水杨酸受体NPR1负调控枝顶孢霉Acremonium sp. D212与宿主植物共生
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摘要
[目的]主要探究内生真菌Acremoniumsp.D212(枝顶孢霉D212)与植物激素水杨酸相互作用调控植物发育的机制。[方法]用枝顶孢霉D212与三七共培养后,通过转录组测序及PCR方法,检测基因表达;将枝顶孢霉D212接种水稻培养后,进行免疫反应,观察枝顶孢霉D212在水稻根中的定殖及水稻根系的发育情况;利用免疫组化及细胞学的方法观察生长素输出蛋白的细胞定位。[结果]枝顶孢霉D212与三七共生时,引起了水杨酸途径及病程防御相关基因的大量表达,但没诱导病程防御反应基因表达水平的提高。但是,降低了独脚金内酯中PnDAD2d,PnD14c及PnD27基因的表达。枝顶孢霉D212可以同水稻共生,并可大量定殖于水杨酸受体突变体OsNPR1-RNAi植株中,促进根系的生长。枝顶孢霉D212与水稻OsNPR1-RNAi突变体的共生降低了生长素输出蛋白OsPIN1与OsPIN2在根表皮细胞中的细胞膜定位。[结论]枝顶孢霉D212与宿主植物共生需要水杨酸受体NPR1,并改变生长素的运输而影响根系生长。
[Objective]This study aimed to detect the mechanism of endophytic fungus Acremonium sp.D212 and phytohormone salicylic acid interaction in regulating plant development.[Method]The genes expression levels were detected by transcriptome sequencing and PCR method after Panax notoginseng inoculated with Acremonium sp.D212 The colonization of Acremonium sp.D212 and rice root development were observed,after rice inoculated with Acremonium sp.D212.Immunohistochemical and cytological methods were used to detect the cellular localization of auxin efflux protein.[Result]Symbiosis of Acremonium sp.D212 and P.notoginseng induced gene expression,and most of these genes involved in salicylic acid pathway and defence reaction,but did not increased the expression levels of pathogenesis-related genes.However,the expression levels of strigolactone genes PnDAD2 d,PnD14 c and PnD27 were decreased.Acremonium sp.D212 could form symbiont with rice and effectively colonized in salicylic acid receptor mutant OsNPR1-RNAi to promote root growth.The localization of auxin efflux protein OsPIN1 and OsPIN2 in plasmamembrane of root epidermal cell of rice mutant OsNPR1-RNAi inoculated with Acremonium sp.D212 were decreased.[Conclusion]The salicylic acid receptor NPR1 was required for the symbiosis of Acremonium sp.D212 and host plant in regulating root growth by disruption of auxin transport.
[Objective]This study aimed to detect the mechanism of endophytic fungus Acremonium sp.D212 and phytohormone salicylic acid interaction in regulating plant development.[Method]The genes expression levels were detected by transcriptome sequencing and PCR method after Panax notoginseng inoculated with Acremonium sp.D212 The colonization of Acremonium sp.D212 and rice root development were observed,after rice inoculated with Acremonium sp.D212.Immunohistochemical and cytological methods were used to detect the cellular localization of auxin efflux protein.[Result]Symbiosis of Acremonium sp.D212 and P.notoginseng induced gene expression,and most of these genes involved in salicylic acid pathway and defence reaction,but did not increased the expression levels of pathogenesis-related genes.However,the expression levels of strigolactone genes PnDAD2 d,PnD14 c and PnD27 were decreased.Acremonium sp.D212 could form symbiont with rice and effectively colonized in salicylic acid receptor mutant OsNPR1-RNAi to promote root growth.The localization of auxin efflux protein OsPIN1 and OsPIN2 in plasmamembrane of root epidermal cell of rice mutant OsNPR1-RNAi inoculated with Acremonium sp.D212 were decreased.[Conclusion]The salicylic acid receptor NPR1 was required for the symbiosis of Acremonium sp.D212 and host plant in regulating root growth by disruption of auxin transport.
引文
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[11]赵宜婷,武丽霞,詹晓平,等.水杨酸抑制生长素运输而调节Acune水稻根的生长[J].西南农业学报,2019,32(4):770-775.
[12]Sun C,Shao Y,Vahabi K,et al.The beneficial fungus Piriformospora indica protects Arabidopsis from Verticillium dahliae infection by downregulation plant defense responses[J].BMC Plant Biol,2014,14:268.
[13]Sauer M,Paciorek T,Benkova E,et al.Immunocytochemical techniques for whole-mount in situ protein localization in plants[J].Nat Protoc,2006(1):98-103.
