, the most economical and effective, which laid a foundation for deep
fermentation of WQ-6 strain and large-scale industrial production, and provided new
biocontrol resources for the prevention and control of soil-borne diseases of facility
vegetables.
13
Morphology and Glucosinolate Metabolism of Chinese Kale
in Response to Supplementary Far-red Light
Yamin Li1 and Houcheng Liu1
1 College of Horticulture, South China Agricultural University, Guangzhou 510642, China
Objectives Chinese kale (Brassica alboglabra Bailey) is one of the most popular
Brassicaceae vegetables in South China, which is known for high nutritional value
and abundant glucosinolates (GSL). GSL are an important group of secondary
metabolites that participate in plant stress responses and flavor formation (pungency
and bitterness). Although far-red light is beyond the visible light, it has been proven
to greatly influence plant growth, development, as well as metabolite accumulation.
This study aims to explore how Chinese kale in response to far-red light.
Methods The Chinese kale cv. ‘Lv Bao’ were cultivated under 250 μmol m–1 s–1
white light-emitting diodes (6:00 - 18:00) in the plant factory. There were three
supplemental far-red (735 nm) light treatments: 0 W/m2 (CK), 3 W/m2 (FR-3) and 6
W/m2 (FR-6). The transcriptomics profiles, agronomic traits, and individual GLs
were combined to investigate the regulation mechanism of Chinese kale in response
to far-red light.
Results (1) Along with the increased far-red light the plant display, plant height,
internode length, leaf length, leaf width, and leaf area of Chinese kale significantly
enlarged, while the leaf angle narrowed. (2) Higher supplemental far-red light
obviously accelerated the transformation from vegetative growth to reproductive
growth in Chinese kale by increasing the flower budding rate at 60 days after sown.
(3) Nine individual GSL were detected in Chinese kale. The total GSL, aliphatic GSL,
and indolic GSL were significantly decreased by far-red light, especially FR-6. Eight
kinds of GLs had the similar decreased trend with total GSL, while the content of
4-Hydroxyglucobrassicin were increased by FR-3. (4) Based on Brassica database,
210 genes were identified as GSL biosynthesis genes homologous to Arabidopsis
thaliana. In the chain elongation step, genes annotated MAM1/2, IPMDH and BAT5
were down-regulated by far-red light. In the core structure formation step, genes
coding CYP79F1, CYP83A1, SUR1, UGT74B1, UGT74C1, GSTU20, and GSTF9/10
were also down-regulated by far-red light.
14
Conclusions In conclude, higher far-red light supplementation showed stronger
influence on the growth and metabolism of Chinese kale. The results indicate that
far-red light supplementation could be used strategically to modify a larger and more
compact appearance of Chinese kale. Meanwhile, far-red light supplementation is
efficient in alleviating the pungency and bitterness flavor by suppressing the GSL
biosynthesis and consequently reducing the GSL contents of Chinese kale.
Acknowledgements This work was supported by Key Research and Development
Program of Guangdong (2019B020214005, 2019B020222003).
Figure 1. Effect of supplementary far-red light on Chinese kale morphology.
The morphological indices (a-k) of Chinese kale 60 days after sown.
15
Figure 2. Effect of supplementary far-red light on the contents of glucosinolates (GSL) in Chinese kale.
Relative abundance of (a) aliphatic and (b) indolic GSL is indicated from blue (low) to orange (high). (c) Relative
expression of genes involved in GSL biosynthesis is indicated from green (low) to red (high). PRO = progoitrin,
RAA=glucoraphanin, SIN=sinigrin, GNA=gluconapin, GBN=glucobrassicanapin, 4OH=4-hydroxyglucobrassicin,
GBC=glucobrassicin, 4ME=4-methoxyglucobrassicin, NEO=neoglucobrassicin.
PRO
RAA
SIN
GNA
GBN
4OH
GBC
4M E
NEO
CKFR-3
FR-6
-1.20
-0.90
-0.60
-0.30
0.00
0.30
0.60
0.90
1.20
M ethionine
RAA GBNSIN
GNA
Aliphatic GSL
PRO
(a)
Desulfo GSL
PRO
RAA
SIN
GNA
GBN
4OH
GBC
4M E
NEO
CK FR-3
FR-6
-1.20
-0.90
-0.60
-0.30
0.00
0.30
0.60
0.90
1.20
PRO
RAA
SIN
GNA
GBN
4OH
GBC
4M E
NEOCK
FR-3
FR-6
-1.20
-0.90
-0.60
-0.30
0.00
0.30
0.60
0.90
1.20
PRO
RAA
SIN
GNA
GBN
4OH
GBC
4M E
NEO
CK
FR-3
FR-6
-1.20
-0.90
-0.60
-0.30
0.00
0.30
0.60
0.90
1.20
PRO
RAA
SIN
GNA
GBN
4OH
GBC
4M E
NEO
CK
FR-3
FR-6
-1.20
-0.90
-0.60
-0.30
0.00
0.30
0.60
0.90
1.20
PRO
RAA
SIN
GNA
GBN
4OH
GBC
4M E
NEO
CKFR-3
FR-6
-1.20
-0.90
-0.60
-0.30
0.00
0.30
0.60
0.90
1.20
PRO
RAA
SIN
GNA
GBN
4OH
GBC
4M E
NEO
CK
FR-3
FR-6
-1.20
-0.90
-0.60
-0.30
0.00
0.30
0.60
0.90
1.20
Tryptophan
NEO4OH
Indolic GSL
GBC
4M E
(b)
Desulfo GSL
PRO
RAA
SIN
GNA
GBN
4OH
GBC
4M E
NEO
CK FR-3
FR-6
-1.20
-0.90
-0.60
-0.30
0.00
0.30
0.60
0.90
1.20
PRO
RAA
SIN
GNA
GBN
4OH
GBC
4M E
NEO
CKFR-3
FR-6
-1.20
-0.90
-0.60
-0.30
0.00
0.30
0.60
0.90
1.20
PRO
RAA
SIN
GNA
GBN
4OH
GBC
4M E
NEO
CK FR-3
FR-6
-1.20
-0.90
-0.60
-0.30
0.00
0.30
0.60
0.90
1.20
Core structure formation
Side chain modification
Chain elongation
Core structure formation
MYB28
IQD1
MAM1/2
MAM1/2##1
IPMDH1
BAT5
CYP79F1
CYP83A1
CYP83A1##1
CYP83A1##2
CYP83A1##3
SUR1
UGT74B1
UGT74C1
GSTU20
GSTF9
GSTF10
FMOGS-OX1
FMOGS-OX5
AOP3
CYP81F3
IGMT1
APK1
APK2
CK FR3
FR6
-1.20
-0.90
-0.60
-0.30
0.00
0.30
0.60
0.90
1.20
Transcription factor
Chain elongation
Core structure form ation
Side chain m odification
Co-substrate pathways
PRO
RAA
SIN
GNA
GBN
4OH
GBC
4M E
NEO
CK
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FR-6
-1.20
-0.90
-0.60
-0.30
0.00
0.30
0.60
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1.20
(c)
Transcription factor
M YB28
IQD1
M AM 1/2
M AM 1/2-1
IPM DH1
BAT5
CYP79F1
CYP83A1
CYP83A1-1
CYP83A1-2
CYP83A1-3
SUR1
UGT74B1
UGT74C1
GSTU20
GSTF9
GSTF10
FM OGS-OX1
FM OGS-OX5
AOP3
CYP81F3
IGM T1
APK1
APK2
CK FR3
FR6
-1.20-0.90-0.60-0.300.000.300.600.901.20
Transcription factor
Chain elongation
Core structure form ation
Side chain m odification
Co-substrate pathways
PRO
RAA
SIN
GNA
GBN
4OH
GBC
4ME
NEO
CK
FR-3
FR-6
-1.20
-0.90
-0.60
-0.30
0.00
0.30
0.60
0.90
1.20
Transcription factor
16
Effects of Zeolitic Urea on Growth and Yield of Wheat
Ayaz Ahmad1, Shahzada Sohail Ijaz1, Khalid Saif Ullah Khan1,
Muhammad Ansar1, Mahnoor Azad1
1 Institute of Soil Science, PMAS-Arid Agriculture University, Rawalpindi, Pakistan
Objectives Urea is the most commonly used fertilizer in agriculture but its losses are
high (40-70%) due to quick release of Nitrogen. Negatively charged zeolites are
known to act as sorbent of the nutrients. The adsorbed nutrients are released gradually.
Objectives of the study were to test nutrient release pattern of the Zeolite based
fertilizer and its effect on growth and yield of wheat.
Methods Two year field experiment was conducted on medium texture soil.
Treatments were: Control, N @ 114 kg ha-1 from urea, N @ 114 kg ha-1 from zeolite
based fertilizer, N @ 85.5 kg ha-1 from zeolite based fertilizer, N @ 57 kg ha-1 from
zeolite based fertilizer. Soil samples were collected at tillering, booting and maturity
stages of wheat crop and analyzed for total N, NO3- N and NH4-N. Plant samples
were collected at each stage and analyzed for total N. At crop maturity biomass and
grain yields were measured.
