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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 pYLTAC17 and pYLTAC747H/sacB,which contain about 165000 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 pYLTAC17 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

Node ordinal encoded genetic algorithm for the optimal allocation of water resources

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Progress in Natural Science:Materials International 2005年第5期15卷 448-452页

作者： YANG Xiaohua1*, YANG Zhifeng 1,2, SheN Zhenyao1 and LI Jianqiang3(1. State Key Laboratory of Water Environment Simulation, School of Environment, Beijing normal university, Beijing 100875, china 2. Key Laboratory for Water and Sediment Sciences Ministry of Education, School of Environment, Beijing normal university, Beijing 100875, china 3. Water Resources and Hydropower Planning and Design general institute, MWR, Beijing 100011, china) State Key Laboratory of Water Environment Simulation School of Environment Beijing Normal University Beijing 100875 China Key Laboratory for Water and Sediment Sciences Ministry of Education School of Environment Beijing Normal University Beijing 100875 China Water Resources and Hydropower Planning and Design general institute MWR Beijing 100011 China

A new method, node ordinal encoded genetic algorithm (NOEGA), is proposed for solving water resources optimal allocation problems, in which the capacity of water resources is split into a number of smaller parts so that successive operations can be overlapped. Our objective is to maximize the whole benefit function. To overcome the “dimensionality and algorithm complexity curse” while searching for solutions and looking for an optimal solution, the operations of one-point crossover operator, gene exchange operator, gene random operator, gene shift operator and node ordinal strings are established. It is proved to be an effective optimal method in searching for global solutions. The NOEGA does not need a diversity of initial population, and it does not have the problem of immature convergence. The results of two cases show that using NOEGA to solve the optimal allocation model is very efficient and robust. In addition, the algorithm complexity of NOEGA is discussed.

关键词： water resources optimal allocation node ordinal code genetic algorithm.

Cloning and sequencing of nifBHDKENX genes of Paenibacillus massiliensis T7 and its nif promoter analysis

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Science china(Life Sciences) 2006年第2期49卷 115-122页

作者： ZHAO Hongxin1,2,3, XIE Baoen2 & CheN Sanfeng1,2 1. State Key Laboratory for Agrobiotechnology and Biological School, china Agricultural university, Beijing 100094, china 2. Key Laboratory for Agro-Microbial Resource and Application of Agriculture Ministry, china Agricultural university, Beijing 100094, china 3. Chemical and Biological School, ShenYang normal university, Shenyang 110034, china 1. State Key Laboratory for Agrobiotechnology and Biological School China Agricultural University Beijing 100094 China 2. Key Laboratory for Agro-Microbial Resource and Application of Agriculture Ministry China Agricultural University Beijing 100094 China 3. Chemical and Biological School ShenYang Normal University Shenyang 110034 China

A 324 bp of nifH fragment was PCR amplified from Paenibacillus massiliensis T7 using the universal degenerate primers. The PCR-amplified nifH fragment was labeled with DIG; then used as a probe in Southern blot analysis. Southern blot result showed that there were two positive signals, indicating that there might be two copies of nifH in P. massiliensis T7. A total of 10254 bp DNA sequence containing purD; nifBHDKENX was obtained by five rounds of inverse-PCR amplification. The predicted proteins of nifBHDKENX had high homology with those from other nitrogen-fixing bacteria. Only one putative σ[54]-dependent promoter sequence was detected upstream of the nifB gene; nifBHDKENX were likely to be organized in one operon. Assays of β-galactosidase activity of P. massiliensis T7PB carrying a nifB-lacZ fusion under different concentrations of NH4 [+]; O2 showed that the expression of nifB-lacZ was strongly inhibited by O2.

关键词： Paenibacillus massiliensis T7, nifBHDKENX, nifB-lacZ, inverse-PCR.

Isolation, characterization and functional analysis of a cdc48 homologue from tobacco BY-2 cells

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Progress in Natural Science:Materials International 2007年第2期17卷 156-162页

作者： CheN Zhiling, YU Yi, LIU Lina and XIA Guixian ( College of Life Science, Capital normal university, Beijing 100037, china institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, china) [a]College of Life Science Capital Normal University Beijing China [b]institute of Microbiology Chinese Academy of Sciences Beijing China

The fission yeast Schizosaccharomyces pombe was used to identify genes from tobacco BY-2 cells that may play roles in cell cycle regulation. A cDNA encoding a protein homologous to the yeast CDC48 was isolated and the gene was designated as NtCDC48 . The cDNA contains an open reading frame coding for a predicted protein of 808 amino acids which comprises of two typical ATPase modules (aa 245-374 and aa 518-646) . Overexpression of NtCDC48 in tobacco BY-2 cells led to an increase in the mitotic index as well as to the formation of diffused mitotic spindles. NtCDC48-GFP fusion proteins are distributed ubiquitously through Gl to M phases, yet their subcellular localization varied regularly along with the cell cycle progression. These results indicate that NtCDC48 may play an important role in the regulation of cell cycle in BY-2 cells.

