MiAsg分子克隆及与南方根结线虫病害的关系

根结线虫(Meloidogyne spp.)是一种世界性的植物病害。在前期研究中,通过利用生物信息学方法在线虫全基因组中预测了一些功能基因。本研究以预测到的线粒体ATP合成酶g亚基基因(Asg)序列设计特异引物克隆了南方根结线虫(M. incognita)的Asg基因(MiAsg),对其序列进行了特征分析后,利用病毒诱导的基因沉默(VIGS)技术,将其导入番茄植株,然后接种M. incognita,研究MiAsg基因与根结线虫病害的关系。结果表明,克隆到的MiAsg基因与预测到的MiAsg基因相似性高达100%。接种M. incognita 60 d后,MiAsg基因沉默的番茄植株根结数分别比空载体对照减少了59.6%,比清水对照降低了59.5%。结果表明,MiAsg基因沉默对根结线虫病害具有很好的防控效果,也说明MiAsg基因可能参与线虫的致病性。     

The use of infrared spectroscopy and machine learning tools for detection of Meloidogyne infestations

Plant parasitic nematodes are generally soilborne pathogens that attack plants and cause economic losses in many crops. The infested plants show nonspecific symptoms or, often, are symptomless; therefore, diagnosis is performed by taking soil and root tissue samples. Here, we show that a combination of different infrared spectra analysis and machine learning algorithms can be used to detect plant parasitic nematode infestations before symptoms become visible, using leaves instead of roots and soil as samples. We found that tomato and guava plants infested with Meloidogyne enterorlobii produced different spectral patterns compared to uninfested plants. Using partial spectra from 1,450 to 900/cm as the "fingerprint region", principal component analyses indicated that after 5 (tomatoes) or 8 weeks (guava), plants with no visible symptoms of infestations were positively diagnosed. To improve the early detection response, we used machine learning modelling. A support vector machine (SVM) was used to obtain more robust, accurate models. The SVM model contained 34 support vectors, 17 for each level. The overall performance of the model was >97% and the total accuracy was significantly higher, demonstrating the absence of chance prediction. The best prediction of infestation was obtained at the second and fourth weeks for tomatoes and guavas, respectively, reducing the diagnostic time by half. The combined application of these techniques reduces the processing time from field to laboratory and shows enormous advantages by avoiding root and soil sampling.     

南方根结线虫寄生真菌的分离和鉴定

 1992年和1993年,在广东和广西采集南方根结线虫样品44个,分离129个菌株,它们分属9属18种.     

酯酶在根结线虫分类上应用的研究

应用聚丙烯酰胺凝胶电泳技术,对采自我国14个省市的5种根结线虫(Meloid-ogyne spp.),共47个样本作了正向极酯酶的分析;并研究了一些可能影响测定的因素。结果表明,根结线虫种间的酯酶谱存在明显而稳定的特异性,在种内也是稳定的。对接种寄主、样本来源、生理小种、室内培养与田间自然寄生、营养等不同条件下的对比分析表明,它们均不影响种的特征性酶谱。因此将酯酶作为根结线虫分类的辅助手段是有意义的。但线虫的发育阶段不同,酯酶谱有差异。采用产卵期的雌成虫进行分析则简便、可靠。     

Reduction in biological efficacy of ethoprophos in a soil from Greece due to enhanced biodegradation: comparing bioassay with laboratory incubation data

Soils were collected from a potato-growing area in Serres, Northern Greece, where the nematicide ethoprophos was reported to have lost its effectiveness against cyst nematodes following 30 years of regular use. Incubation studies with ethoprophos and two bioassays using root-knot nematodes demonstrated that, in this heavily treated soil, the nematicide was degraded rapidly and nematicidal activity persisted only up to 14 days. In soil from an adjacent field with no known history of nematicide use during the preceding 14 years, ethoprophos was degraded more slowly and retained its nematicidal activity for more than 35 days. Ethoprophos efficacy was extended when the soil that had been treated in the field was autoclaved, although the effect was only transitory. The addition of ‘pre-conditioned’ soil from the previously treated field to samples of soil from the previously untreated field resulted in a significant acceleration of ethoprophos degradation compared with that observed in unamended soil from the previously untreated field. © 1999 Society of Chemical Industry     

Cucumis metuliferus is resistant to root‐knot nematode Mi1.2 gene (a)virulent isolates and a promising melon rootstock

