古巴斯氏线虫生物学特性的研究

对采自热带地区的古巴斯氏线虫的生物学特性进行了研究,结果表明： 该线虫侵染大蜡螟幼虫的温度范围为１５～３５℃;在２５℃下,线虫在大蜡螟幼虫体内完成一个侵染周期需２００～２１２小时;对大蜡螟幼虫的侵入率为４２．８％ 。在常温（２０～３３℃）下贮存６个月后其存活率为９３．５％ ;侵入率为３１．３％ 。古巴斯氏线虫的耐高温能力和运动能力均优于小卷蛾线虫Ａ２４品系,但耐干燥能力较差。古巴斯氏线虫的耐高温及常温下耐贮存的特性显示了其在防治害虫上的应用潜力     

Electron Microscopy and Molecular Characterization of Phytoplasmas Associated with Strawflower Yellows in the Czech Republic

Strawflower (Helichrysum bracteatum) with symptoms resembling those associated to phytoplasma infection were observed in several areas in the Czech Republic during the period 1994–2001. Plants with leaf bronzing, reddening and necrosis, proliferation of secondary shoots, flower abnormalities and dwarfing died in advanced stages of the disease. The disease incidence ranged from 2% to 70% and caused significant loss to the flower and seed production. Transmission electron microscopy showed phytoplasmas in sieve cells of affected plants, but not in healthy ones. Association of phytoplasmas with the disease was confirmed by polymerase chain reaction using phytoplasma universal ribosomal primers R16F2n/R16R2. An amplification product of the expected size (1.2 kb) was observed in all samples of the symptomatic strawflowers. The restriction profiles obtained following separate digestion with three endonucleases (AluI, HhaI, MseI) showed that phytoplasmas infecting strawflowers from different localities in the Czech Republic were uniform and undistinguishable from aster yellows (subgroup 16SrI-B). Sequence analysis of 1771 bp of the ribosomal operon amplified with primers P1/U3, R16F2n/R2 and 16R758/P7 indicated that the closest related phytoplasmas were those associated with 'Rehmannia glutinosa var. purpurea', both originating from Bohemia. This is the first report on the occurrence of a phytoplasma-associated disease of strawflower in the Czech Republic.     

寡聚糖在农作物病害防治中应用研究

为研究寡聚糖在农作物病害防治中的作用机制(直接抑菌或诱导抗病)，采用生长速率法测定了寡聚糖与3种致病真菌的互作关系，结果表明，寡聚糖对3种病原菌均存在不同程度的抑制作用，EC50分别为：立枯丝核菌0．01％、腐霉菌0．1％、镰刀菌0．1％：从寡聚糖对3种离体病原菌的抑制程度来分析，它的田间病害防治中的应用价值，应归因于三方面的因素：它对病原菌的抑制作用、对农作物的生长调节作用及诱导植物抗病性。     

耐草甘膦菜豆耐性分子学机理研究

用耐性菜豆根尖为材料，以λgt10为载体获得了4.8×105个重组噬菌体的cDNA文库。以植物EPSP合成酶基因保守序列合成二段探针，经噬菌体原位杂交筛选出了阳性克隆，并进行了酶切鉴定。测定 cDNA 序列长度为2024个核苷酸，其中编码长1569个核苷酸，共编码523个氨基酸和一个终止密码子，与已发表的其它植物成熟EPSP合成酶氨基酸序列相比具有很高的同源性（≥84.8%），但进入叶绿体运输肽的氨基酸同源性低。以耐性菜豆EPSP合成酶的cDNA序列为模板，用RT-PCR方法扩增感性菜豆EPSP合成酶cDNA片段，并克隆于pUC18质粒上，经测定序列并比较发现:感性菜豆EPSP合成酶cDNA核苷酸序列1737位碱基为G，而耐性的为C，从而表达出的513位氨基酸残基感性的为E，耐性的为Q，表明两种菜豆EPSP合成酶cDNA核苷酸序列一位点的差异是它们对草甘膦耐性不同的原因。     

Diversity of Phytoplasmas Infecting Fruit Trees and Their Vectors in Croatia

Journal of Plant Diseases and Protection - A survey started in 2002 (Križanac et al. 2008) was continued to determine the diversity of phytoplasmas infecting fruit trees and their vectors in...     

