Sequencing of Aspergillus nidulans and comparative analysis with A. fumigatus and A. oryzae

The aspergilli comprise a diverse group of filamentous fungi spanning over 200 million years of evolution. Here we report the genome sequence of the model organism Aspergillus nidulans, and a comparative study with Aspergillus fumigatus, a serious human pathogen, and Aspergillus oryzae, used in the production of sake, miso and soy sauce. Our analysis of genome structure provided a quantitative evaluation of forces driving long-term eukaryotic genome evolution. It also led to an experimentally validated model of mating-type locus evolution, suggesting the potential for sexual reproduction in A. fumigatus and A. oryzae. Our analysis of sequence conservation revealed over 5,000 non-coding regions actively conserved across all three species. Within these regions, we identified potential functional elements including a previously uncharacterized TPP riboswitch and motifs suggesting regulation in filamentous fungi by Puf family genes. We further obtained comparative and experimental evidence indicating widespread translational regulation by upstream open reading frames. These results enhance our understanding of these widely studied fungi as well as provide new insight into eukaryotic genome evolution and gene regulation.     

链霉菌TD-1代谢产物对饲料中污染霉菌的 抑制作用及稳定性

采用对峙培养法,检测分离于土壤中的链霉菌TD-1代谢产物对饲料中污染霉菌构巢曲霉(Aspergillus nidulans)、纯绿青霉(Penicillium verrucosum)、赭曲霉(Aspergillus ochraceus)、黄曲霉(Aspergillus flavus)、黑曲霉(Aspergillus niger)、米曲霉(Aspergillus oryzae)、茄腐镰刀菌(Fusarium solani)、桔青霉(Penicillum citrinum)、串珠镰刀菌(Fusarium moniliforme)的抑菌活性,结果表明:TD-1菌株代谢产物对以上霉菌具有较强的抑制作用,抑菌带宽度为10 mm左右.通过TD-1菌株的代谢过程曲线发现,该菌株发酵至第6 d抑菌率达90%左右.稳定性试验结果显示:该菌株所产生的活性物质具有较好的耐热性、耐酸碱性、抗紫外辐射能力.     

Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus

Aspergillus fumigatus is exceptional among microorganisms in being both a primary and opportunistic pathogen as well as a major allergen. Its conidia production is prolific, and so human respiratory tract exposure is almost constant. A. fumigatus is isolated from human habitats and vegetable compost heaps. In immunocompromised individuals, the incidence of invasive infection can be as high as 50% and the mortality rate is often about 50% (ref. 2). The interaction of A. fumigatus and other airborne fungi with the immune system is increasingly linked to severe asthma and sinusitis. Although the burden of invasive disease caused by A. fumigatus is substantial, the basic biology of the organism is mostly obscure. Here we show the complete 29.4-megabase genome sequence of the clinical isolate Af293, which consists of eight chromosomes containing 9,926 predicted genes. Microarray analysis revealed temperature-dependent expression of distinct sets of genes, as well as 700 A. fumigatus genes not present or significantly diverged in the closely related sexual species Neosartorya fischeri, many of which may have roles in the pathogenicity phenotype. The Af293 genome sequence provides an unparalleled resource for the future understanding of this remarkable fungus.     

微生物对黄曲霉毒素的抑制机制研究进展

部分黄曲霉菌会对农作物及产品造成严重污染。受相关基因的影响,黄曲霉会产生一种对人类健康与畜禽养殖构成重大威胁的真菌毒素,即黄曲霉毒素,在田间、储藏期或运输过程中黄曲霉毒素污染均可能发生。防止黄曲霉毒素进入食物链的策略有很多,其中采用生物法防治黄曲霉污染越来越引起研究者的重视,也逐渐被人们所接受。利用微生物对黄曲霉及毒素进行控制,可有效减轻黄曲霉菌及毒素对粮食作物及产品的污染,发挥功能的物质主要是微生物的代谢产物,包括多肽、小分子有机物、有机酸、抗生素和酶等。黄曲霉毒素污染对农作物产生的重大经济影响及其对人和动物的毒害作用已引起世界各国的高度关注。介绍了对黄曲霉及毒素具有拮抗作用的微生物种类(主要有芽孢杆菌、乳酸菌、酵母菌、链霉菌和一些海洋微生物等),分析了拮抗微生物中所发挥作用的有效成分,阐述了其对黄曲霉的抑制机制及对黄曲霉毒素的降解情况,希望为生物防治黄曲霉及黄曲霉毒素污染提供参考。     