Results Results implied that zeolite based fertilizer enhanced yield of wheat crop
even at 75% of N dose as compared with the full dose of N from urea. Plant uptake of
N was also high with zeolitic urea.
Conclusion It is concluded that zeolitic urea has potential to slow release of nitrogen
and to improve nitrogen use efficiency of urea fertilizer.
17
Changes in soil organic carbon pools and enhanced crop productivity in
sugarcane bagasse-biochar amended Soil under wheat-maize
cropping system of Pakistan
Muhammad Irfan*1, Qaiser Hussain1, Khalid Saifullah Khan1,
Azeem Khalid2, Muhammad Azeem3, Sana Ur Rehman4
1 Institute of Soil Science PMAS-Arid Agriculture University Rawalpindi, 46300, Punjab
Pakistan 2 Department of Environmental Sciences PMAS-Arid Agriculture University Rawalpindi,
46300, Punjab Pakistan 3 Department of Environment Science & Engineering College of Natural Resources and
Environment Northwest A&F University, Yangling, Shaanxi, 712100, P. R. China
4 Land Resources Research Institute, National Agriculture Research Center Islamabad
Corresponding Author: Muhammad Irfan (Ph.D Scholar)
Corresponding Author Email: [email protected]
Abstract Intensive agricultural practices results in soil degradation through loss of
soil organic matter. Amending soils with biochar have multiple environmental benefits,
including improvement in soil physicochemical properties, reduction in leaching losses
of essential nutrients and reduction in greenhouse gas (GHG) emissions. The present
study was conducted to evaluate the direct and residual effect of biochar in
combination of N fertilizer on carbon fractions in wheat-maize cropping system of
arid region of Pakistan. Biochar (sugarcane straw) prepared by conventional
pyrolysis, having with seven treatments in experiment B0N0=control (no biochar and
recommended urea fertilizer), B1N0=0.5% biochar C ha-1, B2N0=1% biochar C ha-1,
B0N1= recommended N fertilizer (100 kg ha-1), B1N1=0.5% biochar C ha-1+
recommended urea fertilizer, B2N1=1% biochar C ha-1+ recommended urea fertilizer.
The biochar was applied before the sowing of wheat and further residual effect was
studied on the other crop (maize). The results of the two years field study showed that
biochar amendment increased soil pH, soil organic carbon (SOC), carbon fractions
(dissolved organic carbon, mineralizable organic carbon hot water extractable carbon
and HCl insoluble carbon) and also on crop productivity. However, results also
indicated that water soluble carbon and soil bulk density was gradually decreased
during the both growing cycles. The soil organic carbon (SOC) and labile fractions
significantly enhanced with increasing biochar application dose over the course of the
experiment suggesting that biochar application has great potential for carbon
sequestration in the arid soil. The results therefore also suggest that addition of
18
biochar with recommended urea fertilization could have significant effect on
biological and grain yield of both crops. The outcomes propose that biochar
amendment to carbon deficient soils had an appropriate management practice to
enhance soil carbon accumulation and sequestration. This whole two year field study
was supported by Institute of Soil Science, PMAS-Arid Agriculture University
Rawalpindi, 46300, Punjab, Pakistan.
19
Nanotechnology for Reduction and Remediation of
Nonpoint Source of Pollution
Zhenli He and Xiaoping Xin
University of Florida, Institute of Food and Agricultural Sciences, Department of Soil
and Water Science/Indian River Research and Education Center, Fort Pierce, FL 34945,
United States
Abstract To meet the ever-increasing demand of world population for food and fiber
and remediate nonpoint source of pollution in agriculture, nanotechnology-enabled
agricultural innovations show great promise in delivering a more sustainable,
efficient and resilient agricultural system, while promoting food security and
reducing environmental pollution. Further exploration of nanotechnology
applications in agriculture is necessary to realize its potential in manufacturing
innovative agrochemicals and novel delivery platforms for enhancing crop
production and quality. Here, we reviewed fundamentals of nanotechnology and
focused on its potential in agricultural applications. Progress has been made in the
development of nano-fertilizers, nano-additives, nano-pesticides/herbicides/bactericides,
nano-filters to improve agrochemical efficiency, reduce run-off, enhance plant growth,
and remediate contaminated water and soils. Especially, nano-delivery systems have
been designed to deliver effective components to the targeted sites within a plant to
provide potential solutions to some devastating crop diseases, and carbon
nanoparticles have been used as fertilizer additives to enhance Seed germination,
plant growth and nutrients uptake. However, nano-enabled agriculture is still in its
infancy and its applications are mostly theoretical. Therefore, more research is
needed to develop biodegradable, cost-effective and safe nanomaterials for future
application. Moreover, systematic studies are crucial to safeguard our food
production system, while making efforts to raise public awareness to nanotechnology.
Keywords Agriculture; environmental remediation; nanomaterials; nutrient use
efficiency; soil pollution
20
Compost Utilization in Horticultural Cropping Systems
Peter J. Stoffella
Professor Emeritus, University of Florida, IFAS, Indian River Research and Education
Center, Fort Pierce, Florida, 34951 USA
Biological degradable materials, such as, municipal solid waste, biosolids, yard
trimmings, animal manures, and food processing by-products are being commercially
developed into horticultural grade compost. Commercial compost and be produced
though aerobic (static pile, window, or in-vessel) or anaerobic methods. These
systems produce a viable product for horticultural crop production while
subsequently reduce landfill material as well as extending landfill longevity. Compost
utilization in horticultural crop systems can be a potential alternative to polyethylene
mulch, fumigation, biological weed control in row alleys, source on macro and micro
nutrients, alternative to peat in potted plant culture, and reduce nutrient leaching in
ground water and loads in surface water. Precautions of compost utilization should be
with variability within and between production facilities, maturity issues, heavy
metal content, and availability of application equipment.
21
Evaluation of nitrate leaching and ammonia volatilization from
carbon based designers’ fertilizers in alkaline calcareous soils
Muhammad Rashid1, Qaiser Hussain2*, Khalid Saifullah Khan1,
Rifat Hayat1, and Sarosh Alvi2
1 Institute of Soil Science, PMAS-Arid Agriculture University, Rawalpindi, Pakistan 2 Soil and Water Testing Laboratory, Rawalpindi, Pakistan *Corresponding author: [email protected]
Abstract Chemical fertilizers are considered as costly inputs for adequate crop
production; however, their ever-increasing demand and prices has provoked serious
efforts for the development of nutrient efficient and climate-smart fertilizers. Biochar
and lignite have marked their significance as efficient soil amendments due to their
multiple agricultural and environmental benefits. But, their potential as a fertilizer
nutrient carrier need further research attention. Considering these research gaps
especially under alkaline calcareous soil, slow-release carbon-based designers’
fertilizers were synthesized and the nitrogen losses from these fertilizers in the form
of nitrate leaching and ammonia volatilization were assessed in two different textured
alkaline calcareous soils in comparison with conventional urea and commercial
slow-release fertilizer. Application of slow-release carbon-based designers’ fertilizers
such as Bio-SRF and Lig-SRF reduced the ammonia volatilization by 38.46 and
47.02% in coarse texture soil while, 39.56 and 44.90% in fine textured soil,
respectively, compared with conventional urea. Similarly, reduction of nitrate
leaching due to application these carbon-based designers’ fertilizers was 49.66 and
57.01% in coarse textured soil while 47.64 and 53.32% in fine textured soil,
respectively, compared with conventional urea. The reduction of N volatilization and
leaching losses imparted by carbon-based designers’ fertilizers could be a viable N
source for crops. However, field-level validation experiments could better assess the
efficacy of these carbon-based designers ‘fertilizers in enhancing crop yield and
nutrient use efficiency.
Keywords Slow-release fertilizer; biochar; lignite; ammonia volatilization; nitrate
leaching; alkaline soil
22
Analysis in Vitro Culture and Light Quality
Effect of Lewisia cotyledon
Tian Tian1, Chun Liu2, Li Zhang1*
1 College of Agriculture, Ningxia University, Yinchuan, 750021; 2 Vegetable and Flower Research Institute, Chinese Academy of Agricultural Sciences,
Beijing, 100086
Background Lewisia cotyledon is a kind of portulaceae flower with thick leaves and
beautiful flowers. It is native to the central mountain area of the west coast in the
United States. Potted plants are often used to decorate restaurants, open bars, private
gardens and rock gardens. 'Elise mix' series is the first hybrid of L. indica. Because of
its several features, such as a long flowering period, the bright color, drought
resistance, heat resistance, and blossoming without low temperature. It is suitable for
potted and open field application .
Methods 1. Callus induction:The young leaves were cut into 0.5 cm× 0.5 cm squares,
and the callus induction medium was inserted into the leaves with the back facing
down. The medium was based on MS, supplemented with sucrose 30 g/L, agar 6 G/L,
6-BA, 2,4-D and NAA.
