不同葡萄品种对霜霉病的抗性

葡萄霜霉病是葡萄生产上重要的病害之一,通过对辽宁省不同葡萄品种进行室内离体叶片接种和田间自然发病情况调查,以期为葡萄抗性品种的选育和葡萄霜霉病的防治提供科学依据。结果表明,在供试品种中,没有对霜霉病完全免疫的品种,室内离体叶片接种和田间调查结果基本一致,不同品种间霜霉病的抗性存在差异。供试的65个品种中,室内离体叶片接种评价高抗品种3个,抗病品种23个,感病品种24个,高感品种15个;田间自然发病调查评价高抗品种1个,抗病品种19个,感病品种35个,高感品种10个。欧美杂交品种(系)相对欧亚杂交品种(系)较抗病。     

鲜食葡萄品种对霜霉病的抗性及抗病机理研究

采用田间自然发病调查，田间接种鉴定和离体葡萄叶圆片室内接种鉴定鲜食葡萄品种对霜霉病的抗性及其抗性机理研究表明，品种间对霜霉病的抗性存在显著差异。可依感病病级分为三类，低感品种有“z工伊豆”、“奥林匹亚”、“山东早红”、“森田尼”、“早玛瑙”。几乎所有的无核品种均高感霜霉病，葡萄对霜霉病的抗性与叶内可滴定酸、还原糖、游离氨基酸含量无显著相关，与叶背气也密度、气孔开度也无相关，而外施高浓度邻苯二酚却显著降低了霜霉病发病程度。     

不同葡萄品种对霜霉病的田间苗期抗性评价

对36个葡萄品种(品系)苗期对霜霉病的田间抗病性进行了评价, 结果表明, 不同葡萄品种(品系) 对葡萄霜霉病的抗性存在明显差异, 绝大多数欧美杂交种的抗性要强于欧亚种葡萄, 供试的10个欧美杂种中, ‘瑞峰无核’对葡萄霜霉病表现为高抗; ‘DEMIR’、‘龙宝’、‘红富士’、‘黑奥林’、‘峰后’5个品种表现为中抗。     

葡萄品种对霜霉病抗性鉴定的生化指标研究

 通过盆栽接种和田间自然发病鉴定,从22个供试葡萄品种中鉴定筛选到1个高抗霜霉病的品种超藤和8个中抗品种饭刚黑、信浓乐、弗雷无核、黑蜜、优选皮奥萘、高妻、京秀、峰后。相关性分析表明,22个供试葡萄品种接种前和接种后2d,PPO、PAL活性和CAT比活性与霜霉病病情指数呈极显著负相关,SOD活性与霜霉病病情指数相关不显著。PPO、PAL活性和CAT比活性与葡萄品种对霜霉病的抗性呈极显著正相关,可利用这3种酶的活性作为葡萄品种霜霉病抗性鉴定的辅助评价指标。     

葡萄霜霉病抗病性鉴定方法及品种抗病性测定

本文通过对葡萄霜霉病抗病性鉴定方法进行优化,建立了一种更为快捷、方便、可靠的鉴定方法,并对32个葡萄品种对霜霉病抗病性进行了鉴定,为葡萄抗病品种的选育和应用提供依据。结果显示:以田间混合的霜霉病菌为接种体采用叶盘法鉴定葡萄品种的抗病性更加快捷、方便。供试的32个葡萄品种对葡萄霜霉病的抗性存在显著差异。其中免疫品种有‘康拜尔早生';高抗的有‘阳光玫瑰'、‘美乐'、‘Ms27-31'等5个品种;中抗的有‘贝达'、‘小芒森'、‘2E-16-2'等6个品种;低抗的有‘瑞都红玉'、‘早黑宝'、‘摩尔多瓦'等10个品种;感病的有‘里扎马特'、‘玫瑰香'、‘香妃'等10个品种。     

