| 生防改良菌株F1-35的GFP标记及其对西瓜幼苗生理及防御系统的影响 |
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| 引用本文： | 甘 良,查顺清,刘继红,蓝星杰,王 阳,宗兆锋.生防改良菌株F1-35的GFP标记及其对西瓜幼苗生理及防御系统的影响[J].西北农业学报,2015,24(12):108-116. |
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| 作者姓名： | 甘 良 查顺清 刘继红 蓝星杰 王 阳 宗兆锋 |
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| 作者单位： | （旱区作物逆境生物学国家重点实验室，西北农林科技大学 植物保护学院，陕西杨凌 712100） |
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| 基金项目： | 国家自然科学基金（31371978）；西北农林科技大学基本科研业务费（QN2012060）；国家公益性行业(农业)科研专项(201103016)；高等学校学科创新引智计划项目(B07049)。 |
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| 摘 要： | 为探究生防改良菌株F1-35的绿色荧光蛋白标记及其在标记后相关性状的改变情况,采用PEGCaCl2法介导原生质体转化,成功地将GFP基因转入到生防改良菌株F1-35中,转化率为每微克质粒DNA6个转化子,转化子经继代培养5代后仍能在含潮霉素B的培养基上正常生长,且其荧光稳定,强度良好;其分生孢子也能表达强烈的荧光。灌根处理西瓜幼苗发现,与原始生防改良菌株F1-35比较,经GFP标记后的菌株F1-35-GFP对西瓜的促生、抗逆作用以及其在西瓜幼苗根部的定殖情况和对西瓜枯萎病的防治效果未发生改变,防治效果仍能达60%以上;通过分析F1-35对西瓜枯萎病的防治效果及其对西瓜幼苗防御酶的影响关系发现,F1-35能提升PPO等防御酶的活性,从而抑制西瓜枯萎病的发生。可见,GFP能稳定存在于生防改良菌株F1-35中,且不对其促生、抗逆作用及防效产生影响,F1-35可通过提升西瓜幼苗防御酶活性来防治西瓜枯萎病的发生。
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| 关 键 词： | 绿色荧光蛋白 生防改良菌株F1-35 原生质体转化 防御系统 |
GFP-expressing in Improved Biocontrol Strain F1-35 and its Effects on Physiological Characteristics and Induction of Defense Responses of Watermelon Seedlings |
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| GAN Liang,ZHA Shunqing,LIU Jihong,LAN Xingjie,WANG Yang and ZONG Zhaofeng.GFP-expressing in Improved Biocontrol Strain F1-35 and its Effects on Physiological Characteristics and Induction of Defense Responses of Watermelon Seedlings[J].Acta Agriculturae Boreali-occidentalis Sinica,2015,24(12):108-116. |
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| Authors: | GAN Liang ZHA Shunqing LIU Jihong LAN Xingjie WANG Yang and ZONG Zhaofeng |
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| Abstract: | To explore the genetic marker of green fluorescent protein (GFP) on improved biocontrol strain F1-35 and the changes of strain F1-35 after marked,we mediated protoplast transformation by the method of PEG-CaCl2,then the gene coding for GFP was successfully transformed into F1-35 strain,the transformation frequency was 6 transformants per microgram DNA.Transformants could grow normally on the medium containing hygromycin B after subcultured 5 generations,the fluorescence remained stable,and the strength of green fluorescence was strong. After drenched with either strain F1-35-GFP or F1-35,the physiological characteristics of watermelon seedlings and the colonization on roots surface of the two strains and the bio-control efficacy were not significantly different,the control efficacy could remain above 60%;on the other hand,we explored the relationship between the bio-control efficacy and the defense-related enzymes activities,and found that F1-35 could showed an excellent bio-control efficacy by improving the activities of defense enzymes. In conclusion,the gene of GFP could exist in strain F1-35 stably,furthermore,there was no influence on its growth promoting and adversity resistance and the bio-control efficacy; F1-35 can effectively control Fusarium wilt through improving activities of defense enzymes. |
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| Keywords: | GFP Improved biocontrol strain F1-35 Protoplast-mediated genetic transformation Defense responses |
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