在广谱抗稻瘟病品种云引与普感稻瘟病品种丽江新团黑谷(Lijiangxintuanheigu,LTH)构建的近等基因系抗、感病子代基因组重测序的差异基因中,从与免疫应答(immune response)相关的分类单元里得到8个基因序列有差异的基因.它们都为编码富...在广谱抗稻瘟病品种云引与普感稻瘟病品种丽江新团黑谷(Lijiangxintuanheigu,LTH)构建的近等基因系抗、感病子代基因组重测序的差异基因中,从与免疫应答(immune response)相关的分类单元里得到8个基因序列有差异的基因.它们都为编码富含亮氨酸重复序列的类受体蛋白激酶基因,并聚集于11号染色体的邻近位置,且与已知的白叶枯病抗性基因Xa21具有一定同源性.为进一步探究这些类受体蛋白激酶类基因在稻瘟病抗性中可能发挥的功能,我们分析了这些基因的结构、进化及抗感子代间的序列差异类型,同时还分别采用离体和喷雾接菌的方式对云引和LTH进行稻瘟病接种并取样.通过实时荧光定量PCR(quantitative real time polymerase chain reaction,q RT-PCR)发现,部分基因能受到稻瘟病菌的诱导,推测它们可能在水稻对抗稻瘟病病菌中发挥一定作用,为进一步挖掘稻瘟病相关抗性基因奠定基础.展开更多
Plants usually keep resistance(R)proteins in a static state under normal conditions to avoid autoimmunity and save energy for growth,but R proteins can be rapidly activated upon perceiving pathogen invasion.Pib,the fi...Plants usually keep resistance(R)proteins in a static state under normal conditions to avoid autoimmunity and save energy for growth,but R proteins can be rapidly activated upon perceiving pathogen invasion.Pib,the first cloned blast disease R gene in rice,encoding a nucleotide-binding leucine-rich repeat(NLR)protein,mediates resistance to the blast fungal(Magnaporthe oryzae)isolates carrying the avirulence gene AvrPib.However,the molecular mechanisms about how Pib recognizes AvrPib and how it is inactivated and activated remain largely unclear.In this study,through map-based cloning and CRISPR-Cas9 gene editing,we proved that Pib contributes to the blast disease resistance of rice cultivar Yunyin(YY).Furthermore,an SH3 domain-containing protein,SH3P2,was found to associate with Pib mainly at clathrin-coated vesicles in rice cells,via direct binding with the coiled-coil(CC)domain of Pib.Interestingly,overexpression of SH3P2 in YY compromised Pib-mediated resistance to M.oryzae isolates carrying AvrPib and Pib-AvrPib recognition-induced cell death.SH3P2 competitively inhibits the self-association of the Pib CC domain in vitro,suggesting that binding of SH3P2 with Pib undermines its homodimerization.Moreover,SH3P2 can also interact with AvrPib and displays higher affinity to AvrPib than to Pib,which leads to dissociation of SH3P2 from Pib in the presence of AvrPib.Taken together,our results suggest that SH3P2 functions as a“protector”to keep Pib in a static state by direct interaction during normal growth but could be triggered off by the invasion of AvrPib-carrying M.oryzae isolates.Our study reveals a new mechanism about how an NLR protein is inactivated under normal conditions but is activated upon pathogen infection.展开更多
文摘在广谱抗稻瘟病品种云引与普感稻瘟病品种丽江新团黑谷(Lijiangxintuanheigu,LTH)构建的近等基因系抗、感病子代基因组重测序的差异基因中,从与免疫应答(immune response)相关的分类单元里得到8个基因序列有差异的基因.它们都为编码富含亮氨酸重复序列的类受体蛋白激酶基因,并聚集于11号染色体的邻近位置,且与已知的白叶枯病抗性基因Xa21具有一定同源性.为进一步探究这些类受体蛋白激酶类基因在稻瘟病抗性中可能发挥的功能,我们分析了这些基因的结构、进化及抗感子代间的序列差异类型,同时还分别采用离体和喷雾接菌的方式对云引和LTH进行稻瘟病接种并取样.通过实时荧光定量PCR(quantitative real time polymerase chain reaction,q RT-PCR)发现,部分基因能受到稻瘟病菌的诱导,推测它们可能在水稻对抗稻瘟病病菌中发挥一定作用,为进一步挖掘稻瘟病相关抗性基因奠定基础.
基金This work was supported by the National Key R&D Program Foundation of China(grant no.2016YFD0300508)the National Rice Industry Technology System of Modern Agriculture for China(grant no.CARS-01-20)+2 种基金the“5511”Collaborative Innovation Project for High-Quality Development and Surpasses of Agriculture between Government of Fujian and Chinese Academy of Agricultural Sciences(grant no.XTCXGC2021001)Key Program of Science and Technology in Fujian province,China(no.2020NZ08016)the Special Foundation of Non-Profit Research Institutes of Fujian Province(grant no.2018R1021-5).
文摘Plants usually keep resistance(R)proteins in a static state under normal conditions to avoid autoimmunity and save energy for growth,but R proteins can be rapidly activated upon perceiving pathogen invasion.Pib,the first cloned blast disease R gene in rice,encoding a nucleotide-binding leucine-rich repeat(NLR)protein,mediates resistance to the blast fungal(Magnaporthe oryzae)isolates carrying the avirulence gene AvrPib.However,the molecular mechanisms about how Pib recognizes AvrPib and how it is inactivated and activated remain largely unclear.In this study,through map-based cloning and CRISPR-Cas9 gene editing,we proved that Pib contributes to the blast disease resistance of rice cultivar Yunyin(YY).Furthermore,an SH3 domain-containing protein,SH3P2,was found to associate with Pib mainly at clathrin-coated vesicles in rice cells,via direct binding with the coiled-coil(CC)domain of Pib.Interestingly,overexpression of SH3P2 in YY compromised Pib-mediated resistance to M.oryzae isolates carrying AvrPib and Pib-AvrPib recognition-induced cell death.SH3P2 competitively inhibits the self-association of the Pib CC domain in vitro,suggesting that binding of SH3P2 with Pib undermines its homodimerization.Moreover,SH3P2 can also interact with AvrPib and displays higher affinity to AvrPib than to Pib,which leads to dissociation of SH3P2 from Pib in the presence of AvrPib.Taken together,our results suggest that SH3P2 functions as a“protector”to keep Pib in a static state by direct interaction during normal growth but could be triggered off by the invasion of AvrPib-carrying M.oryzae isolates.Our study reveals a new mechanism about how an NLR protein is inactivated under normal conditions but is activated upon pathogen infection.
