In Vivo Metabolism Study of Xiamenmycin A in Mouse Plasma by UPLC-QTOF-MS and LC-MS/MS

Xiamenmycin A is an antifibrotic leading compound with a benzopyran skeleton that is isolated from mangrove-derived Streptomyces xiamenensis. As a promising small molecule for fibrotic diseases, less information is known about its metabolic characteristics in vivo. In this study, the time-course of xiamenmycin A in mouse plasma was investigated by relative quantification. After two types of administration of xiamenmycin A at a single dose of 10 mg/kg, the plasma concentrations were measured quantitatively by LC-MS/MS. The dynamic changes in the xiamenmycin A concentration showed rapid absorption and quick elimination in plasma post-administration. Four metabolites (M1–M4) were identified in blood by UPLC-QTOF-MS, and xiamenmycin B (M3) is the principal metabolite in vivo, as verified by comparison of the authentic standard sample. The structures of other metabolites were identified based on the characteristics of their MS and MS/MS data. The newly identified metabolites are useful for understanding the metabolism of xiamenmycin A in vivo, aiming at the development of an anti-fibrotic drug candidate for the therapeutic treatment of excessive fibrotic diseases.     

2株放线菌的抗菌活性及分类学地位

旨在对2株放线菌HD-103和HD-109的抗菌活性和发酵液稳定性进行比较分析,并明确2株菌的分类学地位。以13种植物病原真菌为靶标菌,利用琼脂柱法分析2株生防菌的抗菌谱;以尖孢镰刀菌(Fusariumoxysporumf.sp.glycines)为指示菌,利用杯碟法检测2株生防菌发酵液的热稳定性、pH稳定性及紫外稳定性,并进行比较分析;在此基础上采用多相分类法对2株菌进行鉴定。研究结果显示：2株菌的抗真菌谱均较广,其中HD-103对禾谷镰刀菌(Fusariumgraminearumf.sp.Glycines)的抑制效果最显著,抑菌圈直径达到23.7mm,HD-109对烟草赤星病菌(Alternariaalternate)抑制效果最好,抑菌圈直径达25.0mm;HD-103的抗菌活性物质较HD-109的活性物质更为稳定;分类学鉴定结果表明,菌株HD-103与HD-109为微白黄链霉菌(Streptomycesalbidoflavus)的2个菌株。菌株HD-103与HD-109能够抑制多种植物病原真菌,其发酵液具有较好的热稳定性和酸碱稳定性,具有开发为新型生物农药的潜力。     

产纳他霉素生防菌A01和A02与已知产生菌的RAPD比较

为了明确自主分离的天然抗真菌活性产物——纳他霉素产生菌A01和A02与已知纳他霉素产生菌的遗传相似性,采用RAPD技术对这2株菌和3个产纳他霉素的标准菌株进行了比较分析。利用筛选出的7个随机引物对5个菌株的PCR扩增共得到154条清晰稳定的DNA条带,其中同源性条带86条,多态性条带68条,分别占总条带数的55.8%和44.2%,平均每个引物扩增出9.7个多态性条带,得到了丰富的DNA指纹图谱。所得数据经NTSYS pc 2.10e软件聚类分析,表明5个菌株间的遗传距离为0.0991~0.4738,其中A02与其它菌株间的遗传距离均较远。结合前期的分类研究结果,确证了菌株A02为新的纳他霉素产生菌。     

井冈霉素高产菌株原生质体诱变筛选研究

以吸水链霉菌井冈变种k-121-5为出发菌株制备原生质体,并对其进行紫外诱变,通过筛选得到5株较为高产的菌株,经连续传代、摇瓶发酵,井冈霉素A产率比出发菌株提高14%～26%。     

Survey of Bacterial and Fungal Seedborne Diseases in Imported and Domestic Potato Seed Tubers

