Evaluation of Streptomyces sp. strain g10 for suppression of Fusarium wilt and rhizosphere colonization in pot-grown banana plantlets

Streptomyces sp. strain g10 exhibited strong antagonism towards Fusarium oxysporum f.sp. cubense (Foc) races 1, 2 and 4 in plate assays by producing extracellular antifungal metabolites. Treating the planting hole and roots of 4-week-old tissue-culture-derived Novaria banana plantlets with strain g10 suspension (10⁸ cfu/ml), significantly (P<0.05) reduced wilt severity when the plantlets were inoculated with 10⁴ spores/ml Foc race 4. The final disease severity index for leaf symptom (LSI) and rhizome discoloration (RDI) was reduced about 47 and 53%, respectively, in strain g10-treated plantlets compared to untreated plantlets. Reduction in disease incidence was not significant (P<0.05) when plantlets were inoculated with a higher concentration (10⁶ spores/ml) of Foc race 4. Rhizosphere population of strain g10 showed significant (P=0.05) increase of more than 2-fold at the end of the 3rd week compared to the 2nd week after soil amendment with the antagonist. Although the level dropped, the rhizosphere population at the end of the 6th week was still nearly 2-fold higher than the level detected after 2 weeks. In contrast, the root-free population declined significantly (P=0.05), nearly 4-fold after 6 weeks when compared to the level detected after 2 weeks. Neither growth-inhibiting nor growth-stimulating effects were observed in plantlets grown in strain g10-amended soil.     

抗生素AGPM生物合成途径的初步研究

采用前体添加实验法、静息细胞培养法以及酶抑制剂法对藤黄灰链霉菌中抗生素AG PM生物合成途径进行了初步探讨。研究表明能转化成聚酮合成所需活性前体的氨基酸如异亮氨酸、缬氨酸、蛋氨酸、谷氨酸等以及短链脂肪酸乙酸、丙酸、丁酸盐对抗生素AGPM合成均有明显促进作用 ;另外 ,在培养基中添加脂肪酸和聚酮生物合成途径的专一性抑制剂浅蓝菌素 (2 5μg mL)或脂肪酸合成抑制剂碘乙酰胺 (0 5mmol L)时 ,菌体生长不受影响 ,而抗生素AGPM合成受到强烈抑制 ,分别为对照的 35 3 %和 2 6 2 % ;     

产诺加霉素链霉菌中失活snogA基因的研究

诺加霉素是重要的蒽环类抗肿瘤抗生素,由黑胡桃链霉菌ATCC27451发酵产生。本研究从诺加霉素产生茵中克隆得到560bp的氨基甲基化酶（snogA）编码基因片段,并将其插入基因整合型质粒pKCll39的多克隆位点,构建得到基因中断质粒pLMX-3-58。通过接合转移和同源重组,构建得到氨基甲基化酶编码基因被中断的重组菌株删-3-59。基因重组突变株基因型验证结果表明,中断质粒以正确方式整合入基因组,将氨基甲基化酶编码基因中断。发酵验证结果表明,重组茵株发酵产物中不含有诺加霉素。本研究表明snogA基因在诺加霉素生物合成途径中是必需的。这为进一步阐明诺加霉素生物合成途径和组合生物合成改造诺加霉素提供了参考。     

Inoculum Density and Population Dynamics of Suppressive and PathogenicStreptomycesStrains and Their Relationship to Biological Control of Potato Scab

SeveralStreptomycesstrains are capable of suppressing potato scab caused byStreptomyces scabies.Although these strains have been successful in the biocontrol of potato scab in the field, little is known about how populations of pathogenicStreptomycesin the potato rhizosphere are influenced by inoculation of the suppressive strains. The effects of inoculum densities of pathogenic and suppressiveStreptomycesstrains on their respective populations on roots and in rhizosphere soil were examined during the growing season. The relationships between inoculum density or rhizosphere population densities and disease severity were also investigated. Populations of suppressiveStreptomycesstrain 93 increased significantly on roots with increasing inoculum dose. At its highest inoculum dose, the suppressive strain reached a population density greater than 10⁶CFU/g root 14 weeks after planting. The ability of the suppressive strain to increase its populations with increasing inoculum density was hindered at high inoculum doses of the pathogen, suggesting that density-dependent competitive interactions may be occurring between the two antagonists. Strain 93 was most effective at preventing scab early in the growing season (8 weeks after planting), when tubers were most susceptible to the scab disease. Population densities of the suppressive strain in soil were more highly negatively correlated with scab severity than were populations on roots, suggesting that rhizosphere soil rather than potato roots may be the primary source of inoculum of the suppressive strain for tubers.     

