Abstract

Cross talking between natural senescence and cell death in response to pathogen attack is an interesting topic; however, its action mechanism is kept open. In this study, 33 OsWRKY genes were obtained by screening with leaf aging procedure through RNA-seq dataset, and 11 of them were confirmed a significant altered expression level in the flag leaves during aging by using the reverse transcript quantitative PCR (RT-qPCR). Among them, the OsWRKY2, OsWRKY14, OsWRKY26, OsWRKY69, and OsWRKY93 members exhibited short-term alteration in transcriptional levels in response to Magnaporthe grisea infection. The CRISPR/Cas9-edited mutants of five genes were developed and confirmed, and a significant sensitivity to M. oryzae infection was observed in CRISPR OsWRKY93-edited lines; on the other hand, a significant resistance to M. oryzae infection was shown in the enhanced expression OsWRKY93 plants compared to mock plants; however, enhanced expression of other four genes have no significant affection. Interestingly, ROS accumulation was also increased in OsWRKY93 enhanced plants after flg22 treatment, compared with the controls, suggesting that OsWRKY93 is involved in PAMP-triggered immune response in rice. It indicated that OsWRKY93 was involved in both flag leaf senescence and in response to fungi attack.

Highlights

  • Rice is the main food crop of the developing world

  • The first sample of the flag leaf was collected at the heading stage when the flag leaf was fully expanded [0 weeks after heading (WAH) and named 0W]; chlorophyll content is higher in 1w than 0w, and it is gradually decreased from 1w to 5w; the following five flag leaf samples were collected every week

  • We have identified 11 OsWRKYs that were differentially expressed during the senescence of flag leaves through RNA-Seq together with the reverse transcript quantitative PCR (RT-qPCR) analysis

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Summary

Introduction

The increase of yield is seriously restricted by flag leaf senescence in rice. The flag leaf, the uppermost leaf in the rice plant, is thought to contribute highly to what is accumulated in grain (Ghosh et al, 1990; Li et al, 1998). Studying the mechanism of flag leaf senescence is essential to improving the yield of rice grain. Leaf senescence is the final stage of leaf development. As an organ level senescence, leaf senescence is a crucial means for plants to reallocate nutrients and valuable substances from senescent leaves to reproducing seeds, eventually maximizing reproductive success (Himelblau and Amasino, 2001). Leaf senescence is a strictly organized process finely governed by developmental age. Leaf senescence is influenced by various internal and environmental signals that

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