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作 者 ：王艳;曾幼玲;贺宾;秦丽;李金耀;高燕;张富春

期 刊 ：作物学报 2006年 32卷 02期 页码：278-282

关键词：根癌农杆菌；带柄子叶；预培养；菌液浓度；浸染时间；转基因油菜；

Keywords:Agrobacterium tumefaciens, Cotyledons with petiole, Preculture, Concentration of bacterium, Immerged time, Transgenic oilseed,

摘 要 ：

以甘蓝型油菜带柄子叶为转化受体，通过根癌农杆菌EHA105（Agrobacterium tumefaciens）介导将盐地碱蓬（Suaeda heteroptera）的Na⁺/H⁺反向运输体基因NHX导入新疆主栽油菜1khp11品系中，获得了抗卡那霉素的再生植株，并对影响遗传转化的一些关键因素进行了研究。结果表明,带柄子叶在含有2,4-D的培养基上经过3 d预培养后于浓度OD₆₀₀为0.3～0.4的菌液中浸染7 min, 卡那抗性绿苗率可达8%～9%，经过对抗性植株的PCR检测、点杂交及PCR-Southern杂交证明外源基因已整合到油菜基因组中，为新疆盐碱土上油菜产量的提高提供了新的途径。

Abstract:

The crop yield is affected seriously by the salty soil in Xinjiang. Cotyledons with petiole of the line of Brassica napus 1khp11 were transformed with strain of Agrobacterium tumefaciens EHA105, harboring Na⁺/H⁺ antiporter gene of Suaeda heteroptera in order to select the salt tolerant oilseed. The regeneratedly kan-resistant plants were obtained. In the research, major factors of transformation were discussed detailed. The results showed that the best physiological and transforming condition was preculture for 3 days with 1 mg/L 2,4-D for cotyledons with petiole and immersion in OD₆₀₀ 0.3–0.4 liquid bacterium for 7 min, 8%–9% kan–resistant green seedlings were obtained. It was proved that NHX gene was inserted into the genome of oilseed by the methods of PCR, Dot blot and PCR-Southern.

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