Effect of Conditioning Treatments on the Survival of Radopholus similis at High Temperatures

Heat treatments are an environmentally safe method for eliminating quarantine pests from tropical foliage. Conditioning heat treatments can induce thermotolerance against subsequent and otherwise phytotoxic temperatures in tropical foliage, allowing heat treatments to be even more effective. However, if thermotolerance is also induced in nematodes of quarantine significance like Radopholus similis, heat treatments would be rendered ineffective. A lethal thermal death point (LT_99.9) was established for R. similis by recording mortality at 25 (control temperature), 43°C, 45°C, 47°C, or 49°C after a 0, 1-, 2-, 4-, 6-, 8-, 10-, 12-, or 15-minute exposure. In a second experiment, nematodes were conditioned at 35, 40, or 45°C for 0, 15, 30, 60, 120, and 180 minutes, allowed to rest for 3 hours, and then challenged at 47°C for 5 minutes. No nematodes survived the challenge heat treatment; rather, nematode mortality was hastened by the conditioning treatment itself. In a third experiment, R. similis inside anthurium roots were conditioned at 25°C or 40°C for 15 minutes and then treated at 45°C for up to 8 minutes. Mortality of conditioned and unconditioned nematodes was similar (P > 0.1). Conditioning treatments increase plant thermotolerance but do not induce thermotolerance in R. similis. Heat treatments have promise as disinfection protocols for quarantines.     

Development of species‐specific primers for rapid diagnosis of Tetranychus urticae,T. kanzawai,T. phaselus and T. truncatus (Acari: Tetranychidae)

Species diagnosis is of the utmost importance to both pest management and plant quarantine services. Because of difficulties in the morphological diagnosis of spider mites, molecular techniques are of great value to rapidly and accurately diagnose closely related species. We examined four species of genus Tetranychus (the green and red forms of T. urticae, and T. kanzawai, T. phaselus and T. truncatus), which are found in Korea and are of significance to plant quarantine services. DNA samples isolated from a single egg, larva or adult weighed 64–188 ng. We designed species‐specific primers by performing sequence alignment for 107 sequences of the ribosomal internal transcribed spacer 2 (ITS2) region, which we obtained from GenBank, and sequences generated in this study. Specific nucleotides of each species were selected for designing primers specific for each species. Each species‐specific primer pair, when used to perform PCR analyses, detected only the species from which it originated. However, a T. urticae‐specific primer pair did not discriminate between the green and red forms of this species. These species‐specific primers can be applied in practice for the rapid and accurate diagnosis of spider mite species in plant quarantine and in agricultural fields.     

欧洲球螋的检疫

北京出入境检验检疫局从美国进境旅客携带的石榴中检出欧洲球螋。了解欧洲球螋的基本特性,能为相关部门对其检疫鉴定提供参考依据。对欧洲球螋的相关文献进行了查阅和整理,详细介绍了欧洲球螋的分类地位、分布、形态特征及近似种等信息,我国出入境检验检疫部门对球螋属的截获情况,讨论了欧洲球螋的检疫意义。认为欧洲球螋在我国无分布且具有一定的入侵风险,口岸应对该虫引起足够的重视。     

Expanding radiation quarantine treatments beyond fruit flies

1 The potential of ionizing radiation as a disinfestation treatment for insects other than tephritid fruit flies is discussed. Radiation quarantine treatments are unique in that insects are not killed immediately but rendered sterile or incapable of completing development. 2 The most tolerant insect stage to radiation is that which is most developed. Female insects, but not always mites, are sterilized with equal or lower doses than males. 3 Insects irradiated with sterilizing doses usually have shorter longevities than non‐irradiated ones. Low oxygen conditions often increase tolerance to radiation. 4 Insects in diapause are not more tolerant of radiation than non‐diapausing ones. 5 Some pests of several groups, such as aphids, whiteflies, weevils, scarab beetles, and fruit flies, may be controlled with doses ≤ 100 Gy. Some lepidopterous pests and most mites require about 300 Gy. Stored product moths may require as much as 1 kGy to sterilize, and nematodes could need > 4 kGy. 6 Even though application of irradiation to pallet‐loads of produce could mean that up to three times the minimum required dose is applied to the perimeter of the pallet, many fresh commodities tolerate doses required for quarantine security against many quarantined pests. Irradiation is arguably the most widely applicable quarantine treatment from the standpoint of commodity quality.     

Early detection of grapevine leafroll virus in <Emphasis Type="Italic">Vitis vinifera</Emphasis> using <Emphasis Type="Italic">in vitro</Emphasis> micrografting

Micrografting of grapevine was investigated for its use as a tool in virus indexing of grapevine stock. Cabernet franc and Cabernet sauvignon scions infected with grapevine leafroll-associated closterovirus III (GLRaVIII) were grafted on to virus-free indicator rootstocks of LN 33 and Cabernet sauvignon growing in tissue culture. The two rootstocks and two scions were grafted in all four possible combinations along with two control grafts (virus-free scion on virus-free rootstock). A modified MS Murashige and Skoog (1962) tissue culture medium supplemented with 0.5 mg l^–1 6-benzylaminopurine was sufficient to induce multiple shoots. Shoots and micrografts readily produced roots in the basal medium. Micrografting gave an overall success rate of 77.8%, with no significant difference between LN 33 rootstock and Cabernet sauvignon. When leafroll infected scion material was micrografted on to virus-free rootstock, the rootstock leaf turned red (23.5% in LN 33 and 63.9% in Cabernet sauvignon) or it showed leafrolling (28.5%, no significant difference between LN 33 and Cabernet sauvignon) within 2–3 weeks. After 12 weeks in culture, the extent of viral symptoms in the micrografted material was high (81.3%), with no significant difference between LN 33 and Cabernet sauvignon; however, the expression of symptoms was more severe on Cabernet sauvignon than on LN 33 rootstock. Double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) was used to validate the visual symptoms and the presence of virus was confirmed in 80% of the rootstock with visual symptoms of infection. Results indicate that micrografting is an effective method for viral indexing of grapevines. The method can be used in conjunction with wood indexing for post-entry quarantine to identify infected material and reject it much earlier than is currently possible.     