Potato seed tubers are imported to Israel from northern Europe and planted in spring; tubers harvested early from the spring crop are used as seed for the autumn crop. Although only seed lots registered as certified are imported, a previous survey (1984–1994) indicated that most imported lots were affected by latent or active infections caused byErwinia carotovora,Streptomyces scabies, Rhizoctonia solani, Fusarium spp. andSpongospora subterranae. The survey was extended until 1998, and included additional pathogens:Ralstonia solanacearum,Helminthosporium solani, Colletotrichum coccodes andVerticillium dahliae. Most of these pathogens were also monitored in domestic seed tubers, and are reported for the first time. Brown rot was not observed in any of the imported lots. Blackleg and soft rot caused byErwinia spp. were detected in most of the imported lots; however, less than 7% of the lots were contaminated at high levels, while approximately 65% were contaminated at moderate levels. Common scab was detected in most of the imported lots; 51% of the imported lots were contaminated at moderate or high levels, whereas only 6.5% of the domestic seed lots were contaminated at these levels. Black scurf was detected in most of the imported lots; on average, 47.3%, 44.2% and 1.4% of the lots were contaminated at low, moderate and high levels, respectively, and only 7.1% were disease-free. In contrast, most of the domestic lots were either disease-free (45.4%) or had a low disease incidence (37.3%). Only 16.7% of the lots were moderately infected and 0.2% were highly contaminated. Silver scurf was observed in most of the imported lots during all years of the survey, with no differences among the producing countries; on average, 22.7%, 66.1% and 7.5% of the lots were contaminated at low, moderate and high levels, respectively, and only 3.7% were disease-free. Most of the domestic lots (76%) were disease-free and only 6.6% were infected at moderate or high levels. Black dot was observed in a considerable portion of the shipments from Holland during all years of the survey, particularly in 1998, when 34% of the lots were infected. The shipments from France and Germany were infected at low levels, except in 1998, when 19% and 11% of the lots, respectively, arrived infected. In shipments from Scotland and Ireland low incidences of the disease were observed in 1994 and 1995. In the domestic lots, black dot incidence was low (<2.4%) except in 1996, when 11% of the lots were infected.V. dahliae was monitored only in domestic seed tubers. The incidence of disease-free lots was 56–64%, whereas in 20–30% of the lots the level of infection was <5%, and in 6–16% of the lots the level was >5%. The survey findings demonstrate transmission of seedborne pathogens; most of these pathogens can become established in the soil and eventually cause severe outbreaks of disease in potatoes grown in Israel. http://www.phytoparasitica.org posting May 16, 1999.     

Comprehensive Genomic Analysis of Marine Strain Streptomyces sp. 891, an Excellent Producer of Chrysomycin A with Therapeutic Potential

Chrysomycin A is one of the most promising therapeutic candidates for treating infections caused by multidrug-resistant Gram-positive bacteria. By hybridizing next-step generation (Illumina) and third-generation (PacBio) sequencing technologies, a high-quality chromosome-level genome together with a plasmid was firstly assembled for chrysomycin A-producing marine strain 891. Phylogenetic analysis of the 16S rRNA gene and genome sequences revealed that this strain unambiguously belonged to the genus Streptomyces, and its genomic features and functional genes were comprehensively analyzed and annotated. AntiSMASH analysis of this strain unveiled one key biosynthetic gene cluster, T2PKS, responsible for the biosynthesis of chrysomycin, the biosynthesis pathway of which was putatively proposed. These findings definitely shed light on further investigation for construction of a robust industrial strain with high-yield chrysomycin A production using genetic engineering techniques and combinatorial biology approaches.     