关键词： tobacco BY-2 suspension cells NtCDC48 spindle cell cycle.

Excessive manure application stimulates nitrogen cycling but only weakly promotes crop yields in an acidic Ultisol:Results from a 20-year field experiment

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Journal of Integrative Agriculture 2024年第7期23卷 2434-2445页

作者： Song Wan Yongxin Lin Hangwei Hu Milin Deng Jianbo Fan Jizheng he Key Laboratory for Humid Subtropical Eco-geographical Processes of Ministry of Education School of Geographical Sciences/School of Carbon Neutrality Future TechnologyFujian Normal UniversityFuzhou 350007China School of Agriculture and Food Faculty of ScienceThe University of MelbourneParkvilleVictoria 3010Australia State Key Laboratory of Soil and Sustainable Agriculture Institute of Soil ScienceChinese Academy of SciencesNanjing 210008China

Population growth and growing demand for livestock products produce large amounts of manure,which can be harnessed to maintain soil sustainability and crop ***,the impacts of excessive manure application on crop yields,nitrogen(N)-cycling processes and microorganisms remain ***,we explored the effects of 20-year of excessive rates(18 and 27 Mg ha^(–1)yr^(–1))of pig manure application on peanut crop yields,soil nutrient contents,N-cycling processes and the abundance of N-cycling microorganisms in an acidic Ultisol in summer and winter,compared with none and a regular rate(9 Mg ha^(–1)yr^(–1))of pig manure ***-term excessive pig manure application,especially at the high-rate,significantly increased soil nutrient contents,the abundance of N-cycling functional genes,potential nitrification and denitrification activity,while it had a weaker effect on peanut yield and plant *** with manure application,seasonality had a much weaker effect on N-cycling gene *** forest analysis showed that available phosphorus(AP)content was the primary predictor for N-cycling gene abundance,with significant and positive associations with all tested N-cycling *** study clearly illustrated that excessive manure application would increase N-cycling gene abundance and potential N loss with relatively weak promotion of crop yields,providing significant implications for sustainable agriculture in the acidic Ultisols.

关键词： denitrification manure amendment nitrification N-cycling functional genes peanut yields

Environmentally persistent free radicals in PM_(2.5)from a typical Chinese industrial city during COVID-19 lockdown:The unexpected contamination level variation

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Journal of Environmental Sciences 2024年第1期135卷 424-432页

作者： Lingyun Wang Wuduo Zhao Peiru Luo Qingyun he Wenfen Zhang Chuan Dong Yanhao Zhang College of Chemistry Zhengzhou UniversityZhengzhou 450001China Center of Advanced Analysis and gene Sequencing Key Laboratory of Molecular Sensing and Harmful Substances Detection TechnologyZhengzhou UniversityZhengzhou 450001China State Key Laboratory of Environmental and Biological Analysis Department of ChemistryHong Kong Baptist UniversityHong Kong 999077China institute of Environmental Science Shanxi UniversityTaiyuan 030006China

The outbreak of COVID-19 has caused concerns *** reduce the rapid transmission of the virus,strict city lockdown measures were conducted in different *** is the country that takes the earliest home-based quarantine for *** normal industrial and social activities were suspended,the spread of virus was efficiently ***,another merit of the city lockdown measure was noticed,which is the improvement of the air *** levels of multiple atmospheric pollutants were ***,in this work,24 and 14 air fine particulate matter(PM_(2.5))samples were continuously collected before and during COVID-19 city lockdown in Linfen(a typical heavy industrial city in china),and intriguingly,the unreduced concentration was found for environmentally persistent free radicals(EPFRs)in PM_(2.5)after normal life *** primary non-stopped coal combustion source and secondary Cu-related atmospheric reaction may have impacts on this *** cigarette-based assessment model also indicated possible exposure risks of PM_(2.5)-bound EPFRs during lockdown of *** study revealed not all the contaminants in the atmosphere had an apparent concentration decrease during city lockdown,suggesting the pollutants with complicated sources and formation mechanisms,like EPFRs in PM_(2.5),still should not be ignored.

关键词： PM_(2.5) EPFRs COVID-19 City lockdown health risks

genetic analysis and gene mapping of a new rolled-leaf mutant in rice (Oryza sativa L.)