基于CLIMEX和ArcGIS的湿地松粉蚧在中国的适生性分析

地松粉蚧(Oracella acuta (Lobdell) Ferris)是我国植物检疫性有害生物之一,危害马尾松、湿地松、火炬松等松属植物。本文基于中国820个气象站点的气象数据、湿地松粉蚧的生物学数据和已知分布资料,结合CLIMEX模型与ArcGIS软件对湿地松粉蚧在我国的潜在分布区进行了预测。结果表明,湿地松粉蚧在我国的适生区主要分布在3.85°N-39.9°N之间,适生面积占全国总面积的38.79%。寄主植物分布的省份均为湿地松粉蚧的中高度适生区。高风险区主要包括台湾、海南、贵州、重庆、江苏和上海;广东、广西、云南、陕西、河南、安徽、湖北和河南的大部分地区以及福建南部、山东南部、湖南北部、浙江东部和四川东部。基于本研究中湿地松粉蚧在我国的适生性分析结果,尤其针对中高风险区域,应加强湿地松粉蚧检疫措施,完善监测体系,防止其进一步的扩散传播。     

澳大利亚输华农林产品口岸有害生物截获概况

【目的】口岸截获有害生物的数据分析可为进境农林产品检疫监管提供依据。【方法】对澳大利亚自2010至2015年输华农林产品及其入境口岸有害生物截获情况做综合分析。【结果】2010—2015年从澳大利亚输华农林产品货物中截获各类检疫性有害生物115种。其中,粮谷中以截获的杂草种类和截获频次最多,分别达到59种和31801批次;木材及木制品中以截获的蠹虫最多,达43种,共2456批次;从水果和种苗中截获病原菌6种,共79批次。另外,从入境旅客携带物中共截获检疫性有害生物9种,共66批次。【结论】澳大利亚输华农林产品可携带的检疫性有害生物自口岸传入的风险较高,相关部门应加强口岸检疫工作。     

我国首次检出双色多刺蚁及多刺蚁属介绍

【目的】2016年7月北京出入境检验检疫局从泰国进境商品山竹果中检出双色多刺蚁,经查询,该种为我国进出境检疫部门首次检出。了解多刺蚁属情况可为该类昆虫的检疫鉴定提供依据。【方法】笔者通过对双色多刺蚁及多刺蚁属相关文献进行翻译和整理,详细介绍了双色多刺蚁的分类地位、分布、形态特征及近似种,以及多刺蚁属在中国的种类,并简要分析了我国检疫系统对多刺蚁属的截获情况。【结果】双色多刺蚁(膜翅目:蚁科)主要分布于东南亚和澳大利亚地区,在我国仅分布于云南省。多刺蚁属在我国已知45种1亚种。2003—2016年7月我国检疫系统共检出多刺蚁属2097批次。【结论】多刺蚁属随货物包装携带入境的概率较高,各进出境检疫部门应加强对该类昆虫的检疫工作。     

进境美国高粱截获杂草种子现状分析

【目的】近年来,美国高粱开始大量进入我国,其携带的杂草种子状况尚未有相关研究。通过对进境美国高粱携带的杂草种子现状进行分析,可为出入境检验检疫机构的检疫监管和后续监测提供依据。【方法】通过对2014—2016年进境美国高粱截获的杂草种子的研究,了解其携带的杂草种子状况。【结果】黄埔检验检疫局和南沙检验检疫局从进境的美国高粱中截获的杂草种子种类共涉及19个科106种。主要包括禾本科27种、菊科14种、大戟科3种、茄科2种、苋科15种、豆科10种、蓼科7种、锦葵科4种、旋花科7种、十字花科4种、藜科4种等,其中检疫性杂草共涉及5科25种,检出率高。【结论】美国高粱携带的杂草种子数量大,种类丰富,检疫性杂草含量大,应予以高度重视。     

不同DNA抽提方法对普通PCR和实时荧光定量PCR方法检测家蚕微孢子虫的影响

为了优选快速、 灵敏、 特异的家蚕微孢子虫Nosema bombycis分子检测方法和DNA抽提方法, 本文通过对家蚕微孢子虫TaqMan探针荧光定量PCR检测方法和SYBR Green荧光定量PCR检测方法的建立以及反应体系优化, 并与普通PCR方法进行比较; 再采用4种不同DNA抽提方法分别对PCR和实时荧光定量PCR方法检测家蚕微孢子虫悬浮液的效果评价。结果显示: 不经过DNA抽提, 直接将家蚕微孢子虫发芽液进行PCR反应的效果优于其他方法, 检测灵敏度由高到低依次为直接法、 酚/氯仿抽提法、 动物组织DNA试剂盒抽提法和植物组织DNA试剂盒抽提法; TaqMan探针法检测家蚕微孢子虫发芽液的灵敏度和SYBR Green法相近, 达到微孢子10²个/mL, 两者均优于普通PCR方法。实验表明, 直接采用发芽液结合荧光定量PCR方法检测家蚕微孢子虫最为简便、 快速、 灵敏。该研究结果将有助于提高家蚕微粒子病监控技术和检疫能力, 对家蚕微粒子病的检疫和防治具有积极意义。