2024年12月10日 星期二
米槠雄花发育相关的转录组和MADS-box 家族基因表达特性分析
Analysis on transcriptomes and expression characteristics of MADS-box family genes related to male flower development of Castanopsis carlesii
2022年 第31卷 第2期 页码[39-48]    下载全文[5.5MB]  
摘要

 以米槠〔Castanopsis carlesii (Hemsl.) Hayata〕花芽期、半开期和盛花期的雄花为研究材料,利用高通量测序平台构建米槠雄花发育过程的转录组数据库。分别对花芽期与半开期、半开期与盛花期以及花芽期与盛花期进行比较,筛选与雄花发育相关的差异表达unigenes,并对差异表达unigenes进行功能注释。同时对米槠MADS-box家族基因进行鉴定,分析其在米槠雄花发育3个时期的表达量变化。结果显示:在获得的转录组中,clean reads数为628 800 000~643 700 000,高质量总碱基量为6.29~6.44 Gb,Q30碱基百分比均不小于90.67%,GC含量为45.09%~45.39%。花芽期与半开期、半开期与盛花期以及花芽期与盛花期间分别有15 086、6 770和16 150个差异表达unigenes。GO功能注释结果显示:差异表达unigenes主要注释在催化活性、连接和膜等相关的生物过程。KEGG代谢通路注释结果显示:差异表达unigenes主要注释在全局及概要图、碳水化合物代谢、环境适应性和信号转录等相关的代谢通路。说明催化酶在米槠雄花发育过程有重要作用,注释在细胞组分中的差异表达unigenes主要与花粉成熟相关,同时雄花发育受碳水化合物代谢和转录因子的调控。表达量分析发现,米槠type-Ⅱ型MADS-box家族基因在雄花发育过程中的表达具有差异性。SHP和AG基因表达趋势相似,在花芽期表达量较低,半开期和盛花期表达量升高;AP3基因随着雄花发育表达量逐渐升高;2个MIKC*类同源基因主要在半开期表达;3个AP1/FUL类同源基因在雄花发育3个时期中均有表达,但表达趋势有差异;AGL6基因在雄花发育3个时期表达量均较高;4个SEP类同源基因在雄花发育中表达量较高,其中,SEP3基因表达量最高,在雄花发育3个时期的表达量变化不大,其余3个SEP类同源基因均在盛花期表达量升至最高。综合分析结果表明:在米槠雄花转录组中,多数差异表达unigenes的功能与花粉发育和催化酶的作用相关,且参与碳水化合物代谢和转录等代谢通路。MADS-box家族基因在雄花发育的3个时期差异表达,与雄花发育密切相关。

Abstract

 Taking male flowers of Castanopsis carlesii (Hemsl.) Hayata in flower bud stage, half flowering stage and flowering stage as research materials, the transcriptome database of the development process of male flowers of C. carlesii was constructed by using high-throughput sequencing platform. The flower bud stage and half flowering stage, half flowering stage and flowering stage, and flower bud stage and flowering stage were compared, the differentially expressed unigenes related to male flower development were screened, and their functions were annotated. Meanwhile, the MADS-box family genes of C. carlesii were identified, and the variations of expression levels of these genes in three stages of male flower development of C. carlesii were analyzed. The results show that in the obtained transcriptome, the number of clean reads are 628 800 000-643 700 000, the total amount of clean bases are 6.29-6.44 Gb, the percentages of Q30 base are all no less than 90.67%, and the GC contents are 45.09%-45.39%. There are 15 086, 6 770 and 16 150 differentially expressed unigenes between flower bud stage and half flowering stage, half flowering stage and flowering stage, and flower bud stage and flowering stage, respectively. The GO functional annotation result shows that the differentially expressed unigenes are mainly annotated in the biological processes related to catalytic activity, binding and membrane, etc. The KEGG metabolic pathway annotation result shows that the differentially expressed unigenes are mainly annotated in the metabolic pathways related to global and overview map, carbohydrate metabolism, environmental adaptation and signal transcription, etc. These results suggest that catalytic enzymes play an important role in the development process of male flowers of C. carlesii, the differentially expressed unigenes annotated in cell components are mainly related to pollen maturation, and the male flower development of C. carlesii is regulated by carbohydrate metabolism and transcription factors. The expression level analysis shows that there are differences in expression of type-Ⅱ MADSbox family genes in the development process of male flowers of C. carlesii. The expression trends of SHP and AG genes are similar, which are relatively low in flower bud stage, and increase in half flowering stage and flowering stage; the expression level of AP3 gene gradually increases with  male flower  development; two MIKC* homologous genes are mainly expressed in half flowering stage; three AP1/FUL homologous genes are expressed in three stages of male flower development, but their expression trends are different; the expression levels of AGL6 gene are all relatively high in three stages of male flower development; the expression levels of four SEP homologous genes are relatively high in  male flower development, in which, the expression level of SEP3 gene is the highest, and the variation of expression level in three stages of male flower development is not evident, while the expression levels of the other three SEP homologous genes all increase to the highest levels in flowering stage. The comprehensive analysis result shows that functions of most differentially expressed unigenes are related to pollen development and catalytic enzymes in the transcriptomes of male flowers of C. carlesii, and are involved in metabolic pathways of carbohydrate metabolism and transcription, etc. MADS-box family genes are differentially expressed in three stages of male flower development, which is closely related to male flower development.

关键词米槠; 雄花; 转录组; 差异表达unigenes; MADS-box家族基因
Key wordsCastanopsis carlesii (Hemsl.) Hayata; male flower; transcriptome; differentially expressed unigenes; MADS-box family gene
作者连辉a, 江淑珍a,b, 熊远芳a,b, 沈军a, 陈世品a
所在单位福建农林大学: a. 林学院, b. 园林学院 兰科植物保护与利用国家林业和草原局重点实验室, 福建 福州 350002
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基金项目福建农林大学优秀研究生学位论文资助经费(1122YS01013)