Conservation and rearrangement of mitochondrial structural gene sequences

Mitochondria contain the simplest DNA molecules that are present in eukaryotes. Mitochondrial DNA (mtDNA) is easily purified, and is an important model system for studying eukaryote gene structure and basic molecular processes. The protein sequences of mitochondrial gene products have been shown to be conserved from yeast to man, and there are definite similarities at the DNA sequence level. In contrast, the overall organization of the mitochondrial genome is drastically different in these organisms. To understand this, we need to extend work on mtDNA to a wider range of species. We have chosen to study the mtDNA of Aspergillus nidulans because a particularly comprehensive analysis of this system can be achieved using genetics as well as biochemistry, and like most eukaryotes it is an obligate aerobe, whereas Saccharomyces cerevisiae is not. We have investigated whether defined pieces of particular yeast mitochondrial genes show enough homology to Aspergillus mtDNA fragments to enable the corresponding Aspergillus genes to be located on the physical map. The results reported here show that this is the case for all five genes tested, and present the first data on the physical organization of the structural genes in the mitochondrial genome of A. nidulans.     

3种海藻的粗蛋白对植物病原真菌的抑制作用

以圆弧青霉(Penicillium cyclopium)、绿色木霉(Trichoderma viride)、红色链孢霉(Neurcsporacrassa)、玉米大斑病菌(Helminthosporium turcicum)、稻瘟病菌(Piricularia oryzae)和黄曲霉(Aspergillusflavus)等6种植物病原真菌为供试菌,检测了3种海藻(铁钉菜Ishige okamurae、羊栖菜Sargassum fusiforme、冈村凹顶藻Laurencia okamurai)的粗蛋白对它们的菌丝生长及孢子萌发的影响.实验表明,3种海藻的粗蛋白对6种真菌的菌丝生长及孢子萌发的作用效果不同,其中冈村凹顶藻粗蛋白对红色链孢霉菌丝的生长,羊栖菜粗蛋白对稻瘟病菌孢子的抑制作用最明显,而羊栖菜和铁钉菜粗蛋白对红色链孢霉的菌丝生长,冈村凹顶藻粗蛋白对绿色木霉及黄曲霉的孢子萌发均无抑制效果.3种海藻的粗蛋白能凝集兔血红细胞,表明它们当中含有凝集素.     

稻瘟病菌中己糖载体蛋白的生物信息学分析

己糖载体蛋白是一种重要的糖载体蛋白,主要用于己糖的摄取和运输。本文通过生物信息学的方法,研究发现稻瘟病菌有67个可能的己糖载体蛋白。其中,MGG 06203、MGG 03620、MGG 15700、MGG 00040、MGG 13651、MGG 05946的氨基酸序列分别与Neurosporacrassa、Aspergillusnidulans和Colletrotrichumgramini-cola中已鉴定出的己糖载体蛋白高度同源。此外,稻瘟病菌不同的己糖载体蛋白基因在附着胞形成的各个阶段表达量存在很大的差异,尤其是其中有25个基因在稻瘟病菌侵染水稻48 h后有明显上调表达,表明它们可能参与了稻瘟病菌的致病过程。     

米曲霉自溶作用的研究Ⅱ自溶过程中细胞物质及酶活的变化

在米曲霉最佳的自溶条件下，对其自溶过程中重要的细胞物质及酶进行了测定，结果自溶液中测出１７种氨基酸酸，总氨基酸释放率为６８．７９％，核酸释放率为５０．００％，糖的释放率为２４．５８％，酸性蛋白酶活性高，而５’－磷酸二酯酶活性较低。     

曲霉属产毒真菌DNA的制备及其毒素相关基因aflR的PCR 检测

 研究了用改良的CTAB 法提取曲霉属产毒真菌DNA 的方法,并与试剂盒提取法进行比较。经过改良的CTAB 法能够成功提取出曲霉属产毒真菌的DNA,且浓度和纯度均可达到PCR 扩增的标准。根据合成黄曲霉毒素的aflR 基因设计两对引物,运用PCR 方法进行扩增鉴定,结果表明黄曲霉、寄生曲霉和溜曲霉基因中可检出aflR 基因,烟曲霉和杂色曲霉中并未检出,达到对黄曲霉毒素产生菌进行鉴定的目的。这种方法可为产毒真菌的快速检测提供参考。     

