甘蔗黑穗病菌拮抗菌HAS的筛选和鉴定

采用传统形态学鉴定及16S rRNA基因序列分析,筛选鉴定对甘蔗黑穗病菌具有拮抗作用的细菌的归属,并采用细菌的通用引物P0、P6扩增16S rRNA基因片段.结果表明:得到1 533 bp的DNA片段,其序列与枯草芽孢杆菌序列的同源性高达99％.其形态学特征:菌体为短杆状,能运动;革兰氏阳性反应;3％KOH溶解性实验呈阴性反应;有鞭毛,鞭毛周生;芽孢中生,椭圆形,孢囊稍膨大,初步将该菌株归属为枯草芽孢杆菌.与病原真菌对峙培养的结果表明,该菌对引起甘蔗和其它作物病害的多个植物病原真菌均具有很好的抑制作用.据此,可将分离的菌株HAS确定为对甘蔗黑穗病菌有较好防治效果的枯草芽孢杆菌.     

豆粕生物肽的生产工艺研究

通过菌种筛选、摇瓶发酵条件优化,系统地研究了微生物发酵生产豆粕生物肽的工艺。结果表明:枯草芽胞杆菌为最佳菌株,其豆粕生物肽最佳生产工艺条件为:种子培养时间16 h,豆粕粒度40目,初始pH6.8,葡萄糖2%,豆粕11%,接种量6%,发酵时间48 h;在此工艺条件下蛋白质降解为多肽的得率为41.18%。     

拮抗香蕉枯萎病菌的解淀粉芽孢杆菌LX1菌株的鉴定及其抗菌蛋白基因的克隆

收集海南不同盐碱地的土样进行拮抗香蕉枯萎病菌的微生物分离,对具有较强抑菌作用的菌株进行形态特征、生理生化和分子鉴定,并克隆其抗菌蛋白基因.结果分离到1株具有较强抑制香蕉枯萎病菌能力的细菌LXl,经鉴定为解淀粉芽孢杆菌(Bacillus amyloliquefaciens).抑菌实验发现LX1菌株的发酵上清液中粗蛋白有一定的抑菌作用,经过Sephacryl S-200HR柱层析、DEAE Sepharose Fast Flow离子交换层析分离纯化了其中抑菌作用最强的抗菌蛋白.质谱鉴定结果表明,抗菌蛋白与B.amyloliquefaciens FZB42内切葡聚糖酶同源性最高.根据质谱结果克隆了该抗菌蛋白的编码基因,该基因的核苷酸和氨基酸序列与B.amyloliquefaciens FZB42的内切葡聚糖酶的核苷酸和氨基酸序列同源性分别达99％和100％.拮抗香蕉枯萎病菌的芽孢杆菌LX1可作为潜在的防治香蕉枯萎病的的生防制剂,其抗菌蛋白基因也可通过遗传工程应用于香蕉枯萎病的防治.     

枯草芽孢杆菌YB5对菜豆根腐病菌的抑菌机制测定及应用

腐皮镰孢引起的菜豆根腐病是一种世界性病害,在菜豆生产中危害严重。为对其有效的生物防治,针对菜豆根腐病利用枯草芽孢杆菌进行了拮抗机制及应用的研究。结果表明:YB5能有效抑制菌丝生长、孢子产生和萌发。YB5的无菌滤液对高温敏感。YB5菌液对种子发芽没有抑制作用,且能促进根系的发育。YB5菌液及无菌滤液对菜豆腐皮镰孢根腐病的盆栽均具有较好的防效,YB5菌液防效可达94.6%。     

苏云金芽孢杆菌的概况及其研究进展

苏云金芽孢杆菌(Bacillus thuringiensis,Bt)杀虫剂是对人和环境友好的安全型细菌杀虫剂。目前Bt杀虫剂已被广泛应用,但因使用剂量较大,导致防治害虫时费用过高。若在生物防治中使用超高效的细菌杀虫剂,就可降低其使用剂量。Bt 00-50-5菌株是从美国引进的高效杀虫菌株,其晶体蛋白的特征分析尚未完成,其毒素的作用机理研究也未进行。本项目即将开展如下研究：(1)先用具有特异性杀虫功能的Bt 00-50-5菌株进行发酵,继续完成对防治中国害虫防治效果的评价;再分析细菌的编码杀虫毒素基因和毒素     

Influence of temperature on the fermentation of bambara groundnut (Vigna subterranea) to produce a dawadawa-type product

Bambara groundnut (Vigna subterranea) was fermented to produce a dawadawa-type product using a starter culture of Bacillus licheniformis isolated from naturally fermenting bambara groundnut beans. Fermentation was carried out at 30 and 37 °C for four days and at 45 °C for two days. The pH of the substrate decreased after 24 hours and then rose at 30 and 37 °C but remained constant at 45 °C after the initial drop. Total titratable acidity of the fermenting beans mimicked the pH values. Proximate analyses for moisture, protein and fat of the cotyledons showed an increase in all three constituent at each of the three fermentation temperatures. At the end of fermentation, total available carbohydrate was 55%, 59% and 62% of the original value at 30, 37 and 45 °C, respectively. Fermentation of bambara groundnut at 45 °C for two days is recommended as the ideal fermentation temperature and time.     

