Temporal disease dynamics and relative importance of sexual and asexual reproduction of grape downy mildew (Plasmopara viticola) in an isolated vineyard in the North Georgia Mountains,USA

The North Georgia Mountains are the southernmost region along the United States East Coast where European wine grapes (Vitis vinifera) are grown commercially. Epidemics of downy mildew, caused by Plasmopara viticola, are frequent and severe, but little is known about the epidemiology and population biology of the pathogen in this region. Disease monitoring in an experimental vineyard from 2015 to 2017 indicated that times of disease onset and progress rates were highly variable across years and cultivars. Oospores were observed microscopically, and simulation with a process-based model indicated presence of conditions favourable for oospore germination in the spring and early summer each year. A total of 409 P. viticola isolates collected over three  years were genotyped with seven microsatellite markers, revealing very high genotypic diversity, which when combined with the observation of oospores is indicative of a sexually reproducing population. Among the 409 isolates, 225 multilocus genotypes (MLGs) were identified, of which 164 were detected only once and 61 were repeated (clonal). Eight MLGs (represented by 28 isolates) were detected across years, suggesting the possibility of asexual overwintering of P. viticola in this region. Across sampling dates, the percentage of isolates belonging to nonrepeated (unique) MLGs ranged from 27.3% to 63.2%. Even towards the end of the annual epidemic, the percentage of isolates in nonrepeated MLGs was still relatively high, around 30%. These MLGs may have originated from oospores germinating late during the growing season, although incomplete sampling at earlier dates and contribution by immigration cannot be fully excluded.     

vvi-miR160s介导VvARF18应答赤霉素调控葡萄种子的发育

【目的】研究vvi-miR160家族（vvi-miR160s）及其靶基因在‘魏可’葡萄种子发育过程中的作用,探究其应答赤霉素（GA）调控葡萄果实无核的潜在机理。【方法】采用miR-RACE、生物信息学分析、RT-qPCR、RLM-RACE等方法,鉴定vvi-miR160家族成员及其靶基因,分析vvi-miR160s及其靶基因应答GA的时空表达模式及其潜在功能。【结果】花前GA处理强烈抑制‘魏可’葡萄胚珠及种子发育,高效诱导其无核,且无核率达99.8%。克隆鉴定了VvMIR160s前体基因序列（501 bp）及成熟体序列,且它们在不同物种间具有较高的保守性;基于vvi-miR160s序列,预测到靶基因VvARF18,利用RLM-RACE技术检测到vvi-miR160s对VvARF18的裂解位点在第10和第11位之间,裂解频度9/17,证明VvARF18是vvi-miR160s的真实靶基因。该靶基因编码683个氨基酸,在398—411位存在核定位信号,且该蛋白亚细胞定位于细胞核上。VvARF18与其他物种间序列的同源保守性较高,基因结构相似,其中VvARF18蛋白与茶、烟草、梅花等物种亲缘关系较近。VvARF18启动子中包含4种作用元件,且含有较多激素相关元件。RT-qPCR结果显示,随着葡萄果实的发育,vvi-miR160c/d/e呈现‘V’形表达趋势,在硬核种子发育期表达较低,而VvARF18表现出与前者相反的表达趋势,在硬核种子发育期呈现高表达,表明vvi-miR160c/d/e对VvARF18负调控,但vvi-miR160a/b与VvARF18表达水平却不存在明显负相关。GA处理极显著地上调了vvi-miR160a/b在葡萄硬核种子发育期的表达,同时也显著抑制了VvARF18在同时期的表达,且它们的表达水平呈负相关,表明GA处理促进了vvi-miR160a/b对VvARF18的负调控作用;相反,GA减弱了vvi-miR160c/d/e对VvARF18的负调控作用。【结论】在vvi-miR160家族中,vvi-miR160c/d/e可能介导VvARF18在葡萄种子发育的特定阶段调控种子的发育形成,而vvi-miR160a/b可能主要介导VvARF18参与调控GA诱导葡萄种子败育的过程。     

