农杆菌介导NHX基因转化甘蓝型油菜的研究

作物学报 ›› 2006, Vol. 32 ›› Issue (02): 278-282.

农杆菌介导NHX基因转化甘蓝型油菜的研究

王艳;曾幼玲;贺宾;秦丽;李金耀;高燕;张富春

新疆生物资源基因工程重点实验室, 新疆大学生命科学与技术学院，新疆乌鲁木齐830046

收稿日期:2004-12-28 修回日期:1900-01-01 出版日期:2006-02-12 网络出版日期:2006-02-12
通讯作者: 张富春

Research of NHX Gene Transformation in Brassica napus by Agrobacterium tumefaciens

WANG Yan; ZENG You-Ling; HE Bin; QIN Li; LI Jing-Yao; GAO Yan and ZHANG Fu-Chun

Xinjiang Key Laboratory of Biological Resources and Genetic Engineering，College of Life Science and Technology, Xinjiang University，Urumqi 830046, Xinjiang

Received:2004-12-28 Revised:1900-01-01 Published:2006-02-12 Published online:2006-02-12
Contact: ZHANG Fu-Chun

摘要/Abstract

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。

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

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.

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

中图分类号:

S565

王艳;曾幼玲;贺宾;秦丽;李金耀;高燕;张富春. 农杆菌介导NHX基因转化甘蓝型油菜的研究[J]. 作物学报, 2006, 32(02): 278-282.

WANG Yan; ZENG You-Ling; HE Bin; QIN Li; LI Jing-Yao; GAO Yan and ZHANG Fu-Chun. Research of NHX Gene Transformation in Brassica napus by Agrobacterium tumefaciens[J]. Acta Agron Sin, 2006, 32(02): 278-282.

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