[14]Yoshida S,Kameoka H,Tempo M,et al.The D3 F-box protein is a key component in host strigolactone responses essential for arbuscular mycorrhizal symbiosis[J].New Phytol,2017,196:1208-1216.
[15]Abdalla M A,Matasyoh J C.Endophytes as producers of peptides:an overview about the recently discovered peptides from endophytic microbes[J].Nat Prod Bioprospect,2014(4):457-270.
[16]Martos G G,Teran Mdel M,Diaz Ricci J C.The defence elicitor AsES causes a rapid and transient membrane depolarizatioc,a triphasic oxidative burst and the accumulation of nitric oxide[J].Plant Physiol Biochem,2015,97:443-450.
[17]Duleja S S,Giri R,Saini R,et al.Interaction of endophytic microbes with legumes[J].J Basic Microbiol,2012,52:248-260.
[18]Xu M,Zhu L,Shou H,et al.A PIN1 family gene,OsPIN1,involved in auxin-dependent adventitious root emergence and tillering in rice[J].Plant Cell Physiol,2005,46(10):1674-1681.
[19]Chen Y,Fan X,Song W,et al.Over-expression of OsPIN2 leads to increased tiller numbers,angle and shorter plant height through suppression of OsLAZY1[J].Plant Biotechnol J,2012,10(2):139-149.
[2]张玉洁,和泽高,唐龙山,等.三七内生真菌分离与分子鉴定(I)[J].文山学院学报,2011,24:1-3.
[3]Gutjahr C,Phytohormone signaling in arbuscular mycorhiza development[J].Curr Opin Plant Biol,2014,20:26-34.
[4]Lin L,Xu X.Indole-3-acetic acid production by endophytic Streptomyces sp.En-1 isolated from medicinal plants[J].Curr Microbiol,2013,67:209-217.
[5]崔晋龙,范黎,丁翠,等.栽培黄芩高活性内生真菌菌株的筛选与鉴定[J].西北植物学报,2007,27:1384-1388.
[6]樊美珍,黄勃,李春如,等.枝顶抱属一新记录种[J].菌物系统,1999,18:449.
[7]于慧瑛,吕国忠,孙晓东.不同生长年限人参根际土壤真菌种类及数量的初步研究[J].人参研究,2006(4):9-11.
[8]慕东艳,吕国忠,孙晓东,等.从药用植物根际土壤中分离出的2个中国新记录种[J].菌物研究,2012,10:1-3.
[9]姚磊,徐良雄,薛璟花,等.枝顶孢属真菌的抑菌活性及其代谢产物研究[J].热带亚热带植物学报,2012,20:192-196.
[10]Hael-Conrad V,Abou-Mansour E,Diaz-Ricci J C,et al.The novel elicitor AsES triggers a defense response against Botrntis cinerea in Arabidopsis thaliana[J].Plant Sci,2015,241:120-127.
[11]赵宜婷,武丽霞,詹晓平,等.水杨酸抑制生长素运输而调节Acune水稻根的生长[J].西南农业学报,2019,32(4):770-775.
[12]Sun C,Shao Y,Vahabi K,et al.The beneficial fungus Piriformospora indica protects Arabidopsis from Verticillium dahliae infection by downregulation plant defense responses[J].BMC Plant Biol,2014,14:268.
[13]Sauer M,Paciorek T,Benkova E,et al.Immunocytochemical techniques for whole-mount in situ protein localization in plants[J].Nat Protoc,2006(1):98-103.
[14]Yoshida S,Kameoka H,Tempo M,et al.The D3 F-box protein is a key component in host strigolactone responses essential for arbuscular mycorrhizal symbiosis[J].New Phytol,2017,196:1208-1216.
[15]Abdalla M A,Matasyoh J C.Endophytes as producers of peptides:an overview about the recently discovered peptides from endophytic microbes[J].Nat Prod Bioprospect,2014(4):457-270.
[16]Martos G G,Teran Mdel M,Diaz Ricci J C.The defence elicitor AsES causes a rapid and transient membrane depolarizatioc,a triphasic oxidative burst and the accumulation of nitric oxide[J].Plant Physiol Biochem,2015,97:443-450.
[17]Duleja S S,Giri R,Saini R,et al.Interaction of endophytic microbes with legumes[J].J Basic Microbiol,2012,52:248-260.
[18]Xu M,Zhu L,Shou H,et al.A PIN1 family gene,OsPIN1,involved in auxin-dependent adventitious root emergence and tillering in rice[J].Plant Cell Physiol,2005,46(10):1674-1681.
[19]Chen Y,Fan X,Song W,et al.Over-expression of OsPIN2 leads to increased tiller numbers,angle and shorter plant height through suppression of OsLAZY1[J].Plant Biotechnol J,2012,10(2):139-149.