利用农杆菌介导法获得RNAi转基因枫香的研究

随着我国林业生态工程和退耕还林的开展，木材生产与需求之间的矛盾愈加严重，培育速生优质大径材的研究更加重要。基于LEAFY基因在植物生长发育中的特性，利用PCR和Gateway技术构建了含有LEAFY基因保守序列的RNA干涉植物表达载体LFY-RI，利用根瘤农杆菌Agrobacterium tumefaciems介导法获得转基因植株，利用RNA干涉技术抑制转基因植株中LEAFY基因的表达，从而获得枫香Liquidambar formosana不育转基因植株。并利用聚合酶链式反应（PCR）技术和PCR-Southern杂交技术检测了转基因植株中外源基因整合情况。共获得了5株枫香转基因植株，分子检测表明，其中4株为转基因阳性植株。由于转基因植株还比较幼嫩，对它们生长发育以及木材品质的变化尚处于在观察中。图5参12

The LEAFY gene in RNA interference (RNAi) transgenic Liquidambar formosana mediated by Agrobacterium tumefaciems

It is incompatible between production and need of timber,the aim of the test is breeding fast-growing,super-quality and large-diameter tree cultivar.The LEAFY gene affects plant flowering.Using polymerase chain reaction(PCR) with the primers 5'-CACCTCAGAGAGAGAAAAATAGATTATGGATCC-3' and 5'-GTCGACAGTTCGGGGGGGAAATATAAT-3',a conservation sequence of the LEAFY gene for Liquidambar formosana(Chinese sweetgum) was cloned.An RNA interference(RNAi) pBIN vector of the LEAFY gene was constructed by Gateway technology,and then agrobacterium mediation was used to transfer the aim gene into the the first two to three leaves of Chinese sweetgum seedlings.Then the transgenic leaves were cultured to plants in vitro.These plants' leaves were analyzed with PCR and Southern blotting.It is sure that the foreign gene was integrated into the genome,resulting in four transgenic plants.However,the transgenic four plants' development and timber quality has not been analyzed as they are so young.The study is carried on.Ch,5 fig.12 ref.]