枯草芽孢杆菌植入棉苗体内拮抗黄萎病研究

在棉花无土育苗基质中植入枯草芽孢杆菌,使其优先占领生态位,而拮抗黄萎病。结果表明:枯草芽孢杆菌CD102菌液稀释后植入灭菌基质,能在短期内增殖和持续繁殖;采用浸种、浇灌、针刺和喷雾的方法处理基质培育的棉苗,CD102均能不同程度定植其中;带CD102的基质培育棉苗,移栽大田后对黄萎病具有较好防治效果。     

枯草芽孢杆菌CS16菌株抑菌活性物质的提取分析

枯草芽孢杆菌CS16发酵液经盐酸沉淀,甲醇抽提得到粗提物,稳定性试验结果表明,抑菌物质在pH2～11较稳定,对温度、紫外、有机溶剂耐受力较好.利用中压液相色谱得到较纯的抑菌物质,薄层层析后显色分析表明,该抑菌物质初步可以确定为酚类物质.     

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%。     

枯草芽孢杆菌CS16抑菌活性与胞外产物成分分析

通过测定枯草芽孢杆菌CS16菌株的抑菌活性,对其胞外代谢物活性与成分进行分析,以期开发有效的生防菌剂.结果表明:CS16菌株对多株植物病原真菌具有明显的抑制作用,对病原细菌的抑菌作用相对较弱;CS16发酵液中羧甲基纤维素酶、淀粉酶和木聚糖酶活力分别为509.58 U/mL、7 198.71 U/mL和1 853.68 U/mL;其发酵液经酸沉淀所得粗提物可明显抑制香蕉枯萎病菌丝生长,最小抑菌浓度为0.062 5 mg/mL;利用反相硅胶柱、Sephadex LH-20凝胶柱和正相硅胶柱分离纯化获得较纯的抑菌物质,经薄层层析和HPLC定性分析,初步认为其胞外抑菌物质中含有Iturin A.     

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).     

一株源于红树林根际土壤的枯草芽孢杆菌产淀粉酶条件及酶学性质研究

从三亚红树林保护区中筛选出一株产淀粉酶枯草芽孢杆菌,通过改变培养温度、初始pH值和盐浓度等因素研究该菌株的最适产酶条件,同时也分析了该菌株所产粗酶的热稳定性及pH稳定性。结果表明：该菌株属于中温产酶菌,产淀粉酶的最适温度为40℃,最适pH值为8,盐耐受能力可达到4%。酶学性质分析发现,该菌株产生的淀粉酶在40℃保温150 min后仍有70%以上的相对酶活力,在pH为9的环境中孵育1 h,相对酶活力可达到90%以上。这些结果表明,该产淀粉酶菌株对热带地区海水环境具有较好的适应性,其作为消化酶类饲料添加剂具有广阔的应用前景。     

48株枯草芽孢杆菌对棉花黄萎病防治效果评测

目前,枯草芽孢杆菌已广泛应用于防治棉花黄萎病,但不同菌株的防治效果（防效）参差不齐,有待进一步筛选验证。前期,本课题组从健康棉花植株和根际土壤中分离获得了48株枯草芽孢杆菌。为明确这些菌株对棉花黄萎病的防效,筛选具有优异防效的枯草芽孢杆菌菌株,采取对峙培养法、对扣培养法测定其对棉花黄萎病菌的抑制作用,并通过温室试验验证其对棉花黄萎病的防效。通过对峙培养和对扣培养筛选出18株拮抗效果较好的菌株。温室试验结果显示：无论浸种或灌根处理,EBS03、EBS07、EBS10和BS04菌株对棉花黄萎病的防效均大于55％,其中EBS03防效最好,浸种与灌根处理对棉花黄萎病的防效分别为76.31%、87.10%;此外,播种后60 d EBS03和EBS10菌株浸种处理的棉苗株高、地上部鲜物质质量、总鲜物质质量较对照分别提高78.08%和77.77%、113.33%和108.00%、107.23%和103.61%。综上,EBS03和EBS10菌株具有较好的促生长作用,并对棉花黄萎病有一定的防效,具有进一步开发应用的潜力。     

枯草芽孢杆菌BS-2对脉孢菌的抑菌活性成分分析

枯草芽孢杆菌(Bacillus subtilis)BS-2对脉孢菌(Neurospora)有较强的抑制作用。本文通过对枯草芽孢杆菌BS-2进行紫外诱变,获得了抑菌活性比出发菌株增强的突变株BS-2-E和抑菌活性完全丧失的突变株BS-2-N。用LC-MS分析BS-2、BS-2-E及BS-2-N发酵液的盐酸粗提物,比较这3个菌株的质谱响应峰,初步认为BS-2的抑菌活性物质主要是伊枯草菌素(iturins)和芬枯草菌素B(fengycin B)。     

