4种微生物菌剂对辣椒主要病害的生物防治作用

本文选择4种不同微生物活性成分的生防菌剂，采用盆栽生物量测定、温室和田间小区试验对辣椒主要病害进行抗病效果及促生增产作用评价。结果表明，木霉-芽胞杆菌复合菌剂对辣椒的枯萎病、疫病、炭疽病和灰霉病防效分别达到72.2%、78.3%、71.9%和59.7%，枯草芽胞杆菌-粉红粘帚霉复合菌剂对辣椒4种病害的防效分别达到60.5%、62.1%、60.5%和50.9%；而淡紫紫孢菌菌剂只对辣椒炭疽病具有很好防效（77.5%）。同时，4种微生物菌剂对辣椒植株生物量均有显著促生作用，其中木霉-芽胞杆菌复合菌剂及枯草芽胞杆菌-粉红粘帚霉复合菌剂在盆栽试验中鲜重和干重增幅分别为132.79%、190.35%和211.80%、293.84%；两者在温室及田间小区试验中比对照增产182.3%、152.7%和65.0%、23.9%。本研究表明，生防菌剂可以替代化学药剂用于植物病害的防治，同时还能够促进作物植株生长，有增产作用。     

Administration of probiotic Bacillus licheniformis induces growth,immune and antioxidant enzyme activities,gut microbiota assembly and resistance to Vibrio parahaemolyticus in Litopenaeus vannamei

This study aimed to elucidate the effects of dietary Bacillus licheniformis ATCC 11946 (BL) on the growth, immune and antioxidant activities, intestinal morphology and microbiota, and susceptibility to Vibrio parahaemolyticus (VP_AHPND) in juvenile pacific white shrimp, Litopenaeus vannamei (LV). Juvenile LV (initial weight = 0.63 ± 0.001) were fed diets containing varying BL concentrations (0 (BL0), 10⁶ (BL1), 10⁷ (BL2) and 10⁸ (BL3) CFU/g feed) for 8 weeks. Growth performance, immune and antioxidant enzyme activities, and intestinal morphology significantly improved in the probiotic‐treated groups than the untreated. Regardless of the treatment group, the two dominant phyla were Proteobacteria and Bacteroidetes, whereas the two dominant genera were Ruegeria and Vibrio. Increasing inclusion of probiotics in diets led to significant increase in beneficial bacterial genera (Ruegeria, and Pseudoalteromonas) and a significant decrease in some known opportunistic pathogens (Vibrio, Tenacibaculum, Photobacterium, Kangiella and Spongiimonas) with the BL3 group witnessing the best. A 7‐day challenge study with VP_AHPND showed significantly high protection in the probiotic‐treated groups, with the BL3, BL2 and BL1 obtaining 51%, 34% and 22% relative percentage survival, respectively. In conclusion, BL at 10⁸ CFU/g feed should be used to help in shrimp production since it attained the most significant improvement.     

Bacillus siamensis和Bacillus subtilis中2个β-D-葡萄糖苷酶编码基因克隆、表达及酶学研究

采用RACE技术扩增2株Bacillus属菌株XY18、XY20的β-D-葡萄糖苷酶编码基因bgl全长,构建重组载体pET28a(+)/bgl、pET28b(+)/bgl,导入E. coli BL21诱导表达,采用Ni亲和层析纯化蛋白。经酶学性质测定发现：2个蛋白均为弱酸性蛋白,温度为40 ℃时酶活力达到最大,具有一定耐热性。以上结果表明,为优化2株Bacillus属菌株参与香草兰发酵工艺,可提供适当弱酸性条件、适宜温度。     

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。     

Effect of Bacillus cereus as a water or feed additive on the gut microbiota and immunological parameters of Nile tilapia

We evaluated the effects of Bacillus cereus, as an additive in water and feed, on the gut microbiota and immunological parameters of Nile tilapia (Oreochromis niloticus) fingerlings. Experiments were performed in tanks and net cages respectively. Experiment 1: Tilapia were housed in tanks for 42 days, and B. cereus was added to the water at 1.0 × 10⁴ cfu mL^?1 (Treatment 1) and 1.0 ×10⁵ cfu mL^?1 (Treatment 2) weekly. For the control, no probiotic was added. Experiment 2: Tilapia were housed in cages for 42 days, and the feed was supplemented with B. cereus at 1.0 × 10⁷ cfu g^?1 (Treatment 1) and 1.0 × 10⁸ cfu g^?1 (Treatment 2) weekly. For the control, no probiotic was added. Each treatment contained three replicates, with 50 male tilapias per replicate. The fish from the probiotic treatments in both tank and cage experiments had significantly higher serum lysozyme and peroxidase activities than the control. In the cage experiment, alkaline phosphatase and total superoxide dismutase activities in tilapia were significantly higher in probiotic treatments compared with the control. The results of polymerase chain reaction‐denaturing gradient gel electrophoresis showed that B. cereus supplementation in the feed and water affected the autochthonous gut bacteria community of tilapia and stimulated various potentially beneficial bacteria. Therefore, B. cereus, as a water or feed additive, could enhance the immune status and affect the gut microbiota of tilapia. Bacillus cereus was more effective as a feed supplement rather than a water additive for enhancing the immune status of tilapia.     

