单 位：
华中农业大学生命科学技术学院;阜阳职业技术学院生化工程学院
关键词：
日本百脉根;菌根因子;LcLRK1基因;根内球囊霉;LysM型受体激酶;RNA干扰
摘 要：
从日本百脉根(Lotus corniculatus)中克隆到一个可能与菌根因子识别相关的Lc LRK1基因,通过设计简并引物并利用RACE PCR手段,在百脉根中扩增得到Lc LRK1基因全长序列。通过生物信息学的方法,对Lc LRK1基因进行结构分析和预测;接种Rhizophagus irregularis,检测Lc LRK1基因在百脉根不同组织中的表达水平;构建Pro Lc LRK1∶∶GUS重组载体,观察Lc LRK1基因在根中的表达部位;最后通过RNA干扰(RNAi)技术进行Lc LRK1基因沉默。结果表明:Lc LRK1编码一个Lys M型受体激酶,由3个胞外Lys M结构、1个跨膜结构域与1个胞内激酶结构域组成;Lc LRK1在AM共生早期表达水平明显增强,并在根的表皮细胞及根毛基部表达;Lc LRK1基因沉默后抑制了早期丛枝的形成;相对于对照组,在Lc LRK1RNAi的植株中AM真菌的侵染率明显下降,丛枝数目也明显降低。证实Lc LRK1基因与菌根真菌信号分子识别相关,并且影响AM真菌丛枝的形成。
译 名：
Lys M receptor LcLRK1 from Lotus corniculatus involved in the arbuscule formation
作 者：
HAN Yachao;XIE Xian'an;GENG Cong;ZHAO Bin;College of Biochemical Engineering,Fuyang Vocational and Technical College;College of Life Sciences and Technology,Huazhong Agricultural University;
关键词：
Lotus corniculatus;;Myc-factor;;LcLRK1 gene;;Rhizophagus irregularis;;LysM receptor kinase;;RNA interference(RNAi)
摘 要：
A Lys M receptor Lc LRK1 gene possibly involved in Myc-factor perception was isolated from Lotus corniculatus. The full-length of Lc LRK1 gene was isolated from L. corniculatus by using the degenerate primers and RACE PCR strategies. The structure of Lc LRK1 gene was analyzed with bioinformatics. The expression profiles of Lc LRK1 gene were investigated in variant tissues from L. corniculatus after inoculation Rhizophagus irregularis. A recombinant plasmid harboring Pro Lc LRK1 ∶ ∶ GUS reporter was constucted to localize Lc LRK1 in the roots of L. corniculatus. The Lc LRK1 gene was knock-down with RNA interference. The results showed that Lc LRK1 gene,encoding a Lys M receptor kinase,was comprised of three extracellular Lys M domains,a transmembrane domain and intracellular kinase domain. Lc LRK1 gene was up-regulated in roots at early stage of AM symbiosis and the transcripts of Lc LRK1 were mainly present in the root epidemis and root hairs. The knock-down of the Lc LRK1 gene significantly decreased AM fungal colonization. In RNAi lines,the infection rate and arbuscule numbers were strongly reduced compared to that in the controls. The Lc LRK1 gene may participate in the perception of Myc-factor and down-regulation of Lc LRK1,affecting the formation of AM fungal arbuscule.
