异质生境下山东银莲花的转录组分析及EST-SSR分子标记开发  

Transcriptome analysis and development of EST-SSR molecular markers in Anemone shikokiana under heterogeneous habitats

作　　者：单筱涵 安康 周春霞 张鑫 逄玉娟[1] 李丽霞[1] 卞福花[1] SHAN Xiaohan;AN Kang;ZHOU Chunxia;ZHANG Xin;PANG Yujuan;LI Lixia;BIAN Fuhua(College of Life Sciences,Yantai University,Yantai 264005,Shandong,China)

机构地区：[1]烟台大学生命科学学院,山东烟台264005

出　　处：《广西植物》2025年第1期95-107,共13页Guihaia

基　　金：国家自然科学基金面上项目(31971546)。

摘　　要：为探究稀有物种山东银莲花在全光照的山顶灌丛和阴暗的针阔混交林下两种不同生境中的生态适应机制,并开发其EST-SSR分子标记,该研究利用Illumina高通量测序技术对开花期的山东银莲花叶片进行转录组测序,获取其功能注释和差异表达基因(DEGs)。结果表明:(1)转录组测序共得到53536条Unigenes序列,其中27448条成功获得注释。(2)差异表达基因5635个,1600个在山顶灌丛的山东银莲花中上调表达,其余4035个下调表达。有2460个差异表达基因注释到GO数据库2533个三级条目中,1051个差异表达基因注释到KEGG数据库的113条代谢通路中。(3)山东银莲花适应于异质生境的代谢通路主要涉及光合作用-天线蛋白通路和类黄酮生物合成通路,光合作用-天线蛋白通路中lhca 5基因上调表达,lhcb 1、lhcb 2和lhcb 3基因下调表达,类黄酮生物合成通路中chs、c 4 h、f 3′h、f 3 h、fls、ans、chi、ccoaomt和hct基因均上调表达。(4)从山东银莲花转录组数据中共获得7146个SSR位点分布于6006条Unigenes序列中,共计106种重复基序,优势重复基序为单核苷酸重复。设计合成100对EST-SSR引物中共有68对引物具有有效性,其中11对具有多态性,共扩增24个多态性片段。该研究首次开发了山东银莲花EST-SSR分子标记,为山东银莲花的保护和利用提供了重要的分子标记资源。In order to explore,the ecological adaptation mechanisms of Anemone shikokiana in two distinct habitats,namely full-light hilltop scrub and shady mixed broadleaved-coniferous forest,and to develop its molecular markers,Illumina high-throughput sequencing technology for leaves of A.shikokiana collected during the flowering stage to obtain functional annotation and differentially expressed genes(DEGs).The results were as follows:(1)A total of 53536 Unigenes sequences were obtained,of which 27448 were successfully annotated.(2)A total of 5635 DEGs were obtained,1600 up-regulated and 4035 down-regulated genes comparing A.shikokiana in full-light hilltop scrub and in shady mixed coniferous forest.A total of 2460 DEGs were annotated to 2533 tertiary entries in GO database.In addition,1051 DEGs were involved in 113 KEGG pathways.(3)The metabolic pathways adapted to heterogeneous habits in A.shikokiana mainly involved photosytheis-antenna protein pathway and flavonoid biosynthesis pathway.In the photosynthesis-antenna protein pathway,the expression of lhca 5 was up-regulated,while the expressions of lhcb 1,lhcb 2 and lhcb 3 were down-regulated.Meanwhile,in the flavonoid biosynthesis pathway,the expressions of chs,c 4 h,f 3′h,f 3 h,fls,ans,chi,ccoaomt and hct were all up-regulated.(4)A total of 6006 Unigenes containing 7146 SSRs were obtained from the transcriptome data of A.shikokiana.In the identified SSRs,the dominant repeat motifs were single nucleotide repeats in 106 repetitive motif types.Among the 100 pairs of EST-SSR primers,a total of 68 pairs were effective and 11 pairs with polymorphism,and 24 polymorphic fragments were amplified.Overall,in this paper,for the first time EST-SSR molecular markers were developed,which would provide important molecular marker resource for the conservation and utilization of A.shikokiana.

关 键 词：异质生境 山东银莲花 转录组 EST-SSR 生态适应机制 

分 类 号：Q943[生物学—植物学]