几种植物提取物诱导马铃薯对致病疫霉的抗性

 采用纤维素膜技术研究了大蒜、洋葱、芹菜、丁香、花椒、大黄等24种植物离体组织及其提取物(其中8种中草药为水煎剂,16种其它植物为水浸剂)对马铃薯晚疫病菌(Phytophthora infestans)静止胞萌发、附着胞形成及侵入丝形成的直接影响和提取物诱导块茎对晚疫病病菌侵入机构形成的影响,并在此基础上进一步就提取物免疫处理种薯、诱导成株期抗病性、增加块茎产量的影响做了初步探讨。在洋葱、葱、大蒜等12种植物组织表面,静止胞萌发率明显低于对照(擦镜纸),而且在6种植物组织表面未见附着胞形成,10种植物组织表面未见侵入丝形成;在大蒜、韭菜等13种植物组织提取液中,静止胞萌发率明显低于对照(擦镜纸),其中6种植物组织提取液中附着胞未见形成,7种植物组织提取液中侵入丝未见形成。经8种植物组织提取物诱导处理的块茎切片,不仅保鲜期长,而且表现出较强的诱导抗性;静止胞萌发率、附着胞形成率及侵入丝形成率均明显低于对照;经8种植物组织提取物诱导处理的种薯在出芽、早期生长、抗晚疫病及产量等各方面均优于对照,病害保护率达54%、增产率达31%。     

致病疫霉有性生殖在晚疫病流行中的意义

对致病疫霉有性生殖的发生、有性生殖生理、遗传变异及可能造成的危害进行概述 ,对致病疫霉有性生殖在晚疫病流行中的意义作了探讨 ,为制定致病疫霉可靠的防治策略提供科学依据。     

中国不同地区致病疫霉遗传多样性的RAPD分析

 本文应用RAPD技术检测了我国主要马铃薯产区致病疫霉的遗传分化情况及不同地区菌株间的亲缘关系。用筛选出的10个随机引物对1997-2001年间采自我国9省市的82株及3株来自日本的致病疫霉DNA进行了PCR扩增,获得了79条谱带,其中多态性标记75条,占95%。根据扩增结果,运用UPGMA分析,获得了表现菌株间亲缘关系的树状图。菌株间的最大遗传距离为0.5,以距离0.3为阈值,可将供试菌株划分为10个组(RG1-10)。结果发现:A1交配型菌株群体内的差异大于A1和A2菌株群体之间的;RAPD分组与菌株的地理来源、交配型及对甲霜灵的敏感性无明显相关性。研究结果显示,来自中国北方甘肃、内蒙、吉林、黑龙江地区的菌株与一些来自云南、四川等西南地区的菌株亲缘关系相近。病原菌随种薯的迁移可能是导致这种现象的原因之一。     

马铃薯晚疫病化学防控现状与展望

马铃薯晚疫病是由致病疫霉引发的重大毁灭性病害,长期在我国各马铃薯主产区暴发流行,是马铃薯产业发展的重要瓶颈因素.目前大田防控马铃薯晚疫病仍主要依赖化学防控.本文梳理了近年来我国市场防治晚疫病的杀菌剂产品发展趋势,对马铃薯晚疫病化学防控策略与应用现状进行了归纳.结合本课题组近年来的田间调研结果,探讨了我国马铃薯晚疫病防控...     

辣椒疫霉致病力分化的初步研究

关于辣椒疫霉Phytophthora capsici Leonian致病力的分化研究,Black等曾报道来自台湾的辣椒疫霉对辣椒有一定的寄生专化性,菌株间存在致病力分化现象,可分为3种致病型,国内这方面的研究迄今报道较少。为了明确安徽省辣椒疫霉是否存在致病力分化现象,为辣椒抗病育种及该病的防治提供理论依据,作者对分离自该省不同地区的辣椒疫霉菌株的致病力及其分化进行了初步研究,结果如下。     

