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本研究以高比强度纤维材料0-153和转基因抗虫棉sGK9708为亲本构建的高代重组自交系群体(F6:9)中选育出的高比强度纤维品系(69307)作为材料,利用抑制性消减杂交技术,以15DPA纤维为driver、20DPA纤维为tester,成功构建出陆地棉开花后20d纤维的cDNA消减文库。通过蓝白斑筛选、菌落PCR及反向Northern技术最终筛选出差异表达的阳性克隆340个。通过对阳性克隆测序及序列分析,共得到115个单一序列,其中35个重叠群,80个单拷贝。利用Blast2GO等对差异表达基因进行生物信息学分析,结果表明这些差异表达基因广泛参与糖类、脂类、氨基酸等物质的代谢,以及纤维素生物合成、细胞壁合成与修饰、氧化还原、细胞信号转导等生物学过程。
In this study, a high-yielding fiber line (69307) selected from a high-generation recombinant inbred line population (F6: 9) with high specific strength fiber material 0-153 and transgenic insect-resistant cotton sGK9708 as its parents was used as material Suppression subtractive hybridization (SSH) technique was used to construct the cDNA subtractive library of cotton fiber 20 days after flowering with 15 DPA fiber as driver and 20 DPA fiber as tester. A total of 340 differentially expressed positive clones were finally screened by blue-white screening, colony PCR and reverse Northern blotting. A total of 115 single sequences were obtained by sequencing the positive clones and analyzing the sequences. 35 contigs and 80 single copies were obtained. Bioinformatics analysis of differentially expressed genes using Blast2GO showed that these differentially expressed genes were widely involved in the metabolism of carbohydrates, lipids, amino acids and other substances as well as cellulose biosynthesis, cell wall synthesis and modification, redox, cell signaling Guide and other biological processes.