• ISSN 1008-505X
  • CN 11-3996/S
黄星瑜, 刘小红, 匡志明, 骆乐, 徐国华. 氮素敏感时期水稻分蘖芽的转录分析[J]. 植物营养与肥料学报, 2021, 27(7): 1247-1257. DOI: 10.11674/zwyf.20627
引用本文: 黄星瑜, 刘小红, 匡志明, 骆乐, 徐国华. 氮素敏感时期水稻分蘖芽的转录分析[J]. 植物营养与肥料学报, 2021, 27(7): 1247-1257. DOI: 10.11674/zwyf.20627
HUANG Xing-yu, LIU Xiao-hong, KUANG Zhi-ming, LUO Le, XU Guo-hua. Transcriptional analysis of rice tiller buds at nitrogen sensitive stage[J]. Journal of Plant Nutrition and Fertilizers, 2021, 27(7): 1247-1257. DOI: 10.11674/zwyf.20627
Citation: HUANG Xing-yu, LIU Xiao-hong, KUANG Zhi-ming, LUO Le, XU Guo-hua. Transcriptional analysis of rice tiller buds at nitrogen sensitive stage[J]. Journal of Plant Nutrition and Fertilizers, 2021, 27(7): 1247-1257. DOI: 10.11674/zwyf.20627

氮素敏感时期水稻分蘖芽的转录分析

Transcriptional analysis of rice tiller buds at nitrogen sensitive stage

  • 摘要:
    目的 氮素影响水稻分蘖芽的发育,从而影响水稻株型和产量。探究水稻分蘖芽在氮素敏感时期的基因表达情况,揭示氮素对水稻分蘖芽调控的可能途径。
    方法 以水稻品种‘日本晴’为试验材料,萌发后用1/2MS培养基培养一周,之后用2.5 mmol/L的氮素溶液培养,待幼苗长至三叶期,进行0和2.5 mmol/L氮素溶液处理,培养至五叶期。对不同氮素浓度下分蘖芽的生长进行分析,确认水稻材料的氮素敏感时期,并于根茎结合处取样,提取RNA,进行氮素敏感时期水稻分蘖芽的转录组分析,包括差异表达基因的挖掘以及GO功能富集分析、KEGG通路分析、蛋白互作网络分析。
    结果 缺氮条件下,水稻分蘖芽生长受到抑制,转录分析结果显示,不同供氮条件下842个基因存在显著的表达差异,其中586个基因缺氮时上调,256个基因缺氮时下调。GO功能富集分析发现绝大多数差异表达基因属于胞内 (cell)、胞内成分 (cell part) 和细胞器 (organelle) 的类别。在差异表达基因的KEGG通路分析中,植物激素信号传导途径是最显著富集的通路,氮代谢途径次之,说明植物激素通路和氮代谢通路在水稻分蘖芽的生长过程中具有重要作用。差异表达基因主要涉及生长素、细胞分裂素、脱落酸、水杨酸、茉莉酸等相关激素的合成代谢途径以及参与氮代谢的硝酸还原酶。蛋白互作分析推测硝酸还原酶可能会与植物激素相互作用。
    结论 通过对氮素敏感时期水稻分蘖芽相关基因的转录分析,发现缺氮条件下,激素信号传导途径和氮代谢途径中相关基因的表达量均受到影响。其中,与硝酸还原酶和植物激素脱落酸合成相关的基因表达量上调,而影响分蘖芽生长的细胞分裂素和生长素基因表达均下调。

     

    Abstract:
    Objectives Nitrogen affects the development of tiller buds, thus influencing the architecture and yield of rice. This study explores gene expression of rice tiller buds at the sensitive stage of N supply to reveal the possible pathways of N regulation.
    Methods The rice variety, ‘Nipponbare’, was used as the experimental material and seeds were placed in a culture medium a week after germination. We used a solution containing N 2.5 mmol/L to cultivate the plants until the 3rd leaf became fully developed. The seedlings were divided into two groups and subjected to solutions containing N 0 and 2.5 mmol/L, respectively until the 5th leaf became fully developed. The growth of tiller buds under different N concentrations was observed to confirm rice phenotype during the N sensitive period. Samples were taken at the junction of roots and stems, these samples were used for RNA extraction. RNA samples were collected for transcriptional analysis, including differentially expressed genes. Also, GO analysis, KEGG analysis, and protein-interaction analysis were carried out.
    Results Under N deficient condition, the growth of tiller buds was inhibited, and the N content in the rice plants (P < 0.05) reduced. The results of transcription analysis showed that 842 genes were differentially expressed under different N supply conditions, with 586 genes up-regulated and 256 genes down-regulated. The GO analysis showed that most of the differentially expressed genes belonged to cell, cell part, and organelle categories. KEGG analysis of DEGs revealed that the “plant hormone signal transduction” pathway was the most enriched pathway and the “N metabolism” pathway was the second. This suggests that plant hormones and N metabolism could play essential roles in regulating the growth of rice tiller bud. The differentially expressed genes mainly relates to synthesis and metabolism pathways of auxin, cytokinin, abscisic acid, salicylic acid, jasmonic acid, and nitrate reductase involved in N metabolism. Protein-interaction analysis showed that nitrate reductase possibly interacted with phytohormone.
    Conclusions The transcriptome analysis of the tiller buds indicated N deficiency influenced the phytohormone biosynthesis and transduction and N metabolism at the sensitive stage. Nitrogen deficit up-regulated the expression of nitrate reductase and phytohormone genes, while down-regulated cytokinin and auxin lead to tiller bud outgrowth as a result.

     

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