柑桔溃疡病菌抗药突变体的生理特性研究

 经药剂驯化和紫外光诱导双重作用可获得柑桔溃疡病菌抗拌种灵突变体(XcR),病原菌抗药突变频率低于10-6。Xc R的生长速率与敏感菌株基本一致,对萎锈灵和叶枯唑具有潜在的正交互抗性,与浸种灵没有交互抗性。XcR胞外产物和胞外水解酶活性一般多于或高于敏感菌株,其中以淀粉酶活性变化最为明显。Xc R琥珀酸脱氢酶活性显著降低,并对拌种灵的敏感性亦降低。XcR能够引起烟草的过敏性反应,但失去对寄主的致病性。Xc R的性质表明,柑桔溃疡病菌对拌种灵产生抗药性的风险较低,病原菌获得抗药性状的同时可能导致其在田间的适合度下降。琥珀酸脱氢酶的活性可作为柑桔溃疡病菌抗药性的标记。     

稻瘟病化学防治药剂的历史沿革与研究现状

回顾了稻瘟病化学防治药剂的历史,从早期的无机杀菌剂到农用抗生素、有机氯类、有机磷类与稻瘟灵、烯丙苯噻唑、黑色素合成抑制剂(melanin biosynthesis inhibitors,MBIs)和甲氧基丙烯酸酯类,简要评述了各类药剂的主要优缺点,介绍了这几类化合物目前在稻瘟病防治中应用的主要品种及其抗药性发生情况,重点论述了三环唑的病害防治机制与抗药性的最新研究进展。     

羧酸酰胺类(CAAs)杀菌剂研究进展

羧酸酰胺类(Carboxylic acid amides, CAAs) 化合物是一类对卵菌病害具有优异防治效果的新型杀菌剂。该类杀菌剂由3组化学结构各不相同的化合物组成,包括肉桂酰胺、缬氨酰胺氨基甲酸酯和扁桃酰胺类。由于这3类杀菌剂具有相似的杀菌活性且相互间存在正交互抗药性,国际杀菌剂抗药性行动委员会已于2005年底将这3类具有不同化学结构的杀菌剂归为羧酸酰胺类杀菌剂。对CAA类杀菌剂的化学结构特征、生物学活性、作用机制与抗药性的最新研究进展进行了综述。     

托布津、多菌灵不可交替使用和混合使用

托布津是广谱内吸性杀菌剂,能防治多种作物病害,兼具预防和治疗作用。喷施后在植物体内转化成多菌灵而防治病害,防病谱与多菌灵基本相同。多菌灵的主要杀菌机理是抑制真菌的有丝分裂中纺锤体的形成,作用机制单一,故易诱致病菌产生抗药性,使防治失效。托布津、多菌灵、苯菌灵(苯来特)和噻菌灵(特克多)等药剂都属苯并咪唑类杀菌剂。因此一定要避免长期单一使用本剂或它们彼此间同时混合使用或交替使用,以防止病菌产生抗药性正交互抗药性托布津、多菌灵不可交替使用和混合使用$广西大学农学院!南宁530005@赖传雅     

叶枯唑安全风险管理对策措施及建议

叶枯唑是我国20世纪70年代自主创制研发的杀菌剂,主要登记用于水稻白叶枯病、大白菜软腐病等细菌性病害防治。本文概述了叶枯唑的基本特性和浙江省叶枯唑生产销售情况,分析了叶枯唑在登记使用过程中存在的主要问题,并结合我国叶枯唑登记现状,提出了进一步加强叶枯唑安全风险管理的对策措施。     

内吸杀菌剂拌种灵防治棉苗立枯病、炭疽病的研究

本文是报道内吸杀菌剂拌种灵(2—氨基—4—甲基—5—甲酰苯胺基噻唑)及其复配剂拌种双(拌种灵+福美双)对棉苗立枯病、炭疽病的防治效果的研究。通过室内活性测定及田间试验,证明拌种灵对立枯病具有相当或略高于常用药剂五赛合剂(五氯硝基苯+赛力散)、稻脚青(甲基砷酸锌)的效果,而拌种双除保持了对立枯病与拌种灵相当的效果外,对炭疽病的防效与五赛合剂相近,略低于稻脚青。使用拌种灵或拌种双的棉籽发芽率近于未处理的对照,后期长势良好。拌种灵对立枯病效果优于拌种双,而拌种双对炭疽病效果优于拌种灵。     

