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. 2020 May 19;11:453. doi: 10.3389/fpls.2020.00453

FIGURE 1.

FIGURE 1

The information of NopD gene, detection of NopD protein induction by genistein and nodule traits of soybean plants inoculated with S. fredii wild-type HH103 or mutants HH103ΩNopD and HH103ΩTtsI. (A) Position of the NopD gene and phylogenetic tree of NopD from various rhizobial strains by neighbor-joining method using MEGA6.0 software. (B) Expression analysis of NopD in S. fredii wild-type strain HH103 and mutants HH103ΩNopD and HH103ΩTtsI with (+) or without (−) genistein (3.7 μM). Final expression was calculated relative to the expression of the wild-type strain HH103 in the absence of genistein. All sample harvests were performed with three biological replicates, and the individual values for each RNA sample were analyzed by three technical replications. The sample of the wild-type strain S. fredii HH103 in the absence of genistein was used as the control. Asterisks indicate significant differences at the level α = 1% (P < 0.01). Immunoblot analysis of NopD in extracellular protein extracts of the wild-type strain S. fredii HH103 and indicated mutant derivatives induced with or without genistein (3.7 μM). Immunoblots were performed with anti-NopD antibodies. (C) The analysis of phenotype was performed three times; significant differences were determined by t-tests; * indicates 0.01 ≤ P ≤ 0.05 and **indicates P ≤ 0.01. Soybean varieties (with origins): Heinong35 (Heilongjiang), Qingdou (Shanxi), Zheng9525 (Henan), Baimaodou (Zhejiang), Chidou1 (Inner Mongolia), Suinong14 (Heilongjiang), ZYD00006 (Heilongjiang), Charleston (America), Dongnong594 (Heilongjiang), Kenjian28 (Heilongjiang).