摘要
茎用芥菜（Ｂｒａｓｓｉｃａ ｊｕｎｃｅａ ｖａｒ． ｔｓａｔｓｉ） 品种“泡叶”的倒插叶柄被携带农杆碱型Ｒｉ质粒ｐＲｉ１８５５ 的发根农杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ ｒｈｉｚｏｇｅｎｅｓ）菌株ＬＢＡ９４０２ 感染后获得毛根，出根率为１００％ 。转化根在不含激素的培养基上快速生长，并表现出典型的毛根性状。在含ＢＡ ８．０ｍ ｇ／Ｌ和ＮＡＡ ０．６ ｍ ｇ／Ｌ的ＭＳ培养基上毛根再生成完整转化株。冠瘿碱分析表明ＴＲ－ＤＮＡ已在转化株中整合和表达，ＰＣＲ分析和Ｓｏｕｔｈｅｒｎ 杂交证实内含８６２ ｂｐ ｒｏｌＢ序列的ＴＬ－ＤＮＡ也已整合入转化株的染色体组。
Hairy roots of mustard (Brassica juncea var. tsatsi) cv. “Paoye” were obtained from in vitro inoculation of reversely inserted petioles with Agrobacterium rhizogenes strain LBA9402 harbouring the agropine type Ri plasmid (pRi1855). The root inducing rate was 100%. Transformed roots grew rapidly on hormone free MS medium and showed typical hairy root phenotype. Transformed plantlets regenerated from hairy roots on MS medium supplemented with BA 8.0 mg/L and NAA 0.6 mg/L. Opine analysis evidenced the integration and expression of TR DNA, PCR analysis and Southern hybridization confirmed the integration of TL DNA including 862 bp rol B sequence in the transformed plants.
基金
国家自然科学基金
博士点基金
关键词
发根农杆菌
茎用芥菜
转基因植株
遗传转化
Agrobacterium rhizogenes
Brassica juncea var. tsatsi
Hairy root
Polymerase chain reaction
Transgenic plant
