用于生物防治水稻纹枯病的拮抗细菌的筛选

分别从健康的水稻植株、感染纹枯病植株、根际土壤、秧田水和菌核上分离出能拮抗水稻纹枯病的假单胞菌属(Pseudomonas spp.)和芽胞杆菌属(Bacillus spp.)的细菌菌株。大多数菌株在室内试验中对菌丝生长有抑制作用,有的菌株在温室试验中起抑制病害和对植株有保护作用,有的(1/49)具有促进秧苗生长的作用。纹枯病的病斑相对长度与病害发生率密切相关(r~2=0.33,P=0.01),与抑菌圈大小(r~2=0.03)和离体叶片病斑大小(r~2=0.12)无相关,植株干重与病斑相对长度和发病率均无显著相关(r~2=0和0.01)。     

水稻纹枯病拮抗细菌7-5的鉴定及其生防机制初步研究

【目的】旨在获得对水稻纹枯病有生防效果的拮抗细菌,探索生防细菌作用机理。【方法】通过苗期防病试验从本实验室保存的7株拮抗细菌中筛选对水稻纹枯病有较好防病效果的菌株。结合生理生化性状和16S rDNA基因序列同源性分析,对生防细菌7-5进行初步鉴定,并通过PCR扩增、薄层层析等探讨7-5产生抗生素的种类。【结果】供试7株细菌中7-5、4-74和4-78对水稻纹枯病防效分别可达69.0%、70.8%和75.4%。其中4-74和4-78抑菌谱窄,7-5对水稻纹枯病菌、桃褐腐病菌和草莓灰霉病菌都具有较强抑菌能力。菌株7-5 鉴定为绿针假单胞菌;该菌株可产生吩嗪-1-羧酸、嗜铁素、氢氰酸、蛋白酶、藤黄绿脓菌素和2,4-二乙酰基间苯三酚等抗菌物质,不产生硝吡咯菌素、几丁质酶和纤维素酶。【结论】绿针假单胞菌7-5主要通过产生抗生素防治水稻纹枯病。     

拮抗细菌的分泌物拮抗素JK-91-b对水稻纹枯病菌抗生活性试验

拮抗细菌的分泌物拮抗素ＪＫ—９１—ｂ对水稻纹枯病菌抗生活性试验陈志谊，高先亭，陈毓苓，殷尚智（江苏省农业科学院，南京２１００１４；）（江苏农学院，扬州２２５００１）关键词拮抗素；抗生活性；水稻纹枯病ＡＳｔｕｄｙｏｎｔｈｅＥｘｕｄａｔｅｏｆＡｎｔｉｂｉ...     

水稻纹枯病拮抗细菌的评价与利用

 从稻田土壤、纹枯菌菌核、病斑和水稻植株上共分离获得1437个细菌分离物。在离体条件下测定它们对水稻纹枯菌的拮抗能力。其中拮抗能力强的菌株61个,占4.8％;没有拮抗作用的菌株有486个,占38.1％;其余57％的菌株是略有拮抗作用。24个菌株对水稻纹枯病的防治效果的盆栽生物测定表明,其中有9个菌株的防效在60％以上。挑选了6个菌株在田间进一步测试其防效,结果证实了B916对水稻纹枯病的防治效果最好。1994～1997年连续4年在姜堰、吴江和句容3个基点示范推广使用B916发酵液防治水稻纹枯病,累计面积为2000 hm2以上。大田示范试验结果表明,拮抗菌B916发酵液能够有效地控制水稻纹枯病的为害,防效为50.0%～81.9%。     

拮抗细菌Bacillus subtilis B5423-R 抑制水稻纹枯病的阈值群体数量

 在菲律宾国际水稻研究所实验农场田间条件下，通过人工接种Rhizoctonia solani,采用不同的方法(喷雾次数、间隔期)喷雾B5423-R（B5423的标记菌株，为利福平Rifampicin抗性的突变体），在接种1 d后，采用2 d喷1次、连续3～4次喷雾的方法，使B5423 R在稻株上的群体数量在6～7 d内维持在1×10[sup]6[/sup]cfu/g以上，在25 d内显著地降低相对病斑高率；而采用5 d喷1次、连续3～4次喷雾的方法，B5423-R在稻株上的群体数量在15 d内维持在1×10[sup]5[/sup]～106cfu/g，在25 d内相对病斑高率未能显著降低。表明在纹枯病侵染早期的6～7 d内，在稻株上B5423 R维持在1×10[sup]6[/sup]cfu/g以上是抑制纹枯病的阈值群体数量。     

