陆地棉中赤霉素合成途径关键酶基因的时空表达变化

doi:10.7668/hbnxb.2018.04.002

摘要/Abstract

摘要： 为揭示赤霉素合成途径关键酶基因在棉花生长发育过程中的调控作用，利用实时荧光定量PCR方法中的相对定量方法，对中棉所49材料植株体内的赤霉素合成途径相关基因表达量进行了检测分析。结果显示，幼苗期茎组织内除GA2ox1基因外，其余各基因表达量相对较高。开花期根中GA2ox1、GA3ox1、GID1B，茎中CPS1、GA20ox1、GA3ox1、GID1B和叶中GA3ox1基因表达量上调；吐絮期根中GA2ox1和GA3ox1基因表达量上调，其余基因下调，茎中KS、GID1B基因表达量下调，其余基因上调，其中以GA2ox1上调幅度最大，叶中GA2ox1基因表达量下调，其余基因上调。结果表明，开花期，茎中GA20ox1基因的高表达为植株茎的伸长及果枝发育提供必要的活性赤霉素水平，GA3ox1基因可能与开花成铃有关；吐絮期，随着根茎组织的木质化，根茎中GA2ox1基因表达量逐渐升高以降低活性GAs合成，而叶组织中GA3ox1和GA20ox1基因表达量上调，则为棉桃生长发育与脱水成熟提供必要的激素水平。棉花通过改变植株体内赤霉素合成代谢途径关键酶基因的表达量来调控植株的生长发育，且随着生长发育的进行，各目的基因的表达量变化趋势存在一定差异。为深入研究赤霉素对陆地棉的调控机理提供了理论基础，有助于加快利用赤霉素进行品种改良与种质创新。

关键词: 棉花, 赤霉素, 基因表达量, 不同组织, 生长发育

Abstract: To reveal the relationship between the key enzyme gene expression of gibberellin synthesis pathway and different tissues or stages of growth, the CCRI49 cotton was selected as material, and relative quantification Real-time PCR was developed to determine the gibberellin key gene expression of different cotton organizations in different stages of development. The results showed that, in seedling, the expression level of all gibberellin synthesis pathway genes was higher in stem, except GA2ox1 gene. In full-blooming stage, the expression of GA2ox1, GA3ox1, GID1B in root, CPS1, GA20ox1, GA3ox1, GID1B in stem and GA3ox1 in leaf had an up-regulation. In boll opening stage, the expression levels of GA2ox1 and GA3ox1 were up-regulated in root, while the remaining genes were down-regulated. The expression levels of KS and GID1B in stem were down-regulated, while the other genes were up-regulated, with GA2ox1 rise in maximum. And GA2ox1 in the leaf was down-regulated and the rest were up-regulated. In full-blooming stage, the high expression of GA20ox1 gene provided the necessary active gibberellin level for the elongation and growth of the plant stem, and the GA3ox1 gene might be associated with flowering and bell. In boll opening stage, the expression of GA2ox1 gene in root and stem gradually increased to reduce the synthesis of active, gibberellic acid. The gene expression levels of GA3ox1 and GA20ox1 in leaf tissues were raised to provide the necessary hormone levels for cotton boll growth and dehydration. The results indicated that cotton plant by changing the key enzymes gene expression of gibberellin synthesis metabolic pathway to regulate plant growth and development, and the purpose gene expression volume trends exist a certain difference along with the growth and development. It provides a theoretical basis for studying the regulation mechanism of gibberellin on upland cotton, which can help accelerate the improvement of variety and germplasm innovation.

Key words: Cotton, Gibberellin, Gene expression, Different tissues, Growth and development

中图分类号:

Q786

石建斌, 王宁, 周红, 许庆华, 乔文青, 严根土. 陆地棉中赤霉素合成途径关键酶基因的时空表达变化[J]. 华北农学报, 2018, 33(4): 9-16. doi: 10.7668/hbnxb.2018.04.002.

SHI Jianbin, WANG Ning, ZHOU Hong, XU Qinghua, QIAO Wenqing, YAN Gentu. The Temporal Spatial Expression of Gibberell in Key Metabolic Enzyme in Upland Cotton[J]. Acta Agriculturae Boreali-Sinica, 2018, 33(4): 9-16. doi: 10.7668/hbnxb.2018.04.002.