葡萄对霜霉病抗性机理初探

经田间自然鉴定、接种鉴定及室内离体接种鉴定结果表明：葡萄对霜霉病抗性与品种叶内的含水量、可滴定酸、还原糖、淀粉、水解氨基酸、游离氨基酸含量无显著相关，与叶背气孔密度、气孔开闭程度也无相关，而外施高浓度蔗糖及邻苯二酚却显著降低霜霉菌的致病力。     

葡萄对霜霉病抗性机理初探

经田间自然鉴定、接种鉴定及室内离体接种鉴定结果表明：葡萄对霜霉病抗性与品种叶内的含水量、可滴定酸、还原糖、淀粉、水解氨基酸、游离氨基酸含量无显著相关，与叶背气孔密度、气孔开闭程度也无相关，而外施高浓度蔗糖及邻苯二酚却显著降低霜霉菌的致病力。     

宁夏葡萄霜霉病菌致病型鉴定及葡萄品种抗性评价

为合理利用葡萄抗性品种以及田间病害综合治理,采用离体叶盘接种法分别对2018年和2019年自宁夏回族自治区银川市、石嘴山市和吴忠市采集的43株葡萄霜霉病菌Plasmopara viticola菌株进行致病型鉴定及聚类分析,并对17个主栽葡萄品种进行抗性评价及聚类分析。结果表明：2018、2019年供试菌株致病力和供试葡萄品种抗性之间均存在显著差异,菌株平均病情指数与发病率呈显著正相关,皮尔逊相关系数r和回归系数R²分别介于0.45~0.96和0.20~0.92之间;2018、2019年供试菌株的强致病力菌株和弱致病力菌株分别为CS-YL和VI-BYDD、RG-JSJG和CS-YM;2018年和2019年供试菌株致病型被划分为强、中、弱3种类群,2年所占比例分别为10.00%、15.00%、75.00%和34.78%、39.13%、26.09%;供试17个葡萄品种的抗性类型可被划分为感病、抗病、高抗3种类群,2年所占比例均为17.65%、29.41%和52.94%。不同葡萄品种对不同菌株的抗性类型多样化。表明宁夏回族自治区葡萄霜霉病菌菌株致病力分化明显,不同菌株的致病型与其地域来源无关,不同葡萄品种的抗感程度不同。     

辽宁花生主栽品种(系)对褐斑病和网斑病抗性鉴定

本研究旨在明确辽宁花生主栽品种(系)对花生褐斑病和网斑病的抗性,以期为抗病育种及田间病害防治提供理论依据。试验采用田间自然病圃法,对辽宁主栽的16个花生品种(系)进行了褐斑病和网斑病田间抗性鉴定。结果表明,供试花生品种(系)对两种叶部病害抗性存在显著差异,但缺乏免疫和高抗品种。对花生褐斑病抗性鉴定结果表明,‘阜花17’属抗病类型,‘鲁花11’、‘铁引花2号’、‘良青8号’、‘新花2号’和‘锦花15’属中抗类型,其余属高感或感病类型。对花生网斑病抗性鉴定结果显示:‘鲁花11’、‘铁引花2号’和‘白花生’属抗病类型,‘黑花生’、‘新花2号’、‘花育20’、‘锦花14’、‘新花1号’、‘锦花15’和‘良青8号’属中抗类型,其余属高感或感病类型。多数花生品种(系)对花生叶斑病综合抗性较差。     

部分鲜食葡萄品种霜霉病抗性田间调查

以54个鲜食葡萄品种为试材,调查统计不同葡萄品种的霜霉病发病程度、发病速率、落叶率和新梢成熟度。结果表明,不同葡萄品种对霜霉病的抗性存在明显差异,欧美杂种品种群的葡萄对霜霉病的抗性显著高于欧亚品种群;病情指数与落叶率、新梢成熟度呈极显著的相关性,与发病速率之间关系不显著。     

Studies on the resistance of grapevine to powdery mildew

Studies of 68 cultivars of Vitis vinifera and two interspecific hybrids inoculated with four naturally occurring isolates of Uncinula necator demonstrated variation for pathogenicity between isolates and variation in resistance between cultivars. There was no specific interaction between the cultivars and the isolates. The progenies produced either by selling or hybridization of cultivars of V. vinifera always contained some resistant individuals regardless of the parental resistance ratings. However, the proportion of resistant plants was positively correlated to the level of resistance of the parent(s). The results indicate that 'minor' resistance genes exist in the cultivars of V. vinifera , and that resistance to U. necator could be improved by recurrent selection.     

