茂原链霉菌DSM 40847全基因组测序及序列分析

茂原链霉菌（Streptomyces mobaraensis）是一种高GC含量的革兰氏阳性放线菌,它广泛用于生产转谷氨酰胺酶、博来霉素、莫能霉素等。目前,还没有相关研究报道茂原链霉菌的全基因组序列,这限制了基于功能基因挖掘和利用的定向育种以及分子遗传研究。本研究首次通过高通量测序技术对茂原链霉菌DSM40847进行全基因组测序,使用Velvet软件进行拼接得到266 contigs,整个基因组大小约7.5 Mb,GC含量为73.2%,序列已提交至美国国立生物技术信息中心（NCBI）的GenBank数据库。通过对基因组序列进行分析,成功鉴定转谷氨酰胺酶酶原激活相关蛋白基因,同时对该菌株的次级代谢产物合成基因簇进行了预测,相关研究结果将为茂原链霉菌的功能基因组学研究提供基础数据。     

烬灰吸水链霉菌突变体的诱导研究

烬灰吸水链霉菌对稻瘟病菌、西瓜枯萎病菌等真菌病害均具有不同程度的抑制作用。用物理诱变、化学诱变、复合诱变3种方式对菌株孢子进行处理,最高变异率分别为25·2%,22·2%,24·1%。实验结果表明,物理诱变和复合诱变对菌株孢子有较好的诱变作用,而化学诱变效果不明显。诱变处理时间分别为30s和2min,可诱变产生较多的变异菌株。分别以2%诱变剂诱变处理1·5h以及60s诱变处理筛选的变异菌株对稻瘟病和西瓜枯萎病都有很好的抑制作用。     

Antagonistic activity against rice blast disease and elicitation of host-defence response capability of an endophytic Streptomyces albidoflavus OsiLf-2

Rice blast, caused by the fungus Magnaporthe oryzae, can result in notable yield losses in rice production. The objective of this study was to investigate the potential of a rice endophytic isolate, Streptomyces albidoflavus OsiLf-2, on the control of rice blast and the possible mechanisms involved. In vitro assays displayed a variety of antagonistic effects of OsiLf-2 against different physiological races of M. oryzae, with peak mycelial growth inhibition ranging from 74.1% to 83.0%. In vivo tests of OsiLf-2 showed 18.0% and 19.6% reduction in disease index in greenhouse and field conditions, respectively. The stable active metabolites in its cell-free culture filtrate inhibited the mycelial growth, spore germination and appressorial formation of M. oryzae in a dose-dependent manner. They also possessed strong antifungal capacities toward various phytopathogens in vitro. OsiLf-2 secreted multiple antimicrobial compounds, cell wall degradation enzymes, siderophore, plant hormones, and 1-aminocyclopropane-1-carboxylate deaminase, which might function in direct or indirect resistance to M. oryzae. In addition, a variety of defence responses were induced in OsiLf-2-treated rice, including hydrogen peroxide (H₂O₂) accumulation, callose deposition, defence-related enzymes activation, and elevated expression of salicylic acid (SA) and jasmonic acid (JA) pathways genes, which might contribute in resisting pathogen attack. The significant biological control activity and host defence-stimulation ability of OsiLf-2 suggest that this endophytic actinobacterial strain could be a promising candidate in the management of rice blast disease.     

Acylated Aminooligosaccharides from the Yellow Sea Streptomyces sp. HO1518 as Both α-Glucosidase and Lipase Inhibitors

Three new acylated aminooligosaccharide (1–3), along with five known congeners (4–8), were isolated from the marine-derived Streptomyces sp. HO1518. Their structures were fully elucidated by extensive spectroscopic analysis, mainly based on 1D-selective and 2D TOCSY, HSQC-TOCSY, and HRESIMS spectrometry measurements, and by chemical transformations. All of the compounds were evaluated for their α-glucosidase and pancreatic lipase inhibitory activities. Among the isolates, D6-O-isobutyryl-acarviostatin II03 (3) and D6-O-acetyl-acarviostatin II03 (8), sharing acarviostatin II03-type structure, showed the most potent α-glucosidase and lipase inhibitory effects, far stronger than the antidiabetic acarbose towards α-glucosidase and almost equal to the anti-obesity orlistat towards lipase in vitro. This is the first report on inhibitory activities against the two major digestive enzymes for acylated aminooligosaccharides. The results from our investigation highlight the potential of acylated aminooligosaccharides for the future development of multi-target anti-diabetic drug.     