Synbiotic effect of Bacillus mycoides and organic selenium on immunity and growth of marron,Cherax cainii (Austin, 2002)

The present feeding trial examined the effect of synbiotic use of Bacillus mycoides and organic selenium (OS) as Sel‐Plex on marron immunity, growth and survival. The marron were cultured in recirculated tanks and fed test diets consisting of a basal diet; basal diet supplemented with B. mycoides (10⁸ CFU g^?1 of feed); basal diet supplemented with OS (Sel‐Plex) (0.2 g kg ^{? 1} of feed) and basal diet supplemented with synbiotic (B. mycoides at 10⁸ CFU g^?1 and OS 0.2 g kg ^{? 1} feed) diet, in triplicate. The effect of the prebiotic OS (Sel‐Plex) on the growth rate of B. mycoides was also studied in vitro. The results suggested that synbiotic use of B. mycoides and OS significantly improved some immune parameters of marron, particularly the glutathione peroxidase, and to some extent total haemocyte counts. However, the synbiotic feed did not synergistically improve marron growth, in fact the use of B. mycoides‐supplemented diet alone demonstrated significantly higher growth in marron compared with the growth of marron fed on other test diets. Supplementation of the basal diet with host origin B. mycoides significantly increased the intestinal bacterial population (3.399 ± 825 CFU g ^{? 1} of gut) in marron compared with other diets. Organic selenium as Sel‐Plex in Trypticase Soya Broth also confirmed that OS did not increase the amount of growth of B. mycoides and resulted in a lower intestinal bacterial population in the synbiotic diet‐fed marron. In conclusion, synbiotic of OS and B. mycoides may improve a particular immune parameters of marron and to a lesser extent their growth.     

解淀粉芽胞杆菌YU-1对水稻纹枯病的防治作用

本文旨在筛选对水稻纹枯病有生防作用的菌株,并初步探索其生防作用机理。收集水稻、甘蓝、黄瓜等不同植物根际土壤,采用稀释分离和对峙培养法筛选对水稻纹枯病菌Rhizoctonia solani Kühn有抑菌作用的菌株;通过离体接种防效、盆栽防效、抑菌谱、对水稻纹枯病菌菌核萌发及形成的抑制作用等方面评价其生防潜力,并对生防菌株进行鉴定。结果表明,从采集的37份根际土壤中共分离获得细菌297株,其中4株对纹枯病菌具有较好的拮抗作用,菌株YU-1对水稻纹枯病菌菌丝生长的抑制率达89.8%;对西瓜枯萎病菌、草莓灰霉病菌、番茄早疫病菌、油菜菌核病菌、小麦赤霉病菌菌丝生长的抑制率均在85%以上;对水稻纹枯病的离体和盆栽防效分别为73.1%和66.3%;对水稻纹枯病菌菌核萌发的抑菌率在92%以上;经生理生化和分子鉴定为解淀粉芽胞杆菌Bacillus amyloliquefaciens。由此可看出,菌株YU-1对水稻纹枯病的防治具有较强的应用价值,具有进一步开发成生物农药的潜力。     

贝莱斯芽孢杆菌XC1的筛选、鉴定及其对苹果连作障碍的影响

从连作苹果园健康苹果树根际土壤中筛选出1株对苹果连作障碍镰孢属病原菌具有较强拮抗作用的细菌XC1,该菌对尖孢镰孢菌的抑菌率最高,达到79.83%。根据其形态、生理生化特征和基于16S rDNA、gyrA基因的系统发育分析,鉴定该菌为贝莱斯芽孢杆菌(Bacillus velezensis)。目前菌株XC1已保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏号为CGMCC No.20057。苹果连作土盆栽试验表明,XC1菌肥处理能显著促进平邑甜茶(Malus hupehensis Rehd.)幼苗的生长,其株高、地径、鲜质量和干质量分别比对照提高了54.8%、34.8%、121.1%和96.0%;显著提高连作土壤中的蔗糖酶、脲酶、过氧化氢酶和中性磷酸酶的活性,分别为对照的1.57倍、2.71倍、1.21倍和1.79倍;显著减少连作土壤中真菌的数量,提高细菌和放线菌的数量,且对尖孢镰孢菌(Fusarium oxysporum)有很好的抑制作用,尖孢镰孢菌的基因拷贝数比对照降低了76.4%。这说明贝莱斯芽孢杆菌抑制连作障碍病原菌的效果明显,可改善土壤环境,促进平邑甜茶幼苗的生长,能在一定程度上减轻苹果连作障碍。     

对草地贪夜蛾高毒力的苏云金芽胞杆菌菌株筛选与杀虫活性研究

草地贪夜蛾是一种世界性重大农业害虫，给玉米等多种主要粮食和经济作物造成严重危害。为实现对草地贪夜蛾的高效、绿色、持续防控，亟需筛选高毒力苏云金芽胞杆菌菌株资源。本研究以前期实验室储备的对斜纹夜蛾、小地老虎具有杀虫活性的363株野生菌株库为候选菌株，基于菌株杀虫基因差异及进化关系，去除冗余菌株后获得172株候选菌株。通过培养获取这些菌株胞晶混合物，进行杀虫蛋白定量后，测定杀虫活性，获得27株对草地贪夜蛾初孵幼虫具有较高杀虫活性的菌株，其中菌株B14-D2的杀虫活性最高，LC₅₀为0.155 μg/g。克隆了该菌株中的cry1Ea3基因并在HD73^-无晶体突变株中表达，Cry1Ea3蛋白对草地贪夜蛾初孵幼虫LC₅₀为1.789 μg/g。本研究为新的Bt产品和杀虫基因的开发利用提供了丰富资源。