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	1. Synthesis and high-expression of chitin deacetylase from Colletotrichum lindemuthianum in Pichia pastoris GS115
	Kang Lixin,Chen Xiaomei,Zhai Chao,Ma Lixin
	Biology 16 May 2012
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	Abstract:A gene, ClCDA, encoding chitin deacetylase from Colletotrichum lindemuthianum, was optimized according to the codon usage bias of Pichia pastoris and synthesized in vitro by overlap extension PCR. It was secretorily expressed in P. pastoris GS115 using constitutive expression vector pHMB905A. The expression level reached the highest with 110 mg/l culture supernatant after 72 h methanol induction, which comprises of 77.27 U/mg chitin deacetylase activities. SDS-PAGE, mass spectrometry and deglycosylation assays demonstrated that part of recombinant protein was glycosylated with an apparent molecular mass of 33 kDa. The amino acid sequences of recombinant proteins were confirmed by mass spectrometry.
	TO cite this article:Kang Lixin,Chen Xiaomei,Zhai Chao, et al. Synthesis and high-expression of chitin deacetylase from Colletotrichum lindemuthianum in Pichia pastoris GS115[OL].[16 May 2012] http://en.paper.edu.cn/en_releasepaper/content/4478455


	2. MaKatG1, a bifunctional catalase-peroxidase gene, contributes to virulence and UV tolerance of Metarhizium acridum
	FAN Anni,PENG Guoxiong,XIA Yuxian
	Biology 16 April 2012
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	Abstract:Entomopathogenic fungi are exposed to reactive oxygen species (ROS) produced either as a secondary products derived from a chemical reaction due to exposure to stress (UV, heat) or by the host. However, the mechanism by which fungal pathogens overcome the toxicity of ROS is unclear. In this study, a bifunctional catalase-peroxidase homolog, KatG1, was identified from Metarhizium acridum, a specific pathogenic fungus against locust. The knockout mutant of MaKatG1 exhibited reduced catalase and peroxidase activity and increased susceptibility to H2O2 and menadione, indicating that MaKatG1 has a bifunctional catalase-peroxidase acctivity and plays a role in the oxidative stress response. The germination and appressorium formation rates of the knockout mutant were decreased on locust wings, but were not affected in vitro. Compared with the wild-type, the virulence of the mutant was signifcantly reduced following topical inoculation, but was not affected following injection. These results demonstrated that the MaKatG1 gene plays a role in the early stages of fungal infection including germination and appressorium formation. Furthermore, UV-B tolerance of the mutant was reduced. This study demonstrated that MaKatG1 contributes to virulence and UV-B tolerance.
	TO cite this article:FAN Anni,PENG Guoxiong,XIA Yuxian. MaKatG1, a bifunctional catalase-peroxidase gene, contributes to virulence and UV tolerance of Metarhizium acridum[OL].[16 April 2012] http://en.paper.edu.cn/en_releasepaper/content/4475570

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	3. Ophiocordyceps sinensis, the flagship fungus of China: terminology, life strategy and ecology
	Zhang Yongjie,Li Erwei,Wang Chengshu,Li Yuling,Liu Xingzhong
	Biology 16 March 2012
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	Abstract:The Chinese caterpillar fungus, Ophiocordyceps sinensis (syn. Cordyceps sinensis) is one of the most famous and perhaps the most expensive fungal species in the world. Its biology largely remains a secret, and its commercial cultivation is still a dream. Because of its medicinal, economic, social, and ecological importance, and its limited distribution mainly in China, O. sinensis is herein nominated as the national fungus of China and the fungus of the year (2012) for the journal Mycology. To clarify the confusions, a standard nomenclature is proposed and recommended in this paper for a comprehensive understanding of the terms related to Cordyceps sensu lato and O. sinensis. We also review the recent researches on the life cycle and distribution of this fungus, parasitism of ghost moths by the fungus, the microbial community of natural Chinese cordyceps and its insect hosts, as well as the secondary metabolites produced by the fungi isolated from natural Chinese cordyceps. By taking advantage of different expertise as well as the government support, we believe that the biological secrets of O. sinensis will be unraveled gradually, and the sustainable development and utilization of this traditional medicine will be fully achieved in the future.
	TO cite this article:Zhang Yongjie,Li Erwei,Wang Chengshu, et al. Ophiocordyceps sinensis, the flagship fungus of China: terminology, life strategy and ecology[OL].[16 March 2012] http://en.paper.edu.cn/en_releasepaper/content/4469150