几种杀菌剂在四川的使用情况

为了不断开发更新杀菌剂农药品种,提高药剂防治农作物病害效果,近年来我省对近40个国内外杀菌剂新品进行了田间试验示范,筛选出一批对我省主要病害防效好的新品种,已大面积推广应用的有:三环唑、克瘟灵,速克灵、多菌灵3号、叶枯宁、甲霜铜、多效霉素、特克多、井岗霉素高浓度粉剂等。     

新杀菌剂“噻菌铜”的市场前景和发展潜力

噻菌铜,化学名称为2-氨基-5-巯基-1,3,4-噻二唑铜,英文名称为Thiodiazole-copper,中文商品名称为“龙克菌”,属于低毒的噻唑类有机铜杀菌剂,在我国的市场发展前景看好。1目前国内外对有效防治农作物细菌性病害的药剂还比较缺乏1.1防治水稻细菌性病害常使用的药剂为叶枯唑。而南京农业大学的研究结果表明,水稻白叶枯病菌易对该药产生抗药性,细菌性条斑病也存在类似的情况,发病后病情得不到有效的控制。1.2防治果树细菌性病害特别是柑橘,我国常用国产和进口的无机铜杀菌剂,如氢氧化铜、氧化亚铜、硫酸铜和氧氯化铜等,用量难以掌握,存在着用量…     

新杀菌剂“噻菌铜”的市场前景和发展潜力

噻菌铜,化学名称为2-氨基-5-巯基-1,3,4-噻二唑铜,英文名称为Thiodiazole-copper,中文商品名称为“龙克菌”,属于低毒的噻唑类有机铜杀菌剂,在我国的市场发展前景看好。1目前国内外对有效防治农作物细菌性病害的药剂还比较缺乏1.1防治水稻细菌性病害常使用的药剂为叶枯唑。而南京农业大学的研究结果表明,水稻白叶枯病菌易对该药产生抗药性,细菌性条斑病也存在类似的情况,发病后病情得不到有效的控制。1.2防治果树细菌性病害特别是柑橘,我国常用国产和进口的无机铜杀菌剂,如氢氧化铜、氧化亚铜、硫酸铜和氧氯化铜等,用量难以掌握,存在着用量…     

2011年我国主要麦区小麦白粉病菌群体对三唑酮和苯锈啶的敏感性

 小麦白粉病(Wheat Powdery Mildew)是我国小麦生产上常发性病害之一^[1]。小麦白粉病的防治主要采用抗病品种和化学药剂,辅之以栽培措施的综合防治技术。由于目前生产上抗病品种相对缺乏,药剂防治成为我国小麦白粉病防治的主要措施之一^[2]。自20世纪80年代以来三唑类杀菌剂一直是我国防治小麦白粉病的主要药剂,由于长期、大范围、单一的使用导致小麦白粉病菌对三唑类杀菌剂的抗药性大大提高。监测结果表明,2009年我国小麦白粉病菌群体对三唑酮的平均抗性水平已经达到56.58倍,抗性频率达到99.09%,其中高抗菌株占49.09%^[3]。目前小麦白粉病菌对三唑酮的抗性形势十分严峻,寻找三唑类杀菌剂的替代药剂成为控制该病害的迫切需求。     

噻枯唑对水稻白叶枯病菌作用机制研究初报

 透射电镜观察,噻枯唑在离体条件下能使水稻白叶枯病菌菌体内形成许多颗粒状物质,并出现大的空泡结构。噻枯唑在浓度较低时能刺激菌体的呼吸作用,对菌体胞外多糖的产生有抑制作用,但对菌体胞外水解酶的活性没有影响。噻枯唑能和金属Cu⁺⁺形成螯合物,但这种螯合作用没有改变噻枯唑的抑菌活性。     

水稻白叶枯病菌对噻枯唑的抗药性监测

采用活体寄主法和离体含药平板法 ,测定了 1999年安徽省滁州市田间水稻白叶枯病菌对噻枯唑的抗药性。结果表明 ,在水稻活体上存在噻枯唑抗药突变体 ,其比例为67.65%,比1997、1998年有较大程度升高 ,但是离体含药平板上未能监测到抗药突变体。     

Involvement of Phosphoglucose Isomerase in Pathogenicity of Xanthomonas oryzae pv. oryzae

