两个不同高羊茅品系叶片差异表达基因研究  

Study on Differentially Expressed Genes in Leaf of Two Different Festuca Arundinacea Strains

作　　者：谢健[1,2] 牛熙 许瑶[1] 喻昌燕[1] 冉雪琴 王嘉福[1,3] XIE Jian;NIU Xi;XU Yao;YU Chang-Yan;RAN Xue-Qin;WANG Jia-Fu(Institute of Agro-Bioengineering/College of Animal Science, Guizhou University, Guiyang 550025, China;Tongren University, Tongren 554300, China;Department of medical genetics, Zunyi University, Zunyi 563003, China)

机构地区：[1]贵州大学农业生物工程研究院/动物科学学院,贵阳550025 [2]遵义医学院医学遗传学教研室,遵义553003 [3]铜仁学院,铜仁554300

出　　处：《农业生物技术学报》2018年第5期793-800,共8页Journal of Agricultural Biotechnology

基　　金：贵州省农业科技攻关项目(黔科合NY字2009-3082,黔科合NY字[2012]3009号);贵州省“百”层次创新型人才培养项目(黔科合人才(2016)4012号)

摘　　要：高羊茅(Festuca arundinacea)抗逆、抗病性强,是目前世界上广泛种植的牧草之一。由贵州省草业科学研究所选育的黔草二号高羊茅有两个不同的品系,分别为细叶和宽叶型。为了研究两个品系的高羊茅在叶片形成、抗逆性等方面差异的形成机理,本研究采用反转录差异显示技术(differential display reverse transcript-PCR,DDRT-PCR)对宽叶和细叶型高羊茅叶片mRNA差异表达的基因进行分离和鉴定,共分离得到50条差异条带。回收差异条带并对之进行克隆测序,相关生物信息学分析的结果表明,差异基因主要涉及光合作用、生长调控、抗逆三个方面。通过qRT-PCR进一步验证,得到的基因除了18S核糖核蛋白体基因在宽叶中表达量上调之外,其余基因在细叶中的表达量上调,因而推测叶面积较大的植物往往分配较多的生物量于细胞壁,以增强叶片的韧性,同时积累较多的光合产物,为生长发育做准备,而分配到光合细胞中的较少,导致其单位面积光合效率降低;而叶面积较小的植物分配较多的有机物质于类囊体和羧化酶中,具有较高的光合能力,植物生长快,尤其是地上部分生长迅速,使得其在自然环境条件下的竞争能力较强。本研究为进一步研究植物叶片形态发生的相关基因及阐明黔草二号高羊茅宽叶和细叶品系之间表达调控的分子机理、抗逆性等研究提供了理论基础。Festuca arundinacea is a widely cultivated forage in the world with excellent stress and disease resistance. Qiancao No.2 was cultivated by Guizhou Institute of Prataculture, which has two different strains （broad-leaved and fine-leaved） with obvious differences in the leaf shape. In order to recover the mechanism of leaf formation and resistance of Qiancao No.2, in this study, a differential display reverse transcription PCR （DDRT-PCR） was performed to screen the differential expression genes between the broad-leaved and fine-leaved strains. Finally, 50 differential bands were separated and recovered. The DNA fragments were cloned and sequenced. Most of different expressed genes were enriched into photosynthesis, growth regulation and stress resistance. After further verification by qRT-PCR, it was found that only the genes of 18S ribosome were up-regulated in broad-leaved, and the rest of them were up-regulated in fine-leaved. These results suggested that plants with large leaves usually allocated more biomass to the cell wall while to enhance the toughness of leaves, and accumulated more photosynthetic products to prepare for the growth and development, this situation would lead to lower photosynthetic efficiency in unit area of leaves. However, plants with narrow leaves allocated more organic in chloroplast which made them had higher photosynthetic capacity to accelerate growth, and with strong competition ability in natural environment. In particular, the growth of aboveground parts made them with strong competition ability in natural environment. This study provides a theoretical foundation for the further research on the functional genes of leaf morphogenesis, gene expression and the stress resistance.

关 键 词：反转录差异显示技术(DDRT-PCR)  叶片性状  高羊茅  基因表达 

分 类 号：S184[农业科学—农业基础科学]