水稻瘤矮病毒P12蛋白抗血清制备及其应用

为了进一步研究水稻瘤矮病毒(Rice gall dwarf virus,RGDV)P12的功能,将RGDV-GX P12编码区亚克隆至原核表达载体,并导入大肠杆菌诱导表达;重组的P12融合蛋白经Ni-NTA His·Bind Resins纯化后用于免疫小鼠制备抗血清,并进行Western blotting分析。结果显示,约41 kD大小的RGDV P12融合蛋白可在大肠杆菌中高效表达;利用该重组蛋白所制备的抗血清可特异地与融合表达和非融合的P12蛋白发生强烈的免疫学反应。利用该特异性抗血清在病毒粒子样品中检测不到P12蛋白,而在病株总蛋白样品中可检测到1条与预期大小一致的明显条带,表明RGDV P12是一种非结构蛋白。     

一种危害杧果幼果的瘿蚊科害虫

杧幼果普瘿蚊(Procontarinia frugivora Gagné)是目前已知唯一危害杧果果实的地区性害虫,国内尚未发生分布。本文综述了其分类地位、地理分布、成虫形态特征、生物学特性、传播途径等,讨论了其经济重要性及防治对策,提出了进境检疫鉴定方法,以期为进境检疫工作提供参考。     

The Pathogen of Crown Gall Disease on Flowering Cherry and its Sensitivity to Strain K1026

Six strains of crown gall bacteria were isolated from flowering cherry. It was revealed by Otten paper electrophoresis that of the six strains, only BYH18-4 possessed the octopine type Ti plasmid, the remainder having nopaline type Ti plasmid. BYH5-1 was identified by physiological and biochemical tests to be Agrobacterium tumefaciens (originally biovar 1). The other five were A. rhizogenes (originally biovar 2). It was demonstrated with Stonier's method of double layer medium that flowering cherry crown gall bacteria exhibited different sensitivities to agrocin produced by biocontrol strain K1026. Strain K1026 on greenhouse-grown sunflower seedlings exerted a relatively potent inhibitory action on flowering cherry crown gall bacteria. Artificial inoculation showed that K1026 produced 67–99% inhibition of flowering cherry crown gall disease, compared with the treatment of inoculation with crown gall bacteria only.     

Detection of arboviruses in Culicoides(Diptera: Ceratopogonidae) collected from animal farms in the border areas of Yunnan Province,China

Biting midges of the genus Culicoides(order Diptera, family Ceratopogonidae) are potential biological vectors for the transmission of certain arboviruses among humans, livestock, and wild animals. This study collected a total of 405 Culicoides individuals from seven animal farms located in five counties in the border areas of Yunnan Province, China, and examined the Culicoides species composition and the major arboviruses carried by the Culicoides species. The collected Culicoides were classified into seven species with variable abundances: Culicoides arakawae(5.43%, 22/405), Culicoides homotomus(1.23%, 5/405), Culicoides obsoletus(19.75%, 80/405), Culicoides orientalis(17.28%, 70/405), Culicoides oxystoma(29.38%, 119/405), Culicoides peregrinus(5.68%, 23/405), and Culicoides nipponensis(21.23%, 86/405). Among the seven species, C. oxystoma and C. nipponensis were distributed in all the five counties with abundances of 13.33–44.87% and 10.00–46.83%, respectively, suggesting that these were the dominant species of Culicoides widespread on animal farms in the border areas. PCR was used to detect major arboviruses in the collected Culicoides specimens, including bluetongue virus(BTV), Japanese encephalitis virus, Dengue virus, Zika virus, African swine fever virus, and African horse sickness virus. Among the tested viruses, only BTV serotype 1 was tested positive in C. oxystoma specimens collected from a buffalo farm. Culicoides oxystoma was the dominant species on animal farms in the sampled areas, but it has not previously been documented as positive for BTV in China. The current results thus suggest that C. oxystoma could be an important vector for BTV transmission in these border areas, which, however, needs to be confirmed by further comprehensive experiments. Overall, the present study provides the first profile of Culicoides species on animal farms in the China, Vietnam, and Myanmar border areas, establishes the prevalence of arboviruses carried by these Culicoides species, and suggests the vector potential of C. oxystoma species for the transmission of BTV.     

早园竹林2种造瘿小蜂及其形成的虫瘿的研究

对浙江省德清县早园竹林进行造瘿昆虫危害和虫瘿类型调查,室内解剖和饲养观测虫瘿中的昆虫幼虫及成虫羽化情况。结果表明:(1)营林方式显著地影响造瘿昆虫2013年的造瘿率,砻糠覆盖样地造瘿率显著低于未覆盖砻糠的样地。(2)整株竹子上共获得349个虫瘿,其中新瘿291个,枯瘿58个;依据虫瘿的形态特征分成联瘿型、叶片型、叶鞘型、光瘿型和顶生型5种类型,其在整株竹子上所占比例分别为40.12%、30.95%、19.77%、6.01%和3.15%。(3)单虫瘿解剖最多可获得6头昆虫的幼虫,而联瘿型、叶片型、叶鞘型、光瘿型和顶生型虫瘿的平均幼虫数分别为1.83、2.88、2.77、1.00和2.27。(4)联瘿型、叶片型和叶鞘型虫瘿羽化出5种301头小蜂,包括竹泰广肩小蜂和刚竹泰广肩小蜂2种造瘿昆虫,栗瘿旋小蜂、点腹刻腹小蜂和纹黄枝瘿金小蜂3种寄生蜂;竹泰广肩小蜂、刚竹泰广肩小蜂和寄生蜂在3种不同类型虫瘿中的比率分别为59:18:1、70:31:2和83:31:6。研究结果将有助于加深对早园竹虫瘿的认识,并为下一步综合防治和生产实践提供理论依据。     