Sequence and analysis of rice chromosome 4

Rice is the principal food for over half of the population of the world. With its genome size of 430 megabase pairs (Mb), the cultivated rice species Oryza sativa is a model plant for genome research. Here we report the sequence analysis of chromosome 4 of O. sativa, one of the first two rice chromosomes to be sequenced completely. The finished sequence spans 34.6 Mb and represents 97.3% of the chromosome. In addition, we report the longest known sequence for a plant centromere, a completely sequenced contig of 1.16 Mb corresponding to the centromeric region of chromosome 4. We predict 4,658 protein coding genes and 70 transfer RNA genes. A total of 1,681 predicted genes match available unique rice expressed sequence tags. Transposable elements have a pronounced bias towards the euchromatic regions, indicating a close correlation of their distributions to genes along the chromosome. Comparative genome analysis between cultivated rice subspecies shows that there is an overall syntenic relationship between the chromosomes and divergence at the level of single-nucleotide polymorphisms and insertions and deletions. By contrast, there is little conservation in gene order between rice and Arabidopsis.     

复合菌群发酵酿造酱油的初步研究

应用沪酿3.042 米曲霉和实验室选育的黑曲霉混合制曲,在低盐固态发酵后期添加乳酸杆菌、生香酵母等耐受性较强的菌种进行复合菌群发酵酿制酱油。结果表明:在短周期发酵过程中,不但氨基酸态氮产量得到了明显提高,而且通过感官评定,产品风味也有了较大改善。     

水稻白叶枯病菌luxR同源基因的克隆和表达

luxR基因是革兰氏阴性菌感应反应（Quorum Sensing,QS）LuxR／Luxl环路中起重要作用的基因．从水稻白叶枯病菌中分离到一个luxR同源基因,命名为xooR．xooR全长765bp,编码一个由254个氨基酸残基组成的转录因子,分子质量和等电点分别为28．3ku和8．72．XooR蛋白的N一端15AA-171AA是一个Autoindbind结构域,C-端191AA-245AA是一个HTH（helix-turn-helix）结合DNA结构域利用原核表达载体pET-24a（＋）和gfp基因构建融合蛋白表达系统,Western blot分析表明XooR-GFP融合蛋白大小为54ku,xooR基因成功地在E．coliB1．21中实现了表达．     

The genome of the social amoeba Dictyostelium discoideum

The social amoebae are exceptional in their ability to alternate between unicellular and multicellular forms. Here we describe the genome of the best-studied member of this group, Dictyostelium discoideum. The gene-dense chromosomes of this organism encode approximately 12,500 predicted proteins, a high proportion of which have long, repetitive amino acid tracts. There are many genes for polyketide synthases and ABC transporters, suggesting an extensive secondary metabolism for producing and exporting small molecules. The genome is rich in complex repeats, one class of which is clustered and may serve as centromeres. Partial copies of the extrachromosomal ribosomal DNA (rDNA) element are found at the ends of each chromosome, suggesting a novel telomere structure and the use of a common mechanism to maintain both the rDNA and chromosomal termini. A proteome-based phylogeny shows that the amoebozoa diverged from the animal-fungal lineage after the plant-animal split, but Dictyostelium seems to have retained more of the diversity of the ancestral genome than have plants, animals or fungi.     

多菌联合固态发酵生产酒糟菌体蛋白饲料的实验研究

利用米曲霉、黑曲霉和酵母组成的三菌发酵体系对啤酒糟进行高蛋白菌体饲料化生物转化.方法为以预处理过的酒糟粉为主要培养基,添加少许辅料,接种上述三种菌种进行固态发酵.该方法的培养周期为6d,所得酒精菌体蛋白饲料的粗蛋白含量高达35.9%,糖化酶活力达7564.1u/g,演粉酶活力达4824.4.     

