稻瘟病拮抗菌NK413的鉴定及抑菌效果

菌株NK413是从黑龙江省农田土壤中分离得到的一株放线菌,为明确其分类地位及在植物病害生物防治中的应用潜力,采用多相分类法对其进行鉴定,并采用孢子萌发抑制法、菌丝生长速率法、杯碟法考察了NK413菌株及其代谢产物对稻瘟病菌的抑菌效果及作用方式。结果表明:菌株NK413的形态特征、生理生化特征与小白链霉菌(Streptomyces albulus)接近,细胞壁类型为Ⅰ型、磷脂脂肪酸成分符合链霉菌属特征。16S rDNA全长为1 614 bp,与小白链霉菌亲缘关系最近,因此将NK413定名为小白链霉菌(S.albulusNK413)。NK413对稻瘟病菌孢子萌发和菌丝生长的抑制作用较强,原液对稻瘟病菌的抑菌圈直径达30 mm,无菌发酵滤液稀释1倍对孢子萌发的抑制率为100%,稀释5倍时对菌丝生长的抑制率为100%。抗菌谱广,具有较好的开发前景。     

A single amino acid substitution in the SdhB protein of succinate dehydrogenase determines resistance to amicarthiazol in Xanthomonas oryzae pv. oryzae

BACKGROUND: Xanthomonas oryzae pv. Oryzae Ishiyama, a causal agent of rice bacterial leaf blight, was found to be sensitive in vitro to the systemic fungicide amicarthiazol (2‐amino‐4‐methylthiazole ‐5‐carboxanilide), which is a potent inhibitor of succinate dehydrogenase (SDH, EC 1.3.99.1). This paper aimed to determine the molecular resistance mechanism of X. oryzae pv. oryzae to amicarthiazol. RESULTS: UV‐induced resistant mutants of X. oryzae pv. oryzae to amicarthiazol were isolated. The activity of SDH in wild‐type X. oryzae pv. oryzae was strongly inhibited by amicarthiazol, while that in resistant mutants was insensitive, although their SDH activity was decreased compared with the wild‐type sensitive strain without amicarthiazol. A mutation of Histidine₂₂₉ (CAC) to Tyrosine₂₂₉ (TAC) was identified in sdhB, which encoded the iron–sulfur protein subunit of SDH. The sdhB from the mutant was ligated into a cosmid, pUFR034, to generate pUFR034RAX, which conferred resistance to amicarthiazol when transformed into the wild‐type sensitive strain. CONCLUSION: A mutation of His₂₂₉ (CAC) to Tyr₂₂₉ (TAC) in SdhB was responsible for determining amicarthiazol resistance. Copyright © 2010 Society of Chemical Industry     

Efficacy of vaporised ethyl formate/carbon dioxide formulation against stored‐grain insects: effect of fumigant concentration,exposure time and two grain temperatures

BACKGROUND: The ethyl formate/carbon dioxide (CO₂) formulation Vapormate? is a rapid‐acting fumigant being developed for the control of stored‐grain insects. The effects have been investigated of concentration, exposure times of 1, 3, 24 and 72 h and two grain temperatures, 15 and 25 °C, on its efficacy against mixed‐stage cultures of Sitophilus oryzae (L.) Tribolium castaneum (Herbst) and strongly phosphine‐resistant Rhyzopertha dominica (F.) strain QRD569. RESULTS: High mortalities (≥92%) of mixed‐stage cultures of all three species were obtained when grain was fumigated with the formulation (193 g m^?3 ethyl formate) for 1 h. Complete control of R. dominica QRD569 and T. castaneum was achieved with 63 and 76 g m^?3 ethyl formate respectively, with exposure for 24 h, whereas mean mortality of S. oryzae was 86% under the same conditions. Mortalities of S. oryzae juvenile stages were significantly lower than adults under the conditions tested, which was due to pronounced tolerance of mid‐stage pupae to the fumigant. Reducing grain temperature from 25 to 15 °C had no effect on insect mortality. CONCLUSION: Ethyl formate/CO₂ formulation is highly effective against stored‐grain insects over a range of concentrations and exposure times. Efficacious fumigations were conducted in as little as 1 h, and a strongly phosphine‐resistant R. dominica strain was readily controlled with the fumigant. Copyright © 2009 CSIRO, Australia. Published by John Wiley & Sons, Ltd     