Osmotin and thaumatin from grape: a putative general defense mechanism against pathogenic fungi

ABSTRACT Little information is available concerning the expression of pathogenesis-related (PR) proteins in grapevine (Vitis vinifera) and their effect properties on the major fungal pathogens of grape. A systematic study was performed on the effect of total or individual grape proteins on mycelial growth, spore germination, and germ tube growth of Uncinula necator, Phomopsis viticola, and Botrytis cinerea. Two proteins, identified as PR proteins by immunological methods and by N-terminal sequencing as osmotin and thaumatin-like protein, exhibited strong antifungal activities in vitro, blocking the growth of Phomopsis viticola and Botrytis cinerea mycelia. In addition, they inhibited spore germination and germ tube growth of U. necator, Phomopsis viticola, and Botrytis cinerea. The presence of both proteins displayed a synergistic effect. The expression of osmotin and thaumatin-like protein was induced in grapevine leaves and berries infected with U. necator and Phomopsis viticola. Thaumatin previously was thought to occur exclusively in berries. Immunoblot analyses revealed the accumulation of the two PR proteins in infected leaves and berries, supporting a role in vivo in increasing the resistance of grapevine to fungal attack.     

Identification of race-specific resistance in North American Vitis spp. limiting Erysiphe necator hyphal growth

Race-specific resistance against powdery mildews is well documented in small grains but, in other crops such as grapevine, controlled analysis of host-pathogen interactions on resistant plants is uncommon. In the current study, we attempted to confirm powdery mildew resistance phenotypes through vineyard, greenhouse, and in vitro inoculations for test cross-mapping populations for two resistance sources: (i) a complex hybrid breeding line, 'Bloodworth 81-107-11', of at least Vitis rotundifolia, V. vinifera, V. berlandieri, V. rupestris, V. labrusca, and V. aestivalis background; and (ii) Vitis hybrid 'Tamiami' of V. aestivalis and V. vinifera origin. Statistical analysis of vineyard resistance data suggested the segregation of two and three race-specific resistance genes from the two sources, respectively. However, in each population, some resistant progeny were susceptible in greenhouse or in vitro screens, which suggested the presence of Erysiphe necator isolates virulent on progeny segregating for one or more resistance genes. Controlled inoculation of resistant and susceptible progeny with a diverse set of E. necator isolates clearly demonstrated the presence of fungal races differentially interacting with race-specific resistance genes, providing proof of race specificity in the grape powdery mildew pathosystem. Consistent with known race-specific resistance mechanisms, both resistance sources were characterized by programmed cell death of host epidermal cells under appressoria, which arrested or slowed hyphal growth; this response was also accompanied by collapse of conidia, germ tubes, appressoria, and secondary hyphae. The observation of prevalent isolates virulent on progeny with multiple race-specific resistance genes before resistance gene deployment has implications for grape breeding strategies. We suggest that grape breeders should characterize the mechanisms of resistance and pyramid multiple resistance genes with different mechanisms for improved durability.     

Development of a High-Throughput Method for Quantification of Plasmopara viticola DNA in Grapevine Leaves by Means of Quantitative Real-Time Polymerase Chain Reaction

