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根瘤农杆菌介导的枸杞转化及转化植物的获得 总被引:5,自引:0,他引:5
本文报道一种重要的中药——枸杞(Lycium barbarum L.)的转化、再生系统的建立。以根瘤农杆菌(Agrobaterium tumefaciens)C58cl(pGV3850::neo1103)感染枸杞的幼茎外植体,在含卡那霉素50μg/ml的诱培养基上选择转化的愈伤组织,在含卡那霉素25μg/ml分化培养基上得到再生小芽,再生小芽中有30%外观形态正常。形态正常的小芽转到生根培养基后可长出根从而得到完整的植株。胭脂碱检测。NPT-Ⅱ活性测定及分子杂交的结果均表明,外源基因已导入枸杞并得以表达。在实验过程中还发现:(1) 经过一定预处理后,形成愈伤组织较快的外植体易被转化。(2) 转化后愈伤组织再生正常植株的频率比未转化的愈伤组织高。 相似文献
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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: :ineo1130), and the transformed calli selected from the callus induction medium containing 50 fig/ml kanamycin could regenerate buds on differentiation medium containing 25 μg/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. Nopa-line 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 hig 相似文献
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In anther culture of Panax ginseng, its callus formation showed a wide adaptability to culture media. Large numbers of calli were induced on media exhibiting better effects of induction. Supplemeres of 5 mg 2,4-D/1 and 1 mg KT/1 to the media proved to be much effective. Regeneration of the whole plantlets from anther culture of Panax ginseng is usually quite difficult. During the past three years, however, sixteen of the 100 medium formulae tested were proved to be suitable. The formulae of MS+0.5 mg BA/1+2 mg GA/1+1000 mg LH/1+3% sucrose were considered good and effective. A visibly differentiated body, which was light-milky white and later turned into a light green spot, was formed 40 days after the callus was transferred to the differentiation media. This body differentiated subsequently into buds, roots and, eventually, seedlings. The embryoid clones have been established in order to maintain its ability of continual differentiation into plantlets through successive culturings of many generations. 相似文献
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