玫瑰黄链霉菌活性代谢产物RM诱导黄瓜抗病性的信号转导途径

为明确玫瑰黄链霉菌Streptomyces roseoflavus菌株men-myco-93-63活性代谢产物roflamycoin和men-myco-A(简称RM)诱导黄瓜产生抗病性的信号转导途径,采用高效液相色谱法、分光光度法和实时荧光定量PCR法,测定喷施玫瑰黄链霉菌代谢产物RM后黄瓜叶片中水杨酸含量和脂氧合酶活性,以及水杨酸信号转导途径和乙烯茉莉酸信号转导途径的标志基因——NPR1、PR-1a、CTR1的表达量。结果表明,喷施玫瑰黄链霉菌活性代谢产物RM后120 h,黄瓜叶片内水杨酸含量达到最高值,为7.22μg/g,是对照的1.75倍;喷施玫瑰黄链霉菌活性代谢产物RM后12 h,黄瓜叶片内脂氧合酶活性达到最高值,为52.69 U/g,是对照的1.32倍,并且在120 h内整体活性均高于对照;喷施玫瑰黄链霉菌代谢产物RM后,NPR1和PR-1a基因的相对表达量上调,其最大值分别为0.99和1.35,分别是对照的2.24倍和12.09倍,但抑制CTR1基因的表达。推测玫瑰黄链霉菌代谢产物RM通过水杨酸通路、乙烯通路和茉莉酸通路共同诱导黄瓜抗病性信号转导途径。     

生防菌株Men-myco-93-63发酵培养基优化条件的研究

研究了碳源、氮源和无机盐对玫瑰黄链霉菌Men-myco-93-63发酵的影响,并用正交的方法优化了发酵培养基配方。最适碳源为葡萄糖和淀粉组合;最适氮源为花生饼粉和玉米浆组合;试验的8种无机盐中KH2PO4、CaCO3、NaCl为可提高发酵单位的3种无机盐。正交试验优化后确定的发酵培养基的配方为葡萄糖2.4%,淀粉0.8%,花生饼粉1.5%,玉米浆0.8%,KH2PO40.02%,NaCl 0.4%,CaCO30.3%,为大规模发酵生产奠定了基础。     

天蓝色链霉菌M145中bldA依赖性突变的遗传定位与分析

bldA编码天蓝色链霉菌中识别UUA密码的tRNA,大部分菌株的生长不需要bldA功能。M145菌株发生了突变,使bldA成为必需基因。本研究以M145和1258等菌株为亲本,通过3轮杂交筛选构建了菌株LY3,作为遗传定位bldA依赖性(bldA-dependent,bad)突变的亲本之一。用LY3与1258杂交,筛选了217个重组子并鉴定其基因型。对217个重组子的基因型进行遗传分析,将bad突变定位于天蓝色链霉菌染色体SCO3934与SCO4659基因之间的758 kb区域。     

链霉菌FB1的发酵产物对5种豆科植物种子萌发的影响

以黄大豆、菜豆、青大豆、黑大豆和红小豆等5种豆科植物的种子为材料,研究了链霉菌FB1的发酵产物对种子萌发的影响.结果表明,链霉菌FB1的发酵产物可以提高4种种子的萌发率,促进后3种种子胚根的伸长,提高所有5种豆科植物种子的简明活力,而且以10倍稀释液的作用最为显著.     

Deoxyvasicinone with Anti-Melanogenic Activity from Marine-Derived Streptomyces sp. CNQ-617

The tricyclic quinazoline alkaloid deoxyvasicinone (DOV, 1) was isolated from a marine-derived Streptomyces sp. CNQ-617, and its anti-melanogenic effects were investigated. Deoxyvasicinone was shown to decrease the melanin content of B16F10 and MNT-1 cells that have been stimulated by α-melanocyte-stimulating hormone (α-MSH). In addition, microscopic images of the cells showed that deoxyvasicinone attenuated melanocyte activation. Although, deoxyvasicinone did not directly inhibit tyrosinase (TYR) enzymatic activity, real-time PCR showed that it inhibited the mRNA expression of TYR, tyrosinase-related protein 1 (TRP-1), and tyrosinase-related protein 2 (TRP-2). In the artificial 3D pigmented skin model Melanoderm^TM, deoxyvasicinone brightened the skin significantly, as confirmed by histological examination. In conclusion, this study demonstrated that the marine microbial natural product deoxyvascinone has an anti-melanogenic effect through downregulation of melanogenic enzymes.