根癌农杆菌介导转化诸葛菜获得转基因植株

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。

Agrobacterium-Mediated Transformation of Orychopheagmus Violaceus and Regeneration of Transgenic Plants

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