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

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

作者： GUO LiQiong1.2, liu Yong1. {1. ShuXian1. liu ErXian1.& LIN JunFang1.2.1 college of Food Sciences, South china agricultural university, guangzhou 51.640, china 2 Institute of Biomass Research, South china agricultural university, guangzhou 51.640, china 1. college of Food Sciences South China Agricultural University Guangzhou 510640 China 2. Institute of Biomass Research South China Agricultural University Guangzhou 510640 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, 1.0×1.8 cells/mL of blastospores, 200 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 277 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

Molecular characterization and expression analysis of caveolin-1.in pig tissues

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Science china(Life Sciences) 2008年第7期51卷 655-661页

作者： WANG Chong1.MEI YingJie 1.LI Li 1.MO DeLin 2,LI JiaQi 1.ZHANG Hao 1.TIAN XingGuo 1.&CHEN YaoSheng2.1{1. of Animal Science,South china agricultural university,guangzhou 51.642.china 2School of Life Science,Sun Yat-Sen university,guangzhou 51.006,china 1. college of Animal Science South China Agricultural University Guangzhou 510642 China 2. School of Life Science Sun Yat-Sen University Guangzhou 510006 China

Members of the caveolin family played important roles during fundamental cellular processes,such as regulation of cell morphology,migration,and gene expression in muscle *** this study,caveolin-1.(Cav-1.,one of the caveolins,was identified from longissimus dorsi muscle of Large Yorkshire pig and Chinese indigenous Lantang pig based on the results of mRNA differential display *** deduced amino acids sequence of the porcine Cav-1.contained a caveolin domain,and was very conservative among different *** Cav-1.mRNA was widely expressed in the eight tissues in this study,including heart,liver,kidney,encephalon,spleen,lung,longissimus dorsi muscle,and back fat, and the highest expression quantity was found in back fat of the two pig *** expression quantity of porcine Cav-1.in back fat and longissimus dorsi muscle of Lantang pig was significantly higher than that of Large Yorkshire(P1.and P1.might be a candidate gene for carcass traits,and might provide valuable information for understanding the mechanism of caveolae signaling in fat deposition by using the animal model of pig.

关键词： caveolin family clone expression analysis real-time quantify PCR

Host-Induced Gene Silencing of Effector AGLIP1.Enhanced Resistance of Rice to Rhizoctonia solani AG1.IA

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Rice science 2024年第4期31卷 463-474,I0031-I0034页

作者： zhao Mei {1. Xiaoxue WAN Jun ZHOU Erxun SHU Canwei Pesticide State Key Laboratory of Green Pesticide Guangdong Province Key Laboratory of Microbial Signals and Disease ControlCollege of Plant ProtectionSouth China Agricultural UniversityGuangzhou 510642China college of Life Science and Engineering Henan University of Urban ConstructionPingdingshan 467036China

Rice sheath blight, caused by Rhizoctonia solani AG1.IA, is a major disease in rice-growing areas worldwide. Effectors of phytopathogenic fungi play important roles during the infection process of fungal pathogens onto their host plants. However, the molecular mechanisms by which R. solani effectors regulate rice immunity are not well understood. Through prediction, 78 candidate effector molecules were identified. Using the tobacco rattle virus-host induced gene silencing(TRV-HIGS) system, 45 RNAi constructs of effector genes were infiltrated into Nicotiana benthamiana leaves. The results revealed that eight of these constructs resulted in a significant reduction in necrosis caused by infection with the AG1.IA strain GD-1.8. Additionally, stable rice transformants carrying the double-stranded RNA construct for one of the effector genes, AGLIP1. were generated to further verify the function of this gene. The suppression of the AGLIP1.gene increased the resistance of both N. benthamiana and rice against GD-1.8, and also affected the growth rate of GD-1.8, indicating that AGLIP1.is a key pathogenic factor. Small RNA sequencing showed that the HIGS vectors were processed into si RNAs within the plants and then translocated to the fungi, leading to the silencing of the target genes. As a result, AGLIP1.might be an excellent candidate for HIGS, thereby enhancing crop resistance against the pathogen and contributing to the control of R. solani infection.

