GmEXPA11 facilitates nodule enlargement and nitrogen fixation via interaction with GmNOD20 under regulation of GmPTF1 in soybean

Abstract

Biological nitrogen fixation (BNF) provides 50–60 % of the nitrogen for plant growth and development, while its application is restricted for the deficiency of functional gene in biological breeding. Expansin can enlarge the plant cells through loosening the cell wall, which has a great breeding potential for legumes BNF improvement. In the present study, a cell wall α-subfamily expansin, GmEXPA11, was isolated and analyzed in soybean nodule growth and nitrogen fixation process. GmEXPA11 was highly induced by rhizobial infection and appeared high expressions in the whole process of soybean nodulation and nitrogen fixation. The overexpression of GmEXPA11 facilitated nodule cell enlargement and generated much more big nodules, with an increase of 37.6 % on nodule cell length, 14.7 % on cell width, 25.8 % on big nodule number, 25.6 % on nodule weight, while the RNAi nodules were opposite. Moreover, GmEXPA11 overexpression enhanced nodule nitrogen fixation ability, with the increases of 22.9 %, 6.7 % and 11.7 % on nitrogenase activity, nitrogen content and hairy root nitrogen content, while the RNAi decreased by 11.9 %, 10.7 % and 7.8 %, respectively. Further analysis demonstrated that GmEXPA11 affected nodules enlargement and nitrogen fixation via interacting with nodulin GmNOD20 under the regulation of transcription factor GmPTF1. The expression of GmEXPA11 was significantly increased in the transgenic nodules with GmPTF1 over-expressed. In addition, by analyzing soybean resequencing accessions, four upstream SNPs were found in the promoter of GmEXPA11 and formed two haplotypes with significantly different soybean nodulation and nitrogen fixation characters, which demonstrated the close relationship between GmEXPA11-SNPs and BNF.