ABSTRACT Plasmopara viticola is a strictly biotrophic oomycete that causes downy mildew, which is one of the most important grapevine diseases. Control of the disease is most often achieved by fungicide applications, which may have severe environmental consequences. Therefore, alternative control strategies based on biocontrol agents (BCAs) are currently in development. Thousands of potential BCAs have to be screened for their antagonist efficacy against Plasmopara viticola. Evaluation of their effect on the pathogen can be achieved by detecting the amount of P. viticola DNA in leaves treated with potential antagonists and infected with the pathogen. In this study, a rapid high-throughput method was developed for relative quantification of P. viticola DNA directly from Vitis vinifera leaves by means of multiplex real-time quantitative polymerase chain reaction (PCR) with TaqMan chemistry. This method allows simultaneous amplification, but independent detection, of pathogen and host DNA by using species-specific primers and TaqMan probes that are labeled with different fluorescent dyes. Including detection of V. vinifera DNA in the tests is fundamental because it provides an endogenous reference and allows normalization for variations caused by sample-to-sample differences in DNA extraction, PCR efficiencies, and pipetting volumes. The developed method allows highly sensitive and specific detection of P. viticola DNA (minimal detectable quantity of 0.1 pg). Moreover, high precision and reproducibility of TaqMan assays were observed over a linear range of four orders of magnitude, confirming the reliability of the developed PCR assay. Potential applications range from screening for BCA efficiency to evaluation of fungicide efficacy, or assessment of host resistance.     

Ontogenic resistance to powdery mildew in grape berries

ABSTRACT Berries of Vitis vinifera are reported to be susceptible to infection by Uncinula necator until soluble solids levels (brix) reach 8%, and established colonies are reported to sporulate until brix reach 15%. However, our analysis of disease progress on fruit of selected V. vinifera cultivars indicated that severity became asymptotic several weeks earlier in fruit development. When mildew-free fruit clusters of V. vinifera 'Chardonnay', 'Riesling', 'Gewürztraminer', and 'Pinot Noir' were inoculated at stages ranging from prebloom to 6 weeks postbloom, only fruit inoculated within 2 weeks of bloom developed severe powdery mildew. Substantial ontogenic resistance to infection was expressed in fruit nearly 6 weeks before fruit brix reached 8% and over 2 months before they reached 15%. Rachises of 'Chardonnay' and 'Riesling' fruit clusters developed severe powdery mildew when inoculated at bloom, and disease increased steadily over the next 60 days. The rachis of fruit clusters inoculated 31 days after bloom developed only trace levels of powdery mildew. Berry weight of all four cultivars at harvest was reduced when fruit clusters were inoculated at bloom or 16 days postbloom, primarily by splitting, rotting, and dehydration of mildewed berries, but the weight of later-inoculated berries was not reduced. Inoculation of berries just as ontogenic resistance increased markedly, approximately 3 to 4 weeks postbloom, resulted in the development of inconspicuous, diffuse, non-sporulating mildew colonies on berries, sometimes associated with a network of necrotic epidermal cells. Rather than a protracted and relatively static period of berry susceptibility lasting 3 months, fruit of V. vinifera appear to acquire ontogenic resistance rapidly after fruit set. A refocusing of disease management on this critical period of high fruit susceptibility should greatly improve the efficacy of fungicides directed against powdery mildew.     

Seasonal development of ontogenic resistance to downy mildew in grape berries and rachises

ABSTRACT Clusters of Vitis vinifera and V. labrusca are reported to become resistant to Plasmopara viticola at stages of development ranging from 1 to 6 weeks postbloom. It has been suggested that resistance is associated with loss of the infection court as stomata are converted to lenticels, but the time of onset, cultivar variation, and seasonal variation in ontogenic resistance has remained uncertain, as has the comparative susceptibility of stem tissue within the fruit cluster. In New York, we inoculated clusters of V. vinifera cvs. Chardonnay and Riesling and V. labrusca cvs. Concord and Niagara at stages from prebloom until 5 to 6 weeks postbloom. Berries were infected and supported profuse sporulation until 2 weeks postbloom, and pedicel tissue remained susceptible until 4 weeks postbloom. Although berries on later-inoculated clusters failed to support sporulation, discoloration and necrosis of berry tissues was often noted, and necrosis of the pedicel within such clusters often led to further discoloration, shriveling, reduced size, or loss of berries. When the epidermis of discolored berries that initially failed to support sporulation was cut, the pathogen emerged and sporulated through incisions, indicating that lack of sporulation on older symptomatic berries was due to infection at an early stage of berry development followed by conversion of functional stomata to lenticels during latency. We repeated the study on Chardonnay and Riesling vines in South Australia and found that the period of berry and rachis susceptibility was greatly increased. The protracted susceptibility of the host was related to the increased duration and phenological heterogeneity of bloom and berry development in the warmer climate of South Australia. The time of onset and subsequent expression of ontogenic resistance to P. viticola may thus be modified by climate and should be weighed in transposing results from one climatic area to another. Our results can be used to refine forecast models for grapevine downy mildew to account for changes in berry and rachis susceptibility, and to focus fungicide application schedules upon the most critical periods for protection of fruit.     