	4. Isolation, identification and characterization of human intestinal bacteria with the ability to utilize chloramphenicol as the sole source of carbon and energy
	Zhao Xin,Tian Fengwei,Wang gang,Liu Xiaoming,Zhang Qiuxiang,Zhang Hao,Chen Wei
	Biology 14 March 2012
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	Abstract:Five aerobic intestinal bacterial strains that utilized chloramphenicol (CAP) as sole carbon and energy source were isolated from fecal samples collected from healthy volunteers. Based on their morphology, physiological characters, and 16S rDNA sequence analysis, four of the five strains were identified as Klebsiella pneumonia and one as Escherichia fergusonii. The degradation rate of strain I-10-CHL (E. fergusonii) varied with the initial concentration of CAP and the maximum specific substrate removal rate was observed at 25 μg CAP/ml. The pH value also had an effect on the degradation rate of CAP and bacterial growth. A pH of 6.5 was optimal for CAP degradation and growth of strain I-10-CHL (E. fergusonii). In mixed substrate batch cultivations, where CAP was one of the components, glucose, acetamide and CAP were simultaneously utilized. The presence of glucose and acetamide increased the growth and substrate degradation rates of CAP. These results indicate that CAP degrading enzymes are inducible in nature. During incubation with E. fergusonii cells, reduction intermediates (1-p-nitrophenyl -2-amino-1, 3-propanediol) were observed. The products of CAP metabolism by species other than E. fergusonii have not yet been characterized. The strains reducing the antibiotic were chloramphenicol susceptible, indicating that the pathway does not appear to mediate chloramphenicol resistance. The role of this CAP reduction pathway in the physiology of Klebsiella pneumonia and E. fergusonii is unknown. Further understanding of the E. fergusonii CAP reduction pathway will contribute to our knowledge of the diversity of prokaryotic aromatic compounds degradation mechanisms.
	TO cite this article:Zhao Xin,Tian Fengwei,Wang gang, et al. Isolation, identification and characterization of human intestinal bacteria with the ability to utilize chloramphenicol as the sole source of carbon and energy[OL].[14 March 2012] http://en.paper.edu.cn/en_releasepaper/content/4471643


	5. The novel mosquitocidal Bacillus thuringiensis strain LLP29 provides potential for use in practice
	ZHANG Linngling,WU Songqing,PENG Yan,Li Mingwei,SUN Linying,GUAN Xiong
	Biology 25 August 2011
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	Abstract:LLP29 was a Bacillus thuringiensis strain isolated from leaves of Magnolia denudata. According to the results of preliminary analysis, it was highly toxic to mosquitoes and took less time in incubation. In order to characterize this promising strain, the growth rate, lagellar antigen serology, biochemical profile, antibiotic sensitivity and mosquitocidal activities against Aedes aegypti, Ae. albopictus, and Culex quinquefasciatus were tested. It was shown that LLP29 belonged to B. thuringiensis ssp. israelensis (H14). Its mosquitocidal activity was more twofold higher than the control strain IPS82, a Bti mosquitocidal strain used for production. Meanwhile, it had a higher growth rate than IPS82, which would reduce the cost by shortening the production cycle. It was indicated that LLP29 might be a potential biocontrol agent in the control and prevention of mosquitoes and mosquito-borne diseases.
	TO cite this article:ZHANG Linngling,WU Songqing,PENG Yan, et al. The novel mosquitocidal Bacillus thuringiensis strain LLP29 provides potential for use in practice[OL].[25 August 2011] http://en.paper.edu.cn/en_releasepaper/content/4440005