葡萄籽原花青素的提取工艺优化及其抗氧化活性研究

以新疆地产赤霞珠葡萄籽为原料，采用响应面法对微波辅助提取葡萄籽原花青素的工艺进行优化，并对其体外抗氧化活性进行评价。结果表明，最优提取工艺条件为：液料比25∶1（mL/g），乙醇浓度60%，微波功率427 W，微波时间3 min，在该条件下葡萄籽原花青素得率为8.66%±0.25%；抗氧化试验结果表明：葡萄籽原花青素具有较强的清除1,1-二苯基-2-三硝基苯肼（DPPH）、羟基及2,2-联氮-双(3-乙基-苯并噻唑-6-磺酸)二铵盐（ABTS）自由基的能力，其在三种体系中的IC50值分别为3.71、4.05和3.78 μg/mL，是一种极具开发潜力的天然抗氧化剂。     

11个葡萄品种在乐山地区的花芽分化规律

研究了11个葡萄品种3年生树在四川乐山地区避雨栽培条件下的花芽分化表现。结果表明,结果母枝的花芽分化率,魏可、维多利亚、黑芭拉多、金手指、金优2号和巨玫瑰6个品种的都高于50%,较易形成花芽;户太8号、巨峰、夏黑的在43%~48%;美人指和红指较难形成花芽。结果母枝不同节位的花芽分化率,多数品种在第3节位≥50%;结果枝上花穗着生节位在第3.8~4.9节,其中双花率以巨玫瑰、巨峰和户太8号较高,分别为83.45%、73.61%和73.06%。以结果母枝的花芽分化率≥50%节位作为冬剪的最低节位,则金优2号、户太8号、巨玫瑰、金手指、维多利亚5个品种在第3~4节,黑芭拉多和巨峰2个品种在第2~3节,魏可在1~2节。夏黑表现花芽分化率较低,为43.33%,可能与夏黑不耐弱光有关,设施栽培时应加强透光性。     

酿酒山葡萄新品种‘紫晶甘露’

‘紫晶甘露’是以优质酿酒山葡萄酒品种‘左山二’为母本，‘哈桑’为父本杂交选育而成的能够单品种酿造干红酒的山葡萄新品种。果穗圆锥形，有岐肩或副穗。果实圆球形，果皮黑色，果肉绿色，无肉囊，种子2 ~ 4粒。生长势强，抗寒、抗病性极强，4年丰产，4 ~ 6年平均产量1 876 kg • km-2。果实可溶性固形物含量达到21.0%，总酸11.6 g • L-1，出汁率65.5%。     

利用天敌防治葡萄园害虫研究进展

葡萄是一种重要的经济水果,除鲜食外还可用于酿酒、制干、制汁等。在葡萄挂果到收获期间采用化学药剂来防治园内害虫极有可能造成农药残留,危害人体健康。利用天敌防治葡萄园内害虫是确保葡萄及其制品质量安全的重要途径。总结了国内外葡萄园常见害虫的主要捕食性和寄生性天敌,以及利用天敌防治葡萄园重要虫害的策略,并探讨了该领域目前存在的问题及未来的研究方向。     

Comparative analysis of genetic effects of wheat‐Dasypyrum villosum translocations T6V#2S·6AL and T6V#4S·6DL

Wheat‐Dasypyrum villosum translocations T6V#2S·6AL and T6V#4S·6DL, carriers of Pm21 and PmV, respectively, confer high resistance to wheat powdery mildew. For better understanding of the difference in genetic effect between them, a RIL population was constructed based on the cross between “Yangmai 18” carrying T6V#2S·6AL and “Yangmai 22” carrying T6V#4S·6DL. Analysis of distribution of the translocations showed that T6V#2S·6AL is much more transmittable than T6V#4S·6DL. By comparing their effects on main agronomic traits, we firstly found that T6V#2S·6AL contributes greatly to top spikelet fecundity, but causes a decrease of 6.7%–10.5% of spike number. No stable effects of T6V#4S·6DL on agronomic traits were found, except for positive effect on plant height. Excitingly, a new recombinant, T6V#4S‐6V#2S·6AL carrying PmV, was screened and proved to have a higher transmission rate than the original translocation T6V#4S·6DL, which will greatly promote the utilization of PmV. The above conclusions of this research will provide important guidance for utilization of Pm21 and PmV more effectively, in wheat powdery mildew resistance breeding.     

