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Highly efficient transformation of intact yeast-like conidium cells of Tremella fuciformis by electroporation

Science china(Life Sciences) 2008年第10期51卷 932-940页

作者： GUO LiQiong1,2. LIU Yong1, ZHAO ShuXian1, LIU ErXian1 & LIN JunFang1,2.1 College of Food Sciences, south china agricultural university, Guangzhou 510640, china {2., south china agricultural university, Guangzhou 510640, china College of Food Sciences South China Agricultural University Guangzhou China institute of biomass {2. South China Agricultural University Guangzhou China

Tremella fuciformis is one of higher basidiomycetes. Its basidiospore can reproduce yeast-like conidia, also called the blastospore by budding. The yeast-like conidia of T. fuciformis is monokaryotic and easy to culture by submerged fermentation similar to yeast. So it is a good recipient cell for exogenous gene expression. In this study, two expression vectors pGlg-gfp containing gpd-Gl promoter and gfp gene and pGlg-hph containing gpd-Gl promoter and hph gene were constructed. The lowest sensitive concentration of hygromycin for the blastospore was determined on three types of media. Our ex- periments showed that the lowest sensitive concentration of hygromycin for the blastospore was 5 μg/mL on MA medium. The intact blastospores were transformed with the expression vector pGlg-hph by electroporation. The putative transformants were obtained by the MA selective medium. Experi- mental results showed that the most effective parameters for the electroporation of intact blastospores were obtained by using STM buffer, 1.0×108 cells/mL of blastospores, 2.0 μL in transformation volume, 6 μg plasmid, 2.0 kV/cm of electric pulse voltage, stillness culturing on MB liquid medium for 48 h after electroporation. In these transformation conditions, the efficiency reached 2.7 colonies/μg DNA. Co-transformation of plasmid pGlg-gfp and pGlg-hph with ratio of 1:1 was performed by electroporation with the optimal parameters. The putative co-transformants were obtained by the MA selective medium. Eight randomly selected colonies from the vast putative co-transformants were analyzed by PCR de- tection and southern blotting. The experiments showed that the gfp was integrated into the genomes of three transformants. The co-transformation efficiency was 37.5%. Green fluorescence was observed under laser scanning confocal microscope in these gfp positive transformants. This indicates that the exogenous gfp can be expressed effectively in the yeast-like conidia of T. fuciformis.

关键词： Tremella fuciformis electroporation yeast-like conidia intact cells gene expression hph gene gfp gene

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Normality and shared values concerning differential polynomials

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Science china Mathematics 2010年第3期53卷 749-754页

作者： LEI ChunLin 1 & FANG MingLiang 2. 1 Department of Applied Mathematics, south china agricultural university, Guangzhou 510642. china {2. of Applied Mathematics, south china agricultural university, Guangzhou 510642. china Department of Applied Mathematics South China Agricultural University Guangzhou China institute of Applied Mathematics South China Agricultural University Guangzhou China

Let F be a family of functions meromorphic in a domain D, let P be a polynomial with either deg P≥3 or deg P = 2.and P having only one distinct zero, and let b be a finite nonzero complex number. If, each pair of fun... 详细信息

关键词： meromorphic function normality shared value

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Pattern phenomena in an rf discharge dusty plasma system

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Science china(Physics,Mechanics & Astronomy) 2006年第5期49卷 588-596页

作者： HUANG Feng1,2. YE Maofu2.& WANG Long2.1. College of Science, china agricultural {2., Beijing 100083, china 2. institute of Physics, Chinese Academy of Sciences, Beijing 100080, china College of Science China Agricultural University Beijing China institute of Physics Chinese Academy of Sciences Beijing China

Various dust patterns are observed in an rf discharge dusty plasma system. According to the dust growth process from small to large in size, the formation of different dust patterns can be divided into two stages: the small-particle stage (or dust cloud stage), and the large-particle stage (or dust crystal stage). The evolution relations between different dust patterns with gas pressure changing are investigated. Dust voids, dust acoustic waves and strong turbulence modes are presented at the small-particle stage. The self-organized dust lattices and dust clusters are investigated at the large-particle stage. The static structure of a dust lattice is characterized by means of the pair correlation function. Dust clusters formed by particles with different numbers and the regular evolution of the clusters with gas pressure are also investigated. The packing sequences of dust clusters are verified through two-dimensional confined molecular dynamics simulations.

