根瘤农杆菌介导的枸杞转化及转化植物的获得

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。     

Genetic Transformation of Lycium barbarum L. via A. tumefaciens

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     

转基因红豆草植株的再生

红豆草子叶外植体能被含非致瘤性的Ti质粒的根癌农杆菌感染.该载体质粒含一个npt-Ⅱ基因和一个胭脂碱合成酶基因.对卡那霉素抗性植株的胭脂碱检测、NPT-Ⅱ酶活性测定和DNA分子杂交试验表明,外源的npt-Ⅱ基因和胭脂碱合成酶基因已导入了红豆草细胞并能在植株水平表达相应的性状.     

转基因 红豆草植株的再生

红豆草 子 叶外植体 能被 含非致 瘤性 的 i T 质粒 的根癌 农杆 菌感染.该载体 质 粒含 一个nt P一I基 因和一个胭脂 碱合 成酶基因.对 卡那霉素抗性 植株的胭脂碱检测、N P T一I酶 活性测定和 D N A 分子 杂交 试验表 明,外 源的 n Pt一,基 因和胭脂 碱合成酶基 因已 导入 了红豆 草细胞并能在植株水平表达相应 的性状.     

籼稻幼胚组织培养高频率植株再生及其基因转化的研究

以籼稻品种特青的幼胚作为外植体,研究了激素对愈伤组织诱导及植株再生的影响。ＭＳ＋２,４－Ｄ２ｍｇ／Ｌ作培养基,愈伤组织诱导率达１００％;愈伤组织在ＭＳ＋ＫＴ３ｍｇ／Ｌ＋ＮＡＡ０．５ｍｇ／Ｌ的培养基上,分化频率为６７．３％,在此基础上利用基因枪转化技术,将外源的ｂａｒ基因导入了水稻基因组,转基因植株表现出对除草剂Ｂａｓｔａ很强的抗性。