稻负泥虫生物学特性与防治指标研究

稻负泥虫［Ｏｕｌｅｍａｏｒｙｚａｅ（Ｋｕｗａｙｍａ）］是大别山区中稻田的重要害虫之一,６０～７０年代基本上控制了为害,但自８０年代以来,随着杂交水稻的推广,耕作栽培制度的变化,以及六六六等有机氯农药的停用,虫口迅速回升,以至每隔２～３ａ大发生１次,造...     

Biological and Molecular Properties of a Begomovirus from Dicliptera sexangularis

ABSTRACT Sixangle foldwing, Dicliptera sexangularis (Acanthaceae), showing severe yellow mottle and leaf distortion symptoms was collected from the shoreline of Calusa Island (Lee County, FL). The putative virus was transmitted from infected D. sexangularis to healthy seedlings by mechanical, whitefly (Bemisia tabaci biotype B), and graft-inoculations. Different forms of geminivirus-like DNAs were detected in total DNA extracted from infected plants by Southern blot hybridization analyses using DNA-A and -B of Bean golden mosaic virus (BGMV) from Guatemala as probes. Preliminary polymerase chain reaction experiments and sequence comparisons indicated that the virus was a distinct bipartite begomovirus. The virus was designated Dicliptera yellow mottle virus (DiYMV). Replicative dsDNAs of DiYMV were extracted, digested with selected restriction enzymes, and cloned into a plasmid vector. Both DNA-A and -B were sequenced and compared with those of other begomoviruses. Phylogenetic analyses using AV1, AC1, and BV1 nucleotide sequences indicated that DiYMV has a close relationship with the New World begomoviruses, especially those distributed in the nearby geographic areas of the Florida coast and the Caribbean Basin. However, different percent nucleotide sequence identities and phylogenetic relationships were detected when different open reading frames (ORFs) of DiYMV were compared with their counterparts from begomoviruses from the Caribbean Basin. Based on phylogenetic analyses of the AC1 and BV1 ORFs, DiYMV was closely related to BGMV type II isolates, whereas sequence comparisons of the common region and the AC4-derived amino acid sequences indicated its close relationship with Potato yellow mosaic virus from Venezuela.     

secA gene suitability for fast and easy identification of Phytoplasmas by RFLP analysis

European Journal of Plant Pathology - In this study we investigated the secA gene as possible marker to supplement the classification scheme based on 16S rRNA sequences. We cloned and sequenced...     

稻瘟病菌生理小种鉴定的影响因子研究

选用特特普、珍龙１３、四丰４３、东农３６３、关东５１、合江１８和丽江新团黑谷等７个中国鉴别品种，对相同菌株在不同条件下的小种鉴定作了研究，结果表明接种体浓度和施肥水平对小种鉴定结果有较大影响，在小种鉴定中应多施氮肥以保稻苗嫩绿，同时接种的孢子浓度应在２０万／ｍｌ左右，以保证稻瘟病菌生理小种鉴定的准确性和可靠性。     

分子标记技术在绿僵菌研究中的应用

分子标记技术在真菌研究中被广泛应用,本文就其在昆虫病原真菌绿僵菌的分类、菌株鉴定等方面的研究、应用和存在的问题进行综述,并对其应用前景进行展望。     

Biological and Molecular Characterization of a New Carlavirus Isolated from an Aconitum sp

ABSTRACT A new virus was isolated from symptomless Aconitum napellus plants. The virus, for which the name Aconitum latent virus (AcLV) is proposed, has flexuous particles 640 nm in length. The experimental host range was limited to Nicotiana clevelandii. Electron microscopy studies of ultrathin sections of infected A. napellus tissues revealed the presence of elongated virus particles. No inclusion bodies characteristic of potyvirus infection were observed. AcLV was purified from naturally infected A. napellus by cesium chloride step gradient centrifugation. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis of dissociated purified virus preparations, a major protein component with a molecular mass of 35 kDa was observed. Diagnostic antibodies that could specifically bind to virus particles were produced. The 5' terminus (620 nucleotides) of the viral RNA was cloned and sequenced. It comprised 71 nucleotides from the untranslated 5' terminus and 549 nucleotides of an open reading frame encoding 183 amino acids. Comparison of the predicted amino acid sequence with those of other plant viruses revealed 40 to 60% identity with several carlaviruses. Based on particle morphology, absence of inclusion bodies in ultrathin sections, the relative molecular weight of the coat protein, the nucleotide sequence, and predicted amino acid homology, it is suggested that this virus belongs to the carlavirus group.     