关键词： dusty plasma dust patterns

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Roles of Salicylic Acid-responsive Cis-acting Elements and W-boxes in Salicylic Acid Induction of VCH3 Promoter in Transgenic Tobaccos

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Acta Biochimica et Biophysica Sinica 2006年第1期38卷 46-52页

作者： Hai-Yan LI~(1,2.,Wei WEI~(2.),and Yu LI~(1*) ~1 institute of Mycology,Jilin agricultural {2.,Changchun 130118,china ~2.College of Biotechnology,Jilin agricultural {2.,Changchun 130118,china institute of Mycology Jilin Agricultural University Changchun 130118 China College of Biotechnology Jilin Agricultural University Changchun 130118 China

A salicylic acid (SA)-inducible VCH3 promoter was recently identified from grapevine (Vitisarnurensis) that contains two inverse SA-responsive cis-acting elements and four *** furtherdemonstrate the roles of these elements,four fragments with lengths from-1187,-892.-589,-2.6 to+7 bp were fused with the β-glucuronidase (GUS) reporter géne and transferred to Nicotiana tobacum,together with another four VCH3 promoter fragments with mutation in the two inverse *** functions,of each promoter fragment were-examined by analysis of GUS activity in thetransgenic tobacco root treated with *** GUS activity was shown in the roots of transgenictobaccos with the VCH3 (-1187)-GUS construct containing two SA-responsive cis-acting elements andfour ***,GUS activity directed by the VCH3 (-892.-GUS construct,containing one SA cis-acting element and four W-boxes,was reduced by up to 35% compared with that in tobaccos transformedwith the VCH3 (-1187)-GUS construct,indicating that the SA cis-acting element plays an important role inSA induction of the VCH3 *** the m2.CH3 (-1187)-GUS nor the mVCH3 (-892.-GUSconstruct,with mutation on the SA-responsive elements,abolished the expression of GUS activity,demon-strating that the W-boxes in the VCH3 promoter are also involved in SA *** arialysis ofGUS activity directed by each of the eight VCH3 promoter fragments showed that GUS was expressedspecifically in vascular *** was concluded that both the SA-responsive cis-acting elements and the W-boxes are important for the SA induction of the VCH3 *** promoter might have a potential usein plant genetic engineering.

关键词： VCH3 promoter salicylic acid(SA)-responsive cis-acting element W-box site-directed mutagenesis SA induction transgenic tobacco

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Computational theory of cavitating flows for hydraulic turbomachinery with consideration of influence of water quality

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Science china(Technological Sciences) 2010年第12期53卷 3341-3348页

作者： WANG Lei1,2.& CHANG JinShi2.1 Huadian Electric Power research institute, Hangzhou 310030,{2. 2.College of Water Conservancy and Civil Engineering,{2. agricultural university, Beijing 100083,{2. Huadian Electric Power research institute Hangzhou China College of Water Conservancy and Civil Engineering China Agricultural University Beijing China

Previously it was assumed that the pressure within the cavity or on the cavity surface remained constant and the vapor pressure of clean water at 2.°C and 0 m altitude was utilized as the computational boundary for cavitating flows in hydraulic turbomachinery. Cavitation was confused with vaporization, and the effect of water quality on cavitation pressure characteristics was not taken into account. In recent years, lots of experiments of cavitation pressure characteristics of different water qualities including different sand concentrations of sand water and different altitudes of clean water have been performed by the authors, and the important influences of water quality on cavitation pressure characteristic have been validated. Thus the water quality should be involved in the cavitating flows computation. In the present paper, the effect of water quality on the cavitation pressure characteristic is analyzed and the computational method and theory of cavitating flows for hydraulic turbomachinery that considers the influence of water quality are proposed. The theory is suitable for both the potential flow method and the two-phase flow method for cavitating flows simulation. Finally, the validation results for cavitating flows in a hydraulic tur- bine indicate the significant influences of water quality on the cavitating flow performance.

