Detection of the virulent form of AVR3a from Phytophthora infestans following artificial evolution of potato resistance gene R3a.
Detection of the virulent form of AVR3a from Phytophthora infestans following artificial evolution of potato resistance gene R3a.
复制标题
DOI：
10.1371/journal.pone.0110158
复制
发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Hein I
中科院分区：
文献类型：
--
作者：
Chapman S;Stevens LJ;Boevink PC;Engelhardt S;Alexander CJ;Harrower B;Champouret N;McGeachy K;Van Weymers PS;Chen X;Birch PR;Hein I
工程抗性基因以获得效应器识别是获得广泛、持久抗性的重要步骤。我们设计了马铃薯抗性基因R3a，以获得对疫霉毒力强的AVR3aEM效应形式的识别。对R3a进行随机诱变、基因改组和定点诱变，获得对AVR3aEM识别的R3a*变异。获得识别后的程序性细胞死亡在迭代的人工进化中增强，并且接近R3a对AVR3aKI的识别水平。我们证明了R3a*介导的识别反应，像R3a一样，依赖于SGT1和HSP90。此外，当与AVR3aKI或AVR3aEM共表达时，这种反应的增加与R3a*变异体从细胞质重新定位到晚期内体有关，这一机制以前只在R3a与AVR3aKI共浸润时被发现。同样，AVR3aEM在被R3a*变体识别后特异性地重新定位到相同的囊泡上，而不是被R3a识别。R3a和R3a*对表达AVR3aKI的P. infestans分离株具有抗性，但对表达AVR3aEM的纯合子不具有抗性。
Engineering resistance genes to gain effector recognition is emerging as an important step in attaining broad, durable resistance. We engineered potato resistance gene R3a to gain recognition of the virulent AVR3aEM effector form of Phytophthora infestans. Random mutagenesis, gene shuffling and site-directed mutagenesis of R3a were conducted to produce R3a* variants with gain of recognition towards AVR3aEM. Programmed cell death following gain of recognition was enhanced in iterative rounds of artificial evolution and neared levels observed for recognition of AVR3aKI by R3a. We demonstrated that R3a*-mediated recognition responses, like for R3a, are dependent on SGT1 and HSP90. In addition, this gain of response is associated with re-localisation of R3a* variants from the cytoplasm to late endosomes when co-expressed with either AVR3aKI or AVR3aEM a mechanism that was previously only seen for R3a upon co-infiltration with AVR3aKI. Similarly, AVR3aEM specifically re-localised to the same vesicles upon recognition by R3a* variants, but not with R3a. R3a and R3a* provide resistance to P. infestans isolates expressing AVR3aKI but not those homozygous for AVR3aEM.
登录
查看更多内容
DOI：
10.1111/tpj.12307
发表时间：
2013-11
期刊：
The Plant journal : for cell and molecular biology
影响因子：
--
作者：
Jupe F;Witek K;Verweij W;Sliwka J;Pritchard L;Etherington GJ;Maclean D;Cock PJ;Leggett RM;Bryan GJ;Cardle L;Hein I;Jones JD
通讯作者：
Jones JD
影响因子：
7.2
作者：
Huang, SW;van der Vossen, EAG;Visser, RGF
通讯作者：
Visser, RGF
影响因子：
3
作者：
Duan, Hui;Richael, Craig;Rommens, Caius M.
通讯作者：
Rommens, Caius M.
影响因子：
3.5
作者：
Foster, Simon J.;Park, Tae-Ho;Jones, Jonathan D. G.
通讯作者：
Jones, Jonathan D. G.
影响因子：
7.2
作者：
Gilroy, Eleanor M.;Hein, Ingo;Birch, Paul R. J.
通讯作者：
Birch, Paul R. J.