2. Effects of different light quality and proportion on callus proliferation:The
callus was cut into 0.3 g callus pieces and inoculated into the selected proliferation
medium (MS+30 g/L sucrose+6 g/L agar+2.5 g/L 6-BA+0.1 mg/L NAA, pH=5.8).
After 20 days of culture, the weight gain and the rate of callus weight gain were
counted.In this experiment, eight light quality ratios were set up for LED, which were
100% blue light, 100% red light , 20% blue light + 80% red light , 80% blue light +
20% red light , 30% blue light + 70% red light , 70% blue light + 30% red light , 50%
blue light + 50% red light , PGF (fluorescent lamp for plant tissue culture).
3.Embryogenic callus induction under different light quality:0.3 g calli were
inoculated in the selected callus induction medium, and five kinds of light quality
treatments were set: 100% blue light , 100% red light , 20% blue light + 80% red
light , 70% blue light + 30% red light, PGF . Each treatment was repeated for 10
times, and each bottle was recorded as a repetition. Two calli were grafted into each
bottle. After 30 days of culture, embryogenic callus induction was observed.
Results The results showed that the optimum explants for callus induction were
23
leaves, MS+6-BA 1.0 mg/L+2,4-D 0.2 mg/L+NAA 0.1 mg/L, 20% blue light+80%
red light (B:R=2:8) and MS+6-BA 2.5 mg/L+NAA 0.1 mg/L. The optimum medium
for embryogenic callus induction was MS+6-BA 2.5 mg/L+NAA 0.1 mg/L. The light
quality was 20% blue light + 80% red light (B:R=2:8).
Conclusions At present, the problems of low germination rate, uneven emergence,
unstable flowering period and so on are faced by the sowing and propagation of
Lagerstroemia indica. Through this study, we found that there are two ways of flower
bud regeneration of Lagerstroemia indica. One is that the explants directly form buds
without callus differentiation, and the other is that the explants are induced to form
callus, and finally form buds to complete plant regeneration.When the concentration
of 6-BA, 2,4-D is certain, the induction rate increases first and then decreases with
the increase of NAA concentration, which indicates that auxin in the appropriate
concentration range is beneficial to plant growth.
Acknowledgements This research is supported by the key R & D project of
Ningxia Hui Autonomous Region (special project of modern agricultural science and
technology innovation demonstration area) "new excellent and special flowers
introduction and selection and supporting cultivation technology integration
demonstration".
24
Nutrient Use Efficiency in Crops: A Comprehensive Approach
Virupax C. Baligar1, Zhenli He2, Yuncong C. Li3
1 USDA-ARS, Beltsville Agricultural Research Center, Beltsville MD, U.S.A 2 Indian River Research Education Center -IFAS-Univ of Florida Fort Pierce, FL, U.S.A. 3 Tropical Research Education Center-IFAS -Univ of Florida Homestead, FL, U.S.A.
Abstract This presentation is an over view of significance of improving nutrient use
efficiency in crop to enhance food production for meeting demands of ever-growing
world population and global constraints that could affect achieving such an advance.
In the 21st century, nutrient efficient plants will play a major role in increasing crop
yields. Higher demands for food and fiber by increasing world populations expected
to approach 10 billons by year 2050 further enhances the importance of nutrient
efficient cultivars that are also higher producers in a reduced input system. However,
for achiving needed NUE in crop plants in the future is greatly affected by global
climatic changes (precipitation patterns, drought intensities, light quality/quantity,
greenhouse gases), decline in soil health (deficiency of essential nutrients and
increased toxic elements) and increased occurrences of abiotic (drought, high T and
CO2) and biotic stresses (insect and diseases. Inter- and intra-specific variation for
plant growth and mineral nutrient use efficiency (NUE) are known to be under
genetic and physiological control and are modified by plant interactions with
environmental variables such as soil moisture and deficiency of essential and toxicity
of non-essential elements, temperature, light quality and duration and levels of
atmospheric CO2. Identification of traits such as nutrient absorption transport,
utilization. and mobilization in plant cultivars and their interactions with
environmental variables should greatly enhance nutrient use efficiency in crop plants.
The NUE in the literature is defined in several ways., nutrient efficient plant is
defined; a plant that absorbs, translocate, or utilizes more of a specific nutrient than
another plant under conditions of relatively low nutrient availability in the soil or
growth media. During the last three decades, very limited research has been
conducted to identify and/or breed nutrient efficient plant species or
genotypes/cultivars within species. Complexity of genes involved in NUE for micro
and macronutrients under abiotic and biotic stresses are not well understood. Further,
success in releasing nutrient efficient cultivars has been limited, mainly because of
limited understanding of interactions between plant genetics, environmental variables
25
and adapted management practices. Increased collaborative efforts are needed
between breeders/molecular scientists, physiologists, soil scientists, and agronomists
to evaluate plant nutrient efficiency issues on holistic bases in order to develop
nutrient efficient plant cultivars that could adapt to various types of ecosystems in a
changing climatic conditions and management systems. Hence, during the 21st
century, agricultural scientists have tremendous challenges, as well as opportunities,
to develop nutrient efficient crop plants that can increase plant efficiency for uptake
of applied nutrients and produce higher yields simultaneously adapt to changing
climate and soil fertility and further reduces degradation of soil, water and air quality
and subsequently costs of production.
26
Proliferation and Flowering in vitro of Tissue Culture Seedlings of
Lewisia cotyledon based on Orthogonal Design
Cunyi Ma1; Li Zhang1
College of Agriculture, Ningxia University, Yinchuan, China
Background Lewisia cotyledon, belongs to portulaceae, native to the mountains of
the central west coast of the United States. In recent years, Lewisia cotyledon began
to enter the Chinese market, because of its long flowering period and bright color, it
is popular with the public. However, in the actual production, the seed germination
rate is low and the growth cycle is long, which seriously affects the potted flower
production. Tissue culture and rapid propagation technology has been widely used in
flower production, and the effect is remarkable. The application of tissue culture and
rapid propagation technology in the flower production can promote the rapid
development of its industry.
In bottle flowering refers to the use of in vitro culture method, so that the
flowering process of plants is completed in the container. Flowering in the bottle is
not restricted by climate conditions, which is convenient for cultivation and can
greatly reduce the management cost. However, there are only a few reports on the in
vitro Rapid Propagation of Lewisia cotyledon, and there is no report on the in vase
flowering of Lewisia cotyledon. Therefore, in order to improve the success rate of
Lewisia cotyledon flower cultivation, the rapid proliferation of aseptic seedlings and
in bottle flowering were studied on the basis of in vitro culture of Lewisia cotyledon.
Methods The same size and good growth condition of the aseptic seedlings of
Lewisia cotyledon were selected as experimental materials. Using the orthogonal
design of three factors and three levels L9(33) method, and different concentrations of
sucrose, 6-BA and NAA were added to MS basic medium in order to screen the
optimal proliferation conditions for the aseptic seedling of Lewisia cotyledon.
Different concentrations of 6-BA, NAA and GA3 were added to MS basic medium to
select the optimal conditions for flowering in vitro.
Results The L9(33) orthogonal experiment was designed to select the best conditions
for the proliferation of Lewisia cotyledon. Among the three levels of three different
plant growth regulators, the ninth treatment (A3B3C2) had the best effect. The leaves
27
of Lewisia cotyledon seedlings were thick, the plants were strong, and the average
proliferation multiple reached 3.17 times. L9(33) orthogonal experiment was designed
to select the optimum conditions for flowering in bottles. Among the three levels of
hormone combinations with three different concentrations, the fourth treatment
(D2E1F2) had the most flowering number and the average flowering rate could reach
70%. After inoculation for about 20 days, the plants grew well and began to grow
buds. After 30 days, the plants blossomed.
Conclusions The results showed that MS+0.5mg/LNAA+1.0mg/L6-BA(sucrose
concentration 30g/L) was the best proliferation condition, the highest proliferation
multiple was 3.17 times, the most suitable Flowering in vitro condition was
MS+1.0mg/L 6-BA + 3.0mg/LGA3 + 0.5mg/L NAA, and the average flowering rate
could reach 70%. This research provides theoretical basis and technical guidance for
the efficient propagation and the development of test tube flower in Lewisia
cotyledon.
28
Plant growth regulators and EDTA improve phytoremediation
potential and antioxidant response of Chenopodium
ambrosioides L. in a Cd-spiked soil
Amin Ullah Jan1, Allah Ditta2,3
1 Department of Biotechnology, Faculty of Science, Shaheed Benazir Bhutto University
Sheringal Dir Upper, 18000, Khyber Pakhtunkhwa, Pakistan 2 Department of Environmental Sciences, Shaheed Benazir Bhutto University Sheringal,
Dir Upper, 18000, Khyber Pakhtunkhwa, Pakistan 3 School of Biological Sciences, The University of Western Australia, 35 Stirling
Highway, Perth, WA 6009, Australia
Objectives Soil pollution due to potentially toxic elements is a worldwide challenge
for health and food security. Chelate assisted phytoextraction along with the
application of plant growth regulators (PGRs) could increase the phytoremediation
efficiency of metal contaminated soils. The present study was conducted to
investigate the effect of different PGRs (GA3 and IAA) and synthetic chelator
(EDTA) on growth parameters and Cd phytoextraction potential of Chenopodium
ambrosioides grown under Cd-spiked soil.
