Agrobacterium-mediated transformation of Eupatorium adenophorum
Agrobacterium-mediated transformation of Eupatorium adenophorum
复制标题
农杆菌介导的紫茎泽兰转化
DOI：
10.1007/s11240-010-9784-7
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发表时间：
2010-06
期刊：
影响因子：
--
通讯作者：
Yongqiang Zhang
中科院分区：
文献类型：
--
作者：
Hongmei Cheng;Huiming Guo;Fanghao Wan;Yongqiang Zhang
建立了一种简便、高效的根癌农杆菌介导紫茎泽兰遗传转化体系。茎段在MS培养基上预培养24 h后与A.将携带pBI 121质粒的根癌农杆菌菌株LBA 4404在相同培养基上黑暗培养3天。共培养后，将外植体转移到补充有1.0 mg l−1萘乙酸（NAA）、1.0 mg l−1 6-苄基腺嘌呤（BA）、150 mg l−1卡那霉素（Kan）和200 mg l−1头孢噻肟（Cef）的MS培养基中，并在25 ± 1°C下在16小时光周期下孵育。诱导愈伤组织形成3-4周。将诱导的愈伤组织转移到新鲜的选择培养基中，每15天传代培养。胚性愈伤组织培养6-8周后，可观察到再生植株。将1cm长的芽转移到含有150 mg l-1 Kan和200 mg l-1 Cef的半强度MS培养基中，4周后获得生根的小植株。利用GUS组织化学检测、聚合酶链反应（PCR）和逆转录酶（RT）-PCR对转基因植株进行了鉴定。获得了2%的转化效率。
A simple and efficient protocol for Agrobacterium tumefaciens-mediated genetic transformation of Eupatorium adenophorum has been developed. Stem segments were pre-cultured on Murashige and Skoog (MS) medium for 24 h and then co-cultured with A. tumefaciens strain LBA4404 harboring the pBI121 plasmid on the same medium in darkness for 3 days. After co-cultivation, explants were transferred to MS medium supplemented with 1.0 mg l−1 naphthaleneacetic acid (NAA), 1.0 mg l−1 6-benzyladenine (BA), 150 mg l−1 kanamycin (Kan), and 200 mg l−1 cefotaxime (Cef), and incubated at 25 ± 1°C under a 16-h photoperiod. Callus formation was induced for 3–4 weeks. Induced calluses were transferred to fresh selection medium and subcultured every 15 days. After 6–8 weeks, regenerated plantlets from embryogenic callus could be observed. Buds of 1 cm in length were transferred to half-strength MS medium containing 150 mg l−1 Kan and 200 mg l−1 Cef, and rooted plantlets were obtained after 4 weeks. Putative transgenic plants were confirmed using GUS histochemical assay, polymerase chain reaction (PCR), and reverse transcriptase (RT)-PCR. A transformation efficiency of 2% was obtained.
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影响因子：
5.2
作者：
A. Hassanein;E. Chevreau;N. Dorion
通讯作者：
A. Hassanein;E. Chevreau;N. Dorion
影响因子：
3
作者：
Flores-Benitez, Silvia;Jimenez-Bremont, Juan F.;Alpuche-Solis, Angel Gabriel
通讯作者：
Alpuche-Solis, Angel Gabriel
DOI：
10.1007/s11240-006-9190-3
发表时间：
2007-01
期刊：
Plant Cell, Tissue and Organ Culture
影响因子：
--
作者：
Guo Qing Song;A. Loskutov;A. Loskutov;K. Sink
通讯作者：
Guo Qing Song;A. Loskutov;A. Loskutov;K. Sink
DOI：
--
发表时间：
1962
期刊：
Int. J. Semantic Web Inf. Syst.
影响因子：
--
作者：
T. Murashige;F. Skoog
通讯作者：
T. Murashige;F. Skoog
影响因子：
5.4
作者：
Zhongchao Yin;Guo-Liang Wang
通讯作者：
Zhongchao Yin;Guo-Liang Wang