Fungitoxicity and synergism of mixtures of fluopicolide and pyraclostrobin against Phytophthora infestans

Fluopicolide and pyraclostrobin were new systemic fungicides with highly inhibitory activity on a broad spectrum of oomycetes, but so far the fungitoxicity of their mixtures on the different developmental stages of Phytophthora infestans and whether synergism exists have not been investigated. The joint-toxicity of the mixtures of fluopicolide with pyraclostrobin was determined against the different developmental stages (mycelial growth, zoospore release, cystospore germination and sporangial germination) of P. infestans and late blight on the leaf discs and the potted plants of potato, and whether the synergistic interaction exists in the mixtures of the two fungicides were evaluated in vitro and in vivo based on the synergistic ratios calculated with the Wadley formulas. The protective activity and curative activity against P. infestans and the efficacy duration in controlling potato late blight of the synergistic mixtures of the two fungicides were examined in this study. The results showed that mixtures of fluopicolide (F) and pyraclostrobin (P) at the ratios of 10:1 and 1:4 (F:P) exhibited synergistic interactions and had excellent inhibitory activity against almost all developmental stages of P. infestans. Synergistic interactions of the mixtures at the ratios of 1:4 and 10:1 (F:P) against the mycelial growth of metalaxyl-resistant isolates were more obvious than against the mycelial growth of metalaxyl-sensitive isolates. Synergistic interactions of the mixtures at the ratio of 1:4 (F:P) against the mycelial growth of the most of the tested isolates were more obvious than those of the mixtures at the ratio of 10:1 (F:P). The highest activity of the mixtures was observed against zoospore release at the ratio of 1:4 (F:P) with EC₅₀ value of 0.0044 μg ml⁻¹. The strongest synergistic interaction of the mixtures was observed against zoospore release at the ratio of 10:1 (F:P) with the synergistic ratio of 5.27. The mixtures of fluopicolide and pyraclostrobin at the ratios of 1:4 and 10:1 (F:P) showed synergistic interactions against P. infestans on the leaf discs and intact plants of potato. The EC₅₀ values for protective activity and curative activity of the mixture at the ratio of 10:1 (F:P) were 0.63 μg ml⁻¹ and 0.87 μg ml⁻¹, and EC₅₀ values of protective activity and curative activity of the mixture at the ratio of 1:4 (F:P) were 0.27 μg ml⁻¹ and 0.44 μg ml⁻¹, respectively. At 14 days after spraying of fungicide solutions, the disease control efficacy of the mixtures at the ratios of 10:1 and 1:4 (F:P) was still over 80% at the dosages of 50 μg ml⁻¹ and 100 μg ml⁻¹, and the disease control efficacy of the two mixtures remained at about 97% at the dosages of 200 μg ml⁻¹, indicating long-lasting effective control efficacy of the two fungicides against potato late blight.     

Rapid detection of potato late blight using a loop-mediated isothermal amplification assay

Potato late blight caused by Phytophthora infestans is one of the most destructive plant diseases that threaten global food security. Early and effective diagnosis of P. infestans is required before disease management decisions are made. Here, we developed a quick protocol to detect P. infestans based on a loop-mediated isothermal amplification(LAMP) assay. The P. infestans specific multiple copy DNA sequences(PiSMC), a transposon-like element, provides an ideal target for molecular detection of this pathogen. We designed highly specific and sensitive primers allowing effective LAMP detection of the pathogen at 64℃ in 70 min. In the validation assay, all 15 P. infestans isolates collected from China, Europe and South America could be positively detected, but results of the other 9 Phytophthora species infecting different plants, fungal and bacterial plant pathogens tested were negative. The detection limit of this assay is 1 pg P. infestans DNA. Moreover, the LAMP-PiSMC assay is able to detect P. infestans from infected leaves, tubers and soil. Taken together, this study reports the development of a specific and sensitive LAMP-PiSMC assay for early diagnosis of potato late blight.     

