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基于转录组测序的多年生黑麦草SSR标记开发及其与耐旱性的关联分析

Development of SSR Markers Based on Transcriptome Sequencing and Association Analysis with Drought Tolerance in Perennial Grass from China.

作者信息

Nie Gang, Tang Lu, Zhang Yajie, Huang Linkai, Ma Xiao, Cao Xin, Pan Ling, Zhang Xu, Zhang Xinquan

机构信息

Department of Grassland Science, Animal Science and Technology College, Sichuan Agricultural UniversitySichuan, China.

出版信息

Front Plant Sci. 2017 May 16;8:801. doi: 10.3389/fpls.2017.00801. eCollection 2017.

Abstract

Drought has become a critical environmental stress affecting on plant in temperate area. As one of the promising bio-energy crops to sustainable biomass production, the genus has been widely studied around the world. However, the most widely used hybrid cultivar among this genus, is proved poor drought tolerance compared to some parental species. Here we mainly focused on , which is one of the progenitors of providing a comparable yield and well abiotic stress tolerance in some places. The main objectives were to characterize the physiological and photosynthetic respond to drought stress and to develop simple sequence repeats (SSRs) markers associated with drought tolerance by transcriptome sequencing within an originally collection of 44 genotypes from southwest China. Significant phenotypic differences were observed among genotypes, and the average of leaf relative water content (RWC) were severely affected by drought stress decreasing from 88.27 to 43.21%, which could well contribute to separating the drought resistant and drought sensitive genotype of . Furthermore, a total of 16,566 gene-associated SSRs markers were identified based on Illumina RNA sequencing under drought conditions, and 93 of them were randomly selected to validate. In total, 70 (75.3%) SSRs were successfully amplified and the generated loci from 30 polymorphic SSRs were used to estimate the genetic differentiation and population structure. Finally, two optimum subgroups of the population were determined by structure analysis and based on association analysis, seven significant associations were identified including two markers with leaf RWC and five markers with photosynthetic traits. With the rich sequencing resources annotation, such associations would serve an efficient tool for drought response mechanism study and facilitate genetic improvement of drought resistant for this species.

摘要

干旱已成为影响温带地区植物的关键环境胁迫。作为可持续生物质生产中有前景的生物能源作物之一,该属植物在全球范围内受到广泛研究。然而,该属中使用最广泛的杂交品种与一些亲本物种相比,被证明耐旱性较差。在这里,我们主要关注[具体品种名称未给出],它是[另一品种名称未给出]的祖先之一,在某些地方具有相当的产量和良好的非生物胁迫耐受性。主要目标是通过对来自中国西南部的44个[品种名称未给出]基因型的原始收集进行转录组测序,来表征其对干旱胁迫的生理和光合响应,并开发与耐旱性相关的简单序列重复(SSR)标记。在基因型之间观察到显著的表型差异,叶片相对含水量(RWC)的平均值受到干旱胁迫的严重影响,从88.27%降至43.21%,这有助于很好地分离[品种名称未给出]的抗旱和干旱敏感基因型。此外,基于干旱条件下的Illumina RNA测序,共鉴定出16566个与基因相关的SSR标记,并随机选择其中93个进行验证。总共成功扩增了70个(75.3%)SSR,并使用来自30个多态性SSR产生的位点来估计遗传分化和群体结构。最后,通过结构分析确定了群体的两个最优亚组,并基于关联分析,鉴定出七个显著关联,包括两个与叶片RWC相关的标记和五个与光合性状相关的标记。凭借丰富的测序资源注释,这些关联将为[品种名称未给出]干旱响应机制研究提供一个有效的工具,并促进该物种耐旱性的遗传改良。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e33/5432562/babc1916db04/fpls-08-00801-g002.jpg

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