【作者】 王芳；

【导师】 谢艺贤； 蒲金基；

【作者基本信息】 海南大学 ， 植物病理学， 2012， 硕士

【摘要】 芒果炭疽病是世界性的芒果重要病害,其主要病原菌为胞炭疽菌(Colletotrchumgloeospordes Penz.)。该病在采前主要引起芒果落花落果、叶片坏死和枝枯等症状。由于具有潜伏的特性,田间侵染未成熟果实一般不表现症状,常导致采后果实在贮藏运输和销售过程中腐烂,严重影响芒果果实的产量和质量,被认为是芒果生产上的主要限制性生物因素之一。化学防治作为芒果炭疽病的主要防治方法,在生产上长期应用,病原菌已对主打杀菌剂苯丙咪唑类农药产生了抗性,无法达到预期的防治效果。同时,随着人民生活水平的提高,人们对食品安全日益重视,利用诱抗剂诱导果蔬自身产生系统获得抗病性逐渐成为果蔬病害防治的发展方向和研究热点,是具有远景的植物病害绿色控制技术之益本实验通过对多种化学诱抗剂的田间筛选、芒果抗病性相关酶活性变化以及对诱导的蛋白质差异性的初步研究,获得诱抗效果较好的诱抗剂,并初步阐明其诱抗机理,为进一步研究利用诱抗技术防治芒果炭疽病奠定了理论基础。研究结果如下：1、本研究分别从诱抗剂的田间诱抗效果测定、系统性诱抗效果测定和诱抗剂的混合诱抗效果测定3个方面进行诱抗剂的筛选,测定结果表明：参试的3种诱抗剂对炭疽病均有一定的诱抗效果。Bion(50%有效成分)和COS(2100Da)以100μg/ml浓度在处理后48h的诱抗效果分别达41%、37%,SA则以50μg/ml的浓度在处理后24h的诱抗效果可达22%；各诱抗剂同时也表现出了诱抗的系统性,诱抗效果可达50%以上；不同比例诱抗剂的混合施用表现出一定的抗抗作用,随着时间的推移诱抗效果缓慢降低。2、抗病性相关酶活性测定结果表明,200μg/mlBTH处理芒果叶片后,不同程度地提高5种与抗病相关的酶活性,其中苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)、过氧化氢酶(CAT)在处理后的24h,叶片酶活分别提高了37%、27%和440%,超氧化物岐化酶(SOD)和过氧化物酶(POD)于处理后的72h酶活性分别提高了83%和330%。3、总酚含量测定结果表明,不同浓度BTH、1000μg/ml多菌灵、接种胶孢炭疽菌分别处理芒果叶片和采后果实,总酚含量均有不同程度的提高。不同处理对芒果果实和叶片总酚含量与对照相比差异显著,并且以100μg/ml的BTH处理果实和叶片总酚含量最高,总酚含量分别提高85%和104%；而果皮总酚含量测定结果表明不同处理后总酚含量在不同时期是有差异的,72h的总酚含量以100μg/m1的BTH处理的最高,总酚含量提高45%,而至黄熟果皮则以病原菌接种处理的最高。4、通过双向电泳及差异蛋白质谱分析,BTH诱抗处理中,芒果果皮16个蛋白质点存在差异,对其中11个蛋白质点的肽指纹分析表明,差异蛋白质点多为热激蛋白,可视为抗逆性的蛋白质,其中3个蛋白质具有平衡活性氧的代谢、参与了抗逆酶的合成的作用,推测这些蛋白质与诱导抗性有密切的关系。更多还原

【Abstract】 The mango anthracnose disease,induced by Colletotrichum gloeosporioides Penz.,cause the leaf spot,yong fruits and flowers falling and shoot die-back.But its more characteristic symptom is fruits rot which usually occurs post harvest during the fruit storage and marketing due to latent infection.The disease has a bad effect on quality and production of mangos.Long-term since chemical control as the main method to control mango ant hracnose disease.but,the Coletotrichum gloeosporioides have a resistable effect o n common fungicide,unable to reach the control effect,on the other hand,with t he improvement of living standards,people paid more attention on food safety,S o use the elicitors inducing fruit and vegetable produce systemic acquired disea se resistance as the decelopment trend and research hotspot,as a long-term ands afe disease control methods.The experiment based on a variety of elicitors field screening,the change of the disease resistance related enzymes and the research of difference protein.Ob tain betteer effect of induced resistance inducer,and clarify the mechanism of in duced resistance, and probide the theoretical foundation for further research on the use of technology to control mango anthracnose disease resistance induced. The results as follows:1.We do three aspects for the following experiment,the determination of resi stance induced in field,induced systemic resistance and the application of the in ducer of mixed.The experimental results show that the tasted elicitors on anthra cnose of induced resistance, the100μg/ml Bion and COS can resistance induce d by41%、37%after treatment in48h,50μg/ml SA can resistance induced by22%after treatment in24h;the tasted inducer also showed resistance induction of systemic induced resistance,more than50%;Different proportions of elicitors mixed application shoued a certain antagonism,and with the passage of time in duced resistance decreased slowly.2. Disease resistance related enzymes of determination results shows:Treatme nt of200μg/mlBTH can increased the activity of disease resistance enzymes,su ch as the antivity of phenylalanine ammonia-lyase(PAL),poly-phenoloxidase(PP O),peroxidase(POD),superoxide dismutase(SOD),and catalase(CAT).3.The results of total phenolic content determination shows:Different concentrations of BTH,Carbendazim,vaccination treatment on mango leaves and postharvest fruit,the experimental results shows that,the total phenol content of mango leaves in different treatment show significant differences in different time,and low concentration of BTH treatment have the highest total phenolic content.for mango peel,the total phenolic content also show significant difference in differenct time.Experimental results show the total phenols content can improve after inducer treatment.4. By two-dimensional electrophoresis and mass spectrometry analysisis,we get16differences of proteins and11peptide mass fingerprint of protein spots in mango peel in this experiment.The detected differential protein spots for hea t shock protein, as resistance protein.The heat shock protein of NO.2,7,14with balanced active oxygen metabolism,involved in the resistance enzyme synthesis,t hereby improving disease r esistance,or even inhibit the disease更多还原