Identification of NIN–like protein (NLP) Genes of Sorghum and SbNLP1 Ectopic Expression in Rice Revealed Improved Low Nitrogen Tolerance at the Seedling Stage

Nitrogen (N) is an essential macronutrient for plant growth. NIN–like protein (NLP) transcription factors play important roles in nitrate signaling and response in plants. However, a comprehensive analysis of the NLP gene family in sorghum is still lacking. In this study, we identified five NLP genes in sorghum, and a high collinearity of NLP was detected in sorghum, rice and maize. N deficiency decreased SbNLP3 and SbNLP4 expression levels in roots, and the expression of SbNLP1 and SbNLP2 declined in roots during nitrate resupply. Subcellular localization analysis revealed that SbNLP1 was mostly detected in nuclei and cytoplasm. Compared with wild–type rice ZH11 plants, SbNLP1 overexpression plants showed improved low nitrogen (LN) tolerance, with longer roots and shoots under LN conditions. Transcriptome analysis between overexpression lines OE1–5 and ZH11 showed that 773 and 967 differentially expressed genes (DEGs) were identified in roots and shoots under LN conditions, respectively. In contrast, 674 and 1283 DEGs were identified in roots and shoots under normal nitrogen (NN) conditions, respectively. Thirty–seven N–related DEGs were identified in roots through GO enrichment under LN conditions, and terms of plant hormone signal transduction, biosynthesis of secondary metabolites and plant–pathogen interaction were identified through KEGG enrichment. WGCNA analysis also revealed plant hormone signal transduction pathways and plant pathogen interaction pathways in OE1–5 under LN conditions. These results provide a basis for N use efficiency (NUE) improvement in sorghum and functional analysis of SbNLPs.