【作者】 李淼；

【导师】 檀根甲； 李瑶；

【作者基本信息】 安徽农业大学 ， 植物病理学， 2003， 硕士

【摘要】 本文以猕猴桃品种为研究对象，通过园间抗性调查的田间自然鉴定法，对不同品种进行抗病性鉴定，并分别从形态组织结构、生理生化及分子生物学三个角度进行较为系统的研究，探讨了猕猴桃品种对溃疡病的抗性及其机理，主要获得如下结果： 1．猕猴桃品种对溃疡病的抗性 采用田间自然鉴定法，通过园间病情调查并结合前人的研究结果初步得出，品种都表现为金魁与两种雄株中华软、美味硬最为抗病，早鲜次之，魁蜜稍抗，华美2号、海沃德中感，秦美、金丰最感病。用最长距离法对安徽省9个猕猴桃品种进行抗溃疡病性系统聚类，可以将9个品种划分为5类。在谱系图上，抗病性相同的品种大致划分在同一个类中。第1类为免疫品种，8、9为一类：第Ⅱ类为高抗品种，6、7为一类：在第Ⅲ类中，5为一小类，为中抗品种。第Ⅳ类为中感品种，又可截分为2小类，即4为一小类，偏抗病，3为一小类，为感病品种。第Ⅴ类则为感病品种，1为一小类，偏感病，为高感品种：2为另一小类，为感病品种。在抗病育种和抗性资源布局方面可以从中选择。9个品种被分成5类，与实际抗性调查情况基本相符，这对于品种的合理布局和抗源利用具有指导意义。该结果可为安徽省猕猴桃生产合理利用品种，减少溃疡病的发生与危害提供理论依据。同时证明了溃疡病大田发病的可控性，栽培抗病品种是控制猕猴桃溃疡病的关键措施，并为进一步深入研究其抗病机制奠定基础。 2．猕猴桃品种对溃疡病抗性的形态学机制 运用冰冻切片法与显微观测摄影技术对猕猴桃品种不同生育期、不同海拔地区一年生枝条皮孔、叶片气孔的数目、密度、大小、排列方式及其显微组织细胞结构进行了研究，并对其进行统计分析，比较不同品种间差异的显著性及其与抗病的相关性。结果表明：猕猴桃一年生枝条皮孔的长度、分布密度、排列方式与抗病性密切相关，角质膜和皮层细胞壁厚度与抗病性有一定相关性。高抗品系金魁一年生枝条皮孔细小，分布稀疏，凸起较少。高感品系金丰一年生枝条皮孔细长，分布密集，较大凸起。冰冻切片发现，高抗品系金魁一年生枝条组织具有厚的角质膜，而高感品系金丰一年生枝条组织角质膜相对薄。猕猴桃叶片气孔的长度和密度与抗病性密切相关。感病品系金丰、秦美叶片气孔密度高，开口大，叶表蜡质少，抗病品系金魁、早鲜叶片气孔密度低，开口小，叶表蜡质分布较多。显微结构观察发现，叶片细胞组织结构特征与抗病性无显著相关。 3．猕猴桃品种对溃疡病抗性的生理生化机制 利用分光光度法对猕猴桃品种不同部位组织、不同海拔地区、不同生育期下的过氧化物酶(POD)、多酚氧化酶(PPO)、超氧化物岐化酶(SOD)、过氧化氢酶(CAT)、苯丙氨酸解氨酶(PAL)、蔗糖酶及α-淀粉酶活性进行了研究，并分析其与抗溃疡病的关系。通过对不同抗性猕猴桃品种体内的POD、PPO、SOD、CAT、PAL、蔗糖酶、α-淀粉酶活性的测定分析，结果表明：感染溃疡病菌前后，抗病品种和感病品种一年生枝条及叶片组织中防御酶系活性变化规律存在一定的差异。自然发病后，供试的6个品种胞内保护酶(POD、PPO、SOD、CAT、PAL)及蔗糖酶、α-淀粉酶活性迅速提高。品种未感染溃疡病菌前，幼芽组织中的 POD、PPO、SOD、C叮、PAL酶活性与品种抗性没有明显的相关性，一年生枝条、叶片组织中的POD、PPO、SOD、CAT、蔗糖酶及a－淀粉酶活性抗病类型品种的酶活性均低于感病品种。品种感染溃疡病菌后，一年生枝条、叶片pOD、ppo、SOD、CAT、pAL、蔗糖酶、Q－淀粉酶的活性均升高，且酶活性的增加幅度是抗病品种中酶活提高倍数高于感病品种，但对于蔗糖酶、Q－淀粉酶活性的增加幅度则是感病品种中酶活提高倍数高于抗病品种。而且这种增量在不同生育期以及不同部位是有差异的。酶的活性变化还与某些生化物质含量变化存在某种内在联系。一年生枝条上病斑越大，POD、PPO、SOD、CAT、PAL酶活性越高，且抗性品种酶活性增幅高。 应用聚丙烯酚胆凝胶电泳同工酶分析技术研究了猕猴桃植株体内POD、PPO、SOD、CAT、EST、O－淀粉酶同工酶谱带的变化，结果表明：自然感染溃疡病前后，此6种同工酶谱带特征在不同抗感品种中表现出一定的差异。未感染溃疡病菌前，抗 （感）品系一年生枝条、叶片POD同工酶均有2条酶带，PPO同工酶有3条酶带，但感病品种酶带颜色深且粗，而抗病品种酶带颜色浅且细，阶1·片酶带颜色深于一年生枝条；SOD、CAT同工酶谱带均为1条，Rf值分别为0．38、0．28，感性品利。较抗耐品种谱带亮度高活性强；自然发病后，抗（感）品系＊OD、凹O问工酶谱带数都增加，分别为4、3条和5、4条，且抗病品种新酶带出现时间较感病品种早且酶带粗颜色深活性强，感病品系虽也有新酶带出现，但出现时间迟且酶带少活性弱，抗病品系 －年生枝条、叶片 POD、PPO同工酶新谱带的盯值分别为口石3、0石7和 0．85、0．87；抗感病品利。SOD、CAT同工酶都被诱导产生了 1条新的 ［d工酶谱带，Rf分别为 0．32和0二7，新酶带显色时间迟，且酶带颜色浅活性弱，但抗耐品种较感性品种谱带亮度；m舌性强；E盯问工酶于自然发病前后变化不?更多还原