基于病菌孢子捕捉和real-time PCR技术的田间空气中小麦白粉病菌孢子动态监测及病情估计模型研究

利用Burkard定容式孢子捕捉器结合real-time PCR定量技术,分别对种植高抗、中感和高感白粉病小麦品种的田间空气中白粉病菌分生孢子浓度进行监测,结果表明,real-time PCR定量与传统的显微观察计数两种方法测得的孢子浓度呈显著正相关(P≤0.01),且两种病菌孢子计数方法在同一抗性品种上监测到的孢子浓度动态相近。此外,两种方法测得的孢子浓度与各气象因子的相关性分析结果一致,空气中的白粉病菌孢子浓度主要与空气相对湿度显著正相关。在此基础上,利用两种方法测定的田间空气中白粉病菌孢子浓度分别建立了基于累积孢子浓度的田间病情估计模型。分析发现,基于两种孢子浓度测定方法建立的病情估计模型间无显著性差异,表明real-time PCR定量技术测定的孢子浓度在构建白粉病病情估计模型上具有一定可行性。该结果为real-time PCR定量技术与病菌孢子捕捉技术相结合用于小麦白粉病的监测和预测提供理论依据。     

麦二叉蚜唾液蛋白基因Sg1655时空表达谱及其抑制小麦防御反应的功能

为进一步明确麦二叉蚜Schizaphis graminum唾液蛋白Sg1655在调控小麦防御反应中的作用,基于麦二叉蚜唾液腺转录组数据,利用PCR技术克隆获取Sg1655开放阅读框,利用生物信息学软件对其序列进行分析;通过实时荧光定量PCR（real-time quantitative PCR,RT-qPCR）技术检测Sg1655在麦二叉蚜不同组织及不同龄期中的表达;利用细菌III型分泌系统将Sg1655导入到小麦叶片内,检测其瞬时表达对小麦胼胝质积累的影响。结果表明,Sg1655开放阅读框全长375 bp,编码125个氨基酸残基,N端1~21位为信号肽序列,预测蛋白分子量为14.90 kD。麦二叉蚜Sg1655氨基酸序列与玉米蚜Rhopalosiphum maidis同源序列相致性最高,为89.52%;麦二叉蚜Sg1655与玉米蚜同源序列聚为一个分支,亲缘关系最近。Sg1655在麦二叉蚜各组织中均有表达,其中在唾液腺中表达量最高,Sg1655在麦二叉蚜各个龄期均有表达,且在不同龄期之间无显著差异。小麦叶片内Sg1655瞬时表达可显著抑制小麦胼胝质积累,表明该蛋白可作为潜在效应子参与抑制小麦防御反应。     

气候变化条件下木瓜秀粉蚧在中国的潜在适生区预测

为明确木瓜秀粉蚧Paracoccus marginatus Williams et Granara de Willink 1922在中国的潜在适生区,基于中国820个气象站点的气象数据和未来气候变化数据,结合木瓜秀粉蚧的生物学特性,利用CLIMEX模型对该虫在中国当前气候以及未来气候条件下的潜在适生区进行预测。结果显示,木瓜秀粉蚧在中国的潜在适生区主要在南方,适生区面积占全国陆地总面积的9.07%。高适生区主要包括海南省、广东省南部、广西壮族自治区南部以及福建省和云南省小部分地区。在未来气候条件下,2050年木瓜秀粉蚧的潜在适生区将进一步扩大,适生区面积所占比例将增加至15.46%,且有向北移动的趋势。表明木瓜秀粉蚧对中国南方地区的农业潜在威胁巨大,建议将该虫增补为检疫对象,并加强检疫监管和种群动态监测。     

Synthetic tetramer of a Phytophthora sojae-inducible fragment from soybean GmaSKTI36 promoter improves its pathogen induction activities

Using pathogen-induced promoters to control expression of the functional genes in transgenic plants may greatly increase the chances of boosting disease resistance. However, the number of the inducible promoters is limited. Here, we found that soybean GmaSKTI36 gene is strongly induced upon Phytophthora sojae infection. Functional analysis showed that its promoter could mediate rapid and strong induction of GUS expression upon pathogen infection in both Nicotiana benthamiana leaves and soybean hairy roots. Then, a 122 bp fragment that was critical to the activity was successfully identified by a progressive 5′ deletion analysis. Importantly, we found that a synthetic promoter by tetramerizing this fragment could confer strong P. sojae induction activities. Overall, the results suggested that the GmaSKTI36 promoter, the 122 bp fragment, and the synthetic promoter are potentially useful pathogen-inducible promoters.     