在水稻生态系统中纹枯病拮抗细菌的分布和种类

 通过对从菲律宾Laguna省水稻植株上分离获得的700个细菌菌株进行离体与生物测定发现,在水稻生态系统中,存在着大量格兰氏阴性、非致病性、对纹枯病有拮抗作用的细菌微生物。根据拮抗细菌分离来源分析结果表明,从病株和水稻生长后期能分离到比健株和生长前期更多的拮抗细菌。用Biolog的方法测定了195个细菌菌株的种类。在147个格兰氏阴性菌株中有13个属39个种,在48个格兰氏阳性菌株中有8个属13个种。在195个被测菌株中有20个菌株对纹枯病菌具有较强的拮抗作用,它们分别归属于7个属12个种。     

5个水稻品种对水稻纹枯病菌鉴别能力的比较

采用苗期接种法和大田成株期接种法,对5个具有不同纹枯病抗性的水稻品种Lemont、武育粳3号、Jasmine85、C418和YSBR1进行纹枯病抗性评价。苗期接种结果表明,30个来源不同的纹枯病菌接种5个品种后,5个品种苗期表现出明显的抗感差异,30个菌株之间致病力差异亦明显,且品种与菌株之间互作显著。根据聚类分析结果,从30个菌株中筛选出5个致病力不同的菌株(分别为GD118、C30、E67、YN7和YN3)进行水稻品种成株期抗性鉴定,发现水稻品种成株期的抗感差异亦显著。     

白叶枯病菌拮抗菌筛选及水稻叶围微生物互作研究初报

 1998年7月～10月, 于田间汕优63水稻叶片上分离采集微生物, 并进行叶围微生物鉴定,以筛选白叶枯病拮抗菌。结果表明:除极少量分布的真菌和细菌外, 主要种群包括真菌类群Alternaria和 Fusarium属; 细菌类群Flavobacterium, Xanthomonas, Pseudomonas和Bacillus属。对白叶枯病菌（Xanthomonas oryzae pv. oryzae）具有明显拮抗作用的微生物为Bacillus属,而且Bacillus属细菌对其余大部分叶围微生物具有程度不同的拮抗作用,可以初步断定其产生的拮抗物质具有一定的广谱性。     

水稻纹枯病菌拮抗芽孢杆菌的筛选及鉴定

为充分发掘利用水稻田已有的有益微生物资源,获得水稻纹枯病菌(Rhizoctonia solani)的拮抗芽孢杆菌菌株,从四川农业大学水稻试验田中采集根际土样,采用稀释平板法在LB培养基上分离得到96株芽孢杆菌菌株。采用平板对峙法,将水稻纹枯病菌标准菌株R.solani AG1 IA与待测芽孢杆菌菌株共培养,得到1株对R.solani AG1 IA具有强拮抗作用的芽孢杆菌菌株Q9-0-1,并以形态学特征、生理生化特性为基础,结合gyr B基因序列分析,将其鉴定为短小芽孢杆菌(Bacillus pumilus)。     

禾长蠕孢菌及其代谢产物Ophiobolin A防治水稻纹枯病

 为充分利用已有杂草生防菌菌种资源，进行了稗草生防菌禾长蠕孢菌(Helminthosporium gramineum Rabenh f. sp.echinochloae，HGE）防治水稻纹枯病的初步研究。菌落对峙实验结果表明HGE对纹枯病菌菌丝生长有较强的抑制作用。HGE菌株的发酵液对纹枯病菌的生长有较强的抑制活性，稀释5倍的发酵液24 h的抑菌率为84.1％。发酵液及菌丝的乙酸乙酯提取粗毒素在200 μg/mL浓度下100％抑制纹枯病菌的生长。从粗毒素中分离纯化的有很强抑菌活性的化合物经过NMR及MS波谱分析鉴定为Ophiobolin A。Ophiobolin A在25 μg/mL浓度下即100％抑制纹枯病菌的生长，浓度1 μg/mL时的抑菌率为70.2%。实验结果显示HGE菌及其代谢物Ophiobolin A具有开发成水稻纹枯病生物杀菌剂的潜力。     

新型杀菌剂对马铃薯黑痣病病菌的室内毒力测定和田间效果分析

本研究采用生长速率法,在室内条件下测定了20%甲基立枯磷乳油、24%噻呋酰胺悬浮剂、2.5%咯菌腈悬浮种衣剂、25%吡唑醚菌酯乳油4种新型杀菌剂对立枯丝核菌(Rhizoctonia solani)的毒力。结果表明,4种杀菌剂对立枯丝核菌都有一定的抑制作用,其中,20%甲基立枯磷乳油对立枯丝核菌的抑制作用最强,EC50值为6.9888×10-9 mg/L;24%噻呋酰胺悬浮剂抑制作用次之,EC50为6.6995×10-7 mg/L;2.5%咯菌腈悬浮种衣剂和25%吡唑醚菌酯乳油的抑制作用稍差,EC50分别为7.6599×10-5 mg/L、1.0060×10-4 mg/L。田间防治黑痣病药效试验表明,4种药剂对马铃薯安全,其中20%甲基立枯磷乳油的防治最好,其次为24%噻呋酰胺悬浮剂,2.5%咯菌腈悬浮种衣剂、25%吡唑醚菌酯乳油防治效果较低,这与室内测定结果一致,因此生产上可用20%甲基立枯磷乳油20 mL拌种100 kg、24%噻呋酰胺悬浮剂每667 m2沟施100 mL来防治马铃薯黑痣病。     