Methods GA3 (10-7 M) and IAA (10-5 M) were applied four times with an interval of
ten days through foliar spray, while EDTA (40 mg kg-1 soil) was once added to the
soil.
Results The results showed that Cd stress significantly decreased fresh biomass, dry
biomass, total water contents, and photosynthetic pigments as compared to control.
Application PGRs significantly enhanced plant growth and Cd phytoextraction.
Combined application of GA3 and IAA with EDTA significantly increased Cd
accumulation (6.72 mg kg-1 DW) and bioconcentration factor (15.21) as compared to
C1 (Cd only). The same treatment significantly increased chlorophyll, proline,
phenolic contents, and antioxidant activities (CAT, SOD, and POD) while MDA
contents were reduced. In roots, Cd accumulation showed a statistically significant
and positive correlation with proline, phenolics, fresh biomass, and dry biomass.
Similarly, Cd accumulation showed a positive correlation with antioxidant enzyme
activities in leaves. Chenopodium ambrosioides showed Cd hyperaccumulator
potential, based on bioconcentration factor (BCF) > 1.
29
Conclusions In conclusion, exogenous application of GA3 and IAA reduces Cd stress
while EDTA application enhances Cd phytoextraction and ultimately
phytoremediation potential of Chenopodium ambrosioides.
Acknowledgements This research did not receive any specific grant from funding
agencies in the public, commercial, or not-for-profit sectors.
30
Plants buffer against high light stress-induced transcript of
CsGA2ox8 via alternative splicing to fine-tune gibberellins
level and maintain hypocotyl elongation
Bin Liu1,2, Shuo Zhao1, Pengli Li1, Yilu Yin1, Qingliang Niu1,
Jinqiang Yan3, 4, Danfeng Huang1
1 School of Agriculture and Biology, Shanghai Jiao Tong University, Key Laboratory of Urban
Agriculture (South), Ministry of Agriculture, Dongchuan Road, Shanghai 200240, China 2 Department of Plant Genomics, Centre for Research in Agricultural Genomics
(CRAG), CSIC-IRTA-UAB-UB, Bellaterra 08193, Spain 3 Vegetable Research Institute, Guangdong Academy of Agricultural Sciences,
Guangzhou 510640, China 4 Guangdong Key Laboratory for New Technology Research of Vegetables, Guangzhou
510640, China.
Background In plants, alternative splicing (AS) is markedly induced in response to
environmental stresses, but it is unclear why plants would generate multiple
transcripts under stress conditions.
Methods In this study, RNA-Seq was performed to identify AS events in cucumber
seedlings grown under different light intensities. We characterized the expression of
target genes by quantitative real-time PCR. Functional analysis was done with
transgenic plants. GA contents were measured by Liquid chromatography-mass
spectrometry.
Results We identified a novel transcript of the gibberellin (GA) deactivation enzyme,
Gibberellin 2-beta-dioxygenase 8 (CsGA2ox8). Compared with canonical
CsGA2ox8.1, the isoform CsGA2ox8.2 had an intron retention between the second
and third exons. Functional analysis proved that the transcript of CsGA2ox8.1, but
not CsGA2ox8.2, played a role in the deactivation of bioactive GAs. Expression
analysis demonstrated that both transcripts were upregulated by increased light
intensity, but the expression level of CsGA2ox8.1 increased slowly when the light
intensity was greater than 400 µmol·m-2·s-1 ppfd (photosynthetic photon flux density),
while the CsGA2ox8.2 transcript increased rapidly when the light intensity was
stronger than 200 µmol·m-2·s-1 ppfd.
Conclusions Our findings provide evidence that plants might fine-tune GA levels by
buffering against the normal transcript of CsGA2ox8 through AS.
31
Acknowledgements This study was supported by The National Key Research and
Development Program of China (2019YFD1000300), the International Postdoctoral
Exchange Fellowship program from China Postdoctoral Council (20170053); the
Technology System Construction of Modern Agricultural Industry of Shanghai
(19Z113040008), and the Presidential Foundation of Guangdong Academy of
Agricultural Sciences (BZ201901).
32
Seeds priming technology as a key strategy to increase crops
production under adverse environmental conditions
Daniel Garcia1, Shuo Zhao1, Samiah Arif1, Yinglei Zhao2,
Lin Chau Ming3, Danfeng Huang1,
1 School of Agriculture and Biology, Shanghai Jiao Tong University (SJTU), Shanghai
-200240, China 2 College of Bio Systems Engineering and Food Science, Zhejiang University, Hangzhou
310058, China 3 Horticulture Department, Faculty of Agronomic Sciences (FCA), São Paulo State
University Julio de Mesquita Filho (UNESP), 18610-307, Botucatu, São Paulo, Brazil
Corresponding author:Tel.:+8618621759810. E-mail address: [email protected]
Objectives Faced with environmental adversities, and the increasing food demand
around the globe, more attention has turned to improve crop production by efficient
strategies. Thus, we aimed to study hydropriming, osmopriming, chemo/biopriming,
thermopriming, hormopriming, nanopriming, and hybrid priming methods on the
germination and growth performance of several crops under environment adversities.
Methods Physiological, biochemical, and molecular changes on seeds subjected to
priming methods and allowed to germinate under environment adversities were
detected in a wide number of reports. The main databases were visited: PubMed,
Elsevier, MDPI, Springer among others.
Results All the seven priming methods demonstrated promising results related to
enhance the crop production traits, alleviation of aging damage, enhance chilling and
heat tolerance, improve phytopathogen tolerance, mitigate the effects of salinity,
heavy metal, and flooding, enhance seeds storage prolongation and increase drought
tolerance. The findings shed light on the use of priming technology as a key strategy
to increase crop production via stress-tolerance faced to the predicted environment
adversities and the ever-increasing food demand.
Conclusions The molecular point of view must be further investigated on seeds
subjected to priming methods, in order to understand the specific influence of each
biological mechanism on the production of crops under stress conditions. Moreover,
we strongly recommend the combined priming methods, called here hybrid priming,
generating thus, new approaches about the complex biological mechanism changes in
the enhancement of crop productivity under environment adversity.
33
Effects of greenhouse new/waste rockwool and nutrient
on growth and quality of pakchoi
Xiaotao Ding, Shuailei Zhang, Yingqi Chu, Lizhong He,
Hong Wang, Qiang Zhou, Jizhu Yu
Shanghai Key Lab of Protected Horticultural Technology, Shanghai Academy of
Agricultural Sciences, Shanghai 201106, China
Objectives The effective utilization of waste rock wool and drain nutrient is an
important problem that perplexes the development of modern greenhouse. The
objective of the experiment was to find method to effective use waste rockwool and
drain nutrients.
Methods the experiment was designed with four treatments: 1. new rockwool with
new nutrient; 2. new rockwool with waste nutrient; 3. waste rockwool with new
nutrient; 4. waste rockwool with waste nutrient. All seedlings were watered with
nutrient of EC1.8 dS/m and pH of 5.8. The four treatments in this study were
arranged in a randomized complete block design. The experiment was replicated
three times with one experimental unit (twenty plants) per treatment. In the
experiment, the growth changes of pakchoi in the cultivation of new and waste
rockwool, and new and waste nutrient solution, as well as chlorophyll content,
photosynthesis, yield and important quality parameters were measured.
Results The results showed that the growth and photosynthesis of pakchoi treated
with waste rockwool were clearly better than that treated with new rockwool, yield of
waste rockwool with new nutrient and waste rockwool with waste nutrient treatments
were significantly higher than new rockwool with new nutrient and new rockwool
with waste nutrient treatments of 13.5% and 20.2%,of 14.4% and 21.2%, which may
be related to the more nutrients in waste rockwool. There was no clear difference in
quality, which the content of soluble sugar, total phenol and flavonoids were not
significant difference for different treatments.
Conclusions The treatments of waste rockwool and waste nutrient solution had not
significantly reduced quality of pakchoi, and the content of nitrate and nitrite did not
show drastic changes. This experiment provides an effective method for the recycling
of waste rockwool and drain nutrients in modern greenhouse.
Acknowledgements This work was supported by Shanghai Agriculture Applied
Technology Development Program, China (Grant No. T20180405).
34
Purification ability from constructed wetlands treating sewage
treatment plants effluent and microbial community
succession in epiphytic biofilms
Zheng Qiu, Songhe Zhang*, Yan Ding, Wenjun Zhang, Lixue Gong,
Qiang Yuan, Xiaoying Mu, Dongwang Fu
Ministry of Education Key Laboratory of Integrated Regulation and Resource Development on
Shallow Lakes, College of Environment, Hohai University, Nanjing, 210098, China.