水稻条斑病菌avrBs3/pthA家族基因敲除体系的建立

水稻细茵性条斑病菌存在至少20个avrBs3/pthA家族成员,但其在水稻上的毒性或无毒性贡献并不清楚.本文依据avrBs3/pthA结构上的保守性,利用同源重组策略,借助自杀性载体pKMS1介导,对水稻条斑病菌中的avrBs3/pthA家族基因进行敲除.结果发现:同源交换可通过基因内和基因间重组实现avrBs3/pthA家族的基因敲除;PCR和Southern杂交结果显示,水稻条斑病菌中有5个avrB5s3/pthA家族基因被成功敲除,表明水稻条斑病菌avrBs3/pthA家族基因敲除体系构建成功.这为逐一评价avrBs3/pthA 家族基因的毒性或/和无毒性功能奠定了基础.     

米曲霉和黑曲霉双菌种制曲对豆酱酶系影响的研究

以黄豆和面粉为主要原料,以淀粉酶、蛋白酶、β-葡萄糖苷酶和纤维素酶的活性为指标,采用单因素试验方法,研究米曲霉和黑曲霉(质量比为1:1)对豆酱制曲酶系的影响,优化制曲工艺参数。试验结果表明,采用添加熟面粉的方式,制曲时间为36h,制曲温度为29～32℃,接种量0.04%,双菌种曲中酶的活性高于单一米曲霉曲。     

水稻白叶枯病菌avrBs3/PthA家族基因的随机缺失及突变体PXO99Δavr功能的研究

利用来源于水稻白叶枯菌株JXOⅢ的无毒基因avrXa3构建的同源序列突变转化单元,同源重组得到PXO99~A菌株的1个缺失突变体PXO99△avr。PXO99~A及其突变体基因组DNA以内切酶BamHⅠ完全消化后进行同源检测,发现突变体PXO99△avr中有3条杂交带消失,有2条带信号减弱。进一步以相同探针检测PXO99~A的基因文库,限制性酶切分析和Southern杂交归类,共获得13个不同的阳性克隆。众多avrBs3/PthA基因在基因组中单独存在,或2个和2个以上串联排列。经比对,克隆p5所含条带与突变体缺失的5条带大小相同。因此认为随机交换产生的突变体中有5个基因被敲除。将此文库克隆(p5)分别导入PXO99~A和突变体PXO99△avr中,苗期接种试验表明:p5互补突变体能使其毒性恢复接近PXO99~A的致病表型,推测此克隆正是突变体缺失的基因。同时也表明:缺失的5个串联基因的综合作用主要表现为毒性功能。     

水稻稻瘟病拮抗细菌WH1G的筛选鉴定及其抑菌活性

采用平板对峙法,从安徽水稻根际土壤样品中筛选对稻瘟病菌具有较强抑菌活性的拮抗细菌,通过菌落形态观察、生理生化特征、gyrB基因序列分析对该菌株进行鉴定,测定抑菌谱,对其抑菌机制进行初步研究。结果表明,拮抗菌株WH1G对稻瘟病菌具有较强的抑制作用,抑制率达91.79%,经鉴定为解淀粉芽胞杆菌(Bacillus amyloliquefaciens)。该菌可产生蛋白酶和纤维素酶,不产生几丁质酶和β-1,3-葡聚糖酶;用代谢产物无菌滤液处理稻瘟病菌,可造成孢子萌发率降低,菌丝扭曲异常、分支明显减少并产生大量泡囊。该菌对常见的20种植物病原真菌及细菌均具有不同程度的拮抗作用,表现出广谱抗菌活性。     

