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Editing rice negative regulation resistance gene OsEDR1 by CRISPR/Cas9 and its function analysis*(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

Issue:
2020 02
Page:
1-10
Research Field:
Articles
Publishing date:

Info

Title:
Editing rice negative regulation resistance gene OsEDR1 by CRISPR/Cas9 and its function analysis*
Author(s):
WU Guangheng1 2** FU Xianyu3
1 Provincial Key Laboratory of Eco-Industrial Green Technology, Wuyishan 354300, China
2 College of Ecology and Resources Engineering, Wuyi University, Wuyishan 354300, China 3 College of Tea and Food Science, Wuyi University, Wuyishan 354300, China
Keywords:
EDR1 protein kinase gene editing over-expression bacterial blight
CLC:
-
PACS:
DOI:
10.19675/j.cnki.1006-687x.2019.05007
DocumentCode:

Abstract:
EDR1(Enhanced Disease Resistance1) in Arabidopsis thaliana is a Raf-like MAPKK protein kinase that negatively regulates salicylic acid-induced defense response and controls plant immune defense response through MKK4/MKK5–MPK3/MPK6. In order to study the function of OsEDR1 gene in rice, we developed a loss-of-function mutant of OsEDR1 using CRISPR/Cas9 technology. Over-expression vectors pTCK303-Ubi-OsEDR1 and pBI1.4t-35S-OsEDR1 were constructed to transform rice and Arabidopsis to obtain transgenic plants. Finally, bacterial blight and powdery mildew were inoculated to analyze the resistance. The results showed that CRISPR/Cas9 technology created loss-of-function mutant of OsEDR1 with single base insertion. The inoculation experiment of Xoo PXO99 showed that the length of lesion of loss-of-function mutant was reduced by about 50% compared with that of Nipponbare, which enhanced resistance of rice to Xoo PXO99. Quantitative PCR and inoculation experiment results showed that over-expression of OsEDR1 in Nipponbare could enhance susceptibility to Xoo PXO86. The heterologous expression of OsEDR1 in Arabidopsis edr1 mutant could not compromise resistance of edr1 mutant to powdery mildew and cell death induced by powdery mildew. In conclusion, OsEDR1 negatively regulates resistance to bacterial blight and spontaneous cell death in rice, but it may be different from defense signaling in Arabidopsis. These results will be helpful to further study the mechanism of EDR1 gene and its application in rice breeding.

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Last Update: 2019-07-30