枳NLP转录因子克隆及其在不同水分条件下的表达

doi:10.3864/j.issn.0578-1752.2016.02.018

Abstract

Abstract: 【Objective】The objective of this study was to analyze the expression of Poncirus trifoliata (L.) Raf. NIN-like transcription factors to discuss the regulation mechanism of nitrogen assimilation under different water conditions in citrus.【Method】 Primers were designed to amplify the sequences of the NLP genes CDS of Poncirus trifoliate (L.) Raf.based on orange genome databases and molecular biology. Alignment of the sequences was performed using ClustalX and phylogenetic analysis of this alignment was conducted using MEGA. Analysis of the Relative NLP genes expression data under different water conditions using Real-Time Quantitative PCR.【Result】Four Poncirus trifoliata (L.) Raf. NLP genes: PtNLP2, PtNLP4, PtNLP7 and PtNLP8 were obtained. The sequence alignment analysis showed that the identity of all NIN-like proteins was 45.13%, and possessed both the RWP-RK and PB1 domain, and all NIN-like proteins were high identity to sweet orange, respectively 97.57%, 96.47%, 99% and 97.33%. Phylogenic analysis has indicated that the 4 Poncirus trifoliata (L.) Raf. NLP genes can be classified into four groups with Arabidopsis (PtNLP2 and AtNLP1/2, PtNLP4 and AtNLP4/5, PtNLP7 and AtNLP6/7, PtNLP8 and AtNLP8/9). There were differences in the expression pattern of NLP genes in Poncirus trifoliata (L.) Raf. leaves and roots. The results indicated that with the decrease of soil water, the expression level of NLP genes in Poncirus trifoliata (L.) Raf. leaves were up-regulated. The PtNLP2, PtNLP4, PtNLP7 and PtNLP8 expression level were the highest when the relative water capacity was 15.4%, they were 2.9, 3.5, 5.9 and 2.8 folds higher respectively compared to the control (the relative water holding capacity was 61.0%). After that, the gene expression level began to down-regulate and had no statistically significant difference between the control and the relative water holding capacity of 9.4%. While the expression level of NLP genes in Poncirus trifoliata (L.) Raf. roots of control were the highest, and the gene expression pattern showed a down-regulated trend accompanied by the loss of soil water and the difference was statistically significant. The PtNLP2, PtNLP4, PtNLP7 and PtNLP8 expression level of roots were maximum down-regulated by 6.7, 2.8, 4.8 and 2.3 folds respectively compared with the control. The expression of NLP genes in Poncirus trifoliata (L.) Raf. leaves and roots remained at the low expression level after rewatering and had statistically significant difference compared with the control.【Conclusion】The expression of Poncirus trifoliata (L.) Raf. NLP genes is closely related to the water condition of soil. The expression level of NLP genes in Poncirus trifoliata (L.) Raf. leaves were up-regulated by drought stress, and then down-regulated when the water deficit was very serious. But the expression level of NLP genes in Poncirus trifoliata (L.) Raf. roots were down-regulated continuously accompanied by the loss of soil water, and expression of the PtNLP2 and PtNLP7 had a great change in roots.

Key words: Poncirus trifoliata (L.) Raf., NLP transcription factors, nitrogen, drought stress, gene expression

CAO Xiong-jun, LU Xiao-peng, XIONG Jiang, LI Jing, WU Qian, ZHOU Fang-fang, XIE Shen-xi. Cloning and Expression of Poncirus Trifoliata (L.) Raf. NIN- Like Transcription Factors under Different Water Conditions[J].Scientia Agricultura Sinica, 2016, 49(2): 381-390.

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Cloning and Expression of Poncirus Trifoliata (L.) Raf. NIN- Like Transcription Factors under Different Water Conditions

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