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通过根癌农杆菌对枸杞进行遗传转化。

Genetic transformation of Lycium barbarum L. via A. tumefaciens.

作者信息

Du L Q, Wang H Z, Huang F C, Li A S, Shao Q Q

机构信息

Institute of Genetics, Academia Sinica, Beijing, PRC.

出版信息

Sci China B. 1994 Mar;37(3):286-92.

PMID:8068193
Abstract

A system for transformation and regeneration of Lycium barbarum L., an important Chinese medical plant, has been established. Young stem segments from Lycium barbarum L. were infected with Agrobacterium tumefaciens C58cl(pGV3850::neo1103), and the transformed calli selected from the callus induction medium containing 50 micrograms/ml kanamycin could regenerate buds on differentiation medium containing 25 micrograms/ml kanamycin. 30% of the regenerated buds were normal in morphology. The normal buds could develop into whole plantlets after they were transferred to the rooting medium to induce roots. Nopaline detection, NPT-II enzyme activity assay and Southern blotting hybridization indicated that the foreign genes had been integrated into the genome of Lycium barbarum L. and expressed in the plant. In the processes of experiments, it was found that (i) after the pre-processes, the explants which formed callus quickly were easy to transform; (ii) the rate of normal regenerated plants from transgenic calli was higher than that from the untransgenic ones.

摘要

已建立了一种用于重要中药材枸杞转化和再生的系统。用根癌农杆菌C58cl(pGV3850::neo1103)感染枸杞的幼嫩茎段,从含有50微克/毫升卡那霉素的愈伤组织诱导培养基中筛选出的转化愈伤组织,在含有25微克/毫升卡那霉素的分化培养基上能够再生芽。30%的再生芽形态正常。正常芽转移到生根培养基上诱导生根后可发育成完整植株。胭脂碱检测、NPT-II酶活性测定和Southern杂交表明,外源基因已整合到枸杞基因组中并在植株中表达。在实验过程中发现:(i) 经过预处理后,快速形成愈伤组织的外植体易于转化;(ii) 转基因愈伤组织正常再生植株的比率高于未转基因愈伤组织。

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