Host Effect on the Molecular and Biological Properties of a Citrus exocortis viroid Isolate from Vicia faba

ABSTRACT Citrus exocortis viroid (CEVd) is the casual agent of citrus exocortis disease, and has been found in naturally infected citrus and noncitrus hosts. Field isolates of CEVd may infect susceptible hosts as a complex of genetically related sequence variants (haplotypes). In the present work, a CEVd isolate recovered from a symptomless broad bean plant was characterized as a heterogeneous population with a nucleotide diversity of 0.026, which did not contain a predominant haplotype. When nucleic acid extracts of this infected broad bean were used to inoculate tomato, the plants displayed symptoms and the CEVd population was more homogeneous, with a nucleotide diversity of 0.007. However, when nucleic acid extracts from this tomato isolate were back inoculated to new broad bean plants, this isolate did not revert to the original population, because it showed low nucleotide diversity (0.001) and induced symptoms in the broad bean plants. Symptomless broad bean plants may act as reservoirs of highly heterogeneous populations of CEVd variants, providing an excellent inoculum source in terms of its potential to infect a broad range of putative hosts. The epidemiological implications are discussed.     

Phytoplasmas associated with disease of coconut in Malaysia: phylogenetic groups and host plant species

Coconut palm ( Cocos nucifera ), oil palm ( Elaeis guineensis ), Bermudagrass ( Cynodon dactylon ) and Madagascar periwinkle ( Catharanthus roseus ) with symptoms indicative of phytoplasma disease were collected from different locations in Malaysia. PCR assays employing phytoplasma universal rRNA gene primers P1/P7 alone or P1/P7 followed by R16F2n/R16R2 detected phytoplasmas in eight out of 20 Malayan Red Dwarf (MRD), nine out of 12 Malayan Yellow Dwarf (MYD) and 12 out of 12 Malayan Tall (MT) coconut palms displaying coconut yellow decline symptoms. Positive detections were also obtained from six out of six oil palm seedlings showing symptoms of yellowing and necrosis, from 10 out of 10 Bermudagrass samples with white leaf symptoms, and from eight out of eight periwinkle plants showing phyllody, virescence, little leaf, proliferation and foliar yellowing. Phytoplasmas were not detected in any of the symptomless plants tested. Sequencing and phylogenetic analysis of PCR products determined that phytoplasmas infecting both MRD and MT coconuts and Bermudagrass in Serdang, Selangor State, were all members of the 16SrXIV ' Candidatus Phytoplasma cynodontis' group, whereas isolates in periwinkle in Serdang were all members of the 16SrI ' Ca. Phytoplasma asteris' group. However, the phytoplasmas detected in MYD coconuts and oil palms from Banting, Selangor State, and in periwinkle from Putrajaya were collectively very similar (99%), but shared <97·5% similarity with 16S rDNA sequences of all other known phytoplasmas, indicating that they represent a novel taxonomic group. Thus, at least two phylogenetically distinct phytoplasmas are associated with the coconut yellow decline syndrome in Malaysia, both of which were also detected in other plant species.     

Identification of Mulberry Dwarf Phytoplasmas in the Genital Organs and Eggs of Leafhopper Hishimonoides sellatiformis

ABSTRACT The presence of mulberry dwarf (MD) phytoplasmas in organs of the inoculative vector insects Hishimonoides sellatiformis and Hishimonus sellatus was determined by means of electron microscopy (EM) and polymerase chain reaction (PCR) assays. Many MD phytoplasmas were detected in genital organs as well as in the intestines, salivary glands, brains, fat bodies, and thoracic ganglia of Hishimonoides sellatiformis, but only in the intestine and salivary glands of Hishimonus sellatus. Many phytoplasmas with characteristic morphology were observed via EM in ovaries, seminal receptacles, and testes, and they were further identified by PCR assays with group I-specific primers. In addition, the organisms were detected by direct or nested PCR assays in eggs (head pigmentation stage of embryos) laid on mulberry shoots by inoculative leafhoppers and in the newly hatched nymphs from these eggs. These findings indicate that transovarial transmission of MD phytoplasmas occurs in Hishimonoides sellatiformis.