枯草芽孢杆菌种子包衣对棉花黄萎病抑制作用的研究

通过田间小区试验,研究不同包衣处理防治黄萎病的效果。结果表明:包衣处理出苗时间缩短1~2d,出苗率有所提高,其中枯草芽孢杆菌包衣处理表现最好,株高增长快,利于真叶数、果枝数增加。在黄萎病防治上,40%卫福悬浮种衣剂包衣处理的效果最好,达到了39.2%。产量表现上,枯草芽孢杆菌包衣处理产量最高,为6791kg·hm~(-2),比对照增产36.92%,增产效果明显。     

小麦赤霉病拮抗性芽孢杆菌生防作用的研究

为探索小麦赤霉病生物防治的有效途径,对分离自中国东北嫩江小麦赤霉病流行地区小麦植株叶面和穗部的8株拮抗性芽孢杆菌进行了室内生物测定和田间小区防治试验.平板对峙培养、菌丝生长速率及孢子萌发测定等试验表明,8个菌株对小麦赤霉病菌的菌丝生长及孢子萌发均具有明显的抑制作用,其中以菌株B08的综合抑菌活性最强,其活菌体抑菌带宽达8.4 mm,无菌发酵液稀释5倍处理后96 h对菌丝生长抑制率为66.2%,处理分生孢子可使其完全丧失萌发能力,10倍稀释液可使孢子刚萌发的芽管膨大畸变而不能伸长,活体菌悬浮液和灭菌后的发酵液田间防效达59.5%～65.9%.发酵液热稳定性好,121℃、30 min加压蒸气灭菌后仍保持良好的抑菌活性.研究结果说明,供试拮抗菌株作为小麦赤霉病的生防材料具有很好的开发应用前景。     

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.     

橡胶树内生拮抗细菌菌株Czk1的发酵条件优化

从橡胶树根部分离出1株枯草芽孢杆菌(Bacillus subtilis)菌株Czk1,通过室内平皿对峙试验结果表明,菌株Czk1对所参试的5种橡胶树不同的根病菌都具有很强的拮抗作用,同时也对8个其他作物上发生的常见病原菌都有非常强的拮抗作用.通过单因子和正交试验法对菌株Czk1的培养基进行优化,获得的最佳培养基配方为:...     

枯草芽孢杆菌JN005胞外抗菌物质及对水稻叶瘟防治效果

【目的】检测枯草芽孢杆菌JN005产生的胞外抗菌物质的稳定性和对稻瘟病菌的生物活性,及其对水稻叶瘟的防治效果。【方法】通过不同培养基检测枯草芽孢杆菌JN005产生的抑菌物质,结合平板对峙法、光学显微镜观察、室内离体和活体接种来检测JN005菌株胞外抗菌物质对稻瘟病菌拮抗活性和对水稻叶瘟防治效果。【结果】枯草芽孢杆菌JN005能分泌蛋白酶、纤维素酶、淀粉酶和β-1, 3-葡聚糖酶,其胞外抗菌物质对稻瘟病菌生长具有抑制作用。胞外抗稻瘟病菌物质在0℃~100℃和pH 2~12范围内活性较为稳定,且经蛋白酶K处理后不影响其活性,紫外照射12 h以及4℃保存3个月仍有活性。胞外抗菌物质能引起稻瘟病菌菌丝形态畸变,并抑制分生孢子的萌发。室内湘晚籼12号稻瘟病离体叶片接种表明,先用100倍胞外抗菌物质稀释液处理水稻叶片,24 h后接种稻瘟病菌分生孢子悬浮液(1×10⁵个/mL),其叶瘟发病率为34.07%,效果接近稀释500倍的40%稻瘟灵EC;而先接种稻瘟病菌分生孢子悬浮液,24 h后用100倍胞外抗菌物质稀释液处理,其发病率为38.52%,与稀释1500倍的75%三环唑(WP)和稀释500倍的40%稻瘟灵(EC)有显著性差异。活体喷雾接种试验表明,于湘晚籼12号秧苗长至3叶1心时,接种稻瘟病菌分生孢子悬浮液前24 h喷施稀释100倍的胞外抗菌物质稀释液,其叶瘟防效为75.32%;接种稻瘟病菌分生孢子悬浮液后24 h喷施稀释100倍的胞外抗菌物质稀释液,其防效为71.49%。【结论】JN005菌株胞外抗菌物质对稻瘟病菌的菌丝生长和孢子萌发有直接抑制作用,且对环境稳定性较好,对稻瘟病有较好的预防和治疗作用,有进一步的开发潜力。     

