Abstract

Nodule development directly affects nitrogen fixation efficiency during soybean growth. Although abundant genome-based information related to nodule development has been released and some studies have reported the molecular mechanisms that regulate nodule development, information on the way nodule genes operate in nodule development at different developmental stages of soybean is limited. In this report, notably different nodulation phenotypes in soybean roots inoculated with Bradyrhizobium japonicum strain 113-2 at five developmental stages (branching stage, flowering stage, fruiting stage, pod stage and harvest stage) were shown, and the expression of nodule genes at these five stages was assessed quantitatively using RNA-Seq. Ten comparisons were made between these developmental periods, and their differentially expressed genes were analysed. Some important genes were identified, primarily encoding symbiotic nitrogen fixation-related proteins, cysteine proteases, cystatins and cysteine-rich proteins, as well as proteins involving plant-pathogen interactions. There were no significant shifts in the distribution of most GO functional annotation terms and KEGG pathway enrichment terms between these five development stages. A cystatin Glyma18g12240 was firstly identified from our RNA-seq, and was likely to promote nodulation and delay nodule senescence. This study provides molecular material for further investigations into the mechanisms of nitrogen fixation at different soybean developmental stages.

Highlights

  • Environmental pollution and other issues[15,16]

  • We investigated ten comparisons between five important developmental stages of soybean and identified a large number of differentially expressed genes (DEGs), including those encoding soybean symbiotic nitrogen fixation-related proteins, cysteine proteases, cystatins and cysteine-rich proteins, as well as proteins involved in plant-pathogen interactions

  • To investigate nodule development during soybean growth, we examined the symbiotic phenotypic of soybean inoculated with B. japonicum 113-2 at five developmental stages, and the results are shown in Fig. 1 and Table S1

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Summary

Introduction

Environmental pollution and other issues[15,16]. Studies on how to reduce the amount of nitrogen fertilizer and increase crop nitrogen utilization efficiency are very important, and there have been some good studies on the high nitrogen utilization efficiency of soybean[17,18,19]. A comparative analysis of genome sequences in six legumes revealed many of the symbiotic genes in soybean[25], and RNA-Seq transcription data predicted several nodulation-related gene regulatory networks[26] This genome-based information is constrained by prior knowledge of gene sequences and limits the patterns of gene expression at various stages of nodule development. We are in a position to significantly improve our understanding of soybean nodule development using RNA-Seq, which is an effective method that produces quantitative data related to transcripts with greater sensitivity, higher reproducibility, and wider dynamic range[27] than other conventional methods This method has relatively little variation between technical replicates for identifying differentially expressed genes[28]. The discovered differentially expressed genes (DEGs) and the results described in this study should aid efforts to understand the mechanisms of soybean nodule development and shed new light on the molecular mechanisms of nitrogen fixation at five important developmental stages

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