反义4CL基因调节烟草木质素生物合成  

Lignin Biosynthesis of Tobacco(Nicotiana tabacum L.) Regulated by the Antisense 4CL Gene

作　　者：孟靖 王成[1] 赵曼琳 李曹娜[1] 茹艺[1] 崔智昕 韩阳[1] 

机构地区：[1]辽宁大学生命科学院,沈阳110036

出　　处：《分子植物育种》2017年第11期4383-4388,共6页Molecular Plant Breeding

基　　金：辽宁省自然科学基金(2012020283)资助

摘　　要：采用根癌农杆菌介导法的叶盘转化法,将紫穗槐反义4CL基因片段导入烟草中,获得T0代阳性转化植株。T0代转基因植株自花授粉,收获种子,种植后获得T1代植株。PCR、RT-PCR(T1代)检测、目的片段测序的结果表明,反义4CL基因已经稳定遗传到T1代中。T1代烟草阳性植株和阴性植株的比例为77:23,接近于3:1。两株T1代烟草转化植株q RT-PCR及4CL酶含量检测表明,叶片4CL1基因表达量较野生植株降低了19.51%;茎4CL酶含量降低了10.50%,说明该反义基因能够干扰烟草4CL基因的表达。T1代烟草茎木质素含量平均下降了11.87%,根木质素含量降低了14.23%。种植60 d的转基因植株与野生型植株生长一致,在株高、株重等方面差异不显著(p>0.05),说明反义4CL基因的转入在降低了木质素含量的同时并没有影响烟草的正常生长。In this research the positive transformation plants of T0 generation were obtained by taking the antisense4 CL gene fragment transformed into tobacco（Nicotiana tabacum L.） with Agrobacterium-mediated method. T1 generation plants were obtained by planting T0 harvested seeds which were transgenic pollinated plants. According to the test of PCR, RT-PCR（only in T1） and purpose fragment sequencing showed that the antigen 4 CL gene had been stably transmit into the T1 generation. The ratio of positive plants and negative plants in of T1 generation tobacco（Nicotiana tabacum L.） was 77：23, which closed to 3：1. The results of q RT-PCR and 4 CL enzyme activity of two T1 transgenic tobacco（Nicotiana tabacum L.） plants showed that compared with the wild-type plants, the expression value of 4 CL1 gene of T1 transgenic plants in leaves decreased by 19.51%, the average contents of 4 CL1 enzyme in stems decreased by 10.50%, which indicated that the antisense gene can interfere with the expression of4 CL gene in tobacco. The stem lignin contents and root lignin contents of T1 transgenic plants of tobacco（Nicotiana tabacum L.） were decreased by 11.87% and 14.23%, respectively. The transgenic plants grown for 60 days were consistent with the growth of wild type plants. the difference of transgenic tobacco and wild-type tobacco in growth was not significant（p〉0.05）, which indicated that the transferred of antisense 4 CL gene had not affect the normal growth of tobacco（Nicotiana tabacum L.）.

关 键 词：木质素 烟草 4-香豆酸辅酶A连接酶(4CL) 反义基因 

分 类 号：S572[农业科学—烟草工业]