inbre research update revised
TRANSCRIPT
![Page 1: INBRE Research Update REVISED](https://reader035.vdocument.in/reader035/viewer/2022070510/58ae44941a28abad338b5855/html5/thumbnails/1.jpg)
INBRE Research UpdateRachel Gross
![Page 2: INBRE Research Update REVISED](https://reader035.vdocument.in/reader035/viewer/2022070510/58ae44941a28abad338b5855/html5/thumbnails/2.jpg)
Project 1: SINA Interacting Proteins through Yeast Two-Hybrid Screening• Understanding Degradation for Cellular Health
![Page 3: INBRE Research Update REVISED](https://reader035.vdocument.in/reader035/viewer/2022070510/58ae44941a28abad338b5855/html5/thumbnails/3.jpg)
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
![Page 4: INBRE Research Update REVISED](https://reader035.vdocument.in/reader035/viewer/2022070510/58ae44941a28abad338b5855/html5/thumbnails/4.jpg)
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
![Page 5: INBRE Research Update REVISED](https://reader035.vdocument.in/reader035/viewer/2022070510/58ae44941a28abad338b5855/html5/thumbnails/5.jpg)
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
![Page 6: INBRE Research Update REVISED](https://reader035.vdocument.in/reader035/viewer/2022070510/58ae44941a28abad338b5855/html5/thumbnails/6.jpg)
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)
![Page 7: INBRE Research Update REVISED](https://reader035.vdocument.in/reader035/viewer/2022070510/58ae44941a28abad338b5855/html5/thumbnails/7.jpg)
Symptoms: Dark stripe patterns appear when tuber is fried
http://www.apsnet.org/publications/apsnetfeatures/Pages/PotatoZebraChip.aspx
![Page 8: INBRE Research Update REVISED](https://reader035.vdocument.in/reader035/viewer/2022070510/58ae44941a28abad338b5855/html5/thumbnails/8.jpg)
Symptoms: Purple tops, leaf roll, necrosis, chlorsis, premature death
http://www.apsnet.org/publications/apsnetfeatures/Pages/PotatoZebraChip.aspx
![Page 9: INBRE Research Update REVISED](https://reader035.vdocument.in/reader035/viewer/2022070510/58ae44941a28abad338b5855/html5/thumbnails/9.jpg)
Vector: potato-tomato psyllid
http://www.apsnet.org/publications/apsnetfeatures/Pages/PotatoZebraChip.aspx
![Page 10: INBRE Research Update REVISED](https://reader035.vdocument.in/reader035/viewer/2022070510/58ae44941a28abad338b5855/html5/thumbnails/10.jpg)
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
![Page 11: INBRE Research Update REVISED](https://reader035.vdocument.in/reader035/viewer/2022070510/58ae44941a28abad338b5855/html5/thumbnails/11.jpg)
Pathogen EffectorsPathogen
Effector Proteins
PRRs (R genes)
Plant Cell
Resistance (HR cell death)
PAMPs
Resistance
Like Prf
![Page 12: INBRE Research Update REVISED](https://reader035.vdocument.in/reader035/viewer/2022070510/58ae44941a28abad338b5855/html5/thumbnails/12.jpg)
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.
![Page 13: INBRE Research Update REVISED](https://reader035.vdocument.in/reader035/viewer/2022070510/58ae44941a28abad338b5855/html5/thumbnails/13.jpg)
#3
+control
#23
#9
#5
+control
#29
#28
#25
#24
![Page 14: INBRE Research Update REVISED](https://reader035.vdocument.in/reader035/viewer/2022070510/58ae44941a28abad338b5855/html5/thumbnails/14.jpg)
#30
#31
+control
#32
#33
#34GFP (negative control)
+controlPrfdv