ABSTRACT Xanthomonas oryzae pv. oryzae, the causal agent of bacterial leaf blight of rice, was subjected to transposon mutagenesis to generate mutants defective in pathogenicity. A novel mutant 74M913 was attenuated in virulence but retained its ability to cause the hypersensitive response in leaf blight-resistant rice and tomato. Cloning and sequence analysis revealed that the transposon in 74M913 was inserted in a gene homologous to the phosphoglucose isomerase (pgi) gene of X. axonopodis pv. citri. Growth of the mutant in a synthetic medium containing fructose or xylose as a sole carbohydrate source was much reduced, indicating the transposon disrupted pgi function. The interaction between expression of pgi and hypersensitive response and pathogenicity (hrp) genes was investigated because we had demonstrated previously that expression of hrp genes of X. oryzae pv. oryzae is induced in a synthetic medium containing xylose. However, pgi and the hrp gene (hrcU) were expressed independently. This study suggests that PGI is involved in pathogenicity of X. oryzae pv. oryzae.     

PCR-mediated Detection of <Emphasis Type="Italic">Xanthomonas oryzae </Emphasis>pv. <Emphasis Type="Italic">oryzae </Emphasis>by Amplification of the 16S–23S rDNA Spacer Region Sequence

A detection method specific for Xanthomonas oryzae pv. oryzae, the pathogen responsible for bacterial blight of rice, was based on the polymerase chain reaction (PCR) and designed by amplifying the 16S–23S rDNA spacer region from this bacterium. The nucleotide sequence of the spacer region between the 16S and 23S rDNA, consisting of approximately 580-bp, from X. oryzae pv. oryzae, X. campestris pv. alfalfae, X. campestris pv. campestris, X. campestris pv. cannabis, X. campestris pv. citri, X. campestris pv. cucurbitae, X. campestris pv. pisi, X. campestris pv. pruni and X. campestris pv. vitians, was determined. The determined sequences had more than 95% identity. Therefore, a pair of primers, XOR-F (5′-GCATGACGTCATCGTCCTGT-3′) and XOR-R2 (5′-CTCGGAGCTATATGCCGTGC-3′) was designed and found to specifically amplify a 470-bp fragment from all strains of X. oryzae pv. oryzae isolated from diverse regions in Japan. No PCR product was amplified from X. campestris pathovars alfalfae, campestris, cannabis, carotae, cucurbitae, dieffenbachiae, glycines, pisi, pruni, vitians or zantedeschiae, except for pathovars citri, incanae and zinniae. The method could also detect the pathogen in infected rice leaves within 3 hr, at a detection limit of 4×10¹ cfu/ml. Received 17 December 1999/ Accepted in revised form 10 April 2000     

Characterization of phenotypically distinct strains of Xanthomonas axonopodis pv. citri from Southwest Asia

Strains of Xanthomonas axonopodis pv. citri were isolated from Mexican lime (Citrus aurantifolia) trees in several countries in southwest Asia. These strains produced typical erumpent bacterial canker lesions on Mexican lime but not on grapefruit (C. paradisi). Lesions on grapefruit were watersoaked and blister-like in contrast to the typical erumpent lesions seen after artificial inoculation with all described pathotypes of X. axonopodis pv. citri. This group of strains hydrolysed gelatin and casein and grew in the presence of 3% NaCl as is typical of X. axonopodis pv. citri pathotype A. RFLP analyses and DNA probe hybridization assays also gave results consistent with X. axonopodis pv. citri pathotype A. Metabolic fingerprints prepared with the Biolog® system showed similarities as well as differences to X. axonopodis pv. citri pathotype A. In spite of the physiological and genetic similarities to pathotype A of X. axonopodis pv. citri, these strains had no or very little affinity for polyclonal antiserum prepared against any of the reference strains of X. axonopodis pv. citri and also did not react with monoclonal antibody A1, an antibody that detects all strains of pathotype A of X. axonopodis pv. citri. These strains were also insensitive to bacteriophage Cp3 like X. axonopodis pv. citri pathotype A and unlike X. axonopodis pv. citri pathotype B. We conclude that these strains, designated Xcc-A*, represent a variant of X. axonopodis pv. citri pathotype-A with pathogenicity limited to C. aurantifolia. The existence of extensive genotypic and phenotypic variation within pathotype A of X. axonopodis pv. citri was unexpected and further complicates the systematics of this species.     

