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作物学报 ›› 2013, Vol. 39 ›› Issue (08): 1345-1351.doi: 10.3724/SP.J.1006.2013.01345

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

蛋白激发子Hrip1基因在拟南芥中表达可提高植株的耐盐耐旱能力

彭学聪,杨秀芬,邱德文,曾洪梅,郭立华,刘峥*   

  1. 中国农业科学院植物保护研究所 / 农业部作物有害生物综合治理重点实验室, 北京 100081
  • 收稿日期:2013-01-15 修回日期:2013-04-22 出版日期:2013-08-12 网络出版日期:2013-05-22
  • 通讯作者: 刘峥, E-mail: liuzheng@caas.cn
  • 基金资助:

    本研究由国家高技术研究发展计划(863计划)项目(2012AA101504)资助。

Overexpression of Elicitor Protein Encoding Gene Hrip1 Improves Tolerance to Drought and Salt Stresses in Arabidopsis

PENG Xue-Cong,YANG Xiu-Fen,QIU De-Wen,ZENG Hong-Mei,GUO Li-Hua,LIU Zheng*   

  1. Key Laboratory of Pest Management in Crops, Ministry of Agriculture / Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2013-01-15 Revised:2013-04-22 Published:2013-08-12 Published online:2013-05-22
  • Contact: 刘峥, E-mail: liuzheng@caas.cn

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摘要：

Hrip1是从极细链格孢(Alternaria tenuissima)代谢物中分离的一种蛋白激发子。将蛋白激发子基因Hrip1转化到拟南芥，对5个T1代转基因拟南芥株系进行分子检测, 证明Hrip1基因能够在拟南芥中转录和表达。转基因植株对盐和干旱胁迫的抗性显著增强, 75 mmol L−1 NaCl和50 mmol L−1甘露醇渗透胁迫2 d, 转基因植株种子平均相对发芽率为32.1%和77.9%, 分别比野生型的增加3.72倍和5.61倍; 150 mmol L−1 NaCl和50 mmol L−1甘露醇处理拟南芥幼苗7 d后, 转基因植株平均相对根长为81.79%和93.25%, 分别是野生型的1.53倍和1.34倍。3周龄的转基因植株在250 mmol L−1 NaCl条件下胁迫20 d, 平均存活率为67%, 显著高于野生型(42%)(P<0.05); 干旱胁迫25 d后, 复水5 d转基因植株平均存活率为72%, 而野生型仅为44%。检测结果显示转基因植株叶片的抗氧化酶活性明显高于野生型, 用200 mmol L−1 NaCl和200 mmol L−1甘露醇处理24 h后, POD活性分别比野生型植株提高1.56倍和1.85倍, CAT活性分别比野生型植株提高1.64和1.86倍。说明蛋白激发子Hrip1基因在拟南芥中的表达能够改善和提高植株的耐盐抗旱能力。

关键词: 蛋白激发子, Hrip1, 转基因拟南芥, 干旱胁迫, 盐胁迫

Abstract:

Hrip1 is a novel hypersensitive response-inducing protein secreted by necrotrophic fungus,Alternaria tenuissima. The Arabidopsis thaliana transgenic plants overexpressing Hrip1 showed increased tolerance to drought and salt stresses. Five transformed lines of Arabidopsis were confirmed by PCR, RT-PCR and Western blot analysis. T4 progenies of transgenic Arabidopsis lines and the wild-type plants were tested. The results showed the seed germination rate and root length of transgenic plants were all significantly higher than those of wild type plants (P<0.05) under stress conditions. The average seed relative germination rate of transgenic Arabidopsis lines was 32.1% and 77.9%, which were 3.72 and 5.61 times higher than those of wild-type plants in the two days incubation with 50 mmol L−1 NaCl and 75 mmol L−1 mannitol, respectively. The average relative root length in transgenic plants was 81.79% and 93.25%, which were 1.53 and 1.34 times of that in wild type plants at 7th day on the medium containing 150 mmol L−1 NaCl and 50 mmol L−1 mannitol, respectively. Stress tolerances were improved in the three-week old soil-grown seedlings. Survival rate of transgenic plants and wild type plants was 67 % and 42 % under 250 mmol L−1 NaCl stress for 20 days, respectively and 72 % of the transgenic plants and 44 % of wild-type plants were survived at 5th day re-watering after dehydration stress for 25 days. Otherwise, POD and CAT activities were increased in transgenic plants compared with three wild type plants under salt and drought stresses. At 24 h of treatment with 200 mmol L−1 NaCl and 200 mmol L−1 mannitol, the average POD activity in transgenic seedlings was 1.56 and 1.85 times, CAT activity was 1.64 and 1.86 times higher than those in wild plants, respectively. These data indicated thatthe expression of Hrip1 gene positively regulates salt and drought tolerance in transgenic Arabidopsis.

Key words: Protein elicitor, Hrip1, transgenic Arabidopsis, Drought stress, Salt stress

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