Pot experiments were carried out to characterize the response of two Cucumis metuliferus accessions (BGV11135 and BGV10762) against Mi1.2 gene (a)virulent Meloidogyne arenaria, M. incognita and M. javanica isolates and to determine the compatibility and the effect on physicochemical properties of fruit melons. In addition, histopathological studies were conducted. One week after transplanting, plants were inoculated with one J2 cm^?3 of sterilized sand (200 cm³ pots) and maintained in a growth chamber at 25 °C for 40 days. The susceptible cucumber cv. Dasher II or melon cv. Paloma were included for comparison. The number of egg masses and number of eggs per plant were assessed, and the reproduction index (RI) was calculated as the percentage of eggs produced on the C. metuliferus accessions compared to those produced on the susceptible cultivars. The compatibility and fruit quality were assessed by grafting three scions, two of Charentais type and one of type piel de sapo, under commercial greenhouse conditions. The resistance level of both C. metuliferus accessions ranged from highly resistant (RI < 1%) to resistant (1% ≤ RI ≤ 10%) irrespective of Meloidogyne isolates. Melon plants grafted onto C. metuliferus accession BGV11135 grew as self‐grafted plants without negatively impacting fruit quality traits. Giant cells induced by Meloidogyne spp. on C. metuliferus were in general poorly developed compared to those on cucumber. Furthermore, necrotic areas surrounding the nematode were observed. Cucumis metuliferus accession BGV11135 could be a promising melon rootstock to manage Meloidogyne spp., irrespective of their Mi1.2 (a)virulence, without melon fruit quality reduction.     

南方、花生和爪哇根结线虫PCR检测方法

为快速、准确、稳定的鉴定南方、花生和爪哇根结线虫,利用已报道的南方和爪哇根结线虫的两对特异性引物,结合本研究设计的花生根结线虫特异性引物,通过优化PCR反应体系,建立了3种根结线虫的PCR检测方法。结果表明,该方法能够特异性扩增以上3种根结线虫,特征片段长度分别为399、335和670 bp,灵敏度达到单条2龄幼虫的水平。研究结果将为以上3种根结线虫的快速鉴定提供技术支持。     

从枝菌根真菌对黄瓜南方根结线虫病害防治效应

在温室盆栽条件下,研究了丛枝菌根真菌Glomus mosseae、Glomus intraradices及混合菌剂对黄瓜苗期与移栽后南方根结线虫病害的防治效应.结果表明:苗期接种AM真菌处理的侵染率低时,其抑制根结线虫侵染的能力也低;移栽缓苗后接种根结线虫后,接种AM真菌处理的黄瓜地上部干重、地下部干重、株高、植株养分吸收量均显著高于只接种南方根结线虫处理,根结指数显著低于只接种南方根结线虫处理.说明AM真菌确实能够通过与黄瓜根系建立共生体来防御南方根结线虫对黄瓜的危害,其中G.intraradices、G.mosseae菌种的促生及防治黄瓜南方根结线虫病害的作用显著.     

大型仪器设备共享管理和成本核算探讨

苦瓜(Momordica charantia)具有丰富的营养价值和药用价值。由于耕地面积有限,苦瓜连作现象严重,根结线虫对苦瓜的为害逐年加重,严重影响苦瓜的产量和品质。针对海南省苦瓜根结线虫病的危害、传播途径、发生规律以及栽培特点,总结提出适合海南省苦瓜根结线虫病的防治对策,以期为海南苦瓜的安全生产提供技术支持。     

三种化合物处理对南方根结线虫生长发育及致病能力的影响

为明确不同浓度NH4HCO3、Cu SO4·5H2O和Fe Cl3·6H2O对南方根结线虫Meloidogyne incognita 2龄幼虫活性的影响,在盆栽条件下将化合物处理后的2龄幼虫接种番茄,通过根染色和根系冰冻切片法观察2龄幼虫的发育状况和根系内巨细胞的形成,并测定了番茄根系各生理指标的变化。结果表明,50mmol/L NH4HCO3、1.4mmol/L Cu SO4·5H2O和1.4 mmol/L Fe Cl3·6H2O处理南方根结线虫后,番茄根系内2龄、3龄及4龄幼虫数量均显著小于对照;根系内丙二醛、可溶性糖和游离氨基酸的含量均显著低于对照,根系活力显著增强;经NH4HCO3处理后,根系内巨细胞形成缓慢,14 d时出现巨型细胞空洞现象。3种化合物处理对南方根结线虫生长发育的抑制作用显著,处理后的线虫对番茄植株致病力减弱,表现为NH4HCO3抑制作用最强,Cu SO4·5H2O次之,Fe Cl3·6H2O最弱。