利用枯草杆菌整合表达嗜铬粒蛋白抗真菌片段

为实现CGA1-76片段基因在枯草杆菌中的稳定表达,将CGA1-76片段基因的表达元件重组到枯草杆菌转座子质粒pHV1249微转座子mini-Tn10内,利用mini-Tn10将CGA1-76片段基因的表达元件整合到枯草杆菌DB1342染色体上,用氯霉素和红霉素筛选枯草杆菌转座工程菌DB1342(TnSVTQ).DB1342(TnSVTQ)在无选择压力条件下经10 d连续传代培养,转座子内氯霉素抗性丢失率为0%,说明转座子稳定整合在DB1342染色体上.SDS-PAGE和免疫印迹分析结果表明,在蔗糖诱导下,CGA1-76片段基因在DB1342(TnSVTQ)中获得表达,产物被分泌到细胞外.孔穴琼脂扩散法测定表达上清的抑制真菌活性,结果表明重组CGA1-76能抑制烟曲霉菌和黄曲霉菌的生长,抑菌圈直径分别为9 mm和8 mm.     

阿特拉津降解菌Enterobacter sp.的基因差异分析

筛选并分析生物降解菌Enterobacter sp.在阿特拉津作用下的差异基因。以Enterobacter sp.BIDMC 29基因组为参考,利用高通量测序技术,对阿特拉津降解菌株和对照菌株进行转录组测序分析。结果表明,共有368个基因表现出显著差异,其中上调基因231个,下调基因137个。在差异基因GO富集分析的前30个条目中,尿嘧啶分解代谢、氮利用、硝酸盐同化、硫化氢生物合成、裂解酶活性和过氧化物酶活性等相关基因表现富集。Pathway分析显示,差异基因涉及70条代谢途径,与氮代谢、氨基酸合成和代谢、ABC转运蛋白、丙酮酸代谢等过程有关。菌株通过调节基因表达来提高对阿特拉津的降解能力,为进一步解析阿特拉津的代谢机制奠定基础。     

Ralstonia metallidurans CH34苯酚、甲苯降解基因的生物信息学分析

采用生物信息学方法对Ralstonia metallidurans CH34基因组序列进行了深入分析,推测了参与降解苯酚、甲苯/二甲苯降解的有关基因及功能,编码序列推测结果显示,R. metallidurans CH34中可能存在苯酚、甲苯/二甲苯间位降解代谢途径的基因操纵子及其调节因子和辅助因子,为后续进一步克隆和研究各基因的准确功能提供了有益的信息,奠定了基础.     

米曲霉木聚糖酶高产菌株选育

以米曲霉C-2为出发菌株,紫外诱变后,利用透明圈初筛,再分别以农业废弃物1％蔗渣-1％麸皮复合物、2％蔗渣、2％碱提蔗渣半纤维素和2％纤维素溶剂分馏法(CSLF)提取的蔗渣半纤维素为碳源摇瓶复筛,得到一株高产木聚糖酶的米曲霉菌株A73.结果表明:米曲霉A73在上述碳源培养基中,产木聚糖酶酶活力比出发菌株分别提高了281...     

中国传统酱油生产用米曲霉菌种研究进展

米曲霉是我国传统酱油生产用关键菌种.关于米曲霉菌株选育的研究主要包括,高产蛋白酶菌株选育技术研究、酶学特性研究、产酶影响因素研究等方面.高产蛋白酶菌株的选育为研究重点.虽然,研究均显示经过改良的菌株性能超过了原始出发菌株,但是成果仍然局限于实验室理论分析.我国科研人员仍需努力,加强对米曲霉菌株在工业化生产中实际应用效果的研究.     

资料室不同年代藏书中霉菌的调查及杀菌方法的比较

调查了湘潭学院生物系资料室不同年代藏书中霉菌的种类1数量和分布情况,并试验几种防腐方法,结果表明：图书出版的时间越早,厚度越大,书中的霉菌数量越多;分离鉴定出灰绿曲霉组（As-pergillus,glaucus)、局限曲霉组（Aspergillus,restrioctus）、黄曲霉（Aspergillus,flavus）、刺孢根霉（Rhipus,echinatus）等4类霉菌,其中灰绿曲霉存在调查的每一本藏书本,且数量最多。紫外线照射、甲醛熏蒸及石灰炭喷雾对上述几种霉菌均有较好的抑制作用。