枯草芽孢杆菌在番茄病害防治上的应用与研究进展

综述了枯草芽孢杆菌在防治植物病害中的生防机制,包括竞争作用、拮抗作用、溶菌作用、诱导植物产生抗性和促进植物生长5个方面,介绍了枯草芽孢杆菌在6种番茄病害即灰霉病、青枯病、早疫病、立枯病、枯萎病、根腐病防治中的应用,并对其应用中存在的主要问题进行了总结、前景进行了展望。     

热带土壤中解淀粉芽孢杆菌HNU1的鉴定及发酵条件优化

重金属毒性强、不可降解，在环境中可持续存在，土壤中的重金属可通过食物链累积，对生物体的健康构成严重威胁。土壤微生物修复重金属污染方法具有经济成本低、周期短、效率高、无二次污染等优点，也存在外源微生物在土壤环境中不易存活和繁殖的缺点。通过形态观察、生理生化试验以及16S rDNA同源性序列分析对前期研究中分离保存的一株铜富集细菌HNU1菌株进行鉴定，鉴定该菌为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。并通过单因素试验和正交试验找出该菌株的最适发酵条件，该菌的最适培养基组成为蔗糖2 g·L-1，酵母粉4 g·L-1，胰蛋白胨6 g·L-1，MgSO4 5 g·L-1。培养条件为初始pH为8.0，温度37 ℃，接种量0.1%，摇床转速180 r·min-1，50 mL三角瓶中装液量20 mL。鉴定的解淀粉芽孢杆菌HNU1菌株对Cu具有良好的富集作用，可进一步开发为微生物肥料用于重金属污染农田土壤的修复，对土壤修复和改良具有重大意义。     

饲粮添加酿酒酵母和地衣芽孢杆菌对绵羊生长性能与瘤胃发酵的影响

【目的】研究饲粮中添加酿酒酵母和地衣芽孢杆菌对绵羊生长性能和瘤胃发酵的影响,为酿酒酵母和地衣芽孢杆菌在肉羊饲粮中的合理应用提供理论依据。【方法】试验选用48只4月龄、体重相近（22.96±2.00）kg、健康的杜泊×小尾寒羊杂交F₁代公羔,根据饲喂添加剂不同随机分为4组：D（对照组,两种菌均不添加）;D1（酿酒酵母,6×10¹⁰CFU/kg）;D2（地衣芽孢杆菌,2×10¹⁰CFU/kg）;D3（酿酒酵母6×10¹⁰CFU/kg+地衣芽孢杆菌2×10¹⁰CFU/kg）,每组12只羊,试验期共75 d,前15 d为适应期,正饲期60d。试验羊每天分别于08:00和18:00进行饲喂,自由采食和饮水。正饲期内每天准确称量记录每只试验羊的喂料量和剩料量,并在第1、30、60 天晨饲前对试验羊进行称重,计算平均日采食量、平均日增重（ADG）及料重比（F/G）。试验结束当天08：00正常饲喂试验羊,3 h后采集瘤胃液,测定发酵参数、消化酶活性以及功能微生物。【结果】1）饲粮中添加酿酒酵母和地衣芽孢杆菌对试验羊的初始体重和终末体重及平均采食量的影响均不显著（P>0.05）,D3组的ADG显著高于D组（P<0.05）,D3组的F/G显著低于D组（P<0.05）,D1和D2组的F/G差异不显著（P>0.05）;2）饲粮中添加酿酒酵母和地衣芽孢杆菌对瘤胃液pH、丁酸浓度及乙丙比的影响均不显著（P>0.05）,D3组的NH₃-N浓度显著低于D组（P<0.05）,TVFA和丙酸浓度显著高于D组（P<0.05）,且D3组与D1和D2组均差异不显著（P>0.05）,D3和D2组的乙酸浓度显著高于D1与D组（P<0.05）;3）D3组的β-葡萄糖苷酶、羟甲基纤维素酶、木聚糖酶及淀粉酶活性均显著高于其他3组（P<0.05）。D1和D2组的β-葡萄糖苷酶、果胶酶、羟甲基纤维素酶及淀粉酶活性与D组无显著差异（P>0.05）。D3组蛋白酶活性显著高于D和D2组（P<0.05）,与D1组差异不显著（P>0.05）,D1和D2组的β-葡萄糖苷酶、果胶酶、羟甲基纤维素酶、木聚糖酶、蛋白酶及淀粉酶活性均差异不显著（P>0.05）;4）添加酿酒酵母和地衣芽孢杆菌对黄色瘤胃球菌、栖瘤胃普雷沃氏菌、嗜淀粉瘤胃杆菌和原虫数量的影响不显著（P>0.05）,D3组的溶纤维丁酸弧菌数显著高于其他3组（P<0.05）,白色瘤胃球菌和产琥珀酸丝状杆菌显著高于D组（P<0.05）,但与D1和D2组差异不显著（P>0.05）。与对照组相比,试验组产甲烷菌数量显著降低（P<0.05）,其中D3组最低。【结论】饲粮中添加6×10¹⁰CFU/kg酿酒酵母和2×10¹⁰CFU/kg地衣芽孢杆菌均会对绵羊瘤胃发酵产生积极影响,提高了瘤胃消化酶的活性,增加了瘤胃有益菌的数量,且二者组合饲喂效果更加显著。     

