解淀粉芽胞杆菌B1619生物摇瓶发酵工艺的正交优化

解淀粉芽胞杆菌Bacillus amyloliquefaciens B1619对设施蔬菜土传病害有较好的防治效果。为了提高菌株B1619生物发酵效率,本文通过单因子水平筛选和正交试验,对菌株B1619发酵培养基和培养条件进行了优化。结果表明,发酵培养基3个主要因子的最佳配比组合为酵母膏0.25%,大豆粉2.00%,生长素0.05%,优化后生物发酵效率比初始培养基提高了21.79%;发酵培养条件3个主要因子的最佳组合为温度28℃、转速180 r/min、装液量60 mL/250 mL,优化后生物发酵效率比初始培养条件提高了35.42%。经验证,优化后的生物发酵工艺获得的发酵菌液含菌量为7.13×10⁹cfu/mL,与初始发酵工艺的发酵菌液含菌量(3.80×10⁹ cfu/mL)相比,提高了87.83%,优化效率十分显著。     

Molecular Weight-Dependent Immunostimulative Activity of Low Molecular Weight Chitosan via Regulating NF-κB and AP-1 Signaling Pathways in RAW264.7 Macrophages

Chitosan and its derivatives such as low molecular weight chitosans (LMWCs) have been found to possess many important biological properties, such as antioxidant and antitumor effects. In our previous study, LMWCs were found to elicit a strong immunomodulatory response in macrophages dependent on molecular weight. Herein we further investigated the molecular weight-dependent immunostimulative activity of LMWCs and elucidated its mechanism of action on RAW264.7 macrophages. LMWCs (3 kDa and 50 kDa of molecular weight) could significantly enhance the mRNA expression levels of COX-2, IL-10 and MCP-1 in a molecular weight and concentration-dependent manner. The results suggested that LMWCs elicited a significant immunomodulatory response, which was dependent on the dose and the molecular weight. Regarding the possible molecular mechanism of action, LMWCs promoted the expression of the genes of key molecules in NF-κB and AP-1 pathways, including IKKβ, TRAF6 and JNK1, and induced the phosphorylation of protein IKBα in RAW264.7 macrophage. Moreover, LMWCs increased nuclear translocation of p65 and activation of activator protein-1 (AP-1, C-Jun and C-Fos) in a molecular weight-dependent manner. Taken together, our findings suggested that LMWCs exert immunostimulative activity via activation of NF-κB and AP-1 pathways in RAW264.7 macrophages in a molecular weight-dependent manner and that 3 kDa LMWC shows great potential as a novel agent for the treatment of immune suppression diseases and in future vaccines.     

生物有机肥育苗防控烟草青枯病

【目的】烟草青枯病是影响贵州烟叶生产的主要土传病害之一。生物防控方法作为防控烟草青枯病体系的有效措施之一,寻求经济可行的途径势在必行。【方法】利用实验室自主筛选到的烟草青枯菌拮抗菌NJL-14,通过二次固体发酵研制成烟草专用拮抗青枯病型生物有机肥 (BIO),选取K326和红花大金元两个烟草品种分别进行育苗和盆栽试验。育苗床基质分别添加BIO 1%、2%、3%、4%和5%,以确定适宜的BIO育苗添加量。盆栽试验两个品种分别设定五个处理:常规基质育苗 + 植烟健康土壤(CKn+CKp）、常规基质育苗?+?青枯病病土（CKn+Rs）、常规基质育苗?+?青枯病病土?+?有机肥(CKn+OFp)、常规基质育苗?+?青枯病病土?+?BIO(CKn+BIOp)、育苗和盆栽病土中都施入BIO(BIOn+BIOp)。分别采用荧光定量PCR和Biolog-ECO研究育苗基质和盆栽土壤中细菌和真菌数量变化,以及微生物功能多样性,特别是烟草青枯菌及其拮抗菌的消长关系。【结果】当BIO浓度在2%以内时,烟草的发芽率与对照无显著差异;与普通漂浮育苗相比,托盘育苗不但能稳定和延长拮抗菌NJL-14在根际的定殖,而且显著提高烟苗生物量,K326经BIO育苗处理的烟苗地上部和地下部的干重分别比常规基质育苗的处理高17.2%和30.8%,而红花大金元分别高14.9%和20.0%;采用育苗与移栽土壤双重使用BIO模式对烟草青枯病的防控效果明显,防控效果可以达到100%,明显优于只在盆栽中使用BIO模式,并且能够显著抑制土壤中病原菌数量在107 cfu/g土以下,同时有效提高土壤微生物生态多样性。【结论】采用育苗与移栽土壤双重使用烟草专用拮抗青枯病型生物有机肥模式可有效防控烟草青枯病,控制土壤中病原菌数量,改善土壤微生物功能多样性,为有效地生物防控烟草青枯病奠定基础。     

防治稻瘟病芽胞杆菌的筛选及效果评价

从232株细菌中筛选获得2株对稻瘟病菌具有明显抑菌活性的菌株S09和S170,对稻瘟病菌Magnaporthe oryzae P131菌丝的抑制率分别为90.44%和92.38%;离体叶片法测定表明,菌株S09和S170抑菌效果分别为74.62%和75.52%,与绿地康1号无显著差异。16S rDNA序列分析比对,菌株S09与短小芽胞杆菌Bacillus pumilus基因序列的同源性最高;gyrA基因测序结果表明,菌株S170与解淀粉芽胞杆菌B.amyloliquefaciens基因序列的同源性最高。结合形态观察,确定菌株S09为短小芽胞杆菌,菌株S170为解淀粉芽胞杆菌。菌株S09和S170对稻瘟病的温室防治效果分别是76.56%和80.57%。田间试验结果表明,菌株S09和S170对水稻有明显的促生和增产作用,分别提高水稻种子发芽17.37%和12.34%、促进根长18.68%和33.85%、增加株高12.44%和28.49%、降低空秕率25.17%和35.59%,还可以有效改善稻米的垩白粒率和垩白度。短小芽胞杆菌S09和解淀粉芽胞杆菌S170对叶瘟的田间防治效果分别是70.59%和73.19%,对穗颈瘟的田间防治效果分别是71.55%和74.82%,菌株S170对稻瘟病的防治效果与75%三环唑可湿性粉剂、绿地康1号处理无显著差异。     