*Corresponding author: Songhe Zhang Email: [email protected]
Abstract Sewage treatment plants effluent is a large source of pollution and it’s
important to treat it in a clean way. In aquatic environments, submerged macrophytes
play important roles in bioremediation of wastewater and microbes in epiphytic
biofilms play crucial roles in the pollutants removal in constructed wetlands.
However, the impacts of treated sewage effluents on nutrients removal and epiphytic
microbial community structures in wetlands dominated by submersed macrophytes
remain to be investigated. In this study, concentrations of nutrients in three kinds of
simulated effluents were used according to the first-level standard A and B and
second-level standard in discharge standard of pollutants for municipal wastewater
treatment plant in China (GB18918-2002). M. Spicatum with the same growth stage
and artificial plants (made by polyvinyl chloride) with similar morphological
characterizations (needle leaves) were used in this experiment. While under
second-level standard, the leaf surface of artificial plants and M. Spicatum was
almost completely covered by microorganisms. Compared to M. Spicatum, artificial
plants had higher NH4+-N, TN, COD and TP removal rates in wetlands receiving
treated sewage effluents. Taken together, artificial plants systems had better
performance in nutrients removal than M. Spicatum systems in general. The
α-diversity values of bacterial and eukaryotic community in attached biofilms were
significantly higher in MS systems than in AP systems, revealing microbial
population richness was higher in epiphytic biofilms on M. Spicatum than artificial
plants. However, there were no significant difference in the alpha diversity values
caused by different water qualities or time variable. Both turnover (species
replacement) and nestedness (species loss) contributed to the construction of
microbial community structure, whereas beta-diversity patterns in constructed
wetlands mainly resulted from species replacement rather than species loss. Organic
35
matter decomposition-related microbes were stimulated in MS systems bacteria in M.
Spicatum systems. Phyla Ciliophora occurred in a higher frequency in M. Spicatum
systems than artificial plants systems, which can be ascribed to lower nutrients
removal rates in M. Spicatum systems. Network analysis revealed that there were
more complex interactions among environmental parameters, bacteria and eukaryotes
in M. Spicatum systems than in artificial plants. Those results demonstrate that AP
can provide surface for biofilm growth and have roles similar to submersed
macrophytes in nutrients removal while treated sewage effluents can cause damage to
submersed macrophytes, highlighting artificial plants can be used in effluent deep
treatments and the strict discharge standards should be taken to reduce the nutrients
in treated sewage effluents, for aims to protect the ecological systems.
Keywords Treated sewage effluents; Beta-diversity; Microbes; Shannon index;
Co-occurrence network
36
Impacts of water flow and suspended sediment on microbial
community in epiphytic biofilm on two
submerged macrophytes
Hezhou Chen, Bing Han, Songhe Zhang*
Ministry of Education Key Laboratory of Integrated Regulation and Resource Development on
Shallow Lakes, College of Environment, Hohai University, Nanjing, 210098, China. *Corresponding author: Dr. Songhe Zhang E-mail: [email protected]
Abstract Understanding the interaction of water flow, suspended sediment (SS) and
submerged macrophytes is of great significance to water ecological restoration and
management. In this lab-scale water tank experiment, the influence mechanism of
different submerged macrophytes and water flow on the removal of SS was studied
with a focus on understanding the effects of water flow and SS on the structure and
microbial composition in the epiphytic biofilm of M. verticillatum and V. natans. It
was found that the decreasing trend of water turbidity can be elucidated by the
logarithmic model (R2>0.93, P<0.01) within one day regardless of water flow and
plant types. The turbidity reduction rate was higher in treatments with plants than
without plants in static water (96.2%>91.6%), but an opposite trend was observed
under flowing water (95%<98.2%). The size of particles was mainly below 500 μm
(72.7%-94.6%) on leaves of two submerged macrophytes, and among four treatments,
particles was the smallest but its amount was the biggest on M. verticillatum leaf
under water flow. Confocal laser scanning microscopy images revealed that the mean
values biofilms thickness ranged from 10.18 to 28.36 μm. Mantel correlation
analyses revealed that water flow was significantly correlated with the relative
abundance of top ten bacterial (r=0.6433, p=0.0284) and eukaryotic (r= 0.6343,
P=0.0285) order taxa in epiphytic biofilm of two submerged macrophytes; SS
decreased such correlation. Principal co-ordinates analysis of bacterial and eukaryotic
showed that the bacterial communities in epiphytic biofilm of M. verticillatum and V.
natans. were significantly different from those in spiked SS in treatment without
water flow, but the eukaryotes in spiked SS and epiphytic biofilm had similar
community composition in treatments with water flow, suggesting that the eukaryotes
on sediment may be transferred to the leaves due to water flow. SS increased
epiphytic biofilm bacterial diversity, but inhibited eukaryotic diversity, except
inhibited bacterial diversity and promoted eukaryotic diversity in V. natans under
37
static water. Results of co-occurrence networks shows that SS increased but water
flow decreased the number of keystone microbes. Our results highlighted that water
flow and SS have different mechanism on microbial community in epiphyte biofilm
on submersed macrophytes.
Keywords Suspended sediment; Water flow; Submerged macrophytes; Epiphytic
biofilm; Microbial community
38
Aeration enhanced nutrient removal and increased microbial
diversity during Hydrilla verticillata decomposition
in wetlands in winter
Xin Lv, Tiantian Zhou, Songhe Zhang, Lisha Zhang,
Xiaoying Mua, Ziqiu Zhang, Qiang Yuan
a Ministry of Education Key Laboratory of Integrated Regulation and Resource
Development on Shallow Lakes, College of Environment, Hohai University, Nanjing
210098, China.
Corresponding author: [email protected] (S.H. Zhang)
Abstract Aquatic environments usually support high macrophyte productivity, while
after senescence, abundant nutrients can be rapidly released, causing the potential
deterioration of the water environment. The decay of aquatic macrophyte causing low
performance in nutrients removal is remain unsolved in wetland. Mechanical aeration
has been reported to reduce the total concentration of nitrate and phosphate in
wastewater. In present study, nutrient release, extracellular activities and microbial
communities were investigated during H. verticillata decomposition in a natural (NA)
or aerated (AA) environment for 206 days. A small aeration pump was placed 5 cm
below the water surface to pump air at 1 L of air/min in the AA groups. The results
showed aeration increased the dissolved oxygen concentration and decreased the
nutrient content in the overlying water, enhanced extracellular activities in biofilms
and decreased the C, N and P content in plant residues in comparison to what was
observed in the NA treatment. Humic acid-like and tryptophan-like substances
significantly increased during H. verticillata decomposition, which were originated
from organic matter containing N and P in the overlying water, especially in the AA
treatment. The bacterial 16S rRNA (V3-V4 region) and eukaryotic 18S rRNA (V4
region) genes in surficial sediments and biofilms were analysed at nine sampling
times. The microbial communities in biofilms generally differed from those in
surficial sediments. The bacterial communities in biofilms shifted and became similar
to those in surficial sediments over time. Aeration promoted relative abundance of
aerobic bacteria but suppressed facultatively and obligate anaerobic bacteria. The
dominant phylum in samples were Stramenopila, Alveolata, Rhizaria, Metazoa, Fungi,
Viridiplantae and Apusozoa. Aeration stimulated relative abundance of Metazoa but
suppressed Viridiplantae in sediments. Network analysis showed complex microbial
interactions in the leaching stage and aeration altered the microbial interactions
39
during decomposition. The eukaryotic community was more stable than the bacterial
community, especially among bacteria in sediments, while aeration obviously
disturbed the bacterial community during decomposition. Our results highlight that
aeration can enhance nutrient removal by stimulating microbial activity and shaping
the microbial community during H. verticillata decomposition.
Keywords Humic acid; nutrients; eukaryotic community; decomppsition
40
Detection of chlorophyll content in tomato leaves based
on portable hyperspectral imaging technique
Minghua DU, Lu Meng, Longguo Wu*
School of Agriculture Ningxia University, Yinchuan Ningxia 750021
Background Nitrogen is the primary factor affecting tomato growth and yield. In
production, excessive nitrogen application and excess nitrogen application can inhibit
the healthy growth of tomatoes. Therefore, it is imperative to adopt scientific
nitrogen nutrition diagnosis technology, establishing a set of reasonable diagnostic
indicators and recommend fertilization technology system. In recent years,
hyperspectral imaging technique has been regarded as one of the most promising
diagnostic methods. As a nondestructive analysis technique, it can be applied to plant
nutrition detection more quickly and effectively. Many scholars at home and abroad
have realized the nondestructive testing of nitrogen, phosphorus, potassium,
chlorophyll and other nutrient elements of crops based on hyperspectral imaging
technology. At present, there are few studies on nitrogen content detection in tomato
leaves by using hyperspectral imaging technology. Therefore, on the basis of
previous studies, portable hyperspectral imaging technology was used to study and
detect nitrogen content in tomato leaves.