天蓝色链霉菌Streptomyces coelicolor中几丁质酶C(Chi C)的生物信息学分析

利用生物信息学的方法,分析天蓝色链霉菌Streptomyces coelicolor中几丁质酶C（Chi C）的一些基本性质,并针对链霉菌属菌种的几个几丁质酶基因做了进化树,进而验证了天蓝色链霉菌中至少8种几丁质酶的分类;同时对天蓝色链霉菌Streptomyces coelicolor中几丁质酶C（Chi C）蛋白的高级结构作出了预测,得到其编码的属于18家族的蛋白质高级结构图谱。     

非核糖体肽合成酶缩合结构域基因片段克隆

从大连渤海海域筛选出1株放线菌L1,结合形态观察、生理生化实验和16S rDNA分子鉴定,确定L1属于链霉菌属球孢链霉菌(Streptomyces globisporus)。根据GenBank发布的非核糖体肽合成酶(NRPS)序列设计引物,从放线菌L1的基因组DNA中扩增获得NRPS基因片段。测序结果及比对分析表明该片段属于NRPS缩合结构域部分序列。三维建模显示其结构呈V型,包含缩合结构域核心序列,与数据库已知结构相一致,可以推断该克隆片段为NRPS缩合结构域基因片段,为后续深入研究缩合结构域特异性与相关NRPS功能提供基础。     

亚硝基胍诱变选育林肯霉素高产菌株

以林肯链霉菌９４７－８（Ｓｔｒｅｐｔｏｍｙｃｅｓｌｉｎｃｏｌｎｅｎｓｉｓ９４７－８）为出发菌株（产林肯霉素９４０γ／ｍｌ）。采用孢子热处理方法处理出发菌株孢子,得到变异株９４７－８ｓ,产林肯霉素１０８０γ／ｍｌ。对９４７－８ｓ菌株进行ＮＴＧ诱变处理,得变异株９４７－８ｘ,产林肯霉素为１２１８γ／ｍｌ,且生产能力稳定。     

高温链霉菌质粒pTSC2的测序分析和滚环复制方式的鉴定

从猪粪堆肥中分离到一株编号为X3-3的可以在50℃高温生长的链霉菌菌株,该菌株含有一个约7kb的环型质粒pTSC2。【目的】克隆、测序和分析pTSC2,以及鉴定质粒的复制方式。【方法】利用分段克隆和引物延伸获得pTSC2的全序列,利用多序列比对寻找复制元件rep、dso和sso,利用中性转移和Southern杂交检测复制中间体。【结果】克隆和测序获得了全长为7516bp的pTSC2序列,预测编码8种蛋白,其中4种蛋白与链霉菌滚环复制的质粒pIJ101中负责复制和接合转移的蛋白非常相似。pTSC2的复制元件rep、dso和sso也与pIJ101的相似。克隆、转化变铅青链霉菌ZX7以及高温链霉菌2C证明了rep和dso为复制所必需元件。Southern杂交检测到pTSC2复制过程中积累了大量的单链DNA。【结论】高温链霉菌质粒pTSC2以滚环方式进行复制。这是首次在高温链霉菌中克隆和测序质粒,以及鉴定其复制方式。     

聚-ε-赖氨酸高产菌株的筛选及其发酵工艺的初步研究

利用亚甲基蓝平板初筛、摇瓶复筛,从土壤中筛选了一株聚-ε-赖氨酸(ε-PL)高产菌株,ε-PL摇瓶产量大于2g/L,经初步鉴定该菌株为白色链霉菌(Streptomyces albulus)。对该菌株发酵生产ε-PL的培养基成分进行初步研究表明:葡萄糖是最好的碳源,酵母粉和硫酸铵是最佳的复合氮源,在优化培养基下,ε-PL摇瓶产量达到3.9g/L。     

吸水链霉菌ATCC 29253产Hygrocin A发酵条件的优化

【背景】Hygrocins是一种萘安莎抗生素,具有良好的新药开发潜能。但在常见培养基及发酵条件下菌体内Hygrocin A含量一般很低,甚至难以直接进行准确检测。【目的】提高吸水链霉菌ATCC 29253发酵物中Hygrocin A的产量。【方法】采用单因素与正交试验设计优化相结合的方法系统考察碳源、氮源、磷酸盐、MgCl_2浓度、NaCl浓度、种子菌龄等因素对吸水链霉菌ATCC 29253产Hygrocin A能力的影响。【结果】最佳发酵条件为(g/L):葡萄糖4.0,黄豆饼粉8.0,麦芽提取物10.0,K_2HPO_4 1.5,KH_2PO_4 1.5,NaCl 1.5,Mg Cl2 1.0;种子最佳活化时间为48 h;培养参数:摇床转速200 r/min,初始pH为6.8-7.0,瓶装量50 m L/250 m L,接种量5%,30°C培养10 d。在优化条件下,Hygrocin A产量与其原始培养基M10相比提高了500%,Rapamycin产量同时下降了95%。【结论】通过培养基优化,可显著提高吸水链霉菌ATCC 29253中Hygrocin A产量,为Hygrocin A合成应用研究奠定基础,同时可使Rapamycin产量明显下降。这说明可通过选择培养条件有目的地调节两种抗生素的代谢通量,进而开展多种抗生素同时表达的代谢调控研究。