《植物生理学报》 2017, 53(8): 1499-1506

蚬壳花椒GID1同源基因克隆及其在种子萌发过程中的表达分析

周韬¹, 王平^2,*, 孙吉康^1,*, 李猛²

中南林业科技大学¹生命科学与技术学院, ²环境科学与工程学院, 长沙410004

通信作者：王平；E-mail: wangping@csuft.edu.cn; 1825856598@qq.com

摘　要：

GID1是赤霉素信号通路关键调控基因, 其通路主要影响种子萌发和植物生长。本文以高通量测序构建的蚬壳花椒(Zanthoxylum dissitum) cDNA文库为参考, 克隆了蚬壳花椒3个GID1同源基因ZaGID1a、ZaGID1b、ZaGID1c。利用生物信息学对蚬壳花椒GID1同源基因的核酸序列及氨基酸序列进行分析, 表明3个同源GID1序列编码的蛋白功能区域、理化性质、系统发育、亲疏水性、亚细胞信号肽定位、三级结构预测等各项生物信息学信息均指向赤霉素受体蛋白功能。同源GID1序列在蚬壳花椒种子人工萌发过程中的实时荧光定量PCR (qRT-PCR)表达量分析研究表明, ZaGID1a、ZaGID1c可能对蚬壳花椒种子萌发起关键调控作用, ZaGID1b与种子萌发过程的关系不明显。

关键词：蚬壳花椒; 克隆; 表达分析; 生物信息学

收稿：2017-03-06   修定：2017-07-05

资助：国家自然科学基金(31370612)和湖南省高校科技成果产业化繁育项目(15CY011)。

Cloning of GID1-homologous genes and expression analysis during seed germination in Zanthoxylum dissitum Hemsl

ZHOU Tao¹, WANG Ping^2,*, SUN Ji-Kang^1,*, LI Meng²

¹College of Life Science, ²College of Environmental Science and Engineering, Central South University of Forestry and Technology, Changsha 410004, China

Corresponding author: WANG Ping; E-mail: wangping@csuft.edu.cn; 1825856598@qq.com

Abstract:

GID1 is the key regulatory gene in gibberellin signaling pathway, which mainly affects seed germination and plant growth. In this study, we cloned three homologous genes of GID1 in Zanthoxylum dissitum, namely ZaGID1a, ZaGID1b and ZaGID1c, by referring to its DNA library established by high throughput sequencing. Bioinformatics was applied to analyze nucleotide and putative amino acid sequences of homologous genes of GID1 in Z. dissitum, and the results indicate the encoded protein functional region, physiochemical property, phylogeny, hydrophobicity-hydrophilicity, subcellular signal peptide location and tertiary structure prediction in three homologous GID1 sequences point to the receptor protein function of gibberellin. Quantitative real time PCR expression analysis of homologous GID1 sequences via Z. dissitum artificial germination indicates ZaGID1a and ZaGID1c played a critical role in the regulation of Z. dissitum seed germination, while the relation between ZaGID1b and seed germination was not evident.

Key words: Zanthoxylum dissitum; cloning; expression analysis; bioinformatics