关键词： Rhizoctonia solani host-induced gene silencing lipase tobacco rattle virus

RETINOBLASTOMA RELATED 1.switches mitosis to meiosis in rice

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Plant Communications 2024年第6期5卷 193-212页

作者： Yongjie Miao Hanli You Huixin liu Yangzi {1. Jiangzhe {1. Yafei Li Yi Shen Ding Tang Baohui liu Kewei Zhang Zhukuan Cheng Guangdong Provincial Key Laboratory of Plant Adaptation and Molecular Design Innovative Center of Molecular Genetics and EvolutionSchool of Life SciencesGuangzhou UniversityGuangzhou 510006China Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding Laboratory/Key Laboratory of Plant Functional Genomics of the Ministry of Education Agricultural College of Yangzhou UniversityYangzhou 225009China State Key Laboratory of Plant Genomics Institute of Genetics and Developmental BiologyInnovation Academy for Seed DesignChinese Academy of SciencesBeijing 100101China Zhejiang Provincial Key Laboratory of Biotechnology on Specialty Economic Plants Department of BiologyCollege of Life SciencesZhejiang Normal UniversityJinhua 321004China Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops/Jiangsu Key Laboratory of Crop Genetics and Physiology Yangzhou UniversityYangzhou 225009China

The transition from mitosis to meiosis is a critical event in the reproductive development of all sexually reproducing ***,the mechanisms that regulate this process in plants remain largely ***,we find that the rice(Oryza sativa L.)protein RETINOBLASTOMA RELATED 1.RBR1.is essential to the transition from mitosis to *** of RBR1.function results in hyper-proliferative sporogenous-celllike cells(SCLs)in the anther locules during early stages of reproductive *** hyperproliferative SCLs are unable to initiate meiosis,eventually stagnating and degrading at late developmental stages to form pollen-free *** results suggest that RBR1.acts as a gatekeeper of entry into ***,cytokinin content is significantly increased in rbr1.mutants,whereas the expression of type-B response factors,particularly LEPTO1.is significantly *** the known close association of cytokinins with cell proliferation,these findings imply that hyper-proliferative germ cells in the anther locules may be attributed to elevated cytokinin concentrations and disruptions in the cytokinin *** a genetic strategy,the association between germ cell hyper-proliferation and disturbed cytokinin signaling in rbr1.has been *** summary,we reveal a unique role of RBR1.in the initiation of meiosis;our results clearly demonstrate that the RBR1.regulatory module is connected to the cytokinin signaling pathway and switches mitosis to meiosis in rice.

关键词： RETINOBLASTOMA RELATED1 meiosis initiation cytokinin rice

An Ontology-Based information Retrieval Model for Vegetables E-Commerce

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Journal of Integrative Agriculture 2012年第5期11卷 800-807页

作者： TAO Teng-yang zhao Ming college of {1. and Electrical Engineering China Agricultural UniversityBeijing 100083P.R.China

With the rapid increment of the information on the web, traditional information retrieval based on the keywords is far from user＇s satisfaction in recall and precision. In order to improve the recall ratio and the precision radio of IR engine in the vegetables e-commerce, an information retrieval model based on the vegetables e-commerce ontology is presented in this paper, vegetables e-commerce ontology was constructed by gathering and the analyzing vegetables e-commerce domain information on the web. The vegetables e-commerce ontology is composed of some kinds of vegetable classes and hierarchy relationship of vegetables classes. In the process of information retrieval, domain ontology helps to index information and information inference. An ontology-based information retrieval model is implemented, and which has more functions than the keyword-based web information retrieval engines. The experiment results show that the recall ratio and the precision ratio of ontology-based information retrieval model are higher than that of the information retrieval engine based on keyword at a certain extent.

关键词： domain-ontology vegetables e-commerce information retrieval

Construction and characterization of the transformation-competent artificial chromosome (TAC) libraries of Leymus multicaulis

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Science china(Life Sciences) 2008年第7期51卷 604-613页

作者： XU YueYu 1,2,3,ZHOU YuLei 4,5 ,SONG LinLin 6 ,ZHANG Yan 3 &zhao MaoLin 1 1 Beijing Agro-Biotechnology Research Center,Beijing Academy of Agriculture and Forestry Science,Beijing 100097,china 2 Department of Chemistry and Chemical Engineering,Hunan Institute of Engineering,Xiangtan 411104,china 3 college of Life Science,Capital Normal university,Beijing 100037,china 4 college of Life Science,Zhongkai university of Agriculture and Technology,guangzhou 510225,china 5 college of Grassland Science,Gansu agricultural university,Lanzhou 730070,china 6 Department of Biology,Henan Institute of Science and Technology,Xinxiang 453003,china 1. Beijing Agro-Biotechnology Research Center Beijing Academy of Agriculture and Forestry Science Beijing 100097 China 2. Department of Chemistry and Chemical Engineering Hunan Institute of Engineering Xiangtan 411104 China 3. college of Life Science Capital Normal University Beijing 100037 China 4. college of Life Science Zhongkai University of Agriculture and Technology Guangzhou 510225 China 5. college of Grassland Science Gansu Agricultural University Lanzhou 730070 China 6. Department of Biology Henan Institute of Science and Technology Xinxiang 453003 China

