摘要
以诸葛菜下胚轴和子叶为材料，在附加BA和NAA的MS培养基上诱导芽再生，在1／2MS培养基上诱导生根，获得完整再生植株，建立了诸葛菜组织培养高频再生体系，再用很癌农杆菌介导转化诸葛菜下胚轴和子叶，在附加一定量的氨苄青霉素、头孢霉素和卡那霉素的相应培养基上进行筛选，并培养再生成苗，获得完整抗性再生植株，移植到盛有土壤的花盆中均可存活，生长正常。将再生植株叶片，进行GUS、NPTⅡ酶活性测定和Southernblot分子杂交，证实外源基因已稳定整合到植物基因组中，并高效表达。
Excised hypeotyls and cotyledons of were used as explants for hssue culture. ably were cultured on MS medium suPPlemented with sa 2 mp/L or with M 3 mp/L and NAA 0. 2 lug/L. Regenerated buds were excised when they were 2 cm long, andtransferred ontO 1/2MS medium with IBA 0. 03 lug/L, then whole PlaniletS were regenerated at a frequency of 100%.Genehc transforrnahon of a aam was studied by coculture with H strain A208se (Pace7, PROA93, be Fig. I ), After 2 - 3 d of cocultUre (Fig. 2) the hypeotyls and cotyledens were transferred ontO selechon mwhum containing kanamycin (Km) 25 lug/L and 6aminopenicillanic acia (AP) 250 mg/L (Tables 1 - 4 ). 8 weeks later, shoots emerged, and were excised and transferred onto the roohng medium containing AP 25 rug/L and Cephazoline 100 rug/L, and roots were formed within 4 - 5 weeks. adn the whole plants were transplanted intO POtS they grew well.aam frequency of Plant regrnerahon of hypeotyls was abet 30%, and that of cotyledens was ahaut 51 %. The regenerated PlantS showed high enzyrnahc achvihes of g-glucuronidase(Fig.3 ) and neomycine PhOSPhotransferase H (Fial. 4, 5). bouthern b1Ot analysis confirmed that NPT 1 bine had been stably integrated into the chomml genome Of a ~ (FigS. 6, 7). The transformation frequency of hypeotyls was 10%, and that of cotyledens was 5. 5%.TIs is the first report about the reecnerahon of tranSgenic Plants of a aam
基金
国家自然科学基金
关键词
根癌农杆菌
诸葛来
转基因植株
Agrobacterium tumefaciens,Orychopheagmus violaceus,hypocotyl,cotyleden,transgenic plants
