| 引用本文: | 单筱涵, 安 康, 周春霞, 张 鑫, 逄玉娟, 李丽霞, 卞福花.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[J].广西植物,2025,45(1):108-120.[点击复制] |
| SHAN Xiaohan, AN Kang, ZHOU Chunxia, ZHANG Xin,
PANG Yujuan, LI Lixia, BIAN Fuhua.Transcriptome analysis and development of EST-SSR molecular markers in Anemone shikokiana under heterogeneous habitats[J].Guihaia,2025,45(1):108-120.[点击复制] |
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| 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 |
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单筱涵, 安 康, 周春霞, 张 鑫, 逄玉娟, 李丽霞, 卞福花*
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烟台大学 生命科学学院, 山东 烟台 264005
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| 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53 536条Unigenes序列,其中27 448条成功获得注释。(2)差异表达基因5 635个,1 600个在山顶灌丛的山东银莲花中上调表达,其余4 035个下调表达。有2 460个差异表达基因注释到GO数据库2 533个三级条目中,1 051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lhca5基因上调表达,lhcb1、lhcb2和lhcb3基因下调表达,类黄酮生物合成通路中chs、c4h、f3'h、f3h、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7 146个SSR位点分布于6 006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。 |
| 关键词: 异质生境, 山东银莲花, 转录组, EST-SSR, 生态适应机制 |
| DOI：10.11931/guihaia.gxzw202304067 |
| 分类号:Q943 |
| 文章编号:1000-3142(2025)01-0108-13 |
| 基金项目:国家自然科学基金面上项目(31971546)。 |
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| Transcriptome analysis and development of EST-SSR molecular markers in Anemone shikokiana under heterogeneous habitats |
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SHAN Xiaohan, AN Kang, ZHOU Chunxia, ZHANG Xin,
PANG Yujuan, LI Lixia, BIAN Fuhua
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College of Life Sciences, Yantai University, Yantai 264005, Shandong, China
College of Life Sciences, Yantai University, Yantai 264005, Shandong, China
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| Abstract: |
| 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 53 536 Unigenes sequences were obtained, of which 27 448 were successfully annotated.(2)A total of 5 635 DEGs were obtained, 1 600 up-regulated and 4 035 down-regulated genes comparing A. shikokiana in full-light hilltop scrub and in shady mixed coniferous forest. A total of 2 460 DEGs were annotated to 2 533 tertiary entries in GO database. In addition, 1 051 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 lhca5 was up-regulated, while theexpressions of lhcb1, lhcb2 and lhcb3 were down-regulated. Meanwhile, in the flavonoid biosynthesis pathway, the expressions of chs, c4h, f3'h, f3h, fls, ans, chi, ccoaomt and hct were all up-regulated.(4)A total of 6 006 Unigenes containing 7 146 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. |
| Key words: heterogeneous habitat, Anemone shikokiana, transcriptome, EST-SSR, ecological adaptation mechanisms |
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