http://www.hbnxb.net/CN/Y2018/V33/I4/9

参考文献

[1] 李四游. 赤霉素对莲株型影响及其相关基因克隆与时空表达分析[D]. 武汉:华中农业大学,2015.
[2] 石建斌. 马铃薯赤霉素合成代谢途径关键酶基因GA2ox1, GA20ox1的克隆与功能分析[D]. 西宁:青海大学,2016.
[3] 虞慧芳,曹家树,王永勤. 植物矮化突变体的激素调控[J]. 生命科学,2002,14(2):85-88,76.
[4] 王海波,邹竹荣,龚明. 小桐子赤霉素20氧化酶的基因克隆及低温下表达分析[J]. 生物技术通报,2015,31(10):140-148.
[5] 孙同玉,祝娟,孙鹏,等. 西洋参赤霉素20-氧化酶基因的克隆与序列分析[J]. 中草药,2015,46(11):1656-1660.
[6] 王增辉. 黄瓜赤霉素氧化酶基因GA20ox、Ga2ox、Ga3ox 的生物信息学及GA20ox1功能分析[D]. 泰安:山东农业大学,2014.
[7] 岳川,曾建明,曹红利,等. 茶树赤霉素受体基因CsGID1a 的克隆与表达分析[J]. 作物学报,2013,39(4):599-608.
[8] 陈敏,马琳,贾聪俊,等. 紫花苜蓿赤霉素受体基因的克隆及表达分析[J]. 西北植物学报,2016,36(11):2159-2166.
[9] 刘斌. 黄瓜赤霉素受体基因CsGID1a的功能验证与表达谱分析[D]. 北京:中国农业大学,2017.
[10] 李强,吴建明,梁和,等. 高等植物赤霉素生物合成及其信号转导途径[J]. 生物技术通报,2014(10):16-22.
[11] 鲁井云,卢龙,邢佳毅,等. 外施赤霉素对峰后葡萄坐果及VvAG 基因表达的影响[J]. 中国农业大学学报,2015,20(1):53-59.
[12] 王西成,任国慧,房经贵,等. 葡萄赤霉素合成相关基因克隆、亚细胞定位和表达分析[J]. 中国农业科学,2012,45(11):2224-2231.
[13] 王西成,吴伟民,房经贵,等. 葡萄赤霉素受体基因VvGID1A 的分离、亚细胞定位及表达分析[J]. 园艺学报,2013,40(5):839-848.
[14] 钟希琼,王惠珍. 高等植物赤霉素生物合成及其调节研究进展[J]. 植物学通报,2001,18(3):303-307.
[15] 王丽. 赤霉素合成抑制剂DPC调控棉花营养生长的分子机制研究[D]. 北京:中国农业大学,2014.
[16] 张国华,张艳洁,丛日晨,等. 赤霉素作用机制研究进展[J]. 西北植物学报,2009,29(2):412-419.
[17] 石建斌,杨永智,王舰. 马铃薯突变体赤霉素代谢关键酶基因差异表达分析[J]. 生物技术通报,2016,32(1):124-130.
[18] Prisic S,Peters R J. Synergistic substrate inhibition of ent-copalyl diphosphate synthase:a potential feed-forward inhibition mechanism limiting gibberellin metabolism[J]. Plant Physiology,2007,144(1):445-454.
[19] 柯贞进. 谷子萌发过程中贮藏物质变化及赤霉素代谢关键酶基因表达的分析[D]. 太谷:山西农业大学,2015.
[20] 李晨晨,侯雷,尹亮,等. 水稻极矮突变体s2-47对赤霉素的响应及基因定位研究[J]. 作物学报,2013,39(10):1766-1774.
[21] Itoh H,Tatsumi T,Sakamoto T,et al. A rice semi-dwarf gene,Tan-Ginbozu(D35),encodes the gibberellin biosynthesis enzyme,ent-kaurene oxidase[J]. Plant Molecular Biology,2004,54(4):533-547.
[22] 郑伶杰,马宏,张媛,等. 苹果矮化砧木GA20氧化酶基因的克隆及其表达分析[J]. 华北农学报,2017,32(1):53-59.
[23] 王成祥,李长生,侯蕾,等. 花生赤霉素2-氧化酶基因的克隆和表达研究[J]. 山东农业科学,2013,45(1):14-18,24.
[24] Spray C R,Kobayashi M,Suzuki Y,et al. The dwarf-1(dt)Mutant of Zea mays blocks three steps in the gibberellin-biosynthetic pathway[J]. Proceedings of the National Academy of Sciences of the United States of America,1996,93(19):10515-10518.
[25] Huang S,Raman A S,Ream J E,et al. Overexpression of 20-oxidase confers a gibberellin-overproduction phenotype in Arabidopsis[J]. Plant Physiology,1998,118(3):773-781.
[26] Chiang H H,Hwang I,Goodman H M. Isolation of the Arabidopsis GA4 locus[J]. The Plant Cell,1995,7(2):195-201.
[27] Itoh H,Tanaka-Ueguchi M,Kawaide H,et al. The gene encoding tobacco gibberellin 3beta-hydroxylase is expressed at the site of GA action during stem elongation and flower organ development[J]. The Plant Journal,1999,20(1):15-24.
[28] Lange T R S. Expression of a gibberellin 2β. 3β-hydroxylase cDNA from pumpkin endosperm[J]. Plant Cell,1997,9(8):1459-1467.
[29] Lee D J,Zeevaart J A. Differential regulation of RNA levels of gibberellin dioxygenases by photoperiod in spinach[J]. Plant Physiology,2002,130(4):2085-2094.
[30] Yamaguchi S,Kamiya Y. Gibberellin biosynthesis:its regulation by endogenous and environmental signals[J]. Plant & Cell Physiology,2000,41(3):251-257.

选择文件类型/文献管理软件名称

选择包含的内容