Insertion sequence- and tandem repeat-based genotyping techniques for Xanthomonas citri pv. mangiferaeindicae

Molecular fingerprinting techniques that have the potential to identify or subtype bacteria at the strain level are needed for improving diagnosis and understanding of the epidemiology of pathogens such as Xanthomonas citri pv. mangiferaeindicae, which causes mango bacterial canker disease. We developed a ligation-mediated polymerase chain reaction targeting the IS1595 insertion sequence as a means to differentiate pv. mangiferaeindicae from the closely related pv. anacardii (responsible for cashew bacterial spot), which has the potential to infect mango but not to cause significant disease. This technique produced weakly polymorphic fingerprints composed of ≈70 amplified fragments per strain for a worldwide collection of X. citri pv. mangiferaeindicae but produced no or very weak amplification for pv. anacardii strains. Together, 12 tandem repeat markers were able to subtype X. citri pv. mangiferaeindicae at the strain level, distinguishing 231 haplotypes from a worldwide collection of 299 strains. Multilocus variable number of tandem repeats analysis (MLVA), IS1595-ligation-mediated polymerase chain reaction, and amplified fragment length polymorphism showed differences in discriminatory power and were congruent in describing the diversity of this strain collection, suggesting low levels of recombination. The potential of the MLVA scheme for molecular epidemiology studies of X. citri pv. mangiferaeindicae is discussed.     

新型含嘧啶环的半乳糖苷类衍生物的合成及杀菌活性

为寻找具有更高杀菌活性的含氮杂环化合物,合成了13个未见文献报道的新型含嘧啶环的半乳糖苷类衍生物,其结构经过核磁共振氢谱 (1H NMR)、碳谱 (13C NMR) 和高分辨质谱 (HRMS) 等方法确证。杀菌活性测试结果表明:该类化合物对马铃薯晚疫病菌Phytophthora infestans、小麦赤霉病菌Gibberella zeae、水稻纹枯病菌Thanatephorus cucumeris、葡萄座腔病菌Botryosphaeria dothidea和拟茎点霉菌Phompsis sp. 均具有较好的杀菌活性,其中化合物 4m 对马铃薯晚疫病菌的EC₅₀值分别为8.67 μg/mL,与对照药剂烯酰吗啉 (EC₅₀值8.34 μg/mL) 活性相当。此外,该类化合物对柑橘溃疡病菌Xanthomonas citri subsp. citri和水稻白叶枯病菌Xanthomonas oryzae pv. oryzae也有一定的杀菌活性。     

Survival of Xanthomonas axonopodis pv. citri in Leaf Lesions Under Tropical Environmental Conditions and Simulated Splash Dispersal of Inoculum

ABSTRACT Asiatic citrus canker (ACC) is a severe disease of several citrus species and hybrids in many tropical and subtropical areas. Populations of Xanthomonas axonopodis pv. citri in leaf and twig lesions are the most important inoculum source for secondary infections. In areas with a marked winter season (e.g., Argentina and Japan), low temperatures induce a decrease of 10(2) to 10(4) in population sizes in lesions, thus creating a discontinuity in the X. axonopodis pv. citri life cycle. The purpose of this study was to evaluate the dynamics of X. axonopodis pv. citri populations in leaf lesions exposed to the mild winter temperatures prevailing in a tropical environment. Internal X. axonopodis pv. citri population levels in Mexican lime leaf lesions reached 10(6) to 10(7) CFU lesion(-1) whatever the lesion size. These densities, however, were not strongly negatively affected by winter temperatures prevailing under experimental conditions. The estimated decrease in internal X. axonopodis pv. citri population sizes was approximately 10-fold. When exposed to 35 mm h(-1) of simulated rainfall, internal population sizes decreased over time by approximately 1 log unit for lesions 1 and 2 months old, but did not for older lesions. A microscopic examination indicated that lignin-like compounds are present in lesions more than 6 months old. The slow decrease over time of X. axonopodis pv. citri population sizes in leaf lesions may be the balanced result of defense reactions by the host at late stages of disease development, and the concomitant multiplication of the pathogen at the margin of old lesions. We conclude that the epidemiological significance of overwintered leaf lesions in the tropics is higher than that reported in other areas.     

在水稻悬浮细胞系中非亲和白叶枯病菌繁殖受到抑制的现象及其机理的研究

 研究了水稻白叶枯病菌在水稻悬浮细胞系中的繁殖动力学特征以及悬浮细胞胞外液中各类物质对白叶枯病菌繁殖的影响。结果表明,水稻-白叶枯病菌非亲和互作与亲和互作相比病原菌繁殖的数量明显减少。胞外液中粗提蛋白对白叶枯病菌繁殖不具有抑制作用。非亲和互作胞外液中二元酚含量显著增高,并且水稻细胞防卫反应基因的转录和翻译活性明显增强。表明非亲和互作中白叶枯病菌繁殖受到抑制与水稻细胞防卫反应的启动相关。