致病疫霉诱导的马铃薯酵母双杂交文库构建及无毒蛋白PiAVR3b寄主靶标筛选

致病疫霉是引起番茄、马铃薯晚疫病的植物病原卵菌,其分泌的效应蛋白在抑制寄主免疫方面发挥重要功能。鉴定效应蛋白的寄主靶标对研究植物与病原菌互作具有重要意义。本研究构建了高效马铃薯酵母双杂交cDNA文库,并通过筛选致病疫霉无毒蛋白PiAVR3b的互作蛋白来评价构建文库质量。分别提取接种致病疫霉后24 h和48 h的马铃薯栽培种‘合作88’叶片RNA,等量混合后构建cDNA文库。次级文库容量为1.6×10~7 cfu/mL,重组率为100%,插入片段平均在1 000 bp以上。以PiAVR3b为诱饵,通过酵母双杂交筛选初步获得10个候选靶标蛋白。经点对点验证,进一步确定了4个与PiAVR3b互作的靶标蛋白,分别是马铃薯MYB-like蛋白、线粒体外膜孔蛋白、碳分解代谢抑制蛋白、肽基脯氨酰异构酶。本研究构建的酵母双杂交文库为致病疫霉效应蛋白靶标筛选及植物与病原菌互作研究奠定了基础。     

2013-2014年北方5省(区)致病疫霉抗药性监测及交配型分析

为明确致病疫霉对甲霜灵的敏感性及其交配型的动态变化,采用菌落生长速率法监测了2013-2014年采自我国北方5省(自治区)的马铃薯致病疫霉对甲霜灵的抗性,并利用标准菌株与待测菌株对峙培养测定了交配型.2013年和2014年检测的致病疫霉均以中抗菌株占主导,分别占当年检测菌株总量的82.7％和75.6％,但高抗菌株从3.3％增加到16.3％.测定甲霜灵对部分敏感、中抗和高抗菌株的EC50,发现该药剂对高抗菌株的EC50分别是中抗菌株和敏感菌株的5倍和400倍.2013年采自内蒙古、黑龙江、辽宁3省(自治区)6个县(市)的92个菌株中,A1交配型占主导地位,还发现了A2和A1A2;2014年采自内蒙古、黑龙江2省(自治区)4个县(市)的52个菌株仍以A1为主,A2所占比例较低,但未检测到A1A2.2013-2014年北方5省(自治区)致病疫霉群体结构呈现复杂化的趋势.     

RXLR类效应分子PITG_21645.2的基因序列分析及功能验证

 马铃薯晚疫病黑龙江菌株效应分子PITG_21645.2在本氏烟(Nicotiana benthamiana)上可以抑制由INF1引起的过敏性细胞坏死(Hypersensitive response, HR)。为验证其为致病疫霉致病的重要效应分子,克隆了10个致病疫霉菌株以及3个同属卵菌菌株中的PITG_21645.2同源基因,它们在氨基酸序列上相似度为93%~100%。与INF1共表达的结果显示,仅有PITG_21645.2^MP903丧失了抑制HR的功能。其中PITG_21645.2^MP903的第31位氨基酸存在特异性,编码丝氨酸,其余同源基因在该位点均编码天冬酰胺。PITG_21645.2^MP903回复突变体(S31N)能够恢复该基因抑制INF1引起HR的功能,说明第31位氨基酸是影响抑制功能的关键位点。另外,PITG_21645.2还能抑制BAX、大豆疫霉PsojNIP和效应分子Avh238在本氏烟上激发的HR,而PITG_21645.2^MP903都丧失抑制功能。本氏烟上表达PITG_21645.2能够促进致病疫霉在寄主上的定殖,从而提示PITG_21645.2在致病疫霉的侵染中发挥致病性功能。     

用纤维素膜技术研究致病疫霉侵入前与植物相互作用的新方法

 纤维素膜置于不同植物组织表面,在膜上接种致病疫霉游动孢子,观察游动孢子的静止、萌发、附着胞形成及侵入。发现不同植物对致病疫霉游动孢子静止的影响无显著差异,但对游动孢子萌发、附着胞形成、侵入的影响不同,表现出不同的相互作用。马铃薯块茎和叶片对孢子萌发、附着胞形成及侵入均有明显的促进作用;日本桔果实等只对孢子萌发有促进作用,对附着胞形成、侵入无明显影响;而洋葱鳞茎和葱茎,即使在有马铃薯组织存在下也明显抑制孢子萌发和附着胞形成。纤维素膜技术是研究致病疫霉侵入前与植物相互作用的有效方法。     

