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)。     

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。     

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...     

一个主栽粳稻品种的不同种植方式与稻纹枯病拮抗细菌的消长及种类的关系

 以晚粳品种丙96-42为试材对杭州临平镇条播、撒播和移栽方式下水稻纹枯病拮抗细菌的群体变化及种类进行了研究，从水稻不同生育期采集叶片、叶鞘和谷粒样本810份，获得细菌菌株7193个。结果显示，在每种种植方式下，稻株上的拮抗细菌数量都是叶鞘中最多，谷粒中最少。3种种植方式下水稻纹枯病拮抗细菌的群体数量基本上是随着稻株成熟而逐渐下降，在水稻6个生育期中，拮抗细菌出现的高峰因不同种植方式而有所差异。条播稻上拮抗细菌占细菌总量的比率从苗期到蜡熟期维持在3.6%～4.8%，各生育期之间无显著差异，但撒播和移栽稻上拮抗细菌所占的比率因不同生育期而有所不同。从539份纹枯病拮抗细菌中鉴定出8个属的14个种，不同种植方式与水稻纹枯病拮抗细菌的种类无明显差异。     

用于生物防治水稻纹拮病的抗细菌的筛选

 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(r²=0.33，P=0.01)， 与抑菌圈大小(r²=0.3)和离体叶片病斑大小(r²=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(r²=0或0.1)。     

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。     

香蕉内在拮抗细菌研究Ⅰ——菌株B215的分离及分子鉴定

在香蕉枯萎病(Fusarium oxysporum f.sp.cubense,FOC)重病园区,从生长正常的香蕉假茎内分离获得1株细菌,编号为B215.经对峙培养和孢子萌发抑制测定,B215对FOC和香焦其他几种主要病原菌的菌丝生长和孢子萌发具有良好抑制效果.对峙培养明显抑制香蕉枯萎病菌菌丝生长,抑菌带宽度0.5-0.6 cm.菌株B215培养液滤液使香蕉枯萎病菌孢子和菌丝生长点膨大成球状畸形,原生质外泄,细胞崩溃十瘪,其EC50为4.12%.形态学、生理生化试验显示B215为芽抱杆菌(Bacillus);进一步对该菌株做16srDNA序列测定与分析,表明其与已报道的枯草芽孢杆菌(Bacillus subtilis Strain KL-073)具有100%的同源性,二者所构建的系统发育树上处于同一个分支.将B215鉴定为枯草芽孢杆菌(B.subtilis).     

水稻纹枯病菌拮抗菌B34分离鉴定及杀菌蛋白的获得

从水稻植株上分离到1株对水稻纹枯病菌有较好拮抗作用编号为B34的菌株,室内对水稻纹枯病菌的抑菌带达到10.3 mm。经生理、生化检测及细胞壁化学成分分析,确定其为致黄假单胞菌（Pseudomonas aureofaciens）。其代谢粗提物经蛋白酶作用,拮抗作用消失。经70%饱和度的（NH4）2SO4沉淀,透析后得到粗杀菌蛋白,分别上样SephadexG-100和DEAE纤维素32LKB柱层析系统,分离纯化了杀菌蛋白,经SDS-PAGE检测其分子量约为33.0 kDa,经低温、真空干燥,得到了该杀菌蛋白的结晶体。     

油菜菌核病拮抗细菌的筛选和高效菌株的鉴定

从油菜根际和叶围分离得到320个细菌分离物,在马铃薯葡萄糖琼脂培养基上的拮抗实验中,18个分离细菌表现对油菜菌核病菌不同程度的拮抗作用,其中Y1菌株对油菜菌核病菌菌丝的生长具有明显的抑制作用.对Y1进行油菜离体叶片、温室盆栽和田间小区接种实验,该菌均表现对油菜菌核病明显的防病效果;在温室盆栽试验和田间小区试验中,防病效果达到92%.经过鉴定,Y1菌株为枯草芽孢杆菌.     

