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林业科学 ›› 2019, Vol. 55 ›› Issue (12): 61-73.doi: 10.11707/j.1001-7488.20191207

• 论文与研究报告 • 上一篇    下一篇

毛竹细胞分裂素相关基因鉴定及在笋中表达特征分析

杨雅倩1,傅鹰1,周明兵1,2,*   

  1. 1. 浙江农林大学 省部共建亚热带森林培育国家重点实验室 杭州 311300
    2. 浙江省竹资源与高效利用协同创新中心 杭州 311300
  • 收稿日期:2018-12-27 出版日期:2019-12-25 发布日期:2020-01-02
  • 通讯作者: 周明兵
  • 基金资助:
    国家自然科学基金项目(31870656);国家自然科学基金项目(31500542);国家自然科学基金项目(31470615);浙江自然科学基金重点项目(LZ19C160001)

Identification of Cytokinin Related Genes and Characterization of Their Expression in Phyllostachys edulis Shoots

Yaqian Yang1,Ying Fu1,Mingbing Zhou1,2,*   

  1. 1. State Key Laboratory of Subtropical Silviculture Zhejiang A&F University Hangzhou 311300
    2. Zhejiang Provincial Collaborative Innovation Center for Bamboo Resources and High-Efficiency Utilization Hangzhou 311300
  • Received:2018-12-27 Online:2019-12-25 Published:2020-01-02
  • Contact: Mingbing Zhou
  • Supported by:
    国家自然科学基金项目(31870656);国家自然科学基金项目(31500542);国家自然科学基金项目(31470615);浙江自然科学基金重点项目(LZ19C160001)

摘要:

目的: 已有研究表明快速拔节的毛竹笋不同节间组织细胞分裂素分布有差异,这可能是影响毛竹快速生长的主要因素之一。本研究探索不同节间细胞分裂素相关基因的表达特征对毛竹快速生长的影响,为进一步阐明毛竹快速生长机理提供理论基础。方法: 利用拟南芥和水稻基因组数据库及KEGG代谢通路等,初步获得与细胞分裂素代谢通路和信号通路相关的基因。通过NCBI、Pfam和SMART软件在线鉴定,最终得到与细胞分裂素相关基因。将毛竹基因组数据库与所得细胞分裂素相关基因家族Blast比对,获得毛竹细胞分裂素关键基因。通过同源蛋白进化发育系统和基序分布,研究它们间的亲缘关系与进化特点。以快速生长毛竹笋不同节间组织为材料,通过qRT-PCR探究了细胞分裂素相关基因表达特征及其对毛竹快速生长的影响。结果: 植物体内与细胞分裂素相关的基因主要有5大家族:合成基因家族、降解基因家族、受体基因家族、转运基因家族及信号转导因子家族。同源蛋白基序分析发现,拟南芥、水稻、毛竹细胞分裂素相关基因同源蛋白的结构域保守性高度一致,毛竹细胞分裂素相关基因同源蛋白在进化上与水稻关系更近;但有部分同源蛋白结构有种内与种间差异,进化上呈现片段丢失与重复现象,引起同源蛋白功能分化。由qRT-PCR具体分析毛竹不同节间组织细胞分裂素相关基因表达模式,结果显示,合成基因PhIPT1表达量在接近顶端的幼嫩节间略有下降趋势,而PhIPT2在幼嫩节间表达量更高;降解基因PhCKX在顶端幼嫩节间表达量降低;受体基因PhHK表达量在毛竹笋不同节间变化不规律,而转运基因PhHP在顶端幼嫩节间表达量略微增加,信号转导因子PhRR表达量在不同节间有增有减。结论: 对毛竹中的细胞分裂素5大类相关基因同源蛋白研究,发现大部分细胞分裂素相关同源蛋白在结构和生物学功能上都具有较高的保守性,但部分同源蛋白在结构与进化上具有物种特异性。通过对毛竹不同节间细胞分裂素相关基因表达模式研究发现,毛竹形态学上端细胞分裂素含量高于形态学下端,推测其在毛竹快速生长阶段发挥重要生物学功能,调节秆茎伸长生长和变粗。

关键词: 毛竹, 细胞分裂素, 快速生长, 进化系统, 表达模式

Abstract:

Objectve: Lots of studies have shown that there are differences in the distribution of cytokinins in Phyllostachys edulis different internodes which are growing fast, which may be one of the main causes of rapid growth of P. edulis. So the study was intended to explore the effect of expression patterns of cytokinin-related key genes in the rapid growth of P. edulis shoot, and provide a theoretical basis for further elucidation of the rapid growth mechanism. Method: We extracted cytokinin-related gene families by the retrieval of Arabidopsis thaliana genome database, Oryza sativa genome database and KEGG metabolic pathway. The cytokinin-related key genes were identified by NCBI, Pfam and SMART online. The P. edulis genome database was blasted with the obtained cytokinin-related key gene family to extract P. edulis related key genes. The evolutionary relationships of homologous proteins were analyzed by constructing phylogenetic trees and motifs of homologous proteins. The expression patterns of cytokinin-related gene and their effects on the rapid growth of P. edulis were investigated by qRT-PCR with different internode tissues of rapidly growing P. edulis. Result: There are five cytokinin-related key gene families including isopentenyl-transferases family, cytokinin oxidase family, histidine kinases family, histidine phosphotransferase family and response regulator family. The cytokinin-related key genes of P. edulis is more closely phylogenetic to rice genes. The motifs of gene families in Arabidopsis, rice, and P. edulis are highly conserved, but some homologous genes show species-specific. The expression of five key gene families were specifically analyzed by qRT-PCR. The results showed that the expression level of the PhIPT2 was higher in young internodes and the PhIPT1 decreased slightly; while the expression level of PhCKXs decreased gradually; the expression level of receptor gene PhHKs was not stable in different internodes of P. edulis; the expression level of the transporter PhHPs increased slightly with the younger internodes; the expression level of the response regulator PhRRs increased and decreased between different internodes. Conclusion: The most cytokinin-related key genes are highly conserved in structure and biological function, but some of them are species-specific in structure and evolution. The expression patterns of cytokinin-related key genes in P. edulis shoot were studied, and the content of cytokinin in the morphologically upper part of P. edulis was higher than that in morphologically lower part. Therefore, we supposed that cytokinin plays a major biological function in the rapid growth of P. edulis, and regulates the growth and thickening of stems.

Key words: Phyllostachys edulis, cytokinin, rapid growth, evolutionary system, expression pattern

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