Artificial diet based investigation on the impact of purified Cry1Ac,Cry1Fa and Cry2Ab on the survival and reproductive performance of adult green lacewing, <Emphasis Type="Italic">Chrysopa pallens</Emphasis> (Rambur) (Neuroptera: Chrysopidae)

The green lacewing Chrysopa pallens (Rambur) (Neuroptera: Chrysopidae) is a common and abundant predator in many cropping systems in palearctic realm and it’s conservation is helpful in sustainable pest management in agro-ecosystem. Prior to commercialization of Bt crops in any agro- ecosystem, it is necessary to evaluate the impact of Cry proteins upon non-target organisms especially biological control agents (BCA). In present study an artificial diet consisting of shrimp, beef, beef liver and egg yolk was developed to mass-rear C. pallens for its use as biological control agents in sustainable pest management. Moreover, an artificial diet based risk assessment protocol was developed to investigate the impact of Cry1Ac, Cry1Fa and Cry2Ab on the survival and reproductive performance of C. pallens adults. C. pallens was fed on diets incorporated with Cry proteins and without addition of Cry proteins (control). The same diet containing boric acid was served as a positive control. Temporal stability, bioactivity and intake of Cry proteins by C. pallens were confirmed using double-antibody sandwich, enzyme-linked immunosorbent assay and bioactivity verification bioassays. Survival and reproductive performance of C. pallens, e.g., pre-oviposition period, daily fecundity, total fecundity and 30-day old adults dry weights, exhibited non-significant differences (p?>?0.05) for the diets containing Cry1Ac, Cry1Fa and Cry2Ab (50 μg/g) against Control. However, significant reduction in survival and reproductive performance (p?<?0.05) was observed in positive control. Our findings reveal that artificial diet is a good source of nutritional requirement with enhanced survival and reproductive performance of C. pallens and can be used for mass rearing of predator in case of natural diet scarcity and Cry proteins are safe for adult C. pallens and Bt crops cultivation help in predators conservation in sustainable agriculture.     

Highly specific assays to detect isolates of Pseudomonas syringae pv. actinidiae biovar 3 and Pseudomonas syringae pv. actinidifoliorum directly from plant material

Pseudomonas syringae pv. actinidiae (Psa) is responsible for bacterial canker of kiwifruit. Biovar 3 of Psa (Psa3) has been causing widespread damage to yellow‐ and green‐fleshed kiwifruit (Actinidia spp.) cultivars in all the major kiwifruit‐producing countries in the world. In some areas, including New Zealand, P. syringae pv. actinidifoliorum (Pfm), another bacterial pathogen of kiwifruit, was initially classified as a low virulence biovar of Psa. Ability to rapidly distinguish between these pathovars is vital to the management of bacterial canker. Whole genome sequencing (WGS) data were used to develop PCR assays to specifically detect Psa3 and Pfm from field‐collected material without the need to culture bacteria. Genomic data from 36 strains of Psa, Pfm or related isolates enabled identification of areas of genomic variation suitable for primer design. The developed assays were tested on 147 non‐target bacterial species including strains likely to be found in kiwifruit orchards. A number of assays did not proceed because although they were able to discriminate between the different Psa biovars and Pfm, they also produced amplicons from other unrelated bacteria. This could have resulted in false positives from environmental samples, and demonstrates the care that is required when applying assays devised for pure cultures to field‐collected samples. The strategy described here for developing assays for distinguishing strains of closely related pathogens could be applied to other diseases with characteristics similar to Psa.     

马铃薯纺锤块茎类病毒的检测和防治

马铃薯纺锤块茎类病毒病(potato spindle tuber viroid,PSTVd)是一种严重为害马铃薯生产的病害,降低产量20%—30%。防治的主要措施是应用无类病毒的种薯。由于目前还没有脱掉类病毒的有效措施,只能从未被饱和侵染的群体中鉴定筛选出未被侵染的个体,再脱掉其它病毒,作为核心繁殖材料。1987年以来,利用自制的电泳设备,以往复聚丙烯酰胺凝胶电泳法(return-polyacrylamide gel electrophoresis,R-PAGE)检测类病毒,筛选出未感病的个体,再用茎尖组织培养法脱掉其它病毒。经用马铃薯卷叶病毒等8种病毒酶标抗体鉴定筛选,获得既无类病毒也无主要马铃薯病毒的克新1、2、3和4号等主栽马铃薯品种的核心种。并已提供给省内外的良种场繁殖推广。1989和1990年抽样检测克山良种场繁殖的原种、一级和二级良种,未检测到类病毒。     

内蒙河套灌区盐碱低产田建立有机质富积层改土培肥效果

通过连续四年在内蒙古河套灌区低产盐碱地定点、定位试验与定时测土分析，明确了土壤耕层增施有机物料建立有机质富积层，能够改善土壤理化性状，尤其是能降低土壤表层含盐量，提高作物保苗率和显著增加作物产出量。     

