大豆根瘤菌效应因子NopA的突变对共生结瘤的影响  

Mutation of typeⅢeffector NopA in rhizobia and its effect on nodulation

作　　者：牟景龙 潘思杨 李元力 张浩然 王锦辉[3] MU Jing-long;PAN Si-yang;LI Yuan-li;ZHANG Hao-ran;WANG Jin-hui(Experimental Training Center of Heilongjiang University of Chinese Medicine,Harbin 150006,China;Agricultural Technology Extension Station of Heilongjiang Province,Harbin 150036,China;College of Agriculture,Northeast Agricultural University,Harbin 150030,China)

机构地区：[1]黑龙江中医药大学实验实训中心,黑龙江哈尔滨150006 [2]黑龙江省农业技术推广站,黑龙江哈尔滨150036 [3]东北农业大学农学院,黑龙江哈尔滨150030

出　　处：《生物技术》2023年第3期269-275,300,共8页Biotechnology

基　　金：中国博士后基金项目(2020M681072)。

摘　　要：[目的]为了探究根瘤菌Ⅲ型效应因子在大豆-根瘤菌共生体系形成过程中作用。[方法]通过三亲杂交的方法进行根瘤菌NopA突变体的构建,并利用PCR和qRT-PCR对突变体进行验证。在染料木苷诱导情况下,纯化根瘤菌胞外蛋白,利用Ⅲ型效应因子NopL及NopT的抗体对纯化蛋白进行检测。将野生型菌株HH103和NopA突变体共同接种大豆,测定NopA突变对根瘤菌结瘤能力的影响。[结果]根瘤菌效应因子NopA突变能够抑制NopL和NopT的分泌。结瘤能力鉴定表明,NopA突变能够显著降低东农50大豆植株根瘤数和根瘤干重,根瘤数由每株31.9下降到21.6,根瘤干重由每株33.9 mg下降到20.5 mg。共生结瘤Marker基因表达量分析表明,在根瘤菌侵染过程中,NopA能够促进GmPR1和PR5的表达。[结论]证明了根瘤菌Ⅲ型效应因子NopA能够影响其他Ⅲ型效应因子的分泌,并且能够正调控大豆-根瘤菌共生结瘤。[Objective]To further explore the function of typeⅢeffect factor of rhizobia,NopA was used as the research object.[MethodJ Three-parent hybridization was used to construct NopA mutants in rhizobia,and PCR and qRT-PCR were used to verify the mutant.The extracellular proteins of rhizobia were purified under the induction of genistin,and the antibodies of typeⅢeffectors NopL and NopT were used to detect the purified proteins.The soybean Dongnong5O were inoculated with wild-type strain HH103 and NopA mutant to determine the effect of NopA mutation on nodulation ability.[Result]The results showed that the mutation of NopA could inhibit the secretion of NopL and NopT.NopA mutation could significantly reduce the nodule number in Dongnong50 from 31.9 to 21.6,and nodule dry-weight from 33.9 mg to 20.5 mg.The marker genes expression analysis showed that NopA could promote the expression of GmPRI and PR5 during rhizobia infection.[Conclusion]NopA can affect the secretion of other typeⅢeffectors,and positively regulate the symbiotic nodulation of soybean-rhizobia.

关 键 词：大豆 根瘤菌 Ⅲ型分泌系统 NopA 结瘤能力鉴定 

分 类 号：Q784[生物学—分子生物学]