Combined application of Trichoderma harzianum SH2303 and difenoconazole-propiconazolein controlling Southern corn leaf blight disease caused by Cochliobolus heterostrophus in maize

Southern corn leaf blight(SCLB) disease caused by Cochliobolus heterostrophus is one of the major threats to corn production worldwide. The synergistic application of low toxic chemical fungicide and biocontrol agents could improve biocontrol stability and efficiency against plant diseases, which ultimately reduce use of chemical fungicide. Trichoderma spp., well-known biocontrol fungi have been used to control some foliar diseases. However, few works have been reported on synergistic application of chemical fungicide and Trichoderma against foliar diseases. This study was aimed to investigate the control effect on the synergistic application of Trichoderma harzianum SH2303 and difenoconazole-propiconazole(DP) against SCLB. Results showed that the synergistic application of DP and SH2303 reduced the leaf spot area compared to the control. The efficacy of synergistic application of DP+SH2303 against SCLB could last for 15–20 d in pot trial under the greenhouse condition. Under the natural field condition, maize treated with DP+DP and DP+SH2303 showed 60% control, which was higher than that of SH2303+DP(45%) and SH2303+SH2303(35%). All these treatments induced the synthesis of defense-related enzymes(phenylalanine ammonia lyase(PAL), catalase(CAT), and superoxide dismutase(SOD)) and the defence-related gene expression of SA pathway(PR1). Taken together the in-vitro leaf test and field trial, the control of SCLB by synergistic application of DP+SH2303 was similar to that of DP+DP. Among synergistic application, the sequential application of DP+SH2303 showed better control than the sequential application of SH2303+DP. It was concluded that the synergistic application of chemical fungicide(DP) and biocontrol agent(T. harzianum SH2303) could be used to reduce the chemical fungicide and to reduce the SCLB diseases in maize, which provided alternative approach to realize an eco-friendly controlling of the foliar disease.     

绿僵菌油剂防治东亚飞蝗田间试验

20 0 2年在天津大港以绿僵菌LA06油剂防治东亚飞蝗进行了田间试验。绿僵菌分生孢子系在实验室采用常规双相产孢系统 ,在大米固体培养基上培养产生。油剂用V(大豆油 )∶V(煤油 ) =3∶7配制。试验区选用蝗蝻相对不易迁出迁入的高密度地带 ,3龄蝗蝻占 90%,成点片状密集分布。超低量喷雾 1hm2 喷施绿僵菌油剂1.5L ,含绿僵菌干粉48g ,其分生孢子含量为1.37×10¹²个。药效调查采用全区密度级别估测法调查试验区虫口密度变化 ,并以笼罩法试验作为旁证。结果表明 ,在施药后 6～9d蝗蝻种群急剧减少 ,第 18天虫口减退率达 81.1%。田间防治与笼罩试验结果一致     

棉蚜茧蜂[Lysiphlebia japonica (Ashmead)]对棉蚜不同虫态寄生选择的研究

研究表明,棉蚜茧蜂[Lysiphlebia japonica(Ashmead)]喜好寄生棉蚜2、3龄若蚜,寄生率分别为78.8％和87.2％;对4龄若蚜的寄生率为10.6％;1龄若蚜的寄生率仅为5％;棉蚜茧蜂一般不选择有翅蚜作为寄主。随着供蚜量的增加,未被寄生的蚜虫百分率从21.3％上升至63％;过寄生率从55.1％下降至8.37％;废卵率从53.1％降至7.9％。棉蚜茧蜂对健康棉蚜和被寄生过的棉蚜不能完全识别,存在过寄生现象。     

喷雾法施用线虫大面积防治荔枝拟木蠹蛾

荔枝拟木蠹蛾(Arbela dea Swinhoe)是南方荔枝、木麻黄等主要的钻蛀害虫,作者利用拟木蠹蛾昼伏夜出的习性,用大蜡螟幼虫诱测和回收喷雾荔枝拟木蠹蛾粪道上的线虫(Steinernema carpocapsae Agriotos),不同温度、不同粪道大小对线虫存活影响的测试结果表明,应用1×10~3/ml线虫剂量喷雾拟木蠢蛾挂在树上的粪道,粪道上便有足够数量的线虫和时间使寄主感染。1995年喷雾法施用线虫防治荔枝拟木蠹蛾260hm~2,防治效果在85％～100％,平均为94.28％,有效地压低了田间的虫口密度。解决了树冠嫩枝受害后人工难以注射及倒置斜垂坑道的虫口防治效果差的矛盾,也使利用病原线虫防治害虫这一有效的生物防治手段更容易为果农所接受。     

Metabolites of some biological control agents

The evidence is reviewed for the structure and biosynthesis of some metabolites of Trichoderma and Gliocladium species that are being considered as fungal biological control agents. The diverse biosynthetic capability of these organisms is proposed as a criterion for their potential application as bio-control agents.