利迪链霉菌A02诱导番茄抗灰霉病作用机理研究——对植株两种PR蛋白和总酚的影响

为了研究新型链霉菌生防菌株A02诱导番茄抗灰霉病的作用机制,利用利迪链霉菌A02的发酵液和番茄灰霉病菌,对番茄植株进行诱导和接种处理,采用分光光度法,测定处理前和处理后1-5 d番茄叶片组织中2种PR蛋白(几丁质酶和β-1,3-葡聚糖酶)和总酚含量的变化。试验结果显示,不接种灰霉病菌而单独诱导处理、接种灰霉病菌后诱导处理和诱导处理后接种灰霉病菌这3种处理,都能显著地提高番茄植株几丁质酶和β-1,3-葡聚糖酶的活性,且能提高总酚的相对含量,其中诱导处理中,酶活性和总酚相对含量提高的较为明显,酶活性峰值和总酚相对含量峰值都较高。通过试验,初步证明了利迪链霉菌A02发酵液诱发了番茄体内2种防御酶活性的提高和酚类物质的积累,增强了番茄植株抗灰霉病的防御体系。     

Unique Cyclized Thiolopyrrolones from the Marine-Derived Streptomyces sp. BTBU20218885

Two new cyclized thiolopyrrolone derivatives, namely, thiolopyrrolone A (1) and 2,2-dioxidothiolutin (2), together with the kn own compound, thiolutin (3) were identified from a marine-derived Streptomyces sp. BTBU20218885, which was isolated from a mud sample collected from the coastal region of Xiamen, China. Their chemical structures were determined using spectroscopic data, including HRESIMS, 1D and 2D NMR techniques. 1 possessed a unique unsymmetrical sulfur-containing thiolopyrrolone structure. All the compounds were tested for bioactivities against Staphylococcus aureus, Escherichia coli, Bacille Calmette–Guérin (BCG), Mycobacterium tuberculosis, and Candida albicans. 1 displayed antibacterial activities against BCG, M. tuberculosis, and S. aureus with minimum inhibitory concentration (MIC) values of 10, 10, and 100 μg/mL, respectively. Thiolutin (3) showed antibacterial activities against E. coli, BCG, M. tuberculosis, and S. aureus with MIC values of 6.25, 0.3125, 0.625, and 3.125 μg/mL, respectively.     

拮抗放线菌A03的生防作用及其分类鉴定

从京郊菜园土壤中分离到一株拮抗放线菌,编号为A03。为了科学评价该菌株在植物病害生防中的应用潜力,采用平板对峙培养法和温室盆栽防治法测定了其抑菌活性,并通过形态学、化学和分子分类的方法研究其分类地位。结果表明,该菌株对供试的多种植物病原真菌均有抑制作用,其中对果蔬灰霉病菌、番茄早疫病菌、小麦纹枯病菌和瓜果腐霉的抑制作用较强,平板抑菌带宽度达11．0～12．0 mm。温室防病试验结果表明,其发酵上清液对甜椒灰霉病和番茄灰霉病具有良好的生防作用,防治效果分别为84．97%和79．51%。根据其菌株形态特征、培养性状、生理生化特性、细胞壁化学组分及16S rDNA序列分析,将菌株A03归入链霉菌属,并鉴定为吸水链霉菌Strep- tomyces hygroscopicus。     

链霉菌ZZSP-7的鉴定及其对草莓炭疽病的防效

为获得草莓炭疽病菌胶孢炭疽菌的优良拮抗菌,以草莓种植基地的土壤为拮抗菌来源,通过室内测定不同分离物对草莓炭疽病菌的抑菌活性,筛选抑菌率最高的菌株,在此基础上对所筛菌株进行分类鉴定,并评价其抑菌谱和防效。结果表明,从土壤中分离获得42株拮抗菌,筛选出5株对胶孢炭疽菌具有明显抑制作用的菌株。其中,菌株ZZSP-7的抑菌作用最强,5 d后的抑菌带宽达17.6 mm。经形态、生理生化及16S rDNA基因系统发育分析,确认菌株ZZSP-7为链霉菌Streptomyces sp.。菌株ZZSP-7抑菌谱广,对多种植物病原菌均具有良好的抑菌效果。盆栽试验结果表明,在盆栽中菌株ZZSP-7对“红颜”和“甜查理”两个品种草莓炭疽病菌的防效分别为78.64%和82.81%,具有进一步开发和利用的前景。