【Abstract】 In this paper.the resistance of km ifruit cultivars to kiwifruit bacterial canker was estimated by natural infection in the field.the results on the resistance mechanisms on the basis of pathology,biochmistry,and molecular.The main results are as follows.l.Thc resistance of several km ifruit culitivars to kiwifruit bacterrial canker caused by Pseuomonas syringae pv. actinidaeResistance of several culitivars of kiwifruit to kiwifruit bacterrial canker were studied by means of natural infection identification in the field.The results showed that the resistance of "Jinkui" and "Zhonghua soft" and "Meiwei hard" were high-resistance, "Zaoxian" was intermediate, "Kuimi" was middle-resistance, "Huamei No.2" and "Harvard" were middle-susceptibility, "Qinmei" and "Jinfeng" were high-susceptibility. The cluster analysis of 9 kiwifruit cultivars resistance to bacterial canker disease in Anhui was studied with the longest distance method. The results showed that 9 kiwifruit cultivars could be divided into 5 groups, and cultivars of the same resistance had been fell into the same group in the pedigree figure. Class I was immunity cultivars.8 and 9 were one group. Class II was high-resistance cultn ars.6 and 7 were one group. Class 111 was middle-resistance cultivars. There were two sub-groups in cultivars of class IV .which mostly were middle-susceptibility cultivars. That is: 4 was one sub-group .belonging to more resistant varieties:3 was another sub-group , susceptible one. then the last class V was susceptible ones.l was formed sub-group, more susceptible one,2 was another susceptible group. They could be chosen in disease resistance in breeding or the distribution of different resistant varieties. The result of dividing 9 varieties into 5 groups was coincident with the facts. It could also be used in guiding disease resistance in breeding or the distribution of different resistant cultivars. and the results were the basis of the resistance mechanisms.2.Thc morphological structures resistance mechanism of several kiwifruit culitivars to kiwifruit bacterrial canker caused by Pseuomonas syringae pv. actinidaeThe adoption of freezing tissue slice method and microscope observation photograph technique to study different growth period and altitude of the number. density,size.array mode, tissue cell microstructure of the annual hvig skin hole and foliage stonia, and processing statistical analysis to compare different cultivars marked character of diversity and the relation with resistance disease. The results indicated that the density and length of branch skin hole and array mode were significant differences in resistance lo kiwifruit bacterial canker caused by Pseuomonas syringae pv. actinidae among the cultivars. the thickness of keratose membrane and cortex cell wall were in some correlation with the disease resistancc.Thc annual twig skin hole of high-resistance cultivar "Jinkui" is thin and small, distributing sparsely, few protuberance. The annual twig skin hole cf high-susceptibility cultivar "Jinfeng" is thin and long, distributing densely, big protuberance. The application of free/ing tissue slice method to the microstructure of twig showed that the keratose membrane of high-resistance cultivar "Jinkui" is thick, oppositely the keratose membrane of high-susceptibility cultivar "Jinfeng" is thin. The density and length of foliage stonia are significant differences in resistance to Pseuomonas syringae pv. actinidae among the cultivars. The density of foliage sloma of the susceptibility cultivar "Jinfeng" and "Qinmei" is high and its rir.gcnt extent is big. the \\axiness of the foliage surface is little And that, the density of foliage stoma of the resistance cultivar "Jinkui" and "Zaoxian" was low and its ringent extent was small, the waxiness of the foliage surface is much more. The microstructure observation discovered that the cell structure character of foliage was in no significant correlation with the disease resistance. 3. The physiological and biochemical resistant mechanism of several kiwifruit cu更多还原