水稻负泥虫对不同水稻品种次生物质的行为反应

本研究以寒地稻田优势害虫水稻负泥虫为研究对象,选取黑龙江省主栽水稻品种提取次生物质,检测了其对水稻负泥虫的驱避作用、拒食作用和产卵驱避作用。结果表明,感虫品种‘稻花香2号’的挥发性物质和非挥发性物质对水稻负泥虫的驱避性弱于‘绥粳4号’;虫害胁迫植株产生的次生物质对水稻负泥虫成虫的驱避性最强,而健康植株的驱避性最弱。两种水稻品种次生物质对水稻负泥虫幼虫和成虫的拒食率均显著高于对照,且抗虫品种的拒食率高于感虫品种。水稻负泥虫成虫在两种水稻品种次生物质作用下,平均卵量与对照相比均减少,但是感虫水稻品种相对比抗虫水稻品种的选择性产卵驱避率低。由此可见,水稻不同品种之间的次生物质的组分和含量不同,导致了昆虫行为反应有差异,为进一步研究和分析抗感水稻品种次生物质的组分、含量及作用提供理论依据。     

Aerial Spraying Trials in West Africa for Blackfly Control

Abstract

Organochlorine, organophosphate (o.p.) and carbamate insecticides have been extensively used in Australia to combat Lucilia cuprina Wied. the main initiator of fly-strike of sheep occurring wherever sheep are run on the continent. The improved mules operation, mid-season crutching and insecticides are important in the management of Merino sheep. Insecticides are particularly valuable against body strike, especially in young sheep, in years with intermittent rain during the warmer months. Resistance to larvicides developed in the blowfly over a period of 10y from 1957. The time between the introduction of various insecticides and development of resistance is considered and a comparison made between the emergence of resistance problems to the newer synthetic insecticides in the housefly in Denmark, and in the blowfly in Australia. To provide a complete history of insecticides against the blowfly reference is made to arsenicals, to which there was no suspicion of resistance until low order cross resistance was diagnosed in o.p.-carbamate resistant strains. Some seven years after their introduction DDT and γ BHC were replaced by cyclodiene insecticides in 1954/1955. Reasons are advanced to explain the non-emergence of resistance to DDT and BHC in that period. Resistance to dieldrin and aldrin developed in late 1957 after which diazinon was introduced. The resistance has a typical BHC/dieldrin resistance spectrum and is due to a semi-dominant gene which has persisted in the field in the absence of pressure from cyclodiene insecticides. Non-specific resistance to o.p. insecticides developed in two steps. Low order resistance, diagnosed in 1965, was supplemented by an additional resistance mechanism in 1966, three alleles on two chromosomes are involved. The carbamate, butacarb, was effective against o.p. resistant strains of the blowfly when introduced in the 1966/67 season. In 1967, resistance to butacarb was diagnosed and rapidly became widespread. For the past three fly seasons larvae have been used to monitor resistance levels to o.p. and carbamate insecticides; resistance factors to both these classes of insecticide are significantly higher in larvae than adult females. Resistance levels to o.p. insecticides have stabilised. By contrast resistance levels to butacarb have doubled. A combined o p.-carbamate resistance generally applies in field samples. Larvae from o.p.-carbamate resistant strains form artificial strikes earlier than susceptible larvae, particularly on sheep treated with butacarb. It is concluded that registered o.p. insecticides, but not butacarb, will still give considerable protection against fly-strike, providing the maximum levels of o.p. resistance, reached in laboratory selection programmes, are not exceeded in the field. Investigation into o.p. resistance in the species suggests that the resistance mechanisms place their carriers at a disadvantage in the absence of selection pressure. Measures to minimise the amount of insecticide used against the blowfly are therefore strongly advocated—these include: the improved mules operation, mid season crutching and good animal husbandry.