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Isolation and Characterization of GmSTY1,a Novel Gene Encoding a Dual-Specificity Protein Kinase in Soybean（ Glycme max L.）

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Journal of Integrative Plant Biology 2006年第7期48卷 857-866页

作者： Zhao-Shi Xu You-Zhi Ma Xian-Guo cheng li-Xia Cao lian-cheng li Ming Chen Key Laboratory of Crop Genetics and Breeding Ministry of Agriculture Institute of Crop Science the Chinese Academy of Agricultural Sciences Beijing 100081 China

Phosphorylation of protein klnases has profound effects on their activity and interaction with other proteins. Tyroslne phosphorylation was reported to be involved in various physiological processes in plants; however, no typical receptor tyrosine kinase has been isolated from plants thus far. Dual-specificity kinases are potentially responsible for the phosphorylation of both tyrosine and serine/threonine of target proteins. A cDNA clone encoding a putative dual-specificity protein kinase was isolated by screening the cDNA GAL4 activation domain （AD） fusion library of soybean （Glycine max L.）, and its entire length was obtained using 5＇-rapid ampUflcatlon of cDNA ends. The predicted polypeptide of 330 amino acid residues, designated as GmSTY1, contains all 11 conserved subdomains, which share common characteristics with both the serine/ threonine and tyroslne protein klnases reported thus far. In addition, three potential N-linked glycosylation sites （NXS/T）, as well as phosphorylation motifs （SXXXS/T）, were observed, suggesting that GmSTY1 may be post-translationally modified. Furthermore, a potential N-myristoylation motif （MGARCSK） was found, suggesting that the GmSTY1 protein could associate with membranes in vivo. Southern blotting analysis revealed a single-copy of GmSTY1 in the genome. Northern blotting analysis showed that this gene was upregulated by drought and salt treatment in a time-dependent manner; however, exogenous abscisic acid （ABA） could not significantly affect the mRNA accumulation of GmSTY1. Interestingly, the transcript of this gene was remarkably downregulated by cold treatment during the early stages of the response, but upregulated later. These results Indicate that the protein kinase was possibly regulated by abiotic stresses in an ABA-independent pathway.

关键词： abiotic stress dual-specificity Glycine max , serine/threonine/tyrosine protein kinase.

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Isolation and Expression Analysis of a Novel Abiotic Stress-Induced Gene W89 from Wheat

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Agricultural Sciences in China 2007年第4期6卷 391-398页

作者： ZHANG Rui-yue XU Zhao-shi li lian-cheng CHEN Ming MA You-zhi National Key Facility of Crop Gene Resources and Genetic Improvement Key Laboratory of Crop Genetics and Breeding Ministry of Agriculture/Institute of Crop Science Chinese Academy of Agricultural Sciences Beijing 100081 P.R.China

Water stress and cold stress are important factors restricting plant growth. However, there is little knowledge on the function of stress-responsive genes in plants. Therefore, it is necessary to clone some important genes to study the mechanism of plant adaptation to abiotic stress for improvement of plant resistance. A putative water stress-induced gene, W89, was cloned from the eDNA library of drought-treated wheat seedlings by phage hybridization in situ, and its entire length was obtained using 5＇-rapid amplification of eDNA ends （RACE） and reverse transcription-polymerase chain reaction （RT-PCR）. The full-length eDNA of W89 consists of 2 392 bp and contains a 1 896 bp open reading frame （ORF） encoding a 631 amino acid protein. Southern blot analysis indicated that W89 was a single-copy gene. RT-PCR analysis revealed that the expression of W89 was upregulated by drought, cold, and abscisic acid （ABA）. Amino acid sequence analysis discovered that W89 had a conserved region of DUF248 （pfam03141）, which contained a methyltransferase domain with a sterile alpha motif （SAM）-binding motif. Phylogenetic analysis showed that W89 was 66% identical to Oryza sativa dehydration-responsive protein （BAD67956）. It was supposed that W89 was a novel dehydration-responsive protein encoding gene. On the basis of the functions of methyltransferase and the SAM-binding motif, the SAM-binding motif of W89 was supposed to be connected with other proteins or transcription factors to transduce stress signals and finally regulate the expression of stress-responsive genes on the early stage of drought stress.