Methods The average spectral reflectance data (400-1000nm) of 82 tomato leaf
samples were extracted and pretreated with 7 kinds of original spectral data (mean
smoothing, gaussian filtering, median filtering, convolution smoothing, normalization,
baseline calibration, and standard normalization(SNV). Feature spectral information
was extracted by weight coefficient method(BW), variable elimination
transformation without information method (UVE), competitive adaptive repeated
weighting method (CARS) and continuous projection algorithm (SPA). Linear
regression (MLR, PCR, PLSR) and nonlinear regression (SVR) were used to
construct the quantitative prediction model of chlorophyll in tomato leaves.
Results In the pretreatment comparison, the prediction ability of SNV pretreatment
model was higher than others.Five kinds of normalized pretreatments were compared
and analyzed, and the PLSR model established by normalized -3(average
normalization) had the best effect. The PLSR model of the original spectrum and
other pretreated spectra were compared and analyzed, and the PLSR model
41
established by SNV was selected. Compared with other feature bands selected,
CARS was the best method for selecting feature wavelengths. Eight characteristic
wavelengths (732, 796, 946, 953, 957, 968, 983 and 994nm) were extracted by CARS
method and applied to establish a quantitative prediction model of chlorophyll in
tomato leaves. The model of MLR, PCR, PLSR and SVR were used to compare the
models of characteristic wavelengths extracted by CARS, and it was found that the
model established by MLR, PCR, PLSR was superior to the model established by
SVR. Among the models established by linear regression, the effect of MLR model is
better than PCR and PLSR. PCR and PLSR models established by linear regression
have basically the same correlation coefficient and root mean square error.
Conclusion Our study shows that the noise interference in spectral information can
be improved to a certain extent by the selection of pretreatment methods. Eight
characteristic wavelengths (732, 796, 946, 953, 957, 968, 983, 994nm) extracted by
CARS for tomato leaf chlorophyll quantitative model is set up, has good prediction
effect. The linear regression model of chlorophyll in tomato leaves obtained a higher
prediction effect. The correlation coefficient and RMS error of MLR of the optimal
prediction model were 0.794 and 3.492, respectively. On this basis, the visual
distribution was carried out, and finally MLR model was selected to predict the
chlorophyll in tomato leaves online, which also provided reference for the rapid
detection of other indicators of tomato plants.
Keywords Hyperspectral imaging; Tomato leaves; Chlorophyll; Nondestructive
testing
42
Research of Volatile Components on Champagne and
Japanese Peppermint Stem and Leaf Tissues
Liu Xuanxuan, Zhang Haoyu, Zhang Guangdi
School of Agriculture Ningxia University, Yinchuan Ningxia 750021
Background Peppermint belongs to the genus Mentha haplocalyx Briq. It is widely
distributed and mainly concentrated in the temperate regions of the northern
hemisphere. Peppermint has the function of clearing heat and reducing fire. It can be
used for wind-heat and cold, sweating and antipyretic, calming and analgesic,
anti-inflammatory and antibacterial, etc. Efficacy, this test aims to accurately locate
the extraction of peppermint essential oil, the use of processing materials such as
flavors and fragrances, condiments, tea beverages, etc. through the specific
characteristic volatile substances in different peppermint stem and leaf tissues, and
provide methods for the identification of peppermint varieties.
Methods In this research, gas chromatography-ion mobility spectrometry (GC-IMS)
technology was used to detect and analyze the volatile components in the stems and
leaves of Champagne and Japanese peppermint. In the two types of peppermint, 39
volatile compounds were identified. It mainly included alcohols, ketones, aldehydes,
furans, esters and terpenes. Among the stem and leaf tissue of Champagne mint, there
are 28 volatile compounds are identified. In Champagne mint leaves, the main
volatile substances were 3 -Octanol eucalyptol, linalool, trans-2-hexen-1-ol,
2-heptanone, etc; In Champagne peppermint stems, the main volatile substances
were 2,3-butanedione, etc.; A total of 38 volatile compounds have been identified in
Japanese mint. The main compounds in the leaves were 3-octanol, 1-octen-3-ol,
3-octanone, ethyl isobutyrate, linalool, and butyraldehyde, Α-pinene, 1-pentanol, etc.
In Japanese peppermint stems, the main volatile substances were ethyl acetate,
2-n-pentyl furan, pentyl acetate, 2-nonanone, isobutyl acetate, etc. Compare between
Champagne peppermint and Japanese peppermint leaves showed that
2,3-butanedione was a unique volatile compound in Champagne peppermint.
Compared the stems of two peppermints, 2-nonanone, 2-n-pentylfuran, and ethyl
hexanoate. Ester, butyl acetate, etc. were unique volatile substances in Japanese
peppermint, but they were not detected in Champagne peppermint.
Results Through PCA and visual heat map cluster analysis, it could be known that
43
the volatile compounds in the stem and leaf tissue of each peppermint were different
and could be obtained. Obviously, this test aimed to accurately locate the extraction
of peppermint essential oil, the use of processing materials, such as flavors and
fragrances, condiments, and tea beverages, and to provide methods for the
identification of peppermint varieties.
Conclusions Through the above analysis, it can be seen that Champagne and Japanese
peppermint, these two types of compounds are the main volatile substances in the
extraction of peppermint essential oil. In addition, there are also some terpene volatile
compounds such as α-pinene, which have certain properties. The anti-inflammatory,
anti-viral, antibacterial and other effects of the peppermint are relatively high in the
peppermint stems and leaves; through the visual heat map and PCA analysis, the
volatile organic compounds in the two peppermint leaves are very different in
composition and content. If passed More samples are modeled, and this result will also
provide a theoretical basis for the identification of mint leaves and stems in organic
matter and the identification of unknown mint samples. Through the above results, the
use of the characteristic volatile compounds in different tissues of mint can provide
scientific and reasonable suggestions and support for the deep processing of mint, such
as food flavors, spices and condiments in the future.
44
Study on Visualization Distribution of Chlorophyll in Tomato
Leaves under brackish irrigation Based on
Hyperspectral Imaging Technology
Lu Meng, Jie Zhang, Tian Yang, LongGuo Wu*
School of Agriculture, Ningxia University,Yinchuan 750021,China
Background Ningxia is located in the arid northwest of China, which riches in
brackish water and saline water resources, using brackish water for irrigation to
alleviate the crisis of freshwater resources. The nitrogen content of crop leaves
reflects the response of brackish water to the organism, and provides guidance for the
rational application of brackish water in the future. Hyperspectral imaging
technology has been widely used in the estimation of crop physiological and
biochemical indicators,which is faster and more non-destructive than the chlorophyll
meter.
Methods The test tomatoes were selected from the "Bomei No.2" variety, with a total
of 108 tomato seedlings. The seedlings were transplanted when 3-4 cotyledons were
expanded, and the cultivation substrate pot was used for planting. Each pot was
planted with 4 tomato seedlings. The experiment was designed with 9 treatments in
follows, control group CK: distilled water; A:0.2 g/L NaCl solution; B:0.4 g/L NaCl
solution; C:0.6 g/L NaCl solution; D:0.8 g/L NaCl solution; E:1.0 g/L NaCl solution ;
F:1.2 g/L NaCl solution; G:1.4 g/L NaCl solution; H:1.6 g/L NaCl solution,and each
treatment was repeated 3 times. In the early stage, the optimum irrigation amount was
determined to be 550ml, after planting, irrigation should be done every 2 days. After
30 days of planting, the canopy leaves of tomato were picked. A total of 150 pieces
were collected in the experiment, and the portable chlorophyll tester and portable
hyperspectral imaging equipment were used for chlorophyll measurement and image
collection immediately.
Results The results show that the outlier elimination optimizes the model and
optimizes the original spectrum for data analysis. PLSR method was used to model
the characteristic wavelength extracted by different methods and to optimize the
characteristic wavelength extracted by SPA. On this basis, the models of MLR, PCR,
PLSR, SVR, ANN were used to model the characteristic wavelengths extracted by
SPA, and the different modeling effects are compared and analyzed. The models of
45
MLR, PCR, PLSR are preferably selected. The optimal characteristic wavelengths
were 392, 465, 686 and 760nm, the prediction correlation coefficient of the optimal
model was 0.896, and the prediction root-mean-square error RMSEP was 1.111.
Finally, the PLSR model was optimized to predict the chlorophyll content of tomato
by visualizing three linear models.
Conclusions 1.Using Euclidean distance and Mahalanobis distance combined with
Monte Carlo method to eliminate abnormal samples, a robust model can be
established. 2.The tomato leaf chlorophyll quantitative model established by the 4
characteristic wavelengths (392, 465, 686, 760nm) extracted by the SPA , the method
has a good predictive effect. 3.The linear regression model of tomato leaf chlorophyll
has better prediction effect than the nonlinear regression model. The correlation
coefficient and root mean square error of the optimal prediction model are 0.896 and
1.111 respectively; on this basis, visualize the distribution, and finally choose the
PLSR model for online prediction of tomato leaf chlorophyll.
