摘要
目的:利用多聚半乳糖醛酸酶(PG)反义基因转化选育耐贮中国樱桃。方法:以2个中国樱桃品种为实验材料,研究了激素浓度、叶片生理状态、光照条件等因素对叶片再生的影响,建立了中国樱桃叶片培养高频再生体系。以继代生长40d左右的组培苗顶部充分展开的叶片为外植体,整片放在含6-BA1.5mg/L、IBA0.5mg/L、KT0.5mg/L和腺嘌呤30mg/L的MS分化培养基上,在适宜的光照和处理条件下,芽分化率高达80%以上。以上述叶片为受体,用含有PG反义基因的农杆菌进行侵染。结果:经PCR和PCR-Southern检测证明PG反义基因已整合进樱桃核基因组中。结论:以培养的叶片为受体,用农杆菌介导法进行转基因在中国樱桃上是可行的。
Objective： To selects the long stomge Prunus pseudocerasus Lindl by anti-polygalacturonase（PG） gene transformation. Methods： Two varieties of P.pseudocerasus were used as the materials in the experiment. The effects of plant hormone, physiological conditions of leaves and lightening condition to the regenerations of cultured leaves were studied. The high-frequency regeneration system was established. The whole leaves that were fully extended at the top of plantlets about 40 days old were cultured on the MS medium plus 6-BA 1.5 mg/L, IBA 0.5 mg/L, KT 0.5 mg/L and Ad 30 mg/ L. On such medium the regeneration ratio was more than 80% under the suitable lightening and cultured temperature. Instead of the leaf disc the whole leaves were used for gene transformation mediated by Agrobacterium tumefaciens. Results： The PCR and PCR-Southern tests showed that the anti-polygalacturonase gene was integrated into P.pseudocerasus genome. Conclusion： The studies indicate that it is possible to transfer the foreign gene into P.pseudocerasus by using cultured leavies as the accepter and mediated by A.tumefaciens.
出处
《生物技术通讯》
CAS
2006年第6期885-887,共3页
Letters in Biotechnology
基金
南京信息工程大学校基金资助
关键词
中国樱桃
多聚半乳糖醛酸酶反义基因
农杆菌介导
叶片培养
Prunus pseudocerasus Lindl
anti-polygalacturonase gene
Agrobacterium tumefaciens mediation
leafculture
