摘要
用５～６ｄ的棉花无菌苗下胚轴切段与农杆菌共培，菌株质粒中Ｇｕｓ基因为标记基因，ＮＰＴⅡ基因为选择基因。在含０．１ｍｇ·Ｌ－１２，４－Ｄ和０．１ｍｇ·Ｌ－１ＫＴ的ＭＳ培养基上共培４８ｈ，转移到添加头孢霉素５００ｍｇ·Ｌ－１，卡那霉素５０ｍｇ·Ｌ－１的ＭＳ培养基中诱导和筛选抗性愈伤组织，７０～８０ｄ时，进行ＧＵＳ检测，选择ＧＵＳ阳性愈伤组织，经体细胞分化生成转基因再生植株。
Hypocotyl segments from 5～6 day old seedlings of Gossypium hirsutum were cocultured with Agrobacterium tumefacience, GUS gene as marker gene, NPTⅡ as selecting gene. After the cultures were cocultured in MS medium contained 0.1mg·L12, 4D and 0.1mg·L1 KT for 48h, they were transferred into the medium described above, and suplemented with 500mg·L1 cefotaxime and 50～100mg·L1 kanamycin to induce and select resistant calli. After 70～80 days, GUS was numbered, tested and selected position, while calli were well cultured on embryonesis medium until globular embryos developed and germinated plantlets forming these embryos.
出处
《棉花学报》
CSCD
北大核心
1998年第4期189-192,共4页
Cotton Science
关键词
棉花
农杆菌
基因转化
再生植株
GUS基因
cotton Agrobacterium tumefaciens gene transformation regenerated plant GUS gene