46
Comparison and Screening of Total RNA Extraction
Methods from Fennel Leaves
Wang Lin1, Wang Xiaomin1,2,3,4*, Li Xia1, Guo Liwen1,
KongWeikang1, PanBingqing1
1 School of Agriculture, Ningxia University 2 Key Laboratory of Modern Molecular Breeding for Dominant and Special Crops in Ningxia 3 Ningxia Facility Horticulture (Ningxia University) Technology Innovation Center 4 Ningxia Modern Facility Horticulture Engineering Technology Research Center
Background Fennel is a kind of 1-year-old and 2-year-old herb of Umbelliferae,
which originated in Mediterranean region. It has been planted all over the world
because of its strong adaptability. It has been cultivated for more than 1000 years in
China and distributed all over the country. In recent years, research on fennel mainly
focused on its physiological active components, inducing plant seed germination,
extracting volatile oil components and so on, but the molecular regulation mechanism
of fennel was still unclear. It is necessary to extract high-quality RNA in order to
explore the related genes and elucidate their expression and regulation. Extracted
high-quality RNA from plant tissues was the premise of molecular biology research,
such as gene cloning, northern analysis, RT-PCR and cDNA library construction,
because of differences in plant tissue composition and environmental stress, different
metabolites are produced during plant growth, such as proteins, polysaccharides and
polyphenols etc. It was easy to coprecipitated with RNA, and interfered with the
extraction and purification of RNA, then the quality of total RNA was affected. So
the methods of total RNA extraction of plants should be screened and compared.
Methods Hexadecyl trimethyl ammonium bromide(CTAB) method, modified CTAB
method, CTAB water saturated phenol method, Tris-boric acid method,
cyanoguanidine isosulfate method, modified cyanoguanidine isosulfate method,
Trizol method, and modified Trizol method were used to extracted RNA from leaves
of fennel seedlings at seven leaves old. The concentration, purity and integrity of
total RNA from fennel leaves were determinated by nucleic acid and protein detector
as well as agarose gel electrophoresis.
Results The quality of RNA extracted by modified CTAB method was the best, and
the concentration was 1481.9 μg/μL, the OD260/280 was 1.82, OD260/230 was 2.12, the
yield was higher, and the strips were clear and intact without tailing. The quality of
47
RNA extracted by Trizol method, modified Trizol method and cyanoguanidine
isosulfate method was higher, but the integrity was poor, and the former two methods
had higher cost, while the latter was lower. Both of the purity and integrity of RNA
extracted by Tris-boric acid method were the lowest. The effects of total RNA
extraction by CTAB method, CTAB water saturated phenol method and modified
cyanoguanidine isosulfate method were poor.
Conclusions Our study indicated that CTAB was selected as the most suitable
method for extracting RNA from fennel leaves, which laid a foundation for
subsequent molecular biology research and provided some reference for RNA
extraction from similar plants.
48
Analysis of Flavor Compounds in Grapefruit Mint
Leaf Stem Based on GC-IMS
Yufen Xie1,2, Haoyu Zhang1,2, Wenli Yang1,2, Kunming Zhang2,4, Yuelin Ma2,4,
Xiaoyun Liu2,4, Xiaoyan Yu3*,and Guangdi Zhang1,2,3*
1 School of Agriculture Ningxia University,Yinchuan 750021, China 2 Ningxia Key Laboratory for FoodMicrobial-applications Technology and Safety
Control,Yinchuan 750021,China 3 Technological Innovation Center of Protected Horticulture (Ningxia University) in
Ningxia, Yinchuan 750021, China 4 School of Food and Wine Ningxia University,Yinchuan 750021, China
Background Grapefruit mint, (Mentha Suaveolens × Piperita) is a perennial herb of
the genus Mentha in the labiform family. It is one of the new varieties of mint and has
a very strong grapefruit taste. As a common Chinese traditional medicine, peppermint
USES whole grass as medicine. It has the functions of dispelling wind and heat,
clearing the head, benefiting the throat, penetrating the rash, soothing the liver and
activating the qi. Volatile essential oil extracted from the stems and leaves of mint has
the effects of fragrance, refreshment, brain awakening, solution table and
antibacterial, etc. It has been widely used in the fields of daily chemicals, food and
drugs. Modern medicine has also proved that peppermint has anti-inflammatory and
analgesic effects, anti-fungus, anti-tumor, anti-oxidation and anti-radiation effects.
Grapefruit mint, as a medicinal and edible plant, has a wide range of pharmacological
effects, but the difference between the volatile components of the stem and the leaf is
not clear.
Methods Grapefruit mint samples were collected from cuttings at the age of 4-5
months and planted on floating plates in the closed loop symbiosis system of
fish-flower (grapefruit mint). The leaves and stems of grapefruit mint samples were
treated separately immediately after harvesting for GC-IMS analysis. In this paper,
the differences of volatile components in grapefruit mint leaves and stems from
fish-flower symbiotic closed-loop system were analyzed by gas chromatography-ion
migration spectrum (GC-IMS).
Results We found a total of 36 qualitative volatile substances, including alcohols,
aldehydes, ketones, esters, terpenes, ethers and heterocycles, were identified in the
leaves and stem tissues of grapefruit mint. There were 12 alcohols, 6 aldehydes, 6
49
esters, 6 ketones, 2 ethers, 1 terpene and 3 heterocycles. In alcohols, aldehydes, esters,
ketones, ethers, heterocyclic substances, trans-2-hexenol, geraniol and linalool,
n-hexanal, methyl 2-methylbutyrate, iso-amyl acetate, acetone, 1-octene-3-ketone,
dimethylsulfur and 2-acetylfuran peak volume is large. PCA analysis showed that the
composition of volatile matter in grapefruit mint leaves and stems was significantly
different. The volume of linalool, dimethyl trithiol and ethyl acetate was larger in
leaves and stems, while the volume of trans-2-hexenol, geraniol and eucalyptus oil
was larger in leaves. The peak volume of iso-valeral and butyraldehyde in stem was
larger.
Conclusions GC-IMS was used to analyze the volatile compounds in the leaves and
stems of grapefruit mint. After removing the indeterminate and dimer, there were 36
kinds of qualitative compounds, including alcohols, aldehydes, ketones, esters,
terpenes, ethers and heterocycles. Among them, the volume of trans-2-hexenol,
geraniol, linalool, n-hexyl aldehyde, methyl 2-methylbutyrate, iso-amyl acetate,
acetone, 1-octene-3-ketone, dimethylsulfur and 2-acetylfuran peaks in grapefruit lotus
leaves and stems was relatively large, while that of trans-2-hexenol, geraniol and
eucalyptus oil were relatively high. The iso-amyl aldehyde and butyraldehyde were
higher in the stem. As the main volatile substance, linalool has antiviral and
bacteriostatic effects. Eucalyptus, acetone, dimethyl-sulfur, pinene, 2-ethyl furan all
have inhibitory effects on bacteria. Geraniol has insect repellent and antibacterial
properties. It is clinically used in the treatment of hepatitis and colitis, and has the
effects of alleviating neuroinflammation, inhibiting tumor growth and lowering blood
sugar. Through this study, the differences of volatile substances in the stem and leaf
tissues of grapefruit mint were preliminarily verified, which provided a theoretical
basis for the better application of the stem and leaf tissues of grapefruit mint in
medicinal materials, and also laid a certain foundation for its further application
research.
50
Comparative Analysis of Volatile Components in Leave
and Stem of the Mentha spicala by GC-IMS
Yang Wenli, Zhang Haoyu, Zhang Guangdi
Background The fish-vegetable symbiosis cycle system has realized the ecological
effects of water saving, emission reduction and resource reuse, and is considered to
be a modern agricultural production model with great innovation and sustainable
development potential [10]. Applying spearmint to the fish-flower closed-loop
symbiosis cycle system, using hydroponics technology, it can quickly and efficiently
utilize the metabolites and various mineral nutrients of fish with water as a carrier,
and at the same time achieve zero emissions to the environment. It is a modern
agricultural model that combines high-efficiency aquaculture and hydroponic
planting, which can fully reflect the maximum utilization of resources and the
optimization of benefits. Gas chromatography-Ion mobility spectrometry (GC-IMS)
is a detection technology that combines ion mobility spectrometry and gas
chromatography. It has the advantages of fast detection speed, convenient operation,
and portable equipment. A convenient tool for chemical analysis of food flavor. At
present, it has been widely used in food flavor analysis, quality inspection and other
fields. Compared with the traditional GC-MS method, the GC-IMS method does not
require vacuum, the sample does not need to be enriched and concentrated, and the
temperature for analysis is lower, and some have better thermal stability. Poor
volatiles can be detected more realistically, and the volatile organic components in
the tested sample can be quickly detected by direct headspace sampling. It is used to
identify the authenticity, freshness, shelf life, variety and quality of samples.