香蕉穿孔线虫在北京地区的适应性研究

在北京顺义隔离试验圃内研究香蕉穿孔线虫在北京地区的适应性,重点在于观察该线虫对北京地区栽培作物种类的寄生性和致病力、对北方低温的抗逆生存能力及对北方碱性土壤环境的适应程度。通过实验室和田间试验研究,其结果表明香蕉穿孔线虫在北京地区自然露地环境条件下不能越冬存活。但在温室、大棚等设施环境中,该线虫均可获得适合的寄主、温度及土壤酸碱度(pH)等条件,并可对作物造成严重危害,因此香蕉穿孔线虫在北京地区温室和大棚等设施条件下有着极高的适应性。     

从尼日利亚进境的芋头中截获最短尾短体线虫

广东检验检疫技术中心植检实验室于2008年4月从尼日利亚进境的芋头中分离到数条短体线虫,经纯培养、形态观察、测量,以及对rDNA-ITS、D2D3区序列的扩增、克隆、测序分析,鉴定为最短尾短体线虫(Pratylenchus brachyurus(Godfrey,1929)Filipjev & Schuurmans Stekhoven,1941)。该线虫在国内局部分布,属于检疫性线虫,在系统内首次截获。     

齿兰环斑病毒与建兰花叶病毒分子检测研究

齿兰环斑病毒（Odontoglossum ringspot virus,ORSV）与建兰花叶病毒（Cymbidium mosaic virus, CyMV）是严重危害兰科植物的两种主要病毒。本研究根据病毒外壳蛋白基因设计特异性引物,应用ELISA、普通RT-PCR、巢式RT-PCR和免疫捕获RT-PCR4种方法进行了检测研究与比较。结果表明：普通RT-PCR与ELISA方法检测灵敏度相当;巢式RT-PCR检测灵敏度要比普通RT-PCR与ELISA方法高出104倍以上;免疫捕获RT-PCR检测灵敏度介于普通RT-PCR和巢式RT-PCR之间。采用巢式RT-PCR方法对我国台湾进境的蝴蝶兰植株样本检测,1号样本出现与阳性对照一致的特异条带。双向测序分析,扩增产物序列与ORSV外壳蛋白基因具有100％的同源性,表明1号蝴蝶兰样本携带ORSV。     

基于物联网的小麦赤霉病自动监测预警系统应用效果

2016年在陕西关中地区的眉县、周至县、杨陵区、兴平市、临渭区和华县安排布点,进行了小麦赤霉病自动监测预警系统的测试,以期明确其在关中地区的应用效果。试验结果表明,该系统可以在小麦扬花期前1周发出预警信息,预测准确度达94.4%,系统运行稳定,自动化程度高,可为小麦赤霉病的科学防治决策提供重要的参考依据,具有一定的应用前景。     

寄生蜂调控寄主分子机制的研究进展

在寄生蜂与寄主相互作用的研究中,揭示寄生蜂攻克寄主防御系统和调控寄主生长发育的分子机制,已成为生物防治领域的研究热点。有关研究表明:寄生蜂通过控制血细胞增殖与分化的转录/信号传导因子、调节血细胞延展/包囊的因子及诱导血细胞凋亡,以破坏寄主细胞免疫系统;干扰与血淋巴黑化相关基因及抗菌肽基因的表达以抑制寄主的体液免疫反应;调节与生长发育相关蛋白和酶类以扰乱寄主生长发育。作者从细胞免疫、体液免疫和对寄主生长发育三个方面综述了寄生蜂调控寄主分子机制的研究进展。     

基于单克隆抗体的南方菜豆花叶病毒血清学检测技术

为建立南方菜豆花叶病毒(southern bean mosaic virus,SBMV)的快速、简便、高通量检测技术,加强该病毒的口岸检验检疫,以提纯的SBMV粒子为免疫原免疫BALB/C小鼠,利用杂交瘤技术获得3株杂交瘤细胞株19C3、19H9和20G4,其分泌的SBMV腹水单抗效价均达到10-7,且3个单抗与感染SBMV大豆叶片组织粗提液有强烈的特异性免疫反应,而不与感染南方豇豆花叶病毒(southern cowpea mosaic virus,SCPMV)的豇豆、健康的大豆、毛豆、豌豆、蚕豆和菜豆叶片组织粗提液发生免疫反应。以制备的单抗为核心,建立了检测植物中SBMV的ACP-ELISA和dotELISA两种血清学方法。3个单抗中19H9单抗的检测灵敏度最高,以其建立的ACP-ELISA和dot-ELISA方法检测大豆病叶粗提液的灵敏度分别达到1∶163 840和1∶10 240稀释浓度。利用建立的dot-ELISA方法可从上海口岸截获的大豆种子中检测出SBMV,且该检测结果得到RT-PCR方法验证。表明制备的SBMV单抗及建立的SBMV血清学检测技术可有效用于我国SBMV的口岸检验检疫。