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林业科学 ›› 2017, Vol. 53 ›› Issue (7): 45-53.doi: 10.11707/j.1001-7488.20170705

• 论文与研究报告 • 上一篇    下一篇

盐胁迫下转复合多基因欧美杨107杨幼苗生长及生理响应

陈盼飞1, 左力辉1, 王桂英2, 王进茂1, 任亚超1, 杨敏生1   

  1. 1. 河北农业大学林学院 保定 071001;
    2. 廊坊市农林科学院 廊坊 065000
  • 收稿日期:2016-04-22 修回日期:2017-02-21 出版日期:2017-07-25 发布日期:2017-08-23
  • 通讯作者: 杨敏生
  • 基金资助:
    国家自然科学基金项目(31370663);国家高技术研究发展计划("863"计划)项目(2013AA102703)。

Growth and Physiological Responses of Transgenic Populus×euramericana cv. ‘4/76’ with Multiple Genes Under Salt Stress

Chen Panfei1, Zuo Lihui1, Wang Guiying2, Wang Jinmao1, Ren Yachao1, Yang Minsheng1   

  1. 1. College of Forestry, Agricultural University of Hebei Baoding 071000;
    2. Langfang Academy of Agricultural and Forestry Sciences Langfang 065000
  • Received:2016-04-22 Revised:2017-02-21 Online:2017-07-25 Published:2017-08-23

摘要: [目的]通过观测转复合多基因欧美杨107杨(转基因107杨)幼苗在不同浓度盐胁迫下生长性状,质膜透性、光合作用等各项指标,以及外源基因表达产物积累量的变化,综合评价转复合多基因杨树在盐胁迫下的适应能力。[方法]以转化成功且已经过验证的转基因107杨为试验材料,未转基因107杨为对照(CK),进行组培扩繁,在幼苗株高10 cm时定植在花盆中,置于人工气候室中培养。分别用0、3、6 g·L-1的NaCl溶液进行盐胁迫处理,胁迫25天后观测2个株系幼苗的生长指标、生理参数变化,以及甜菜碱与Bt毒蛋白的积累量。[结果]2个株系的生长指标观测结果表明:在低盐浓度处理下,转基因107杨幼苗株高增长量显著高于非转基因对照,转基因株系幼苗地径增长量与对照差异不显著;高盐浓度处理下,2个株系幼苗株高与地径均受到盐胁迫的显著影响。2个株系的生理参数观测结果表明:在低盐浓度处理下,转基因107杨幼苗质膜透性显著低于对照107杨,叶绿素含量有一定程度的增加,净光合速率与蒸腾速率显著高于对照,光系统Ⅱ实际光合效率Y(Ⅱ)小幅度降低但显著高于对照,光系统Ⅱ能够保持较高的电子传递速率;Fv/Fm增幅较高,具有较大光合作用潜力。在高盐浓度下,转基因株系幼苗叶绿素含量降幅较小,而质膜透性、净光合速率均显著降低,Fv/FmY(Ⅱ)均呈现较大幅度下降,光系统Ⅱ电子传递速率受到较大影响,光合能力下降,变化趋势与对照相同,2个株系幼苗均受到较大程度的盐害。转基因株系中外源基因表达产物积累量的ELISA研究结果表明,随盐浓度的增加,Cry1Ac毒蛋白、Cry3A毒蛋白及甜菜碱含量都呈现增加趋势;在高盐浓度胁迫下外源基因产物积累量均显著增加。[结论]转基因107杨在低浓度盐胁迫下表现出良好的适应性,耐盐性优于对照107杨。在高浓度盐胁迫下转基因株系与对照107杨均受到较大影响,转基因株系并未显示其优势。盐胁迫可以诱导外源Bt基因、BADH基因表达加强,盐胁迫下转基因107杨表现出良好的耐盐和抗虫潜力。

关键词: 转复合多基因, 欧美杨107杨, 耐盐性, BADH基因, Bt基因

Abstract: [Objective] This paper aimed to comprehensively evaluate the adaptability of transgenic Populus×euramericana cv. ‘74/76’ with multiple genes (Abbreviation:Transgenic poplar 107) under salt stress through measurements of growth traits, membrane permeability and photosynthesis and other physiological parameters of the transgenic polar 107 under different salt stresses, as well as of the change in the accumulation amount of exogenous gene expression products.[Method] The transgenic polar 107 was used as test material, which had been verified successful transformation. The transgenic materials were propagated by tissue culture, and then the obtained seedlings were transplanted in pots when their heights were up to 10 cm. The seedlings were cultured in growth chamber and treated with NaCl solutions which concentrations were 0, 3, and 6 g·L-1, with the nontransgenic poplar 107 served as the control (CK). The growth indexes and physiological parameters, as well as accumulation of betaine and Bt toxin proteins of two lines were measured in 25 days after treatment.[Result] The results showed that under low salt concentration treatment, the height of transgenic poplar 107 was significantly higher than that of poplar 107, and the diameter of the transgenic lines was greater than that of poplar 107. Under high salt concentration, the height and the ground diameter of the seedlings of the two lines all were significantly affected by salt stress. Under low salt concentration, membrane permeability of transgenic poplar 107 was significantly lower than that of polar 107, chlorophyll content increased more greatly, and both net photosynthetic rate and transpiration rate increased and were significantly higher than that of polar 107. The actual photosynthetic efficiency Y(Ⅱ) of photosystem Ⅱ decreased slightly, but significantly higher than that of the control, indicating that photosystem Ⅱ was able to maintain higher electron transfer rate. Fv/Fm had a relatively greater increase, suggesting increased photosynthetic capacity. Under high salt concentration, the chlorophyll content of transgenic lines decreased slightly. However, both the plasmalemma permeability and net photosynthetic rate decreased significantly, Fv/Fm and Y(Ⅱ) had a sharp decline, electron transfer rate of photosystem Ⅱ was affected greatly and photosynthetic capacity decreased. The all change trends were the same as that of control, and the seedlings of two lines suffered to a great degree salt damage. The accumulation amount of exogenous genes expression in transgenic lines, detected by ELISA, showed that with the salt concentration increasing, the contents of Cry1Ac toxin protein, Cry3A toxin protein and betaine all had an increasing trend. The accumulation of exogenous genes products significantly increased under the stress of high salt concentration.[Conclusion] Transgenic polar 107 displayed strong adaptability under low concentration salt stress, and its salt tolerance was stronger than the polar 107. Both transgenic polar 107 with multiple genes and control polar 107 were to a great degree impacted under high concentration salt stress, and transgenic line did not show any advantage. Salt stress induced an expression enhancement of the exogenous Bt gene and BADH gene, suggesting that the transgenic poplar 107 would show a good salt tolerance and insect resistance potential under salt stress.

Key words: transgenic with multiple genes, Populus×euramericana cv. ‘74/76’, salt tolerance, BADH gene, Bt gene

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