Using Bacillus subtilis E20‐fermented soybean meal as replacement for fish meal in the diet of orange‐spotted grouper (Epinephelus coioides,Hamilton)

The potential of Bacillus subtilis E20‐fermented soybean meal (FSBM) as a partial alternative component of fish meal (FM) in fed diets of orange‐spotted grouper (Epinephelus coioides) was evaluated in this study. An FM‐based diet and seven diets containing 10%, 20% and 30% and 10%, 20%, 30% and 40% of FM replaced by soybean meal (SBM) and FSBM, respectively, were fed to grouper for 84 days to evaluate possible substitution levels of FM by tracking growth performance, digestive enzyme activity and morphological changes in the liver and distal intestine. No significant differences in survival and muscle composition of grouper were found between controls and treatments. Growth performance and feed efficiency of fish fed diets with FM replaced by FSBM up to 30% were not significantly different from controls, whereas significantly decreased growth performance and feed efficiency occurred with diets containing >20% of SBM. Based on the feed efficiency, the maximum substituted levels of FM by SBM and FSBM in grouper diets were 18.36% and 29.32%, respectively, based on broken‐line analyses. Histopathological changes in the liver and distal intestine, and significantly lower activity levels of digestive enzymes, including pepsin in the stomach and trypsin, chymotrypsin, amylase and lipase in the distal intestine, were found in fish fed a diet containing 30% of FM replaced by SBM. However, these parameters were improved by the substitution of FSBM. It is therefore believed that FSBM has great potential to be used as a protein source in grouper diets in partial replacement of FM.     

苏云金芽胞杆菌CY1菌株的cry基因分析

通过醋酸钠-抗生素筛选法从河北省土壤中筛选得到了苏云金芽孢杆菌新菌株CY1。镜检可观察到伴孢晶体呈小菱形。SDS-PAGE分析结果表明菌株CY1表达的晶体蛋白分子量为130 kDa。利用25对通用引物检测cry1,cry1I,cry7,cry2,cry3,cry4/cry10,cry5,cry6,cry7,cry8,cry9,cry11,cry13/14,cry16/17,cry18,cry19,cry21,cry22,cry24/25,cry26/28,cry27/29,cry30,cry32,cry34/35,cry40基因,仅从CY1菌株中检测出cry7基因,该cry7基因N末端的1 215 bp片段所编码的氨基酸序列与已发表的Cry7Ab1(Acc No.:U04367)序列同源性达99%,仅有2个氨基酸的差异。用3对cry7Ab特异引物检测,进一步证实该菌株携带有cry7Ab基因。     

土壤微生物膜对风沙土固沙保水特性的影响

土壤微生物膜胞外聚合物具有较强的黏结性,为增强风沙土抗蚀性,提高风沙土保水性提供了新途径。鉴于此,该研究采用室内控制试验设置6个微生物菌剂施用水平(0、1、3、5、7和10 g/kg),以期阐明土壤微生物膜对风沙土特性的影响。研究结果表明:1)施用微生物菌剂可产生土壤微生物膜,并胶结风沙土颗粒。2)试验结束时(第49 d),与对照组相比,不同菌剂施用处理的土壤容重降低0.54%～8.88%,孔隙度提高0.39%～3.91%,含水率显著提高0.11%～0.25%(P0.05),大团聚体质量分数增加5.59%～25.01%,土壤pH值由7.16显著增加至8.32～9.23(P0.05)。3)土壤多糖含量对土壤特性的解释率为47%,是影响土壤特性的关键因素。土壤微生物膜能够有效改善风沙土特性,增强风沙土抗蚀性,提高风沙土保水性,但较高的微生物菌剂施用量会增加土壤pH值,造成土壤盐碱化,建议微生物菌剂施用水平为1～5 g/kg。研究成果有利于深入理解微生物膜固沙保水机制,并可为沙化土地防治新技术研发提供重要理论依据和科技支撑。     

河道淤泥和堆肥蛭石混合发酵制备基质及其育苗效果

农村河道清淤产生的淤泥,体量大、有机物浓度高,处置不当会造成二次污染。现代农业的工厂化育苗需求大量的营养土,就地取土导致耕地退化。该研究利用功能微生物发酵淤泥制备育苗基质,研究不同菌株发酵基质的物理和生物学性状,基质培育西瓜苗的生长、生理参数和抗逆性能。结果表明:微生物处理均能够提升淤泥基质物理和生物学性能,同时能够提升育苗质量。其中Trichoderma harzianum T83(T83)、Bacillus amyloliquefaciens IAE(BIAE)菌株发酵基质性能最好。相较于对照处理基质的最大持水量、总孔隙度、毛管孔隙度、通气孔隙度,T83处理分别增加了64.25%、52.65%、45.05%、56.11%;BIAE处理分别增加了101.17%、45.43%、61.43%、38.14%。相较于对照处理西瓜苗的株高、鲜质量、干质量、叶绿素含量、根系活力、根际真菌、细菌数量,T83处理分别增加了66.85%、52.07%、72.16%、43.13%、54.93%、110.62倍、1.63倍;BIAE处理分别增加了80.40%、57.34%、82.37%、54.88%、46.40%、67.26%、2.60倍、2.94倍。T83和BIAE处理西瓜苗叶片过氧化氢酶和超氧化物歧化酶酶活显著增加,根系丙二醛含量显著降低。真菌菌株T. harzianum T83和细菌菌株B. amyloliquefaciens IAE发酵淤泥,能够显著提升其农用品质,为淤泥高附加值化农用提供一条可行的途径。