关键词： cavitating flows cavitation pressure water quality hydraulic turbomachinery

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Efficient transformation and expression of gfp gene in the edible mushroom Pleurotus nebrodensis

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Progress in Natural Science:Materials International 2008年第7期18卷 819-824页

作者： Junfang Lin a,b, Mingyao Zheng a, Jie Wang a,b, Wei Shu c, Liqiong Guo a,b,* a Department of Biotechnology, College of Food Sciences, south china agricultural university, Guangzhou 510640, china b {2. of biomass research, south china agricultural university, Guangzhou 510640, china c Test and Experimental Center, south china agricultural university, Guangzhou 510640, china Department of Biotechnology College of Food Sciences South China Agricultural University Guangzhou 510640 China institute of biomass {2. South China Agricultural University Guangzhou 510640 China Test and Experimental Center South China Agricultural University Guangzhou 510640 China

An efficient transformation method mediated by PEG-protoplasts was developed for the newly commercial edible mushroom Pleu- rotus nebrodensis. Two plasmids were used to co-transform protoplasts of P. nebrodensis. One plasmid is pAN7-1 containing a positive selectable marker gene hph conferring hygromycin B resistance. Another plasmid is pBlue-GFP containing a reporter gene gfp conferring green fluorescent protein. PCR and southern blot analysis showed that hph gene or/and gfp gene were integrated into the genome of P. nebrodensis transformants. The transformation efficiency of the positive selectable marker gene hph was 3 transformants per microgram of plasmid pAN7-1 DNA, which was about 30 times higher than that previously reported in thoroughly studied Pleurotus species such as Pleurotus ostreatus. The transformation efficiency of the reporter gene gfp was 9 transformants per microgram of plasmid pBlu-GFP DNA. The co-transformation efficiency was 2..68%. This is the first report that a "reporter" gene, green fluorescent protein gene can be successfully stably expressed in this Pleurotus species.

关键词： Pleurotus nebrodensis Protoplast-mediated transformation Green fluorescent protein GFP expression

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Palynological characters of Glycyrrhiza,Glycyrrhizopsis,and Meristotropis(Leguminosae),with special reference to their taxonomic significance

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Journal of Systematics and Evolution 2010年第6期48卷 455-463页

作者： 1,2.ei MENG 2.iang-Yun ZHU 1(College of Biological Sciences,china agricultural {2.,Beijing 100094,china) 2.institute of Botany,Chinese Academy of Sciences,Beijing 100093,china) (College of Biological Sciences China Agricultural University Beijing 100094 China) (institute of Botany Chinese Academy of Sciences Beijing 100093 China)

The pollen morphology of 11 species of the genus Glycyrrhiza *** one from each of the genera Glycyrrhizopsis Boiss.& *** Meristotropis Fisch.& *** investigated by scanning electron *** pollen morphology,the main differences between Glycyrrhizopsis and Glycyrrhiza are:Glycyrrhizopsis-pollen grains 36.63 × 40.42.μm in size,oblate spheroidal in shape;and Glycyrrhiza-pollen grains 2..47-33.18 × 2..82.31.83 μm in size,prolate spheroidal in *** and Glycyrrhiza should be recognized as two distinct genera based on palynological and morphological *** and Glycyrrhiza are similar in many important palynological and morphological characters,suggesting that the two should be *** Glycyrrhiza,two types of pollen grains,3-lobed-circular or subtriangular in polar view,are found in different species,in accordance with morphological differences in the two groups,shedding light on the classification and evolution of the genus.

关键词： exine sculpture morphological character pollen morphology taxonomic revision.

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Genetic analysis of segregation distortion of mating-type factors in Lentinula edodes

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Progress in Natural Science:Materials International 2005年第8期15卷 684-688页

作者： CHENG Shuiming 1,2. LIN Fanxue1, XU Xuefeng1, LI Anzheng1 and LIN Fangcan1*(1. State Key Laboratory of agricultural Microbiology and institute of Applied Mycology, Huazhong agricultural university, Wuhan 430070, china 2. National Engineering research Center For Wheat, Henan agricultural university, Zhengzhou 450002. china) State Key Laboratory of agricultural Microbiology and institute of Applied Mycology Huazhong Agricultural University Wuhan 430070 China National Engineering research Center For Wheat Henan Agricultural University Zhengzhou 450002 China

