关键词: 棉花, GhCAD6基因, Southern杂交, 实时荧光定量PCR

Abstract: To study the expression and function of GhCAD6 gene in transgenic cotton fibers, the transgenic material was obtained by Agrobacterium-mediated transformation into Xinluzao 36, and the T6 generation plant material was obtained after successive generations of selection. The genomic DNA from cotton leaf of the T6 generation material plants was detected by PCR, and the positive plants were identified. The genomic DNA of the positive plants were hybridized by Southern blotting. The results showed that GhCAD6 was integrated into the cotton genome in different genes loci in the form of single copy. Total RNA from cotton fiber was extracted by hot borate-proteinase K method. The method for detecting the expression of GhCAD6 gene in cotton fiber was established by quantitative real-time PCR. The expression of GhCAD6 gene in cotton fiber at different developmental stages was determined by the method. The expression of GhCAD6 in transgenic plant fiber was lower than that in control samples at the same developmental stage. The expression of GhCAD6 was inconsistent with that in the control samples at 15 DPA (day post-anthesis), and the other periods were approximately the same. The change of GhCAD6 gene expression at 15 DPA may be the main reason for the variation of fiber quality of transgenic cotton. The results of this research could provide theoretical basis for further analysis of the mechanism of GhCAD6 gene in improving cotton fiber quality.

Key words: cotton, GhCAD6 gene, Southern blotting, quantitative real-time PCR