欧美杨107杨β-1,3-葡聚糖酶(BG2)基因遗传转化及对溃疡病的抗性分析

doi:10.11707/j.1001-7488.20131108

林业科学 ›› 2013, Vol. 49 ›› Issue (11): 60-66.doi: 10.11707/j.1001-7488.20131108

欧美杨107杨β-1,3-葡聚糖酶(BG₂)基因遗传转化及对溃疡病的抗性分析

牛庆霖¹, 王迎², 罗磊², 黄艳艳², 刘静², 冯殿齐^1,2, 曹帮华¹

1. 山东农业大学农业生态与环境重点实验室泰安 271018;
2. 泰安市泰山林业科学研究院泰安 271000

收稿日期:2013-06-06 修回日期:2013-08-08 出版日期:2013-11-25 发布日期:2013-11-26
通讯作者: 冯殿齐
基金资助:
山东省科技发展计划重点项目（2009GG10009002）。

Transformation of BG₂ Gene into Populus×euramericana cv.‘Neva’and Resistance of the Transgenic Poplar to Canker Disease

Niu Qinglin¹, Wang Ying², Luo Lei², Huang Yanyan², Liu Jing², Feng Dianqi^1,2, Cao Banghua¹

1. Key Laboratory of Agricultural Ecology and Environment, Shandong Agricultural University Tai'an 271018;
2. Taishan Institute of Forestry Science Tai'an 271000

Received:2013-06-06 Revised:2013-08-08 Online:2013-11-25 Published:2013-11-26

摘要/Abstract

摘要：

以欧美杨107杨为受体材料，利用根癌农杆菌介导法转化β-1，3-葡聚糖酶基因（BG₂），经卡那霉素筛选，PCR和Southern检测显示有4个转基因阳性株系，初步证明外源目的基因已整合到107杨基因组中。离体抑菌试验表明转基因107杨（T-107杨）细胞粗提液的抑菌能力明显强于未转基因对照107杨（C-107杨）。在接种杨树溃疡病菌的培养基上培养，T-107杨长势明显强于C-107杨；T-107杨可明显抑制丙二醛（MDA）在植物体内的快速积累，其苯丙氨酸解氨酶（PAL）含量也明显高于C-107杨，PAL含量T-107杨和C-107杨都有先升后降的趋势；15天后C-107杨完全死亡，T-107杨仍能生存。试验结果表明BG₂基因的表达提高了107杨对杨树溃疡病的抗性。

关键词: 欧美杨107杨, 根癌农杆菌, BG₂基因, 基因转化, 溃疡病

Abstract:

The BG₂ gene was transformed into Populus×euramericana cv.‘Neva’via Agrobacterium tumefaciens mediated transformation method. Four transgenic lines were obtained through selection of kanamycin resistance, PCR and Southern analyses. These analyses suggested that the BG₂ gene was integrated into the plant genome. Then, the transgenic plants were tested for resistance to the fungal pathogen Cytospora chrysosperma. Crude extracts of the transgenic T-107 plants exhibited in vitro inhibitory activity against the fungal pathogen. When inoculated with Populus×euramericana canker disease, the T-107 plants grew better than the C-107 plants. The MDA levels of T-107 were lower than that in C-107. The PAL contents of both transgenic and control plants were firstly increased obviously then decreased; However, the PAL content of T-107 plants was always higher than that in C-107 plants. After 15 days, all the C-107 plants dead, but the T-107 was still alive. These results indicated that the transformed BG₂ gene could improve the resistance of T-107 plants to canker disease.

Key words: Populus×, euramericana cv.‘Neva’, Agrobacterium tumefaciens, BG₂ gene, gene transformation, canker disease

中图分类号:

S718.46

牛庆霖, 王迎, 罗磊, 黄艳艳, 刘静, 冯殿齐, 曹帮华. 欧美杨107杨β-1,3-葡聚糖酶(BG₂)基因遗传转化及对溃疡病的抗性分析[J]. 林业科学, 2013, 49(11): 60-66.

Niu Qinglin, Wang Ying, Luo Lei, Huang Yanyan, Liu Jing, Feng Dianqi, Cao Banghua. Transformation of BG₂ Gene into Populus×euramericana cv.‘Neva’and Resistance of the Transgenic Poplar to Canker Disease[J]. Scientia Silvae Sinicae, 2013, 49(11): 60-66.

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欧美杨107杨β-1,3-葡聚糖酶(BG₂)基因遗传转化及对溃疡病的抗性分析

Transformation of BG₂ Gene into Populus×euramericana cv.‘Neva’and Resistance of the Transgenic Poplar to Canker Disease

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