Extracellular Antimicrobial Substances Produced by Bacillus subtilis JN005 and Its Control Efficacy on Rice Leaf Blast

【Objective】The purpose of this research is to assay stability, biological activity and efficacy of the extracellular antimicrobial substances produced by Bacillus subtilis JN005 against Magnaporthe oryzae. 【Method】The antimicrobial substances produced by B. subtilis JN005 was tested using different media. The activity of extracellular antimicrobial substances of JN005 strain against M. oryzae and its efficacy on rice leaf blast were measured by dural culture, optical microscope observation, indoor detached-leaf inoculation and live inoculation.【Result】B. subtilis JN005 secreted protease, cellulase, amylase and β-1,3-glucanase and its extracellular antimicrobial substances showed inhibition activity against M. oryzae. The extracellular antimicrobial substances were stable in a wide range of pH values from 2 to 12 and at temperatures from 0℃ to 100℃. The antifungal activity was retained after treatment with proteinase K. The extracellular antimicrobial substance kept its activity even it was irradiated for 12 h under ultraviolet light or exposed to 4℃ for 3 months. The extracellular antimicrobial substances caused distortion of hyphae of M. oryzae, and hindered the germination of conidia. Indoor detached leaves inoculation of Xiangwanxian 12 showed that disease incidence was 34.07% as rice leaves were treated with 100-fold dilution of the extracellular antimicrobial substances, followed by inoculation with conidia suspension of M. oryzae (after 24 h). The control efficacy was close to that of 500 times-diluted 40% isoprothiolane EC. The disease incidence was 38.52% as rice leaves were inoculated with conidia suspension of M. oryzae and treated with 100-fold dilution of the extracellular antimicrobial substances after 24 h, significantly different from 1500 times-diluted 75% tricyclazole WP and 500 times-diluted 40% isoprothiolane EC. The potted plant experiment showed that efficacy was 75.32% as rice was inoculated with conidia suspension of M. oryzae followed by spraying 100-fold dilution of the extracellular antimicrobial substances before 24 h. The efficacy was 71.49% as rice was inoculated with conidia suspension of M. oryzae and followed by spraying of 100-fold dilution of the extracellular antimicrobial substances after 24 h.【Conclusion】The extracellular antimicrobial substances produced by JN005 strain directly inhibited mycelial growth and conidia germination of M. oryzae and it has good environmental stability. The extracellular antimicrobial substances have better preventative efficacy and therapeutic efficacy on rice blast and should be further investigated and developed as such.     

枯草芽孢杆菌防治大豆菌核病效果初报

本文应用枯草芽孢杆菌ZH-2菌株的发酵菌剂,在桦南县进行了生防菌剂防治大豆菌核病的田间防效试验,结果表明生防菌剂100倍稀释液浸种和喷洒处理对大豆菌核病的田间防效达67.9%。     

枯草芽孢杆菌(Bacillus subtilis)CS27液体发酵条件优化

从提高细胞生物量并降低发酵生产成本的目的出发,本文采用单因素试验和正交试验方法,对已筛选出的枯草芽孢杆菌CS27菌株进行液体发酵培养基和工艺条件优化。得到最佳培养基配方为:1.5%黄豆粉、0.5%糖蜜、0.8%氯化铵、1.0%氯化钠、0.1%柠檬酸钠、0.5%碳酸钙、0.1%七水合硫酸镁;最佳发酵条件为:发酵温度32℃,最佳装液量50 mL/250 mL,最适接种量10%,初始pH7.0。在优化后的培养基发酵条件下细胞生物量可以达到1.75×10~9 cfu/mL,为枯草芽孢杆菌CS27菌株规模化生产及应用提供科学依据。     

枯草芽孢杆菌在农作物生产上的应用研究进展

综述了枯草芽孢杆菌(Bacillus subtilis)在土壤提质改良、促进作物生长发育及产质量的提高、耕作土壤降害与修复等方面的研究成果,并对作物生产上枯草芽孢杆菌与环境相关影响因子间其专业化功能开发研究及其分子生物学水平上的作用机理等进行了展望。     

枯草芽胞杆菌Tu-100对几种作物的促生效果

盆栽试验结果表明,枯草芽胞杆菌Tu-100对油菜、小麦、辣椒、西瓜、大豆和玉米苗期有不同的促生效果,在植物干重上分别比对照增加9.47%、1.37%、2.27%、19.23%、9.21%和6.67%.对水稻的盆栽产量增加12.82%.对大豆的结瘤数和鲜瘤重呈现负增长.植物促生根际细菌对不同的作物保持着不同的亲和性.