	6. The Response of Type 2 Quorum Sensing in Klebsiella pneumoniae to a Fluctuating Culture Environment
	ZHU Hu,LIU Huijun,NING Shoujiao,GAO Yuli
	Biology 29 June 2011
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	Abstract:Quorum sensing in Klebsiella pneumoniae is mediated by the synthesis of interspecies autoinducer 2 (AI-2) which is a non-homoserine lactone signal molecule. The response of Type 2 quorum sensing to environmental cues such as carbon sources, the initial pH of medium, and boracic acid was investigated in the present study by a Vibrio harveyi BB170 reporter assay and quantitative reverse transcription-PCR analysis. The results show that glucose can affect AI-2 synthesis to the greatest extent, and 3.0% glucose can stimulate K.peneumoniae to produce more AI-2, with a four times increase in activity compared with that of the control culture. According to our previous research, Type 2 quorum sensing in K.pneumoniae is luxS dependent. Therefore, the close relation of glucose concentration and luxS transcription level was confirmed with quantitative reverse transcription-PCR technology. The results show that the response of quorum sensing to a fluctuating glucose concentration group was performed by changing the amount of luxS RNA transcripts. A maximum of luxS transcription appeared during the exponential growth phase when the glucose concentration was 30.0 g L-1. At the same time, AI-2 production is also slightly impacted by the low initial pH. It is noteworthy that the addition of boracic acid at microdosage (0.1 g/L) can also induce AI-2 synthesis. Presumably, in K.pneumoniae, the 4, 5-Dihydroxy-2, 3-pentanedione (DPD) cyclizes by the addition of borate and the loss of water, is hydrated, and is converted to the final AI-2 signaling molecule.
	TO cite this article:ZHU Hu,LIU Huijun,NING Shoujiao, et al. The Response of Type 2 Quorum Sensing in Klebsiella pneumoniae to a Fluctuating Culture Environment[OL].[29 June 2011] http://en.paper.edu.cn/en_releasepaper/content/4433902


	7. Cloning and analysis of the MAT1-2-1 gene from the traditional Chinese medicinal fungus Ophiocordyceps sinensis
	Zhang Yongjie,Zhang Shu,Liu Xingzhong,Wen Hua'an,Wang Mu
	Biology 09 June 2011
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	Abstract:The entomopathogenic fungus Ophiocordyceps sinensis is an important species in traditional Chinese medicine but has yet to be commercially cultivated. One bottleneck is the very low frequency of stromata formation from artificially infected moth larvae. The mating system of fungi is the determining factor for sexual reproduction, but mating-type genes of O. sinensis have not been previously investigated. In this study, the putative MAT1-2-1 gene within the MAT1-2 idiomorph was amplified by PCR and was determined to consist of 859 nucleotides that encode 249 amino acids; genes within the MAT1-1 idiomorph, however, were not detected. The MAT1-2-1 gene contained the conserved high-mobility group (HMG) box, and MAT1-2-1 flanking sequences were subsequently obtained. Although no putative open reading frames of the MAT1-1 idiomorph were detected within the ca. 8-kb flanking sequences of MAT1-2-1, a putative DNA lyase gene (which is present next to both idiomorphs in some heterothallic fungi) was found ca. 3.0 kb downstream of MAT1-2-1. The intervening distance between MAT1-2-1 and the DNA lyase gene in O. sinensis is larger than that in Cordyceps militaris and Cordyceps takaomontana. In addition, O. sinensis showed low sequence similarities with C. militaris and C. takaomontana in both MAT1-2-1 and the DNA lyase gene. In phylogenetic analysis, different MAT1-2-1 haplotypes of O. sinensis clustered together with high bootstrap support. MAT1-2-1 was detected as a single-copy gene in all examined O. sinensis isolates including tissue cultures and single ascospore cultures. This report describes, for the first time, a mating-type gene of O. sinensis.
	TO cite this article:Zhang Yongjie,Zhang Shu,Liu Xingzhong, et al. Cloning and analysis of the MAT1-2-1 gene from the traditional Chinese medicinal fungus Ophiocordyceps sinensis[J].


	8. Identification of nitrogen-fixing Paenibacillus from different plant rhizospheres
	JIN Haojie,CHEN Xi,CHEN Sanfeng
	Biology 22 May 2011
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	Abstract:Some Paenibacillus strains could be further applied as plant growth-promoting bacteria (PGPB) to advance plant growth and quality because of their properties of high nitrogen-fixing (NF) ability, strong adversity resistance and good storage stability. A total of more than 534 isolates were selected from different plant rhizosphere. All of them could resist 80 C for 15 min and grow on the nitrogen-free medium. By PCR amplification and acetylene reduction assay (ARA), 23 out of 534 isolates had nifH gene and nitrogenase activities. Based on 16S rDNA sequencing, G+C content assay and DNA-DNA hybridization, all of them were identified. It was probable that two strains Be17 and S27 were identified to be new type trains, respectively; The strain Nz30 was identified as Paenibacillus zanthoxyli and the strain By56 was Paenibacillus sonchi; The strains Gb44, Gz45, Gt48, Gt49, Hz55, Bs63, Hp9 and Hp92 which formed a monophyletic cluster were identified as Paenibacillus stellifer; The largest cluster which contained 11 isolates were identified by 16S rDNA similarities might belong to Paenibacillus jamilae. The results suggested that the NF Paenibacillus distributed in various ecosystems and were dominant in different plant rhizospheres.
	TO cite this article:JIN Haojie,CHEN Xi,CHEN Sanfeng. Identification of nitrogen-fixing Paenibacillus from different plant rhizospheres[OL].[22 May 2011] http://en.paper.edu.cn/en_releasepaper/content/4427289


