根瘤菌转录因子RluD基因突变抑制大豆共生结瘤  

Mutation of transcription factor RluD in rhizobia inhibit soybean nodulation

作　　者：霍婧怡 马胜男 辛大伟[1] 陈庆山[1] 王锦辉[1] HUO Jingyi;MA Shengnan;XIN Dawei;CHEN Qingshan;WANG Jinhui(College of Agriculture,Northeast Agricultural University,Harbin 150030;Heilongjiang Academy of Land Reclamation Sciences,Harbin 150038,China)

机构地区：[1]东北农业大学农学院,黑龙江哈尔滨150030 [2]黑龙江省农垦科学院,黑龙江哈尔滨150038

出　　处：《生物技术》2024年第2期135-143,共9页Biotechnology

基　　金：国家自然科学基金项目(32201809)。

摘　　要：[目的]研究根瘤菌自身转录因子RluD在大豆共生结瘤中的作用,[方法]利用三亲杂交对根瘤菌RluD突变体进行构建,并利用PCR和qRT-PCR对突变体进行验证。利用大豆结瘤实验,测定RluD突变对根瘤菌结瘤能力的影响。最后,利用转录组对RluD突变所影响的信号通路进行了研究。[结果]根瘤菌转录因子RluD突变能够抑制根瘤的产生,显著降低大豆植株根瘤数和根瘤干重,根瘤数由每株17.0下降到12.0,根瘤干重由每株24.0 mg下降到18.0 mg。表达量分析表明,在大豆共生结瘤过程中RluD所调控的基因能够参与到大豆的免疫信号途径。转录组结果表明,转录因子RluD主要参与根瘤菌ABC转运,鞭毛组装,胞外多糖生物合成,细菌分泌系统,碳代谢等通路。[结论]根瘤菌转录因子RluD能够正调控大豆共生结瘤,其突变情况下能够降低29.4%的根瘤数和25.0%的根瘤干重。[Objective]To study the function of transcription factor of RluD in rhizobia.[Method]Three-parent hybridization was used to construct RluD mutants in rhizobia,and PCR and Southern Blot were used to verify the mutant.The soybean plants were inoculated with wild-type strain HH103 and RluD mutant to determine the effect of RluD mutation on nodulation ability.Finally,the signaling pathways affected by RluD were investigated using RNA-seq.[Result]The mutation of RluD could in-hibit the formation of nodules.RluD mutation could significantly reduce the nodule number in soybean plants from 17.O to 12.0,and nodule dry-weight from 24.0 mg to 18.0 mg.The qRT-PCR results indicated that the genes regulated by RluD could be associated with immunity signaling during the formation of nodule.Transcriptome results showed that the transcription factor RluD mainly involved in rhizobia ABC transhipment,flagellum assembly,extracellular polysaccharide biosynthesis,bacte-rial secretion system,carbon metabolic pathways.[Conclusion] RluD positively regulate the symbiotic nodulation of soybean,and rlud mutant could reduce nodule number by 29.4%and nodule dry weight by 25.0%.

关 键 词：大豆 根瘤菌 共生结瘤 转录因子 RluD 结瘤鉴定 转录组 信号途径 

分 类 号：S565.1[农业科学—作物学]