Resistance of barley varieties to Heterodera avenae in the Qinghai–Tibet Plateau,China

The cereal cyst nematode, Heterodera avenae, is one of the most economically important pathogens impacting the worldwide production of cereals and is widely distributed in more than 16 regions in China. The present study used the numbers of nematodes inside the plant roots to evaluate the resistance/susceptibility of different subpopulations of barley Hordeum vulgare (QH2R, QH6R and TB2R) to H. avenae under field and pot conditions. Nematode development in two highly resistant varieties was also evaluated by in vivo experiment and microscopic observation. Analyses of 186 selected varieties showed the numbers of susceptible varieties identified with the number of females/cysts per plant (NFP) method were significantly higher than those identified with the Pf/Pi ratio (PPR) method, which indicated that the NFP method rather than the PPR method is more reliable to evaluate the resistance of barley. The field and pot experiment results indicated that the QH2R subpopulation had lower females/cysts numbers than QH6R and TB2R subpopulations, and eight HR varieties (Sunong 7617, Sunong 7635, Dongyuan 87-14, Rudong 14-46, Rudong 87-57, Rudong 87-8-45, Rudong 88-14-2, and Rudong 88-67-1) were identified in QH2R, with the NFP numbers below 4.2. Further microscopic observation of nematode development suggested that H. avenae often penetrated less into highly resistant varieties (Sunong 7635 and Dongyuan 87-14) and more frequently failed to develop into females than the susceptible barleys. The promising resistant varieties identified in the present research might be helpful for breeders to develop CCN-resistant cultivars and control H. avenae populations effectively at low costs.     

番茄潜叶蛾成虫触角及幼虫头部感受器的超微结构

为明确番茄潜叶蛾Tuta absoluta(Meyrick)成虫触角及幼虫头部感受器的种类、分布和形态结构,采用扫描电镜法观察番茄潜叶蛾雌雄成虫触角和4龄雌雄幼虫触角及口器上的感受器种类和形状。结果显示,番茄潜叶蛾成虫触角上共有8种类型感受器,分别为B?hm氏鬃毛、毛形感受器、鳞形感受器、锥形感受器（Ⅰ型和Ⅱ型）、刺形感受器、腔锥形感受器、栓锥形感受器和耳形感受器。其中,雌成虫的Ⅰ型和Ⅱ型锥形感受器数量、刺形感受器数量、B?hm氏鬃毛长度、鳞形感受器长度及基部直径、Ⅰ型锥形感受器长度及基部直径、Ⅱ型锥形感受器长度、刺形感受器臼状窝直径、腔锥形感受器基部直径和耳形感受器基部直径均显著大于雄成虫;而雄成虫的毛形感受器数量和腔锥形感受器中心锥长度显著大于雌成虫。幼虫头部共计有6种类型感受器,主要着生在触角、下颚和下唇上,分别为毛形感受器、锥形感受器、刺形感受器、栓锥形感受器、指形感受器和感觉锥;幼虫触角、下颚和下唇的感受器类型和数量在雌雄间完全相同。而雌性幼虫触角鞭节上的锥形感受器SB5和下颚须上的锥形感受器SB2的长度显著大于雄性幼虫;雄性幼虫下颚茎节上的刺形感受器Sch1和下颚须上的锥形...     

鹅掌楸组培苗专用菌剂筛选及其作用机理初探

评价了不同菌剂对鹅掌楸组培苗移栽存活和生长势的影响,以筛选出鹅掌楸组培苗专用菌剂,并对其作用机理进行初步探讨。结果表明,微生物菌剂"宁盾"以及AM真菌GZ38处理组移栽存活率分别高达99.67%、96.12%,显著高于对照组,温室促生效果分别达53.71%、52.84%;且田间促生效果分别达26.56%、18.37%,存活率显著提高。机理研究表明,菌株GZ38能够较好地与鹅掌楸形成共生关系;"宁盾"、菌株GZ38不仅能够提高鹅掌楸的根系活力,还能够提高土壤中转化酶、脲酶、中性磷酸酶活性以及有效N、P、K的含量,促进其对N、P、K等营养元素吸收,达到促生效果;另外,菌剂处理后,鹅掌楸的膜损伤程度降低,保护酶活性有较大的提高,抵抗不良环境的能力显著增强。因此,选择微生物菌剂"宁盾"与AM真菌GZ38作为鹅掌楸组培苗专用微生物菌剂。