	9. Symbiotic Efficiency, Genetic Diversity and Phylogeny of the Rhizobia Isolated from Leucaena Leucocephala in Arid-hot River Valley Area in Panxi，Sichuan , China
	Xu Kaiwei,Petri Penttinen,Chen Yuanxue,Chen Qiang,Zhang Xiaoping
	Biology 21 March 2011
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	Abstract:The symbiotic efficiency and diversity of the forty-two indigenous rhizobial isolates obtained from the root nodules of Leucaena leucocephala, one of the pioneer leguminous tree species of planted forests in arid-hot river valley area in Panxi, Sichuan, China, were characterized. The result of the inoculation test on their original host plants showed that the majority (86%) of the tested strains were ineffective. The genetic diversity of these rhizobia and the 17 reference strains were investigated by using PCR-RFLP of the 16S rRNA gene, 23S rRNA gene and the internal transcribed spacer (IGS) region between the 16S rRNA and 23S rRNA genes. The RFLP analyses and phylogenetic analyses based on the16S rRNA gene sequences of representative strains showed that Leucaena leucocephala hosted strains from four rhizobial genera, Sinorhizobium, Bradyrhizobium, Rhizobium and Mesorhizobium. One of the nodulating strains did not cluster with any of the reference strains. Phylogenetic analysis of 16S rRNA gene sequence of the strain indicated that the closest related genus was Herbaspirillum within the β-proteobacteria. This is the first record that Herbaspirillum of β-proteobacteria can be a specific symbiont of leguminous plant.
	TO cite this article:Xu Kaiwei,Petri Penttinen,Chen Yuanxue, et al. Symbiotic Efficiency, Genetic Diversity and Phylogeny of the Rhizobia Isolated from Leucaena Leucocephala in Arid-hot River Valley Area in Panxi，Sichuan , China[OL].[21 March 2011] http://en.paper.edu.cn/en_releasepaper/content/4415719


	10. Pusillimonas oleiphila sp. nov., a new member of the genus Pusillimonas that degrades fuel oils at low temperature
	Ma Ting,Huang Lei,Li Guoqiang
	Biology 07 March 2011
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	Abstract:The taxonomic position of a novel strain, designated T7-7T, was investigated. This strain was isolated at 10oC as a diesel-oil-degrading organism from seabed mud in the Bohai Sea. Previously, a polar lipid pattern with phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, and unknown aminolipids AL1 as the primary components was found. Partial 16S rRNA gene sequence analysis indicated that this strain was related most closely to Pusillimonas ginsengisoli DCY25T (98.4% sequence similarity), Pusillimonas soli MJ07T(97.5%) and Pusillimonas noertemannii BN9T (96.7%). The levels of 16S rRNA gene sequence similarity between strain T7-7T and other recognized species of the family Alcaligenaceae were below 95.3%. This suggested that strain T7-7T represented a member of the genus Pusillimonas. Different methods for the construction of phylogenetic dendrograms separated this strain from other previously known strains of Pusillimonas. Fatty acid analysis demonstrated the presence of high concentrations of C16:0 and C17:0 cyclo. Strain T7-7T exhibited relatively low levels of DNA-DNA relatedness with P. ginsengisoli DCY25T(47.1%), P. soli MJ07T(44.5%) and P. noertemannii BN9T (38.6%). Based on the 16S rRNA gene sequence similarity to the strains of Pusillimonas, and other unique phenotypic properties such as fatty acid composition, polar lipid profiles, and DNA-DNA hybridization, T7-7T was classified as a new species with the name Pusillimonas oleiphila sp. nov. The strain type was designated as T7-7T (=CGMCC 1.6148 T =DSM 18250T).
	TO cite this article:Ma Ting,Huang Lei,Li Guoqiang. Pusillimonas oleiphila sp. nov., a new member of the genus Pusillimonas that degrades fuel oils at low temperature[OL].[ 7 March 2011] http://en.paper.edu.cn/en_releasepaper/content/4414817

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