Co-inoculation of selected nodule endophytic rhizobacterial strains with Rhizobium tropici promotes plant growth and controls damping off in common bean

Production of common bean(Phaseolus vulgaris)is limited by the occurrence of damping off(rhizoctoniosis),which is caused by the fungus Rhizoctonia solani.However,the co-inoculation of plant growth-promoting rhizobacteria(PGPR)involved in biological control along with diatomic nitrogen(N2)-fixing rhizobia can enhance N nutrition and increase production.In this context,finding microorganisms with synergistic effects that perform these two roles is of fundamental importance to ensure adequate yield levels.The aim of this study was to evaluate the effects of co-inoculation of nodule endophytic strains of the genera Bacillus,Paenibacillus,Burkholderia,and Pseudomonas with Rhizobium tropici CIAT 899,an N2-fixing rhizobial strain,on the biocontrol of damping off and growth promotion in common bean plants.Greenhouse experiments were conducted under axenic conditions using the common bean cultivar Pérola.The first experiment evaluated the potential of the 14 rhizobacterial strains,which were inoculated alone or in combination with CIAT 899,for the control of R.solani.The second experiment evaluated the ability of these 14 rhizobacterial strains to promote plant growth with three manners of N supply:co-inoculation with CIAT 899 at low mineral N supply(5.25 mg N mL^-1),low mineral N supply(5.25 mg N mL^-1),and high mineral N supply(52.5 mg N mL^-1).The use of rhizobacteria combined with rhizobia contributed in a synergistic manner to the promotion of growth and the control of damping off in the common bean.Co-inoculation of the strains UFLA 02-281/03-18(Pseudomonas sp.),UFLA 02-286(Bacillus sp.),and UFLA 04-227(Burkholderia fungorum)together with CIAT 899 effectively controlled damping off.For the common bean,mineral N supply can be replaced by the co-inoculation of CIAT 899 with plant growth-promoting strains UFLA 02-281/02-286/02-290/02-293.Nodule endophytes UFLA02-281/02-286 are promising for co-inoculation with CIAT 899 in the common bean,promoting synergy with rhizobial inoculation and protection against disease.     

甲基营养型芽胞杆菌的研究进展

甲基营养型芽胞杆菌是一种无致病性的细菌,对外界环境具有多抗逆性,作为芽胞杆菌属的"后起之秀",被越来越多的应用于各行业中。本文主要从甲基营养型芽胞杆菌的生理生化特征、代谢产物及应用等方面论述其研究进展。     

Changes of Morphological Parameter of Wheat Seed Pretreatment by the Biocontrol Agents Bacillus subtilis QM3 during Germination

Bacillus subtilis QM3, a potential biocontrol agent and useful plant-growth-promoting bacteria (PGRB) from yak dung in China with broad-spectrum and strong antifungal bioactivity, was studied to determine if it either stimulated or inhibited the germination of wheat seeds depending on the amount of bacteria. Germination rate (GR) and length of shoot (LS) increased at 10-fold dilution of QM3 fermentation liquid for 12 h (P < 0.05). Germination potentiality (GP) and root cap ration (RC) decreased at 100-fold dilution of QM3 for 0 h. Length of root (LR) and tips of root (TR) at day 7 remained at control levels at 0-fold dilution of fermentation liquid of QM3 for 24 h. The results obtained suggest that some plant diseases, environmental stresses (such as high salt and heavy metal contents), and difficulties in germination can be controlled or relieved by the use of QM3.     

Toxicity of Bacillus thuringiensis Crystal Toxins to Field Populations of Rice Leaf Folder, Cnaphalocrocis medinalis （Guenee） and Establishment of Baseline Susceptibility to Cry1Ab

Eight insecticidal crystal proteins of Bacillus thuringiensis, CrylAa, CrylAb, CrylAc, CrylB, Cry2Aa, CrylC, CrylDa and Cry 1Ea were assessed for toxicity against 1 st instar larvae of rice leaf folder, Cnaphalocrocis medinalis （Guenee） at 48 HAT and 72 HAT. Bioassay results depicted CrylAa was the most toxic （LCso 2.35 ppm） followed by CrylBa （LCso 8,50 ppm） and CrylAb （LCso 8.73 ppm） at 48 HAT, whereas, at 72 HAT CrylAb proved to be highly toxic （LC50 0.50 ppm） followed by CrylAa （LCso 4.07 ppm）, CrylAc （LCso 4,84 ppm） and CrylBa （LCso 6.42 ppm）. Toxins Cry2Aa, CrylCa, CrylDa and CrylEa did not resulted in any mortality at 48 HAT and 72 HAT, respectively. Baseline estimates for CrylAb against 1st instar larvae of C. medinalis sampled from seven geographical locations revealed variation in LC50＇s from 0.37 ppm to LC50 16.25 ppm at 48 HAT and LC50 0.50 ppm to LC50 6.49 ppm 72 HAT, respectively with relative resistance ratios of 44-fold and 13-fold at 48 HAT and 72 HAT over the susceptible population.     