辽宁花生纹枯病病原鉴定及其生物学研究

本文对辽宁花生纹枯病病原学进行了较为系统的研究。根据病原菌形态学特征及致病性测定,明确辽宁花生纹枯病病原菌为立枯丝核菌(Rhizoctonia solani)。病原菌生物学特性研究发现该菌最适pH值为7.0;该菌能利用供试的6种碳源和5种氮源进行营养生长,在尿素作为唯一氮源的培养基上菌丝不生长并且不产生菌核,其余碳氮源培养基均产生菌核;该菌在玉米粉和PDA培养基上生长最好;光照对菌丝的生长及菌核的产生影响不大。     

5种杀菌剂对马铃薯黑痣病的病菌毒力及田间防效

马铃薯黑痣病(Rhizoctonia solani)是一种土传病害,危害呈逐年加重趋势。本研究进行了5种杀菌剂对病菌的毒力测定和病害田间防效试验。结果表明:抑霉唑、嘧菌酯抑菌作用较好,EC50分别为4.46μg·mL-1、8.32μg·mL-1,相关系数r均大于0.91。5种药剂在田间防病中,噻氟菌胺沟施浓度过高对马铃薯出苗产生药害,相对出苗率为92.3%,与对照有显著性差异;嘧菌酯、噻氟菌胺、百菌清、抑霉唑沟施和拌种对地中茎防病效果均在83%以上;抑霉唑(沟施)、嘧菌酯(沟施和拌种)、百菌清(沟施)对预防因病造成植株死亡效果较好,防效分别为71.6%、69.3%、67.4%、67.3%;嘧菌酯预防薯块带菌效果最好,沟施和拌种防病效果分别为90.7%、86%。     

根系内生细菌与玉米抗纹枯病关系研究

通过对辽宁省14个玉米主栽品种抗性和内生细菌相互关系的研究,明确了苗期不同品种间内生细菌总量差异显著性达到α=0．05水平,而抽丝期品种间差异不显著,苗期与成株期的互作差异亦不显著;品种苗期与成株期内生细菌种群总量不相关;不同品种苗期和成株期纹枯病抗性均存在显著差异（α=0．01）;玉米纹枯病品种抗性与内生细菌总量不相关,而与拮抗内生细菌百分率呈显著正相关。     

Efficacy of Bacillus subtilis MBI 600 Against Sheath Blight Caused by Rhizoctonia solani and on Growth and Yield of Rice

Rice sheath blight disease(ShB),caused by Rhizoctonia solani,gives rise to significant grain yield losses.The present study evaluated the efficacy of Integral?,the commercial liquid formulation of Bacillus subtilis strain MBI 600,against rice ShB and for plant growth promotion.In greenhouse studies,four log concentrations of Integral(from 2.2×106 to 2.2×109 cfu/mL) were used as seed treatment(ST).After 25 d,seedlings were dipped(SD) into Integral prior to transplanting.At 30 d after transplanting(DAT),leaf sheaths were inoculated with immature sclerotia of the pathogen.At 45 DAT,a foliar spray(FS) with Integral was applied to some treatments.The fungicide control was 50% carbendazim at 1.0 g/L,and a nontreated control was also included.Overall,there were 10 treatments,each with five replications.ShB severity was rated at 52 DAT,and seedling height and number of tillers per plant were rated at 60 DAT.In 2009,two field trials evaluated Integral at 2.2×108 and 2.2×109 cfu/mL.Integral was applied as ST,and seedlings were produced in a nursery bed.After 32 d,seedlings were treated with Integral as SD and transplanted into 10 m2 blocks.Foliar sprays were given at 45 and 60 DAT.There were seven treatments,each with eight replications arranged as a factorial randomized complete block design.At 20 DAT,the plots were broadcast inoculated with R.solani produced on rice grains.Seedling height before transplanting,ShB severity at 90 DAT,and grain yield at harvest were recorded.Integral at 2.2×109 cfu/mL provided significant increase of seedling heights over other treatments under greenhouse conditions.The Integral treatments of ST + SD + FS at 2.2×109 cfu/mL significantly suppressed ShB over other treatments.In field studies,Integral provided significant increase of seedling height in nursery,and number of tillers per plant,compared with the control.ShB severity was significantly suppressed with higher concentrations of Integral compared to lower concentrations.Grain yield were the highest at an Integral concentration of 2.2×109 cfu/mL.Overall,Integral significantly reduced ShB severity,enhanced seedling growth,number of tillers per plant and grain yield as ST + SD + FS at the concentration of 2.2×109 cfu/mL under the conditions evaluated.     