苎麻疫霉对棉苗致病力的遗传与变异研究

 以分离自江苏省棉铃疫病病组织的苎麻疫霉(Phytophthora boehmeriae Sawada)野生型菌株JS-5为亲本,采用菌丝块创伤接种法测定了苎麻疫霉对棉苗致病力在游动孢子无性系和卵孢子后代的遗传。结果表明,苎麻疫霉对棉苗的致病力在单游动孢子无性系连续两代稳定遗传,而在单卵孢第1代(OG₁)则发生连续性变异。从OG₁中选致病力强、弱2个单卵孢株为亲本,分别建立单卵孢第2代(OG₂)和单游动孢子无性系,并测定其对棉苗的致病力。结果为上述2个单卵孢株的游动孢子后代对棉苗的致病力均与其各自亲本相似,而在它们的单卵孢株群体(即OG₂)中对棉苗的致病力继续发生复杂的连续性变异。上述结果表明,苎麻疫霉对棉苗的致病力可能由细胞核杂合多基因控制。     

近几年我国马铃薯晚疫病流行特点分析与监测建议

马铃薯晚疫病是马铃薯上的重要病害,是影响马铃薯产业发展的主要制约因素。本文系统总结了2008-2014年我国马铃薯晚疫病的发生概况及特点,分析了其影响因素。该病害年度间、地区间、品种间发生不平衡,北方产区受气候因素影响大,西南及中原产区常年偏重发生。2012年、2013年马铃薯晚疫病在全国范围内偏重发生,局部大发生,发生范围广、流行扩展速度快、危害重。在气候、菌源量、品种抗性及防治因素中,气候是影响马铃薯晚疫病发生流行的最重要因素。     

Differential interaction of Phytophthora infestans on tubers of potato cultivars with different levels of blight resistance

Differential interactions in tuber blight attack between potato cultivars and Phytophthora infestans isolates were studied using whole tuber and tuber slice assays. Tuber blight incidence and severity were studied in a whole tuber assay, whilst necrosis and mycelium coverage were evaluated in a tuber slice assay. The overall defence reaction of the potato cultivars tested varied considerably. Cultivars like Kartel and Producent showed resistant reactions, whilst Bintje and, to a lesser extent, Astarte reacted more susceptibly after inoculation with aggressive strains of P. infestans . A highly significant cultivar by year interaction was observed when tuber blight incidence was evaluated in two successive years. Differential responses were revealed by changing ranked order of cultivars after exposure to aggressive isolates of P. infestans . The results show that cultivar by isolate interactions existed for all components of tuber blight resistance studied. The quantitative nature of the observed resistance responses suggests the presence of quantitative trait loci governing resistance to tuber blight. The consequences of differential interactions in relation to the stability of tuber resistance are discussed.     

马铃薯晚疫病CARAH预警模型在我国的应用及评价

CARAH预警模型引入我国后,在马铃薯晚疫病监测预警中发挥了重要作用。本文对目前各地CARAH模型的应用情况进行了综合分析,总结了模型在马铃薯晚疫病中心病株出现时间预测、发生程度风险评估、防治效果等方面的作用,分析了品种抗性、区域气候特征等因素对模型应用的影响,提出了进一步研究和完善建议。     

