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转录组测序和多态性微卫星标记在樗树(苦木科)中的开发。

Transcriptome sequencing and microsatellite marker discovery in Ailanthus altissima (Mill.) Swingle (Simaroubaceae).

机构信息

Fairy Lake Botanical Garden, Shenzhen & Chinese Academy of Sciences, 518004, Shenzhen, People's Republic of China.

Key Laboratory of Aquatic Botany and Watershed Ecology, Wuhan Botanical Garden, Chinese Academy of Sciences, 430074, Wuhan, People's Republic of China.

出版信息

Mol Biol Rep. 2021 Mar;48(3):2007-2023. doi: 10.1007/s11033-020-05402-w. Epub 2021 Mar 17.

Abstract

Ailanthus altissima Swingle, is a tree species native to East Asia and has a great potential in decorative, bioenergy and industrial applications in many countries. To date, despite its commercial importance, the genomic and genetic resources available for this species are still insufficient. In this study, we characterized the transcriptome of A. altissima and developed thirteen EST-SSRs (expressed sequence tag-simple sequence repeats) based on Illumina paired-end RNA sequencing (RNA-seq). Besides, we developed ten polymorphic chloroplast microsatellite (cpSSR) markers using the available chloroplast genome of A. altissima. The transcriptome data produced 87,797 unigenes, of which 64,891 (73.91%) unigenes were successfully annotated in at least one protein database. For cpSSR markers the number of detected alleles (N) per marker varied from three at cpSSR12 to twelve at cpSSR8, the unbiased haploid diversity indices (uh) varied from 0.111 to 0.485, and haploid diversity indices (h) ranged from 0.101 to 0.444 with an average unbiased haploid diversity index (uh) of 0.274. Overall, a total of 65 different cpSSR alleles were identified at the ten loci among 165 individuals of A. altissima. The allele number per locus for EST-SSRs varied from 2.143 to 9.357, and the values of observed and expected heterozygosity ranged from 0.312 to 1.000 and 0.505 to 0.826, respectively. The molecular markers developed in this study will facilitate future genetic diversity, population structure, long distance-gene transfer and pollen-based gene flow analyses of A. altissima populations from its known distribution ranges in China focusing on planted and natural forest stands.

摘要

樗树,原产于东亚,在许多国家具有很大的装饰、生物能源和工业应用潜力。尽管它具有商业重要性,但迄今为止,该物种的基因组和遗传资源仍然不足。在本研究中,我们对樗树的转录组进行了特征描述,并基于 Illumina 配对末端 RNA 测序(RNA-seq)开发了 13 个 EST-SSR(表达序列标签-简单重复序列)。此外,我们利用已有的樗树叶绿体基因组开发了 10 个多态性叶绿体微卫星(cpSSR)标记。转录组数据产生了 87797 个 unigenes,其中 64891 个(73.91%)unigenes成功注释到至少一个蛋白质数据库中。对于 cpSSR 标记,每个标记的检测等位基因(N)数从 cpSSR12 的 3 个到 cpSSR8 的 12 个不等,无偏单倍体多样性指数(uh)从 0.111 到 0.485 不等,单倍体多样性指数(h)从 0.101 到 0.444 不等,平均无偏单倍体多样性指数(uh)为 0.274。总的来说,在 165 个樗树个体的 10 个位点共鉴定到 65 个不同的 cpSSR 等位基因。EST-SSR 每个位点的等位基因数从 2.143 到 9.357 不等,观察和期望杂合度的值分别从 0.312 到 1.000 和 0.505 到 0.826 不等。本研究开发的分子标记将有助于未来对中国已知分布范围内的樗树种群进行遗传多样性、种群结构、长距离基因转移和基于花粉的基因流分析,重点关注种植林和天然林。

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