Physoderma,not Olpidium,is the true cause of faba bean gall disease of Vicia faba in Ethiopia

Faba bean gall (FBG) is a devastating disease of faba bean (Vicia faba) in Ethiopia. Studies were undertaken first to compare and contrast similarities between FBG disease symptoms and morphology in Ethiopia with those reported earlier in China and, secondly, to identify definitively the FBG causal agent, previously considered as Olpidium viciae, through molecular studies. Morphological studies confirmed an epibiotic phase of zoosporangia for dispersing zoospores, characteristic of Physoderma but not Olpidium, and did not show critical diagnostic characteristics of Olpidium such as presence of numerous short zoosporangial discharging tubes, or binucleate resting sporangia. Recognizing this epibiotic phase is a foundation for comprehending FBG epidemiology and will allow forecasting of zoospore release to highlight best timings for applications of chemical sprays to reduce reinfection cycles. Sequences of partial ITS1-5.8S-partial ITS2, the 18S-ITS1-5.8S-ITS2-part of 28S rRNA, and LSU (28S rRNA) derived from tissue with symptoms confirmed Physoderma, and not Olpidium, as the causal agent. Sample sequences were either close to Physoderma or the contaminant ascochyta pathogen Didymella. From symptom, morphological, and molecular data, the causal agent of FBG disease in Ethiopia is Physoderma. From observations of symptoms that Physoderma can cause, it was determined that this Physoderma crosses over between different legume host genera (e.g., Vicia, Pisum, Trifolium), highlighting the significant biosecurity risk for countries currently free of FBG.     

梨胶锈菌的重寄生菌研究——Ⅱ重寄生菌的生物学特性

 梨胶锈菌(Cymnosporangium asiaticum Miyabe ex Yamada)的重寄生菌(Tuberculina vinosa Sacc.)在PDA、综合马铃薯和梨叶汁培养基上能生长、产孢,以PDA最好,Czapek和硝酸铵蔗糖上生长差且不产孢。菌丝生长和产孢的适温20~30℃,适宜pH6~9,荧光可促进产孢而黑光有抑制作用。对碳源的利用,菌丝生长和干重以甘露醇、葡萄糖等6种碳源为好,甘露醇最佳,淀粉最差;葡萄糖、蔗糖、果糖和甘露醇中均能产孢,以葡萄糖最好。对氮源的利用,菌丝生长及干重以硝酸钾、甘氨酸和蛋白胨为好;产孢以天冬酰胺、蛋白胨和硝酸钾为好,甘氨酸和谷氨酸不产孢。孢子萌发适温15~30℃,相对湿度65~100%均能萌发,在水滴中萌发率最高,低于50%不萌发;最适pH4~6,低于pH3和高于pH8不萌发;光照有利于孢子萌发。     

Suppression of clubroot (Plasmodiophora brassicae) development in Arabidopsis thaliana by the endophytic fungus Acremonium alternatum

This study investigated the ability of an endophytic fungus Acremonium alternatum to reduce clubroot formation in the model plant Arabidopsis thaliana, which is highly susceptible to Plasmodiophora brassicae . Quantitative PCR demonstrated that A. alternatum colonized the P. brassicae -infected roots and shoots of the host plant. When Arabidopsis plants were co-inoculated with P. brassicae and A. alternatum , gall formation was reduced as shown by the reduction of the disease index (DI) by up to 50% compared to plants only infected with P. brassicae, whereas the infection rate was lowered by about 20% only in several, but not all, experiments. Clubroot was similarly suppressed when plants were inoculated with autoclaved A. alternatum spores or spore extracts, showing that viable spores were not needed. However, A. alternatum spores did not inhibit P. brassicae resting spore germination. Compared to the normal root galls, the smaller root galls on A. alternatum -inoculated plants contained fewer resting spores of the clubroot pathogen. It was thus hypothesized that inoculation with A. alternatum delayed the development of P. brassicae . Using quantitative RT-PCR to monitor the expression of P. brassicae genes differentially expressed during the development of the disease, a delayed pathogen development was corroborated. Furthermore, greenhouse experiments identified a time window in which the endophyte had to be administered, where the latest effective time point was 5 days before inoculation with P. brassicae and the optimum treatment was to administer A. alternatum and P. brassicae at the same time. These results indicate that A. alternatum and perhaps similar endophytes could be useful for the management of clubroot disease.     

刺桐姬小蜂在中国的适生区分析

运用CLIMEX的气候相似系数法,对入侵害虫刺桐姬小蜂在中国的适生区进行了分析和预测,结果表明:中国大陆有18个省区45个市、县(地区)适合该虫生存,大体分布于18.14°N～36.00°N,98.30°E～121.36°E范围内,这些地区也是刺桐属植物的主要分布区。