影响五倍子水悬浮液对马铃薯晚疫病田间防治效果的主要气象因素

 将4.0%五倍子水悬浮液喷施田间马铃薯品种Bintje的叶片,于不同处理时间后,剪下叶片带回室内人工接种致病疫霉孢子囊悬浮液,保湿培养后调查各处理的发病情况。结果表明,喷施五倍子水悬浮液后48h内未经历降雨的处理,病情指数仅为7.41,与清水对照相比,防效为92.59%;喷施后96h接种的处理,由于在田间经历了降雨,完全丧失了防效,病情指数达100。室内毒力测定结果显示,0.4%五倍子水悬浮液喷施96h后,对致病疫霉游动孢子释放的抑制率仍在80%以上,在室外避雨不避光条件下五倍子水悬浮液96h内抑菌效果无显著降低,表明日光的影响作用较小。在室内盆栽条件下,通过喷淋水滴模拟田间降雨对药液的冲刷作用。结果表明,冲刷作用可导致五倍子水悬浮液对马铃薯晚疫病的防效显著降低。进一步说明,降雨是导致五倍子水悬浮液对马铃薯晚疫病田间防治效果降低的主要气象因素。     

Evaluation and analysis of reduction of late blight disease in a diploid potato progeny

The rate of late blight disease was analysed for individuals of a diploid Solanum phureja – Solanum tuberosum dihaploid hybrid population (PD), using three different assessment techniques, in the laboratory, screenhouse, and field. These hybrids expressed low disease rates in the field, comparable to resistance based on intact R genes. However, none of the parents of PD expressed any R genes and the pattern of segregation within the PD population was not indicative of R-gene inheritance. The foliage (or leaflet) area diseased had the largest broad-sense heritability of all criteria analysed, in all tests performed. In the field evaluations, the PD population showed intermediate levels of broad-sense heritability for foliage area diseased, relative to the much larger heritability detected for the group of controls possessing R genes. Resistance in the field of the PD hybrids had very little genotype-environment (G × E) interaction, indicating stability of its expression. All genotypes without R genes exhibited heritable, reduced rates of late blight disease in the field, but they were susceptible with low heritability in screenhouse and laboratory tests. This differential expression of disease indicates that the plants' indirect response to unknown environmental stress in the field may have been measured. The value of the PD hybrids for breeding of late blight resistant potato and the use of the disease rate data for detection of the underlying quantitative trait loci are discussed.     

Development of natural late blight epidemics in pure and mixed plots of potato cultivars with different levels of partial resistance

Late blight, caused by Phytophthora infestans , is the most severe disease of potato worldwide. Controlling late blight epidemics is difficult, and resistance of host cultivars is either not effective enough, or too easily overcome by the pathogen to be used alone. In field trials conducted for 3 years under natural epidemics, late blight severity was significantly lower in a susceptible cultivar growing in rows alternating with partially resistant cultivars (mixtures) than in unmixed plots of the susceptible cultivar alone. Partially resistant cultivars behaved similarly in unmixed and mixed plots. Mixtures of cultivars reduced disease progress rates and sometimes delayed disease onset over unmixed plots, but did so significantly only for the slowest epidemic. This suggests that reduction of area under the disease progress curve (AUDPC) in mixtures resulted from the cumulative action of minor effects. Disease distribution was focal in all plots at all dates, as shown by Morisita's index values significantly exceeding 1. Significant yield increases for the susceptible cultivar, and occasionally for the partially resistant ones, were observed in mixed-cultivar plots compared with single-cultivar plots. These results show that cultivar mixtures can significantly reduce natural, polycyclic epidemics in broadleaved plants attacked by pathogens causing rapidly expanding lesions.     

Exploiting host resistance to reduce the use of fungicides to control potato late blight

Field trials in 1996, 1997 and 1998 with six potato cultivars differing in levels of foliar and tuber race-nonspecific resistance to late blight were treated with 100, 50 and 33% of the recommended dose of the fungicide fluazinam at application intervals of 7, 14 and 21 days. Using a mixed inoculum of six or seven indigenous isolates of Phytophthora infestans small potato plots were inoculated via infector plants. A foliar blight model for the relationship between the effects of resistance, fungicide application and disease pressure was developed using multiple regression analysis. Cultivars with a high level of quantitative resistance offered the greatest potential for fungicide reduction. The model showed that the effect of resistance on integrated control increased exponentially with increasing cultivar resistance. Reducing fungicide input by lowering the dose resulted in less foliar disease than extending application intervals. The higher the disease pressure, the greater the risk associated with reducing fungicide input by extension of application intervals. The field resistance of cultivars to tuber blight mainly determined the frequency of tuber infection. Exploiting high foliar resistance to reduce fungicide input carried a high risk when cultivar resistance to tuber blight was low. When field resistance to tuber blight was high, a medium level of resistance in the foliage could be exploited to reduce fungicide dose to c . 50%, provided application was at the right time. At a high level of field resistance to both foliar and tuber blight, application intervals could be extended.     

