Abstract

NODULE-INCEPTION-like proteins (NLPs) are conserved, plant-specific transcription factors that play crucial roles in responses to nitrogen deficiency. However, the evolutionary relationships and characteristics of NLP family genes in Brassica napus are unclear. In this study, we identified 31 NLP genes in B. napus, including 16 genes located in the A subgenome and 15 in the C subgenome. Subcellular localization predictions indicated that most BnaNLP proteins are localized to the nucleus. Phylogenetic analysis suggested that the NLP gene family could be divided into three groups and that at least three ancient copies of NLP genes existed in the ancestor of both monocots and dicots prior to their divergence. The ancestor of group III NLP genes may have experienced duplication more than once in the Brassicaceae species. Three-dimensional structural analysis suggested that 14 amino acids in BnaNLP7-1 protein are involved in DNA binding, whereas no binding sites were identified in the two RWP-RK and PB1 domains conserved in BnaNLP proteins. Expression profile analysis indicated that BnaNLP genes are expressed in most organs but tend to be highly expressed in a single organ. For example, BnaNLP6 subfamily members are primarily expressed in roots, while the four BnaNLP7 subfamily members are highly expressed in leaves. BnaNLP genes also showed different expression patterns in response to nitrogen-deficient conditions. Under nitrogen deficiency, all members of the BnaNLP1/4/5/9 subfamilies were upregulated, all BnaNLP2/6 subfamily members were downregulated, and BnaNLP7/8 subfamily members showed various expression patterns in different organs. These results provide a comprehensive evolutionary history of NLP genes in B. napus, and insight into the biological functions of BnaNLP genes in response to nitrogen deficiency.

Highlights

  • Nitrogen is a primary nutrient that is critical for the survival of all living organisms

  • The BnaNLP gene family is larger than the NODULE-INCEPTION-like proteins (NLPs) gene family in other plant species, because B. napus experienced an extra whole genome triplication (WGT) and merging events [35]

  • Similar results were detected in BnaNLP genes (Table 1), which illustrated that this transcription factors (TFs) prefer to be active in acidic conditions

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Summary

Introduction

Nitrogen is a primary nutrient that is critical for the survival of all living organisms. The absorption and utilization of nitrogen directly affects plant growth and development, as well as crop yields. During their long evolutionary history, plants have established complex and delicate regulatory mechanisms to adapt to changes in nitrogen conditions in the external environment. The major form of nitrogen in soil, plays important physiological and nutritional roles in plant growth and development [1]. Several proteins are involved in nitrogen assimilation in plants exposed to changing nitrogen concentrations, such as ammonium transporters (AMTs) and nitrate reductases (NRs) [5,6,7]. Subsequent studies found that NLP genes widely exist among non-nitrogen fixating plants, such as rice (Oryza sativa) and A. thaliana [10], but the NIN proteins seem to be unique in legumes

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