Cryptic species and population genetic structure of Plasmopara viticola in São Paulo State,Brazil

Downy mildew (Plasmopara viticola) is one of the most important diseases in grape-growing areas worldwide, including Brazil. To examine pathogen population biology and structure, P. viticola was sampled during the 2015/16 growing season from 516 lesions on nine grape cultivars in 11 locations in subtropical areas of São Paulo State, Brazil. For identification of cryptic species, a subsample of 130 isolates was subjected to cleaved amplified polymorphic sequence (CAPS) analysis, and for 91 of these isolates the ITS1 region was sequenced. These analyses suggest that the population of P. viticola in São Paulo State consists of a single cryptic species, P. viticola clade aestivalis. Seven microsatellite markers were used to determine the genetic structure of all 516 P. viticola isolates, identifying 23 alleles and 55 multilocus genotypes (MLGs). Among these MLGs, 34.5% were clonal and represented 93% of the isolates sampled. Four dominant genotypes were present in at least five different locations, corresponding to 65.7% of the isolates sampled. Genotypic diversity (Ĝ = 0.21–0.89) and clonal fraction (0.58–0.96) varied among locations (populations). Most populations showed significant deviation from Hardy–Weinberg expectations; in addition, excess of heterozygosity was verified for many loci. However, principal coordinate analysis revealed no clusters among locations and no significant isolation by distance was found, suggesting high levels of migration. The results indicate that downy mildew epidemics result from multiple clonal infections caused by a few genotypes of P. viticola, and reproduction of P. viticola in São Paulo State is predominantly asexual.     

凉州灌区酿酒葡萄氮肥施用研究

[目的]通过田间试验,研究酿酒葡萄对不同氮肥施用量及施用深度的响应,为甘肃省凉州灌区酿酒葡萄氮肥合理施用提供依据。[方法]在凉州区设置酿酒葡萄氮肥施用量及施用深度试验,研究不同氮肥施用量及施用深度对酿酒葡萄产量、收获期果实、叶片和叶柄含氮量及收获期和第2a萌芽期0—200cm土层硝态氮含量的影响。[结果]10和30cm施肥深度之间酿酒葡萄产量、收获期果实、叶片和叶柄含氮量及收获期和第2a萌芽期0—200cm土层硝态氮含量差异不显著。氮肥施用量对酿酒葡萄产量和叶柄含氮量的影响达到显著水平,对果实和叶片含氮量的影响不明显,其中高氮(300kg/hm~2)和中氮(240kg/hm~2)处理之间酿酒葡萄产量、果实含氮量、叶片含氮量、叶柄含氮量差异不明显,但是高氮和中氮处理与低氮(180kg/hm~2)处理相比,产量增加28.6%和24.1%,叶片含氮量增加17.4%和11.3%,叶柄含氮量增加了40.7%和33.0%,而对于收获期和第2a萌芽期0—200cm土层硝态氮含量,高氮处理相对于中氮和低氮处理增加了53.8%,94.4%和41.8%,76.1%,氮肥施用量和施用深度的交互效应,仅叶片含氮量达到显著水平。[结论]受土壤质地和传统沟灌影响,氮肥施用深度对酿酒葡萄影响效果不明显,240kg/hm~2为酿酒葡萄较为合适的氮肥施用量,但氮素也存在损失风险。所以,凉州灌区酿酒葡萄合理施肥应该和灌水方式进行结合来确定合理的施肥量和施肥方式。     

栽培中赤霉酸处理对葡萄品质和耐藏力的影响

通过对比试验研究了栽培中不同赤霉酸(GA3)处理次数对葡萄品质和耐藏力的影响。结果表明:随着赤霉酸处理次数的增多,葡萄的果梗直径、果形指数、单果粒质量、脱粒率、脱粒指数、空腔率、空腔直径、褐变率和腐烂率升高,果刷长度、可溶性固形物含量和VC含量降低,且外观色泽变淡。综合考虑栽培中赤霉酸处理对葡萄品质和耐藏力的影响,建议用10 mg/kg赤霉酸花前10 d处理1次或花前10 d和花后10 d各处理1次来提高葡萄的单果粒质量,保持果实较低的褐变率和腐烂率。