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利用根癌农杆菌(Agrobacterium tumefaciens)介导法将反义4CL(4-coumarate:CoA ligase,4-香豆酸辅酶A连接酶)基因转入三倍体毛白杨(Populus tomentosa).PCR及Southern检测证实外源基因已整合到转基因植株总基因组中.RT-PCR和Western检测表明反义4CL基因在转基因植物体内已表达.Klason木质素含量测定结果显示,抑制内源4CL基因的表达可显著降低转基因株系的木质素含量,比非转基因对照下降最高达41.73％.但综纤维素含量测定结果表明,转基因植株与对照没有明显区别,推测4CL可能并非木质素与碳水化合物的代谢调节点.转基因植株茎杆剥皮后呈现程度不等的红褐色,而对照为白色,其他生长发育特征与对照无明显差异.Wiesner反应结果表明,木质素含量下降的株系呈红色,而对照呈典型紫红色.
Antisense 4CL (4-coumarate: CoA ligase) gene was transferred into triploid Populus tomentosa by Agrobacterium tumefaciens-mediated method. PCR and Southern detection Confirmed that the foreign gene has been integrated into the genome of transgenic plants.RT-PCR and Western detection showed antisense 4CL gene has been expressed in transgenic plants Klason lignin content determination showed that inhibition of endogenous 4CL gene expression can significantly reduce the transgene The lignin content of the plants decreased by 41.73% compared with the non-transgenic control, but the determination of the content of the holocellulose showed that there was no significant difference between the transgenic plants and the control, suggesting that 4CL might not be the metabolic regulation point of lignin and carbohydrate. The results showed that the lines with decreased lignin were red, while the control showed the typical fuchsia.