Ahmed Mukhtar, Iqbal Adnan, Latif Ayesha, Din Salah Ud, Sarwar Muhammad Bilal, Wang Xuede, Rao Abdul Qayyum, Husnain Tayyab, Ali Shahid Ahmad
Centre of Excellence in Molecular Biology (CEMB), University of the Punjab, Lahore, Pakistan.
Institute of Crop Sciences, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China.
Front Plant Sci. 2020 Nov 12;11:476251. doi: 10.3389/fpls.2020.476251. eCollection 2020.
The study aims to improve fiber traits of local cotton cultivar through genetic transformation of sucrose synthase () gene in cotton. Sucrose synthase (SuS) is an important factor that is involved in the conversion of sucrose to fructose and UDP-glucose, which are essential for the synthesis of cell wall cellulose. In the current study, we expressed a synthetic gene in cotton plants under the control of a CaMV35S promoter. Amplification of an 813-bp fragment using gene-specific primers confirmed the successful introduction of gene into the genome of cotton variety CEMB-00. High mRNA expression was observed in two transgenic cotton plants, MA0023 and MA0034, when compared to the expression in two other transgenic cotton plants, MA0035 and MA0038. Experiments showed that mRNA expression was positively correlated with SuS activity at the vegetative (54%) and reproductive stages (40%). Furthermore, location of transgene was found to be at chromosome no. 9 in the form of single insertion, while no signal was evident in non-transgenic control cotton plant when evaluated through and karyotyping analysis. Fiber analyses of the transgenic cotton plants showed increases of 11.7% fiber length, 18.65% fiber strength, and up to 5% cellulose contents. An improvement in the micronaire value of 4.21 was also observed in the MA0038 transgenic cotton line. Scanning electron microscopy (SEM) revealed that the fibers of the transgenic cotton plants were highly spiral with a greater number of twists per unit length than the fibers of the non-transgenic control plants. These results determined that gene expression influenced cotton fiber structure and quality, suggesting that has great potential for cotton fiber quality improvement.
本研究旨在通过对棉花蔗糖合酶()基因进行遗传转化,来改善当地棉花品种的纤维特性。蔗糖合酶(SuS)是参与蔗糖转化为果糖和UDP-葡萄糖的重要因子,而果糖和UDP-葡萄糖对于细胞壁纤维素的合成至关重要。在本研究中,我们在CaMV35S启动子的控制下,在棉花植株中表达了一个合成基因。使用基因特异性引物扩增出一个813 bp的片段,证实基因已成功导入棉花品种CEMB-00的基因组中。与另外两个转基因棉花植株MA0035和MA0038相比,在两个转基因棉花植株MA0023和MA0034中观察到了较高的mRNA表达。实验表明,在营养生长阶段(54%)和生殖阶段(40%),mRNA表达与SuS活性呈正相关。此外,通过Southern杂交和核型分析评估发现,转基因位于第9号染色体上,呈单插入形式,而在非转基因对照棉花植株中未发现明显信号。转基因棉花植株的纤维分析表明,纤维长度增加了11.7%,纤维强度增加了18.65%,纤维素含量提高了5%。在MA0038转基因棉花品系中,马克隆值也有4.21的改善。扫描电子显微镜(SEM)显示,转基因棉花植株的纤维高度螺旋状,每单位长度的扭曲次数比非转基因对照植株的纤维多。这些结果表明,基因表达影响了棉花纤维的结构和品质,提示该基因在改善棉花纤维品质方面具有巨大潜力。