Transcript
INBRE Research UpdateRachel Gross
Project 1: SINA Interacting Proteins through Yeast Two-Hybrid Screening• Understanding Degradation for Cellular Health
SINAs role in Degradation• SINA is group of E3 ligase proteins that labels
different targets within the cell to be degraded. • Studies have shown that some SINAs are involved in
defense regulation
Defense Protein
No Pathogen
Defense Protein
No Pathogen
SIN
A
ub
Screening for Interacting Proteins• Using cDNA from tomato,
cut into fragments and placed into yeast cells with SINAs.
• Colonies with interacting proteins turn blue on special plates
• Next step: isolate the interacting gene and sequence
Findings so far:• Promoter-binding-like protein: Transcription factor
• Sec61 alpha-like subunit: fungal resistance
• CBSD containing transcription: very little information on this protein
• chitin synthesis: fungal resistance primary defense line
• RALF-like protein: involved in plant growth regulation
• CDC5-like protein: cell cycle regulation
Project 2: Zebra Chip Effectors• Zebra Chip Disease is a relatively new bacterial
infection that was first observed in Mexico in the late 1990s and appeared in Texas in 2000.
• Symptoms: easily confused with other diseases until the processing time
• Signs: very little is known• Vector: Potato-Tomato psyllid (Bactericera
cockerelli)
Symptoms: Dark stripe patterns appear when tuber is fried
http://www.apsnet.org/publications/apsnetfeatures/Pages/PotatoZebraChip.aspx
Symptoms: Purple tops, leaf roll, necrosis, chlorsis, premature death
http://www.apsnet.org/publications/apsnetfeatures/Pages/PotatoZebraChip.aspx
Vector: potato-tomato psyllid
http://www.apsnet.org/publications/apsnetfeatures/Pages/PotatoZebraChip.aspx
Signs• Using PCR scientists were able to identify regions
of the pathogens DNA that are related to citrus greening complex (a bacterial pathogen also vectored by the psyllid) and so they named the new pathogen Candidatus Liberibacter psyllaurous
• Preliminary studies show that Ca. Liberibacter can be transmitted plant to plant or through the psyllid population.
http://www.apsnet.org/publications/apsnetfeatures/Pages/PotatoZebraChip.aspx
Pathogen EffectorsPathogen
Effector Proteins
PRRs (R genes)
Plant Cell
Resistance (HR cell death)
PAMPs
Resistance
Like Prf
Cell Death Assay using Agrobacterium• Mix each effector with mutant defense protein Prf-
turns on HR • Use Prfᵈ¹⁴¹⁶ᵛ with GFP as a positive control• Use GFP alone as negative control• If cell death is delayed or not present, it suggests
the effectors are silencing the HR cell death response-block Prf pathway.
#3
+control
#23
#9
#5
+control
#29
#28
#25
#24
#30
#31
+control
#32
#33
#34GFP (negative control)
+controlPrfdv
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