黄绿木霉菌对大豆根腐病镰刀菌的拮抗作用

利用室内对峙培养与发酵液处理病原菌研究了黄绿木霉(Trichoderma aureoviride)对大豆根腐病几种镰刀菌(Fusarium spp.)的拮抗作用,并利用盆栽试验研究了黄绿木霉对苗期大豆根腐病发生的影响.拮抗试验结果表明,黄绿木霉在与镰刀菌对峙培养中,菌丝交叉、接触与纠结,接触后镰刀菌菌丝被破坏;发酵产物只可延长镰刀菌孢子萌发的时间,但对孢子最终萌发率没有影响,发酵产物对菌丝生长速率的影响以山梨醇、葡萄糖及蔗糖作为碳源时,抑菌效果好,抑菌能力分别可达63%、51%、58%;盆栽试验证明黄绿木霉对大豆根腐病有较好的防治效果,防效可达73.2%,处理后幼苗株高与干重均高于未处理的对照.     

Antimicrobial susceptibility and enrofloxacin resistance of streptococcal bacteria from farmed Nile tilapia,Oreochromis niloticus (Linnaeus 1758) in Thailand

This study examined the antimicrobial susceptibility and mutation(s) in quinolone resistance‐determining regions (QRDRs) in streptococcal pathogens isolated from farmed Nile tilapia Oreochromis niloticus in Thailand. Surveillance of antimicrobial susceptibility in tilapia streptococcal pathogens reveals that Streptococcus agalactiae (n = 97) and Streptococcus iniae (n = 3) from diseased tilapia were susceptible to amoxicillin, florfenicol, sulfamethoxazole/trimethoprim and sulfadimethoxine/ormetoprim, however, only 78 isolates were susceptible to enrofloxacin. Twenty‐two enrofloxacin‐resistant S. agalactiae isolates were further examined for mutations in the QRDRs of gyrA, gyrB, parC and parE genes. Twenty isolates had single base pair changed in the gyrA sequence, C‐242‐T. Point mutations in gyrB, GC‐1135, 1136‐AA and T‐1466‐G, were identified in one isolate. All resistant isolates harboured a mutation in the parC gene, C‐236‐A, while no mutations were observed in the parE gene. The study represented mutations of gyrA and parC genes as marked modification of the enrofloxacin‐resistant S. agalactiae from farmed tilapia. This study is a primary report of the QRDRs mutations associated with fluoroquinolone resistance from streptococcal pathogen in the cultured fish. The phenotypic and genotypic characterization of enrofloxacin resistance S. agalactiae evident in this study has led to an improved regulation of antimicrobial use in Thai aquaculture.     

培养基成分和补料对阿维菌素发酵过程中除虫链霉菌菌丝形态的影响

研究了阿维菌素发酵过程中培养基成分和补料对菌丝形态的影响,以及在这些条件下菌丝形态、菌体代谢以及产量之间的相互关系。结果表明:碳氮源种类和碳氮比与菌体生长密切相关,对除虫链霉菌前期的菌丝形态影响较大。只有当除虫链霉菌为密实的球状时,才能获得较高的阿维菌素产量;进一步通过调控使发酵中后期菌丝形态保持一定的菌球尺寸(球核面积、核区周长和菌丝球面积)对维持阿维菌素合成有利。     

Soybean molasses as an organic carbon source in the farming of Litopenaeus vannamei (Boone, 1931) in a biofloc system