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Science china(Life Sciences) 2009年第9期52卷 885-890页

作者： SHI YongFeng1,2,CheN Jie1,LIU WenQiang1,HUANG QiNa1,2,SheN Bo2,LEUNG hei3 & WU JianLi1 1 Chinese National Center for Rice Improvement/National key Laboratory for Rice Biology,china National Rice Research institute,Hangzhou 310006,china 2 College of Life and Environmental Sciences,Hangzhou normal university,Hangzhou 310016,china 3 International Rice Research institute,Metro Manila DAPO Box 7777,Philippines Chinese National Center for Rice Improvement/National key Laboratory for Rice Biology China National Rice Research Institute Hangzhou 310006 China College of Life and Environmental Sciences Hangzhou Normal University Hangzhou 310016 China International Rice Research institute Metro Manila DAPO Box 7777 Manila Philippines

To understand the development of rice leaf blades,we identified a new rolled-leaf mutant,w32,from indica cultivar IR64 through EMS mutagenesis. The mutant showed a stable rolled-leaf phenotype throughout the life cycle. Two F2 populations were developed by crossing w32 to cultivar IR24 and PA64. genetic analysis showed that the rolled-leaf phenotype was controlled by a single recessive gene. To determine the location of the gene,bulked segregant analysis was carried out using mutant and wild-type DNA pools and 1846 mutant-type F2 individuals derived from the cross w32/PA64 were genotyped to locate the gene on the short arm of chromosome 7. The rolled-leaf gene,tentatively named rl11(t),is likely a new gene as no other rolled-leaf genes have been identified near the region. By developing new SSR and InDel markers,the gene was delimited to a 52 kb region near the end of the short chromosome arm. Further fine mapping and cloning of the gene are currently underway.

关键词： rice (Oryza sativa L.) chemical mutagenesis molecular marker rolled-leaf gene physical map

The feasibility analysis of virus vectors as the VIGS tool for studying the gene function

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The feasibility analysis of virus vectors as the VIGS tool f...

gene flow from transgenic oilseed rape (Brassica napus L.) to cruciferous weeds under mentor pollen inducement

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作者： Xiaofu Zhou~(1,2) Hongwei Xu~(1,2) Xingzhi Wang~1 (1.The institute of genetics and Cytology,Northeast normal university,Changchun,130024,china:2.The institute of genetics and Physiology,jilin normal university,siping,13.000,china)

A method of virus-induced post-transcriptional gene silencing for studying rbcS gene function was established and optimized using tobacco rattle virus vector and potato X virus vector as materials in this *** following analyses were conducted:phenotypic characterization of rbcS gene silenced plants,transcription levels of rbcS gene by RT-PCR,and protein levels of rbcS by the antibodies of *** feasibility assessment of the PVX and TRV vectors on the VIGS for studying gene

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Progress in Natural Science:Materials International 2007年第11期17卷 1284-1289页

作者： Zhao Xiangxiang1,2, Luo Yuming1, Lu Weiping2, Qi Cunkou3, Pu Huiming3 and Wang Youping2(1. Biotechnology Key Laboratory of Eco-agriculture in Hongze Lake Region, Jiangsu Province, Huaiyin normal College, Huai'an 223001, china 2. Jiangsu Provincial Key Laboratory of Crop genetics and Physiology, Yangzhou university, Yangzhou 225009, china 3. institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, china) Biotechnology Key Laboratory of Eco-agriculture in Hongze Lake Region Jiangsu Province Huaiyin Normal College Huai'an 223001 China Jiangsu Provincial Key Laboratory of Crop genetics and Physiology Yangzhou University Yangzhou 225009 China institute of Industrial Crops Jiangsu Academy of Agricultural Sciences Nanjing 210014 China

The alien gene flow between genetically modified glyphosate-resistant rapeseed variety Q3 (Brassica napus L.) and four cruciferous weeds was studied under mentor pollen inducement. The results showed that when Thlaspi arvense L., Capsella bursa-pastoris (L.) Medic, Cardamine hirsuta L. and Rorippa palustris (L.) Besser were pollinated with mentor pollen, the mixed Q3 and the weed, pollen grains aggregated largely and germinated quickly, and the numbers of pollen tubes penetrating into the style and the ovary were greatly increased as compared with corresponding self-pollination groups. Twenty four to forty eight hours after pollination, several pollen tubes were observed to penetrate into the ovule via micropyle in each mentor combination. However, when the mentor progenies were analyzed by PCR, all of them showed negative for the Q3 herbicide-resistant gene. Collectively, these results indicated that crossing between T. arvense, C. bursa-pastoris, C. hirsuta, R. palustris (as female) and Q3 (as male) was highly incompatible and the herbicide-resistant gene could not flow from Q3 to these four weeds.

关键词： herbicide-resistant rapeseed cruciferous weeds mentor pollination aniline blue fluorescence gene flow