Transformation-competent artificial chromosome system is able to clone and transfer genes efficiently in *** order to clone genes highly tolerant to barley yellow dwarf virus(BYDV),Aphids,drought and salt from Leymus multicaulis,the two TAC genomic libraries I and II were constructed in vector pYLTAC1. and pYLTAC747H/sacB,which contain about 1.5000 and 236000 recombinant clones *** genome coverage of the two libraries was totally estimated to be about 3―5 haploid genome equivalents,as size selection of genomic DNA fragments was approximately from 9 to 300 *** of the genomic libraries were collected as bulked pools each containing 500 clones or so,stored in twelve 96-deep-well plates and then were gridding in triplicate onto a high-density colony hybridization filter with a 3×3 pattern using a GeneTAC?G3 arraying robot after being transferred manually into three 384-well *** 2501.and 2890 clones of Library in pYLTAC1. and in pYLTAC747H/sacB were stored individually in fourteen 384-well plates and then were automatically gridding in duplicate onto a high-density colony hybridization filter with a 6×6 pattern after a replication of *** positive clones were detected by using the probe glutahione reductase gene of *** libraries constructed here can be used to isolate genomic clones containing target genes,and to carry out genome walking for positional *** the target TAC clones were isolated,they could be immediately transferred into plant genomes with the Agrobacterium system.

关键词： Leymus multicaulis megabase-size DNA transformation-competent artificial chromosome(TAC) genomic library

Electrochemical and colorimetric dual-signal detection of Staphylococcus aureus enterotoxin B based on AuPt bimetallic nanoparticles loaded Fe-N-C single atom nanocomposite

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Food Science and Human Wellness 2024年第4期13卷 2025-2035页

作者： Huan Liang Hongcheng liu Haojian Lin Guobao Ning Xiaokang Lu Siying Ma Fei liu Hui zhao Canpeng Li School of Chemical Science and Technology Yunnan UniversityKey Laboratory of Medicinal Chemistry for Natural ResourceMinistry of EducationKunming 650091China Institute of Quality Standard and Testing Technology Yunnan Academy of Agricultural ScienceSupervision and Testing Center for Farm Product QualityMinistry of AgricultureKunming 650223China State Key Laboratory of Optoelectronic Materials and Technologies Guangdong Province Key Laboratory of Display Material and TechnologySchool of Electronics and Information TechnologySun Yat-sen UniversityGuangzhou 510275China Laboratory for Conservation and Utilization of Bio-resource Yunnan UniversityKunming 650091China

Sensitive detection of Staphylococcus aureus enterotoxin B(SEB)is of importance for preventing food poisoning from threatening human *** this work,an electrochemical and colorimetric dual-signal detection assay of SEB was *** probe(Ab2/AuPt@Fe-N-C)was bound to SEB captured by Ab1.where the Ab2/AuPt@Fe-N-C triggered methylene blue degradation and resulted in the decrease of electrochemical ***,the probe catalyzed the oxidation of 3,3’,5,5’-tetramethyl biphenyl to generate a colorimetric absorbance at 652 *** the target was captured and formed a sandwich-like complex,the color changed from colorless to *** detection by colorimetric and electrochemical methods showed a linear relationship in the concentration ranges of 0.0002.10.0000 and 0.0005-1..0000 ng/mL,with limits of detection of 0.0667 and 0.1.70 pg/mL,*** dual-signal biosensor was successfully used to detect SEB in milk and water samples,which has great potential in toxin detection in food and the environment.

