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Identification of candidate genes for fiber length quantitative trait loci through RNA-Seq and linkage and physical mapping in cotton

机译:通过RNA-Seq以及棉花的连锁和物理作图鉴定纤维长度定量性状基因座的候选基因

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Background Cotton ( Gossypium spp.) fibers are single-celled elongated trichomes, the molecular aspects of genetic variation in fiber length (FL) among genotypes are currently unknown. In this study, two backcross inbred lines (BILs), i.e., NMGA-062 (“Long”) and NMGA-105 (“Short”) with 32.1 vs. 27.2?mm in FL, respectively, were chosen to perform RNA-Seq on developing fibers at 10?days post anthesis (DPA). The two BILs differed in 4 quantitative trait loci (QTL) for FL and were developed from backcrosses between G. hirsutum as the recurrent parent and G. barbadense . Results In total, 51.7 and 54.3 million reads were obtained and assembled to 49,508 and 49,448 transcripts in the two genotypes, respectively. Of 1?551 differentially expressed genes (DEGs) between the two BILs, 678 were up-regulated and 873 down-regulated in “Long”; and 703 SNPs were identified in 339 DEGs. Further physical mapping showed that 8 DEGs were co-localized with the 4 FL QTL identified in the BIL population containing the two BILs. Four SNP markers in 3 DEGs that showed significant correlations with FL were developed. Among the three candidate genes encoding for proline-rich protein, D-cysteine desulfhydrase, and thaumatin-like protein, a SNP of thaumatin-like protein gene showed consistent correlations with FL across all testing environments. Conclusions This study represents one of the first investigations of positional candidate gene approach of QTL in cotton in integrating transcriptome and SNP identification based on RNA-Seq with linkage and physical mapping of QTL and genes, which will facilitate eventual cloning and identification of genes responsible for FL QTL. The candidate genes may serve as the foundation for further in-depth studies of the molecular mechanism of natural variation in fiber elongation.
机译:背景技术棉(Gossypium spp。)纤维是单细胞细长的毛状体,目前尚不清楚基因型之间纤维长度(FL)遗传变异的分子方面。在这项研究中,选择两个回交自交系(BIL),即分别在FL中具有32.1 vs. 27.2?mm的NMGA-062(“长”)和NMGA-105(“短”)进行RNA-Seq在花后10天(DPA)上发育纤维。这两个BIL在FL的4个数量性状基因座(QTL)上有所不同,并且是从作为复发亲本的G. hirsutum和G. barbadense之间回交形成的。结果总共获得了51.7和5430万个读段,并分别组装成两个基因型的49,508和49,448个转录物。在两个BIL之间的1?551个差异表达基因(DEG)中,“长”中有678个被上调,而873个被下调。在339个DEG中鉴定了703个SNP。进一步的物理作图显示8个DEG与包含两个BIL的BIL群体中鉴定出的4个FL QTL共定位。在3个DEG中开发了4个SNP标记,这些标记与FL显着相关。在编码富含脯氨酸的蛋白,D-半胱氨酸脱硫酶和类索马甜蛋白的三个候选基因中,类索马甜蛋白蛋白基因的SNP在所有测试环境中均与FL相关。结论这项研究代表了棉花中QTL定位候选基因方法的首次研究之一,该方法将基于RNA-Seq的转录组和SNP鉴定与QTL和基因的连锁和物理作图相结合,这将有助于最终克隆和鉴定引起棉花的基因FL QTL。候选基因可以作为进一步深入研究纤维伸长自然变化的分子机制的基础。

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