磷酸二氢钾影响马铃薯对晚疫病抗性的初步研究

本研究以马铃薯品种‘丽薯6号’为试材,对叶面喷施不同浓度磷酸二氢钾4 d后的植株接种晚疫病菌,测定晚疫病发生的严重度,并测定磷酸二氢钾和晚疫病菌处理后14 d内马铃薯植株的PAL、SOD、POD、PPO、Chi、GLU活性及SP含量。结果表明,在一定浓度范围内,磷酸二氢钾可减轻马铃薯晚疫病的发生,随着施用浓度的增加,晚疫病发病逐渐减轻。其中磷酸二氢钾质量浓度为0.6%时,晚疫病发病最轻,接种晚疫病菌后8 d防效达35.64%,12 d时仍超过30%,但浓度超过0.6%,晚疫病发生加重。同时,喷施磷酸二氢钾可不同程度地提升健康和接种晚疫病菌的马铃薯植株体内6种防御酶活性及可溶性蛋白含量。因此,磷酸二氢钾在一定时间段内可诱导马铃薯对晚疫病的抗性,减轻晚疫病的发生。     

2013~2015年黑龙江省马铃薯晚疫病菌群体结构研究

为明确马铃薯北方一季作区致病疫霉对烯酰吗啉和双炔酰菌胺的敏感性时空动态及常用药剂对马铃薯晚疫病的田间防效,采用菌丝生长速率法测定了2011—2019年采自河北、内蒙古和吉林3省 (区) 马铃薯主产区的922个致病疫霉菌株对烯酰吗啉和双炔酰菌胺的敏感性,采用茎叶喷雾法于2018年和2019年评估了烯酰吗啉和双炔酰菌胺等7种杀菌剂在推荐剂量下对马铃薯晚疫病的田间防效。结果表明：在时间上,不同年份采集的922株致病疫霉对烯酰吗啉的抗性频率和抗性指数分别为25.71%~100%和0.31~0.50;890株致病疫霉对双炔酰菌胺的抗性频率为0~1.43%,仅于2013年检测到抗性菌株,历年抗性指数均为0.25。在地域上,3省 (区) 致病疫霉菌株对烯酰吗啉的抗性频率和抗性指数分别为57.19%~66.56%和0.39~0.42;对双炔酰菌胺的抗性频率为0~0.36%,抗性指数均为0.25。田间药效试验结果显示：50%烯酰吗啉可湿性粉剂 (有效成分,下同) 300.0 g/hm²对马铃薯晚疫病的防治效果 (83.3%~85.6%) 高于23.4%双炔酰菌胺悬浮剂140.4 g/hm²的防治效果 (76.9%~78.6%) 和50%氟醚菌酰胺水分散粒剂75.0 g/hm²的防治效果 (70.0%~71.4%)。20%霜脲氰悬浮剂162.0 g/hm²、40%氟醚菌酰胺 • 烯酰吗啉悬浮剂240.0 g/hm²、43%霜脲氰 • 双炔酰菌胺水分散粒剂290.3 g/hm²和687.5 g/L氟吡菌胺 • 霜霉威盐酸盐悬浮剂773.4 g/hm²对马铃薯晚疫病的防治效果为79.8%~86.9%。为此,推荐烯酰吗啉单剂或混剂及双炔酰菌胺混剂在河北、内蒙古和吉林3省 (区) 继续用于马铃薯晚疫病的防治,但是需密切监测致病疫霉对烯酰吗啉和双炔酰菌胺的敏感性变化并注意与不同作用机理的杀菌剂混用或者交替使用。