银杏性别决定相关基因的筛选

doi:10.11707/j.1001-7488.20170209

摘要/Abstract

摘要： [目的] 银杏为典型的雌雄异株裸子植物，成熟雌、雄个体在形态特征和生长习性等方面存在显著差异，这些差异的产生与性别决定机制相关。本文拟通过对银杏性别决定相关基因的筛选为进一步探讨银杏的性别决定机制奠定基础。[方法] 利用RNA-Seq技术对来源于同一家系的25年生银杏雌、雄花芽（CY，XY）及大、小孢子叶球（CH，XH）进行转录组测序及生物信息学分析，以期发现可能参与银杏性别决定过程的相关基因。利用实时荧光定量PCR方法对随机挑选的26个差异表达基因的表达量进行验证。[结果] 通过对8个cDNA文库的测序共得到大约60 Gb的高质量序列，利用高质量序列进行de novo组装共得到108 307条unigene。unigene的平均长度为796 bp。26个随机挑选差异表达基因的实时荧光定量PCR结果与RNA-Seq结果的相关性较高。在所有的unigene中有51 953条（47.97%）unigene在核苷酸或蛋白质公共数据库中得到功能注释。差异表达分析结果表明分别有4 709和9 802个基因在XY/CY和XH/CH中差异表达。11个在XY/CY和XH/CH中共同差异表达参与植物激素信号转导的基因与转录因子（如PYL、SNRK2和EIN3等）以及编码甲基转移酶的基因（如MET1和COMT1等）可能参与了银杏的性别决定过程。[结论] 参与多种调控途径的功能基因可能在银杏的性别决定中发挥作用，性别决定相关基因的筛选可为全面理解银杏的性别决定机制奠定基础。

关键词: 银杏, 性别决定, 基因, 转录组测序

Abstract: [Objective] In plants, sex determination is a comprehensive process of correlated events, which involves genes that are differentially or specially expressed in distinct developmental phases. Ginkgo biloba is a typical dioecious gymnosperm species with high ecological and economical values. The morphological traits and growth habits are significantly different between mature female and male individuals. These differences are likely related to sex determination in G. biloba. In the present study, the screening of sex determination genes of G. biloba will lay a solid foundation for further exploring the mechanism of sex determination in this species.[Method] We conducted RNA-sequencing of female and male buds (CY and XY) as well as ovulate strobilus and staminate strobilus (CH and XH) of G. biloba from the same family to gain insights into the genes potentially related to sex determination in this species. The expression level of 26 randomly selected differentially expressed genes (DEGs) was tested using quantitative real-time PCR.[Result] Approximately 60 Gb of clean reads were obtained from eight cDNA libraries. De novo assembly of the clean reads generated 108 307 unigenes with an average length of 796 bp. A high level of correlation was found between the RNA-Seq and qRT-PCR for the 26 randomly selected DEGs. Among these unigenes, 51 953 (47.97%) had at least one significant match with a gene sequence in one of the public protein and nucleotide databases. A total of 4 709 and 9 802 DEGs were identified in XY vs.CY and XH vs.CH, respectively. 11 commonly DEGs and transcription factors associated with plant hormone signal and transduction (such as PYL, SNRK2, EIN3 etc.) as well as genes encoding methyltransferase (e.g. MET1 and COMT1) were assumed to involve in sex determination in G. biloba.[Conclusion] The transcriptome resources generated allowed us to identify diverse functional genes that may be associated with sex determination in G. biloba. Our results laid a solid foundation for fully exploring the mechanism of sex determination in G. biloba.

Key words: Ginkgo biloba, sex determination, genes, RNA-Seq

中图分类号:

S718.46

唐海霞, 杜淑辉, 邢世岩, 桑亚林, 李际红, 刘晓静, 孙立民. 银杏性别决定相关基因的筛选[J]. 林业科学, 2017, 53(2): 76-82.

Tang Haixia, Du Shuhui, Xing Shiyan, Sang Yalin, Li Jihong, Liu Xiaojing, Sun Limin. Screening of Sex Determination Related Genes in Ginkgo biloba[J]. Scientia Silvae Sinicae, 2017, 53(2): 76-82.

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