水稻纹枯病菌6 磷酸葡萄糖胺合成酶基因的克隆、测序及表达分析

 为筛选新型水稻纹枯病菌GlmS活性抑制物质,采用3′RACE和5′RACE克隆了水稻纹枯病菌GlmS的基因组DNA序列和完整的cDNA序列。GlmS基因组DNA序列全长2 529 bp,含有8个内含子;GlmS cDNA序列全长2094 bp,推测编码一个含有697个氨基酸残基,分子量约为76.7  kD的蛋白质。生物信息学分析表明水稻纹枯病菌GlmS含有1个谷氨酰胺氨基转移酶结构域和2个葡萄糖异构酶结构域。采用大肠杆菌重组融合表达GlmS,重组蛋白的分子量经过葡聚糖凝胶层析和SDS PAGE电泳测得分别为306 kD和77 kD,表明GlmS是由4个相同大小亚基组成的多聚酶复合体。重组蛋白的酶学性质研究表明其最适反应温度为37℃,最适pH为6.4,42℃下的半衰期为1 h,在pH 5.5～7.5时比较稳定。GlmS催化反应能被己糖胺通路末端产物鸟苷氮乙酰葡萄糖胺反馈抑制。     

Morphological and Genetical Variability among Rhizoctonia solani Isolates Causing Sheath Blight Disease of Rice

Eighteen isolates of Rhizoctonia solani collected from infected rice plants in four different locations of Bangladesh were studied by using morphological characters and molecular markers. Anastomosis study with a reference isolate confirmed that all the isolates belonged to R. solani. Significant variation was observed in sclerotial size, shape and distribution. Un-weighted pair group method with arithmetic mean dendrogram constructed based on the Gower's general similarity coefficient showed that these isolates were grouped into four clusters at the 0.68 similarity coefficent according to morphological characters. Cluster I was a major cluster consisting of 13 isolates, while clusters Ⅱ to Ⅳ consisted of 1 or 2 isolates. Analyses by variable number of tandem repeat and amplified fragment length polymorphism markers showed that the isolates were grouped into five and three clusters at a similarity coefficient of 0.64 and 0.69, respectively. Although most of the variability was found between isolates from different regions as expected, significant variation was observed within the isolates collected from similar agro-ecological regions. Our results suggest the presence of different races of R. solani within the same local geographic regions.     

水稻纹枯病菌侵染稻株对褐飞虱发生的影响

病原菌侵染引发植株防卫基因表达的信号传导途径水杨酸途径(SA)与虫害(机械损伤)诱发的植物防御信号传导途径茉莉酸途径(JA)会同时存在,且两种信号途径间存在着相互抑制的拮抗作用。本试验结果表明,水稻植株受到纹枯病菌侵染之后,褐飞虱在RH的感病植株和健康植株中明显倾向于选择感染纹枯病菌的RH水稻植株;但在不同水稻品种上的取食行为以及蜜露排泄量及蜜露氨基酸含量的测定均未发现褐飞虱对于感染纹枯病的抗、感水稻品种有显著的取食趋性。说明抗、感褐飞虱的水稻品种感染纹枯病之后并未显现出更易感虫的性状。     

浙江省水稻纹枯病菌的遗传分化与致病力研究

鉴定了从浙江省各地采集的60株水稻纹枯病菌株的菌丝融合群归属情况。该60个菌株均与AG-1融合群的3个亚群AG-1-ⅠA、AG-1-ⅠB、AG-1-ⅠC的标准菌株融合,当属AG-1融合群。其中,有58株均与标准菌株AG-BⅠ融合,所有菌株与其他标准菌株均无融合现象。通过离体接种对菌株致病力进行鉴定,发现各菌株间致病力差异显著。致病力最强的TTZF-1与其中52个菌株差异极为显著。进一步的转录间隔区(ITS)序列分析显示,系统发育树的结果与融合群鉴定的结果一致,与菌株间致病力差异无相关性。     

Genetic Diversity and Pathogenicity Variation in Rhizoctonia solani Isolates from Rice in Sichuan Province, China

Fifty-five representative samples of Rhizoctonia solani isolates, which were collected and isolated from five different ecological regions in Sichuan Province, China, were purified and analyzed for the pathogenicity and molecular genetic variation. The hyphal fusion test revealed that almost all the isolates belonged to the AG-IIA group except the isolate D42. In addition, some of the isolates were ‘bridging isolates’, which could fuse with several groups simultaneously. The pathogenicity analysis on in vit...