Methods Gas chromatography ion mobility chromatography (GC-IMS) was used to
analyze difference of the volatile components of the leaf and stem of Mentha Spicala
which planting in the fish - flower (Spearmint) closed loop symbiosis cycle
system.The GC-IMS Library Serach software was used for quantitative
analysis,according to the retention time of volatile substances by gas chromatography
and IMS migration time. There were 30 volatile substances in stem and leaf tissues of
Spearmint ,which were alcohols, Ketones, esters, aldehydes, sulfur, heterocyclics and
terpenes, among which 10 kinds of were alcohols, accounting for 33.33%.
Results Accordingly, the main volatile components in the leaves and stems of
51
Spearmint are alcohols, ketones, and aldehydes, with the content of alcohols being
the highest. And the overall concentration of volatile components in the leaves is
higher, such as 2-methylfuran, hexanal, 1-pentanol, butyraldehyde, ethanol,
3-methylbutanal, dimethyl sulfide, 2-Ethyl furan, 2-heptanone, benzaldehyde, α-pinene,
isobutanol, ethyl isobutyrate, 1-hexanol, trans-2-hexen-1-ol, linalool, and 3-octyl, all of
these, the content of leaves is much higher than that of stems. In addition,
2-acetylfuran, n-hexanal, ethyl isobutyrate, 1-pentanol, butyraldehyde, ethanol,
isovaleraldehyde, etc. are unique to leaves. And, only a few substances are more
abundant or unique in the stem.
Conclusions Wang Zhonghua and other studies have shown that the spearmint stem
and leaf powder contains a variety of organic acids and flavonoids, which are
beneficial to improve the body’s immune function. After the oil is extracted, the
spearmint still has residual alcohols, ketones and esters. These components can
promote the secretion of intestinal glands, promote the digestion of nutrients,
promote the appetite of animals, increase feed intake, optimize the utilization and
metabolism of nutrients, and the fish-flower (mint) closed-loop symbiosis cycle The
volatile components of the leaves and stems of spearmint in the hydroponic system
are mainly alcohols, ketones, aldehydes and esters. In order to improve the utilization
value of the leaves and stems of spearmint, the spearmint stem and leaf powder after
oil extraction can be applied to livestock and poultry production to increase the added
value of spearmint planting.
52
The Identity of Pathongen Causing Tomato
Powdery Mildew in Yinchuan
Zhou Pengze1, Wang Xiaomin1, 2, 3, 4, Kong Weikang1, Wang Lin1,
Bai Shengyi1, Gao Yanmin1, 2, 3, 4, Li Jianshe1, 2, 3, 4
1 School of Agriculture, Ningxia University. 2 Ningxia Modern Facility Horticulture Engineering Technology Research Center. 3 Key Laboratory of Modern Molecular Breeding for Dominant and Special Crops in Ningxia. 4 Ningxia Facility Horticulture(Ningxia University)Technology Innovation Center.
Background Tomato powdery mildew is a global fungal disease. After Salmon
discovered in 1900 that the pathogen causing tomato powdery mildew in Italy was
Erysiphe polygoni DC. Then tomato powdery mildew in Japan, the Netherlands, the
United Kingdom, France, Bulgaria, Canada, Hungary, Spain and other countries had
been reported. The earliest occurrence of powdery mildew in China was reported by
Japanese scholar Kanji Sawada in Taiwan Province in 1919 and identified as
Erysiphe Polygoni DC. Then there were reports of tomato powdery mildew in
Xinjiang, Heilongjiang and Liaoning, but the results of pathogen identification of
tomato powdery mildew were different. In recent years, The pathogen of tomato
powdery mildew is Oidium neolycopersici was reported in Jilin, Henan, Taiwan,
Gansu and Heilongjiang provinces. Powdery mildew in tomato is a common disease
in China, but no reports were involved in the identification of powdery mildew in
tomato in the city of Yinchuan, Ningxia Hui Autonomous Region.
Therefore, this study conducted morphological and molecular biological
identification of tomato powdery mildew pathogen in Yinchuan region, in order to
provide theoretical basis and reference for the prevention and treatment of tomato
powdery mildew in Yinchuan.
Methods The pathogen of tomato powdery mildew in Yinchuan city was identified
by morphology and molecular biology. First, The collected tomato powdery mildew
leaves were brushed into sterile water containing 0.05% Tween -20 and the spore
suspension was fully shaken. Add a drop of spore suspension and observe Conidia,
Germ tube, Conidiophore, Hyphae and Appressorium under Olympus SC180
microscope. Molecular biological identification: The general primers ITS1 and ITS4
were used for PCR reaction. PCR products were detected by 1% agarose gel
electrophoresis and the target band was sequenced. Sequences using SeqMan
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software sequencing measured signal peak figure, after get the complete sequence.
Nucleic acid sequence alignment was carried out in NCBI. Sites 18S, ITS1, 5.8S,
ITS2 and 28S were determined respectively. Then, sequences between ITS1 and ITS2
were selected for multi-sequence alignment in DNAMAN, and Mega7 was used to
construct phylogenetic tree for genetic evolution analysis.
Results Morphological identification results: the Conidia were present in solitary and
ovoid (or waist-drum). At the end of the bud tube there was a cleft petal or mastoid
appressorium, and appressorium attached to hyphae. The Conidiophore was erect and
unbranched, Furthermore, there was no closed sac shell, according to the above
characteristics is Oidium neolycopersici. Molecular biological identification
results:After PCR amplification, a single clear band appeared between 500 and 750
bp, and sequencing results showed that the length of the band was 640bp, which was
in line with the size of the target ITS sequence. In the ITS sequence with a total
length of 640bp, the sequence sites between ITS1 and ITS2 were 31-582 and the
length was 552 bp. The sequences of ITS1~ITS2 were selected for multi-sequence
comparison to construct phylogenetic tree. The results showed that Leveillula taurica
and Erysiphe polygoni were significantly different from Oidium lycopersici and
Oidium neolycopersici in base sequence and evolutionary distance, but there was no
significant difference in evolutionary distance between Oidium lycopersici and
Oidium neolycopersici. However, there were differences in the evolutionary distance
between the Oidium neolycopersici reported in France and the Oidium neolycopersici
reported in other regions. By ITS sequence alignment, the consistency with the
Oidium neolycopersici was more than 99%.
Conclusions In this study, the pathogenic bacteria of tomato powdery mildew in
Yinchuan city were identified from morphology and molecular biology. The results
show that the Conidia were present in solitary and ovoid (or waist-drum). At the end
of the bud tube there was a cleft petal or mastoid appressorium, and appressorium
attached to hyphae. The Conidiophore was erect and unbranched, furthermore, there
was no closed sac shell, according to the above characteristics is Oidium
neolycopersici, and the homology with Oidium neolycopersici was more than 99%.
The results showed that the pathogen of tomato powdery mildew in Yinchuan city
was Oidium neolycopersici.
54
Applications of Environmental Economic Policies in the Prevention
and Control of Agricultural Non-point Source Pollution in China
Hanqing Wu1; Baogui Zhang1
1 College of Land Science and Technology, China Agricultural University, Beijing, China
Objectives: To realize the green development, effective prevention and control of
agricultural non-point source pollution (ANSP) is the prerequisite.
Methods: Based on the systematic analysis of the current situation, characteristics
and relevant environmental economic policies about the prevention and control of
ANSP, the study used the case analysis to screen out the environmental economic
policies suitable for the prevention and control of ANSP, and then provided a certain
basis for decision makers.
Results: The results indicated that the source control strategy is the key to the
prevention and control of ANSP. ANSP produces negative externality of economy,
but the ecological compensation policy, a positively stimulating environmental
economic policy, which was suitable for internalizing the economic negative
externalization produced by ANSP. The environmental economic policies suitable for
the prevention and control of ANSP mainly focused on ecological compensation
policy. And the applications of producer responsibility extension policy in
agricultural film residual pollution, pesticide packaging waste and ecological
compensation policy in ANSP caused by excessive use of fertilizers and pesticides
were more feasible. The principal part of the ecological compensation policy should
be diversified, and the relationship between government and market should be
complementary, whose combination will benefit the prevention and control of ANSP.
Moreover, the ecological compensation policy and the three-product one-standard
policy, environmental tax policy and producer responsibility extension policy and
green finance-related policies (such as green security, insurance, bonds, procurement
policies, etc.) can effectively control the quality of agricultural products, the use of
fertilizers and pesticides, the recycling of residual film and pesticide packaging waste,
meanwhile, expand the ecological compensation fund pools, so as to better prevent
and control the generation of ANSP.
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Conclusions: There are still many problems in the application of environmental
economic policies in the prevention and control of ANSP, and how to combine
ecological compensation policy with other related environmental economic policies
efficiently and how to supervise the effects of environmental economic policies still
needs further study.
Acknowledgements: This work was supported by the National Natural Science
Foundation of China (Grants No. 2016YFD0800906).