摘要
鄂花四号、中花四号和豫花一号萌动４ｄ的小叶作外植体，通过器官再生在ＭＳ培养基加３ｍｇ／ＬＢＡＰ和１ｍｇ／ＬＮＡＡ上诱导芽分化，转至ＭＳ加５ｍｇ／ＬＢＡＰ上诱导芽伸长，再生植株。芽诱导率为８６％～１００％，每个外植体平均再生５个植株。在培养基中加入１ｍｇ／ＬＡｇＮＯ３和５００ｍｇ／Ｌ羧苄青霉素能有效抑制愈伤褐化和死亡，并能提高植株再生率。萌动４ｄ的小叶与农杆菌ＡＧＬ１分别共培养２ｄ，通过在再生培养基中加卡那霉素筛选，获得转基因再生植株，转基因植株生根后，移栽网室生长，并已开花结荚。外源基因的表达通过卡那霉素抗性和Ｇｕｓ活性组织化学检测得到证实。
Leaflet explants from 4-day-old seedlings of peanut cultivars Ehua No. 4,Zhonghua No.4 and Yuhua No. 1 produced bud primordia via organagenesis on MS medium supplemented with 3mg L-1 BAP and ling L-1 NAA. Plants were regenerated by elongation of bud primordia on transfer to MS medium supplemented with sing L-1 BAP. The proportion of leaflet explants which produced bud primordia was 86%～100% with an average offive regenerated plants per explant. Supplement of the growth medium with 1mg L-1 AgNO3and 500mg L-1 carbenecillin reduced formation of brown and nonregenerable callus and increased the frequency of plant regeneration. Leaflet explants from 4-day-old seedlings of peanut cv. Ehua No. 4 were co-cultivated with Agrobacterium tumefaciens AGL1 containing the binary vector pBI121 for 2 days and cultured on regeneration medium supplemented with 100mg/L kanamycin to select putatively transformed plants, which formed roots on MS medium supplemented with 0. 2mg/L NAA and 100mg/L kanamycin and were transferred to soil. Transgenes expression was indicated by kanamycin resistance and Gus histochemical assay. One putative transgenic line was regenerated per 50 explants inoculated with Agrobacterium strain AGL1。
出处
《中国油料》
CSCD
北大核心
1996年第4期52-56,共5页
基金
澳大利亚国际农业研究中心资助
关键词
花生
小叶
农杆菌
遗传转化
植株再生
Peanut Leaflet Arobacterium tumefaciens Transformation Plantregeneration