1株降解黄曲霉毒素B1的解淀粉芽胞杆菌的分离鉴定及其体外脱毒效果

旨在筛选出能够高效降解黄曲霉毒素B₁（AFB₁）的菌株,并对其脱毒活性组分的脱毒效果进行研究,本研究以香豆素为唯一碳源对目标细菌进行初筛,以对AFB₁的降解率为指标进行复筛,并从形态学、生理生化试验和16S rDNA序列分析对筛选菌株进行鉴定;通过测定该菌株不同发酵组分对AFB₁的降解率,来定位其脱毒活性组分,同时采用CCK-8法测定活性组分降解AFB₁后对LMH细胞毒性,并研究了热处理、紫外线照射、强酸、强碱及蛋白酶K处理对活性组分脱毒作用的影响。结果显示,从样品中初筛分离到24株能够在初筛培养基生长良好的菌株,经复筛从中筛选到一株能高效降解AFB₁的菌株Y1-B1,其AFB₁降解率达到73.2%;经形态学、生理生化试验和16S rDNA序列分析确定该菌为解淀粉芽胞杆菌（Bacillus amyloliquefaciens）。对菌株Y1-B1培养物的不同组分进行的脱毒活性研究表明,不同组分对AFB₁的降解率不同,上清液的脱毒能力最强,细菌细胞裂解物的脱毒能力下降明显,菌体悬液和菌体裂解物上清的脱毒能力最差,由此确定解淀粉芽胞杆菌Y1-B1的脱毒能力主要来源于上清液中的某种活性物质;细胞毒性结果显示,与AFB₁对LMH细胞的毒性作用相比,AFB₁被上清液降解后其产物的毒性显著降低。该脱毒活性组分对不同理化因素处理呈现出不同的表现,对高温、紫外照射抵抗力较强,对强酸、强碱抵抗力较差,蛋白酶K仅造成脱毒能力部分减弱。本研究将为解决黄曲霉毒素污染问题提供新的微生物种质资源,同时为后续微生物脱毒制剂的开发奠定基础。     

Effects of dietary Bacillus coagulans and yeast hydrolysate supplementation on growth performance,immune response and intestinal barrier function in weaned piglets

The present study investigated the effects of Bacillus coagulans and yeast hydrolysate supplementation on growth performance, immune response and intestinal barrier function of weaned piglets. Twenty-four weaned piglets with an average body weight (BW) of 6.89 ± 0.15 kg were divided into four diets for 28 days. The treatments were basal diet (control), basal diet supplemented with antibiotic (20 mg/kg colistin sulphate and 40 mg/kg bacitracin zinc, AT), probiotics (400 mg/kg Bacillus coagulans ≥5 × 109 CFU/g, BC) or yeast hydrolysate (5000 mg/kg yeast hydrolysate, YH). Average daily gain (ADG) and average daily feed intake (ADFI) were improved by AT and YH diets (p < 0.05), while BC diet only increased ADG (p < 0.05). The complement 3 (C3), lysozyme (LZM) and tumour necrosis factor-α (TNF-α) concentrations in serum were increased in BC diet (p < 0.05). Feeding AT and YH caused the increase of jejunal villus height (p < 0.05), and a higher ratio of villus height/crypt depth was observed in AT, BC and YH groups (p < 0.05). The mRNA expression of zonula occludens-1 (ZO-1) in jejunal mucosa was up-regulated by AT, BC and YH diets (p < 0.05). Dietary AT, BC or YH inclusion decreased the interleukin-1β (IL-1β) concentration and TNF-α mRNA expression (p < 0.05), and YH supplementation even down-regulated toll-like receptor 4 (TLR4) and CD14 expressions (p < 0.05). In summary, the dietary administration of BC or YH both improves growth performance through promoting the intestinal barrier function, indicating both of them can serve as potential alternatives to antibiotics growth promoters for the piglet production.     

地衣芽孢杆菌对热应激公兔甲状腺和甲状旁腺组织的影响

将成年塞北兔公兔随机分成3组:常温组、热应激组(基础日粮)和地衣芽孢杆菌组(基础日粮+1000 mg/kg地衣芽孢杆菌),常温组温度为20~25℃,其他2个组人工模拟夏季温度。在试验期间测定血浆皮质醇含量。试验结束后测定甲状腺质量,计算甲状腺指数;观察光镜下甲状腺组织、透射电镜下甲状腺滤泡上皮细胞和甲状旁腺嗜酸性细胞变化。结果显示,地衣芽孢杆菌可极显著降低热应激公兔皮质醇含量,提高热应激15 d后公兔甲状腺质量和指数(P<0.01);明显改善甲状腺滤泡结构,膜结构较为清晰,降低细胞核萎缩和胞质内细胞器损伤;降低甲状旁腺嗜酸性细胞中线粒体空泡化程度。结果表明,在塞北兔公兔日粮中添加1000 mg/kg地衣芽孢杆菌可有效降低热应激公兔血浆皮质醇含量,缓解对甲状腺和甲状旁腺的损伤,对保证塞北兔公兔正常生殖机能具有重要作用。