YDA和MPK3/MPK6上游的MAPKKK3/MAPKKK5在植物免疫和生长/发育中的重叠功能,Journal of Integrative Plant Biology

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YDA和MPK3/MPK6上游的MAPKKK3/MAPKKK5在植物免疫和生长/发育中的重叠功能
Journal of Integrative Plant Biology ( IF 9.3 ) Pub Date : 2022-06-02 , DOI: 10.1111/jipb.13309
Yidong Liu ₁ , Emma Leary ₂ , Obai Saffaf ₁ , R Frank Baker ₃ , Shuqun Zhang ₁

Affiliation

拟南芥丝裂原活化蛋白激酶 3（MAPK3 或 MPK3）和 MPK6 在植物免疫和生长/发育中发挥重要的信号传导作用。MAPK 激酶4 (MKK4) 和MKK5 在这些过程中在MPK3 和MPK6 上游冗余地发挥作用。YODA（YDA）又名MAPK激酶激酶4（MAPKKK4），位于MKK4/MKK5上游，形成一个完整的MAPK级联（YDA-MKK4/MKK5-MPK3/MPK6）调控植物生长发育。在植物免疫中，MAPKKK3 和 MAPKKK5 在同一 MKK4/MKK5–MPK3/MPK6 模块的上游冗余地发挥作用。然而，在mapkkk3 mapkkk5中 MPK3 /MPK6 的剩余激活响应flg22病原体相关分子模式（PAMP）处理的双突变体表明在该MAPK级联中存在额外的MAPKKK（s）以发出植物免疫信号。为了研究 YDA 是否也参与植物免疫，我们尝试生成mapkkk3 mapkkk5 yda三重突变体。然而，由于胚胎发生失败，不可能恢复双突变体组合（mapkkk5 yda）或三突变体（mapkkk3 mapkkk5 yda）之一。使用成簇的规则间隔短回文重复序列 (CRISPR) – CRISPR 相关蛋白 9 (Cas9) 方法，我们在mapkkk3 mapkkk5中生成了YDA、yda-del的弱 N 端缺失等位基因背景。PAMP 触发的 MPK3/MPK6 激活在mapkkk3 mapkkk5 yda-del突变体中进一步降低，并且三重突变体更容易受到病原体感染，这表明 YDA 在植物免疫信号传导中也起重要作用。此外，发现 MAPKKK5和在较小程度上MAPKKK3与 YDA一起有助于配子功能和胚胎发生。虽然双纯合mapkkk3 yda突变体显示出与yda单突变体相同的生长和发育缺陷，但mapkkk5 yda双突变体和mapkkk3 mapkkk5 yda三突变体是胚胎致死的，类似于mpk3 mpk6双突变体。这些结果表明，YDA、MAPKKK3 和 MAPKKK5 在植物免疫和生长/发育方面在 MKK4/MKK5–MPK3/MPK6 模块上游具有重叠功能。

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Overlapping functions of YDA and MAPKKK3/MAPKKK5 upstream of MPK3/MPK6 in plant immunity and growth/development

Arabidopsis MITOGEN-ACTIVATED PROTEIN KINASE3 (MAPK3 or MPK3) and MPK6 play important signaling roles in plant immunity and growth/development. MAPK KINASE4 (MKK4) and MKK5 function redundantly upstream of MPK3 and MPK6 in these processes. YODA (YDA), also known as MAPK KINASE KINASE4 (MAPKKK4), is upstream of MKK4/MKK5 and forms a complete MAPK cascade (YDA–MKK4/MKK5–MPK3/MPK6) in regulating plant growth and development. In plant immunity, MAPKKK3 and MAPKKK5 function redundantly upstream of the same MKK4/MKK5–MPK3/MPK6 module. However, the residual activation of MPK3/MPK6 in the mapkkk3 mapkkk5 double mutant in response to flg22 pathogen-associated molecular pattern (PAMP) treatment suggests the presence of additional MAPKKK(s) in this MAPK cascade in signaling plant immunity. To investigate whether YDA is also involved in plant immunity, we attempted to generate mapkkk3 mapkkk5 yda triple mutants. However, it was not possible to recover one of the double mutant combinations (mapkkk5 yda) or the triple mutant (mapkkk3 mapkkk5 yda) due to a failure of embryogenesis. Using the clustered regularly interspaced short palindromic repeats (CRISPR) – CRISPR-associated protein 9 (Cas9) approach, we generated weak, N-terminal deletion alleles of YDA, yda-del, in a mapkkk3 mapkkk5 background. PAMP-triggered MPK3/MPK6 activation was further reduced in the mapkkk3 mapkkk5 yda-del mutant, and the triple mutant was more susceptible to pathogen infection, suggesting YDA also plays an important role in plant immune signaling. In addition, MAPKKK5 and, to a lesser extent, MAPKKK3 were found to contribute to gamete function and embryogenesis, together with YDA. While the double homozygous mapkkk3 yda mutant showed the same growth and development defects as the yda single mutant, mapkkk5 yda double mutant and mapkkk3 mapkkk5 yda triple mutants were embryo lethal, similar to the mpk3 mpk6 double mutants. These results demonstrate that YDA, MAPKKK3, and MAPKKK5 have overlapping functions upstream of the MKK4/MKK5–MPK3/MPK6 module in both plant immunity and growth/development.

更新日期：2022-06-02

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