Analyses of the mating-type ratio of protoplast monokaryons to spores in 2. strains of Lentinula edodes were performed through statistical methods. The results showed that the segregation distortion phenomenon commonly exists in both types of Lentinula edodes, 17 out of the 2. samples did not display the expected 1∶1 ratio of protoplast monokaryons; and among them 8 fruiting strains distorted 1∶1∶1∶1 segregation of spores. The degree of distortion in all tested wild strains was significantly lower than that of cultured strains. In fruiting test of positive-negative parent, predominant spores were those with the mating type identical to dikaryotic parent, indicating that the predominance of nuclei mainly depends on the mating type of dikaryotic parent. Data from di-mon mating test verified that monokaryons with acceptor nuclear type are present exclusively in a large amount after dedikaryotization, suggesting that the specificities among B factors and some other unknown factors in cytoplasm are probably responsible for the phenomenon of skewed segregation.

关键词： Lentinula edodes mating-type factors segregation distortion cytoplasm.

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Nanostructured Sulfide Composite Coating Prepared by Atmospheric Plasma Spraying

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材料工程 2006年第Z1期34卷 281-284页

作者： GUAN Yao-hui~1,XU Yang~1,ZHENG Zhong-yu~2.TONG Xiao-hui~2.1 Engineering College of china agricultural {2.,Beijing 100083,china 2.Beijing research institute of Mechanical & Electrical Technology,Beijing 100083,china) Engineering College China Agricultural University Beijing 100083 China Beijing research institute of Mechanical and Electrical Technology Beijing 100083 China

Nanostructured FeS-SiC coating was deposited by atmospheric plasma spraying（APS）.The microstructure and phase composition of the coating were characterized with SEM and XRD,*** addition,the size distribution of the reconstituted powders and the porosity of the coating have been *** was found that the reconstitiuted powers with sizes in the range of 2. to 80 μm had excellent flowability and were suitable for plasma spraying *** as-sprayed FeS-SiC composite coating exhibited a bimodal distribution with small grains（30～80nm） and large grains（100～2.0nm）.The coating was mainly composed of FeS and SiC,a small quantity of Fe1-xS and oxide were also *** porosity of the coating was approximately 19%.

关键词： plasma spraying nanostructured FeS-SiC composite coating

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Satellite RNA-mediated Reduction of Cucumber Mosaic Virus Genomic RNAs Accumulation in Nicotiana tabacum

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Acta Biochimica et Biophysica Sinica 2007年第3期39卷 217-223页

作者： Qiansheng LIAO~(1,2. Liping ZHU~2.Zhiyou DU~2.Rong ZENG~(2.3) Junli FENG~2.Jishuang CHEN~(1,2.) 1 College of Life Sciences,Zhejiang university,Hangzhon 31002.,china {2. of Bioengineering,Zhejiang Sci-Tech university,Hangzhou 310018,china 3 College of Biological Safety Technology Hunan agricultural university,Changsha 41012.,china College of Life Sciences Zhejiang UniversityHangzhou 310029 China institute of Bioengineering Zhejiang Sci-Tech UniversityHangzhou 310018 China College of Biological Safety Technology Hunan Agricultural UniversityChangsha 410128 China

Satellite RNAs(satRNAs)are molecular parasites that interfere with the pathogenesis of thehelper *** this study,the relative accumulation of cucumber mosaic virus(CMV)-Fny genomicRNAs with or without satRNAs were quantitatively analyzed by real-time *** results showed thatsatRs apparently attenuated the symptoms of CMV-Fny on Nicotiana tabacum by depressing the accumu-lation of CMV-Fny genomic RNAs,tested as open reading *** accumulation of CMV-Fny 1a,2.,2.,3a,and CP genes was much higher than that of CMV-Fny with satRs added(CMV-Fsat),at differentinoculation ***-FnyΔ2.,in which the complete 2. gene and 41 amino acids at the C-terminal of the2. gene were deleted,caused only a slight mosaic effect on *** seedlings,similar to that of CMV-Fsat,but the addition of satRs to CMV-FnyΔ2. showed further decrease in the accumulation of CMV-FnyΔ2. genomic *** results indicated that the attenuation of CMV,by adding satRs or deleting the2. gene,was due to the low accumulation of CMV genomic RNAs,and that satRNA-mediated reduction ofCMV genomic RNAs accumulation in *** was possibly related to the 2. gene.

关键词： cucumber mosaic virus satellite RNA deletion of 2. gene relative accumulation

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