Gram-Positive Bacteria Associated with Rice in China and Their Antagonists Against the Pathogens of Sheath Blight and Bakanae Disease in Rice

Rice (Oryza sativa L.) is one of the most important crops, which feeds half of the population in the world. It’s the principle of the Integrated Pest Management (IPM) to maintain the pest damage under an economical permitted level by means of resistant cutivars and biological control. Some of the bacteria from rice plants are pathogenic [1], but most of them are nonpathogenic ones, which provide abundant resources for the biological control to rice diseases. It is a first key step of biolo…     

水稻革兰氏阳性细菌的主要种群结构及对纹枯病和恶苗病菌的拮抗性

于1998~2004年对采自浙江、江苏、福建和云南省的756份稻株和稻种样本进行了革兰氏阳性细菌的分离、鉴定研究。被分离的1015个菌株经致病性测定、菌落形态及部分细菌学特征（革兰氏染色、KMB培养基上的荧光色素及芽孢的染色镜检等）测定后,选出代表菌株74个连同5个对照菌株用Biolog及脂肪酸分析法(FAME)进行测试, 鉴定出Bacillus属5个种及其他3属的革兰氏阳性细菌,并发现枯草芽孢杆菌和巨大芽孢杆菌菌株具有很好的纹枯病和恶苗病拮抗能力,但来自短小芽孢杆菌和巨大芽孢杆菌的极少数菌株在条件适宜时能与其他病原菌一起引起水稻褐斑,从这些种筛选生防菌株时应充分考虑其风险。     

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.     

Resistance Evaluation to Sheath Blight in Transgenic Rice Lines

Resistance of forty-one homozygous rice lines transformed with chitinase gene (RC24) and β-1,3 -glucanse gene (β-1,3-Glu) to sheath blight was analyzed by inoculation. Among different lines, the resistance had significant differences according to the result by cluster analysis. The lines could be categorized into resistant, moderately resistant, moderately susceptible and susceptible types, while 92.1% of which belonging to moderately resistant or moderately susceptible type. For different resistant or susceptible lines, the resistance to rice sheath blight was remarkable correlated with the chitinase activity of transgenic lines except resistant type lines, in which enzyme activity coded by target gene was lower than moderately resistant type. The chitinase activity of transgenic lines tested at different time after inoculation or different organs of the same plant was uniform, which suggested that the expression of chitinase gene was constitutive in nature. Check varieties‘ chitinase activity would change at different time after inoculation and reach a peak at sometime, but it had no difference at various parts of the same plant.     

不同药剂不同施药次数防治水稻纹枯病田间药效试验

通过田间试验,比较分析了不同药剂对水稻纹枯病的防治效果。结果表明,25%嘧菌酯悬浮剂1 350 g/hm~2、24%噻呋酰胺悬浮剂345 g/hm~2、25%丙环唑乳油600 g/hm~2对水稻纹枯病的防效均在87%以上,43%戊唑醇悬浮剂的防效为74.38%;4种药剂的田间药效比生产上常用药剂井冈霉素的防效分别提高0.79倍、0.75倍、0.69倍、0.43倍,头季和再生季每hm~2综合增收6 499元、4 042元、2 051元、1 913元。从防治效果和经济效益来看,25%嘧菌酯悬浮剂、24%噻呋酰胺悬浮剂、25%丙环唑乳油均可以作为本地防治水稻纹枯病的主体药剂进行推广应用。     

水稻纹枯病时间与空间生态位的研究

测定了水稻纹枯病时间、垂直空间和水平面空间生态位的生态位功能指数和生态位宽度指数,结果表明,在倒1叶上水稻纹枯病的时间生态位宽度指数最小,为0.1994,在倒4叶上的生态位宽度指数最大,为0.5463,整体上随着叶（鞘）位的降低,水稻纹枯病的时间生态位宽度指数增大。水稻纹枯病垂直空间生态位宽度指数在6月22日最小,为0.1250,在8月22日最大,为0.7759,整体上随着水稻的生长,水稻纹枯病的垂直空间生态位宽度指数逐渐增大。各调查时期水稻纹枯病的水平空间生态位宽度指数差异不大。讨论了生态位、生态位功能、生态位宽度等概念的内涵。     

Resistance Evaluation to Sheath Blight in Transgenic Rice Lines

Resistance of forty-one homozygous rice lines transformed with chitinase gene (RC24) and β-1,3 -glucanse gene β-1,3-Glu) to sheath blight was analyzed by inoculation. Among different lines, the resistance had significant differences according to the result by cluster analysis. The lines could be categorized into resistant, moderately resistant, moderately susceptible and susceptible types, while 92.1% of which belonging to moderately resistant or moderately susceptible type. For different resistant or susceptible lines, the resistance to rice sheath blight was remarkable correlated with the chitinase activity of transgenic lines except resistant type lines, in which enzyme activity coded by target gene was lower than moderately resistant type. The chitinase activity of transgenic lines tested at different time after inoculation or different organs of the same plant was uniform, which suggested that the expression of chitinase gene was constitutive in nature. Check varieties chitinase activity would change at different time after inoculation and reach a peak at sometime, but it had no difference at various pans of the same plant.