Soybean molasses was evaluated as a partial replacement for sugarcane molasses as a carbon source for biofloc development in the superintensive culture of Pacific white shrimp (Litopenaeus vannamei). A 50‐day study was conducted with juvenile (3.2 g) shrimp stocked in 16 800 L tanks at a stocking density of 250 shrimp m^?3. Control of total ammonia concentration was performed by the addition of combined mixtures of soybean and sugarcane molasses to the culture water. Three different molasses treatments were evaluated using different soybean‐to‐sugarcane molasses ratios: 15–85%, 38–62% and 60–40% respectively. The control group was treated only with sugarcane molasses. Water quality, chlorophyll a concentration, heterotrophic bacterial load, Vibrio spp. concentration and zootechnical indexes were all evaluated. Total ammonia concentration was controlled by heterotrophic and chemotrophic pathways. Biofloc formation, as quantified by measuring the total suspended solids, was not altered. The Vibrio spp. concentration showed a significant reduction in treatments with soybean‐to‐sugarcane molasses ratios of 38–62% and 60–40%. All combined mixtures of soybean and sugarcane molasses could maintain water quality and productivity in the superintensive culture of L. vannamei using the biofloc system. Thus, the potential use of a residue from agroindustry as a carbon source in a biofloc culture is demonstrated.     

玫瑰黄链霉菌Men-myco-93-63有效成分的初步提取与纯化

为了解玫瑰黄链霉菌Men-myco-93-63的代谢产物和其生物活性,对该菌株进行摇瓶培养,从菌丝中提取到一种对棉花黄萎病菌有明显抑制作用的有效成分。经过XAD-1600型大孔吸附树脂,ODS C18硅胶中压柱分离得到黄白色粉末。HPLC检测后,初步断定该物质是一类非常相似的物质组成的混合物,并且极有可能具备互变异构的特性。     

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。     

产类黄酮链霉菌的紫外线诱变选育

诱变选育高产类黄酮链霉菌菌株,采用不同强度的紫外线对出发菌株链霉菌HNS2-2单孢子悬液诱变处理。以出发菌株作为对照,分光光度法测定诱变菌株发酵产物黄酮类物质含量为评价指标,进行反复筛选。在20 W紫外灯、辐射距离30 cm、照射20 s条件下,获得高产菌株251,其黄酮类物质产量达41.87μg/mL,比出发菌株提高15.52%,且连续7次传代稳定性能稳定。通过紫外诱变选育的方法,能提高出发菌株黄酮类物质产量。     

玫瑰黄链霉菌Men-myco-93-63负调控基因nsdAmgh的克隆及序列分析

nsdA (Negative regulator of Streptomyces differentiation)基因是天蓝色链霉菌中发现的与抗生素合成相关的负调控基因.根据天蓝色链霉菌 A3(2)的序列设计引物扩增玫瑰黄链霉菌 Men-myco-93-63 菌株中的该基因,经测序和序列分析表明,由保守序列引物nsdA-2R和nsdA-2L克隆到的Men-myco-93-63 菌株中的该基因 800 bp片段与已知nsdA基因核苷酸保守区域序列同源性达到99%,由全长序列扩增引物nsdA WZ-R和nsdA WZ-L克隆到的该基因包含一个完整的开放阅读框,共编码 369 个氨基酸,与变铅青链霉菌和天蓝色链霉菌A3(2)中的nsdA 基因的核苷酸序列同源性达到100%,氨基酸序列同源性达到99%.该全长基因命名为nsdA_(mgh),其在玫瑰黄链霉菌 Men-myco-93-63 中的发现为阻断该负调控基因以期获得抗生素高产工程菌株提供了重要依据.     

柱花草炭疽病菌的鉴定及生物学特性研究

对柱花草炭疽病菌的鉴定及生物学特性测定结果表明：柱花草炭疽病菌为盘长孢状刺盘孢菌[Colletotrichumgloeosporioides（Penz）Sacc]．病菌生长温度15～36℃;适宜生长温度25～30℃;分生孢子萌发的温度6～40℃,最适萌发温度为28℃;分生孢子在饱和湿度下萌发率为83．1％,在饱和湿度加水滴中达87．9％;分生孢子的致死温度为49℃10min;病菌生长的pH值是2．5～14．0,最适pH值为6．4～7．2;病菌对碳源的利用较氮源好。