海岛棉β-胡萝卜素异构酶GbD27-6基因克隆及表达分析

Cloning and bioinformatics analysis of β-carotene isomerase GbD27-6 gene in sea island cotton

  • 摘要: 【目的】 研究GbD27-6基因在海岛棉分枝和侧枝生长中的影响作用,为分析GbD27基因在胞内运输作用及在纤维发育中参与表皮细胞突起研究提供参考。 【方法】 以海岛棉Pimas-7 在15 d的纤维材料为模板,克隆独脚金内酯(SLs)合成通路中的关键基因GbD27-6,利用生物信息学分析网站分析该基因,并利用qRT-PCR技术分析其基因表达量的差异性。 【结果】 GbD27-6基因全长816 bp,编码271个氨基酸,蛋白分子式为C1335H2134N350O389S24,分子质量大约为30.081 18 kDa,等电点为8.40,属于DUF4033超家族蛋白成员,且为不稳定疏水性蛋白。GbD27-6基因定位在细胞膜,GbD27-6基因在海岛棉花瓣、苞叶、花药中均有表达,其中在苞叶中表达量最高,在花瓣中表达量最低;而在不同发育时期的纤维中均有不同程度的表达,其中在纤维发育20 d的表达量最高,在10 d时表达量最低。 【结论】 GbD27-6基因在纤维中存在差异表达。

     

    Abstract: 【Objective】 To study the effect of GbD27-6 gene on branch and lateral branch growth of sea island cotton. This study provides a basic theory for further understanding the role of GbD27 gene in intracellular transport and its participation in epidermal cell processes during fiber development. 【Methods】 In this study, the key gene GbD27-6 in the synthesis pathway of Strigolactones (SLs) was cloned from the fibrous material of Gossypium barbadense L. Pimas-7 15 d, and the expression level of GbD27-6 was analyzed by using the bioinformatics website and qRT-PCR. 【Results】 GbD27-6 gene 816 bp encoded 271 amino acid protein, molecular formula is was C1335H2134N350O389S24, molecular weight is was about 30.081 18 kDa, isoelectric point is was 8.40, it which belonged to DUF4033 superfamily protein and was unstable hydrophobic protein. The results of subcellular localization showed that GbD27-6 gene was located on cell membrane. Real -time fluorescence quantitative analysis showed that GbD27-6 gene was expressed in petal bracts and anthers of Gossypium barbadense L. and the expression level was the highest in bracts and the lowest in petals. The expression level was the highest on the 20th day of fiber development and the lowest on the 10th day of fiber development. 【Conclusion】 The findings showed that GbD27-6 gene was differentially expressed in fibers.

     

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