The Course of Colonization of Two Different Vitis Genotypes by Plasmopara viticola Indicates Compatible and Incompatible Host-Pathogen Interactions

ABSTRACT The course of colonization of leaf mesophyll by the causal agent of grapevine downy mildew, Plasmopara viticola, in a susceptible and a resistant grapevine genotype was examined in order to characterize the development of the pathogen in compatible and incompatible host-pathogen interactions. Within a few hours after inoculation, the pathogen was established in the susceptible Vitis vinifera cv. Müller-Thurgau and formed primary hyphae with a first haustorium. No further development occurred in the following 10 to 18 h. The next step, in which the hyphae grew and branched to colonize the intercellular space of the host tissue, was observed 1.5 days after inoculation. After 3 days, the intercostal fields were entirely filled with mycelium and sporulation was abundant under favorable environmental conditions. The first infection steps were essentially the same in the resistant V. rupestris. However, the invasive growth of P. viticola was delayed, and further development ceased before the intercostal fields were filled with mycelium.     

Nonhost versus host resistance to the grapevine downy mildew, Plasmopara viticola, studied at the tissue level

Following inoculation of host and nonhost plants with Plasmopara viticola, the grapevine downy mildew, a histological survey was undertaken to identify the stage where its development is contained in nonhosts and in resistant host plants. Three herbaceous nonhost species, Beta vulgaris, Lactuca sativa, and Capsicum annuum, and three grapevine species displaying different level of resistance (Vitis vinifera [susceptible], Vitis riparia [partially resistant] and Muscadinia rotundifolia [totally resistant]) where inoculated by P. viticola using a controlled leaf disk inoculation bioassay. During the early steps of infection, defined as encystment of zoospores on stomata, penetration of the germ tube, and production of the vesicle with the primary hypha, there was no evidence of a clear-cut preference to grapevine tissues that could attest to host specificity. The main difference between host grapevine species and nonhosts was observed during the haustorium formation stage. In nonhost tissues, the infection was stopped by cell wall-associated defense responses before any mature haustorium could appear. Defense responses in resistant grapevines were triggered when haustoria were fully visible and corresponded to hypersensitive responses. These observations illustrate that, for P. viticola, haustorium formation is not only a key stage for the establishment of biotrophy but also for the host specificity and the recognition by grapevine resistance factors.     

大豆花叶病毒(SMV)病显症率的模型预测

 大豆感染SMV系统量症前介体不能传毒。接种稀释10倍SMV病液显症率与介体传毒相似。介体和人工接种SMV于5个感病品种V₁-R₅ 9个生长时期共30余批次,结果表明SMV显症率主要决定于温度。显症起始温度为9℃,最适温度约26℃.V₁-R₂时期的植株显症所需有效积温基本一致;R₃-R₅时期比前者略有增加。累积显症率与累积有效积温的相关点图呈"S"型曲线分布,通过Weibull和Gompertz等8组曲线拟合选出拟合最优模型。V₁-R₂时期显症预测Gompertz拟合最好,得预测式:PP₁₁=Exp[-103021.196×Exp(-0.1329TT₁)]R₃-R₅时期Weibull拟合最好,得预测式:PP₁₂=1-Exp{-[0.02222(TT₁-65)^2.581]}