关键词： Staphylococcus aureus enterotoxin Electrochemical immunosensor Colorimetric assay MOF@borophene composite Dual-functional Fe-N-C signal atom catalyst

Optimization design and experiment of the variable differential gear train planting mechanism

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International Journal of agricultural and Biological Engineering 2024年第2期17卷 85-93页

作者： Xiong zhao Ziwei {1. Yuanwu Jia Xingxiao Ma Pengfei Zhang Jianneng Chen School of Mechanical Engineering Zhejiang Sci-Tech UniversityHangzhou 310018China Xi’an International university Engineering {1. Xi’an 710077China Hangzhou Vocational&Technical college Hangzhou 310018China

In order to improve the adaptability of the planting mechanism for different plant spacings,a variable differential gear train planting mechanism based on precise pose and trajectory control was proposed by combining the open chain 2R rod group and the variable differential gear *** to the pose requirements of receiving seedling point,transporting seedling point and planting point,three precise pose points of constrained planting trajectory were *** the three-position motion generation structural synthesis method,combined with computer-aided optimization design software,a set of mechanism parameters that meet the planting requirements were *** on the optimized mechanism parameters,by only changing the coordinates of two trajectory shape control points,three planting trajectories with key point position information adapted to 300 mm,400 mm and 500 mm plant spacing were obtained by interpolation,and three pairs of total transmission ratio of three groups of variable differential gear trains were *** distributing the total transmission ratio of the mechanism,the fixed axis gear train and the differential gear train are *** fixed axis gear train included a pair of non-circular gear pairs and a pair of positive gear pairs,which were convenient for disassembly and *** former drives the sun gear at variable speed,and the latter drives the planet carrier at uniform *** on this structure,the transmission ratio of the positive gear pair is-1.and the transmission ratio of the differential gear train is *** sub-transmission ratio of the single-stage non-circular gear pair was calculated and the pitch curves of three pairs of noncircular gears were *** pairs of non-circular gear pairs with different transmission ratios were replaced in turn and three sets of planting mechanisms were modeled in three *** virtual prototype motion simulation was completed by ADAMS software,and the p

关键词： plant spacing planting mechanism variable differential gear train precise pose trajectory control parameter optimization

Comparison of the influence factors on NDVI for CCD camera and WFI imager on CBERS-02

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Science china(Technological Sciences) 2005年第S2期48卷 100-115页

作者： YU Tao1.3, LI Xiaoying1.2,3, ZHANG Yong1.2,3, zhao Feng1.2,3, GU Xingfa1.3,4, ZHU Li1.2,3, WANG Pengxin5 & MIN Xiangjun6 1. State Key Laboratory of Remote Sensing Science Institute of Remote Sensing Application, CAS, Bei- jing 1.01.1. {1. 2. Graduate university of Chinese Academy of Sciences, Beijing 1.0039, {1. 3. The Center for National Spaceborne Demonstration, Beijing 1.01.1. {1. 4. Institut National de la Recherche Agronomique, Climat Sol Environnement, 8491. Avignon, France 5. college of information and Electrical Engineering, {1. agricultural university, Beijing 1.0083, {1. 6. {1. Center for Resource Satellite Data and Application, Beijing 1.0830, {1. 1. State Key Laboratory of Remote Sensing Science Institute of Remote Sensing Application CAS 100101 Beijing China 2. The Center for National Spaceborne Demonstration 100101 Beijing China 3. Graduate university of Chinese Academy of Sciences 100039 Beijing China 4. Institut National de la Recherche Agronomique Climat Sol Environnent 84914 Avignon France 5. college of {1. and Electrical Engineering China Agricultural University 100083 Beijing China 6. china Center for Resource Satellite Data and Application 100830 Beijing China

CCD and WFI are payloads in the visible light and near infrared bands on CBERS satellites and they have not been applied widely, especially WFI. This article mainly probes into the potential of these two sensors’ application in the vegetation monitoring and analyses the influences of radiometric calibration, atmosphere conditions and sun-observing geometry, etc. on NDVI obtained from these two sensors respectively. In addition it analyses the effect of the spectral response difference of red and near infrared spectral bands in CCD and WFI sensors on NDVI. Results indicate that radiometric calibration is the most important factor on NDVI, which cannot be applied to vegetation monitoring without radiometric calibration. Results also demonstrate that near surface NDVI is different greatly from that of TOA which change with atmosphere conditions. The study shows that NDVI is also affected by the non-lambertian character of surface and the change of the atmospheric path with the observing geometry. And NDVI of WFI is higher than that of CCD. They are very different because of their spectral difference, but they have a good linear relevant relationship.

关键词： CCD WFI NDVI LAI.