关键词： wheat water stress cold gene cloning

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Isolation and Functional Analysis of the bZIP Transcription Factor Gene TaABP1 from a Chinese Wheat Landrace

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Journal of Integrative Agriculture 2012年第10期11卷 1580-1591页

作者： CAO Xin-you CHEN Ming XU Zhao-shi CHEN Yao-feng li lian-cheng YU Yue-hua liU Yang-na MAYou-zhi National Key Facility for Crop Genetic Resources and Genetic Improvement Key Laboratory of Crop Genetics and BreedingMinistry of Agriculture/Institute of Crop Sciences Chinese Academy of Agricultural Sciences Crop Research Institute Shandong Academy of Agricultural Sciences College of Agronomy Northwest A&F University

In plants, basic leucine zipper （bZIP） transcription factors play important roles in regulatory processes, including stress response, pathogenic defense and light response as well as organ and tissue differentiation. Chinese wheat landrace Pingyaoxiaobaimai （PYXBM）, an original parent of drought tolerant wheat varieties grown in northern China, is significantly tolerant to abiotic stresses such as drought, cold and nutrient deficiencies. In order to isolate key stress-responsive genes and then improve stress tolerances of conventional varieties, a bZIP transcription factor gene was isolated from a cDNA library of drought-treated PYXBM using the in situ plaque hybridization method, and was designated as Triticum aestivum L. abscisic acid （ABA）-responsive element binding protein 1 （TaABP1）. It encodes 372 amino acids, and contains three conserved domains （C1-C3） in the N terminal and a bZIP domain in the C terminal which is a typical protein structure for the group member of bZIP family. Transcriptional activation analysis showed that TaABP1 activated the expression of downstream reporter genes in yeast without ABA application. TaABP1 protein fused with green fluorescent protein （GFP） demonstrated that the localization of TaABP 1 protein is in the nucleus. Expression pattern assays indicated that TaABP1 was strongly induced by ABA, high salt, low temperature and drought, and its expression was stronger in stems and leaves than in the roots of wheat. Furthermore, overexpression of TaABP1 in tobacco showed significant improvement of drought tolerance. Data suggested that TaABP1 may be a good candidate gene for improving stress tolerance of wheat by genetic transformation and elucidation of the role of this gene will be useful for understanding the mechanism underlying drought tolerance of Chinese wheat landrace PYXBM.

关键词： wheat expression pattern stress tolerance sub-cellular localization transcriptional activation

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The NAC-like transcription factor Si NAC110 in foxtail millet(Setaria italica L.) confers tolerance to drought and high salt stress through an ABA independent signaling pathway

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Journal of Integrative Agriculture 2017年第3期16卷 559-571页

作者： XIE li-na CHEN Ming MIN Dong-hong FENG Lu XU Zhao-shi ZHOU Yong-bin XU Dong-bei li lian-cheng MA You-zhi ZHANG Xiao-hong College of Agronomy North west A &F University Yangling 712100 P.R. China National Key Facility for Crop Gene Resources and Genetic Improvement Ministry of Agriculture/Institute of Crop Sciences Chinese Academy of Agricultural Sciences Beijing 100081 P.R. China College of Sciences Northwest A&F University Yangling 712100 P.R. China

Foxtail millet（Setaria italica（L.）***）is a naturally stress tolerant *** to other gramineous crops,it has relatively stronger drought and lower nutrition stress tolerance *** date,the scope of functional genomics research in foxtail millet（*** L.）has been quite ***（NAM,ATAF1/2 and CUC2）-like transcription factors are known to be involved in various biological processes,including abiotic stress *** our previous foxtail millet（*** L.）RNA seq analysis,we found that the expression of a NAC-like transcription factor,SiNAC110,could be induced by drought stress;additionally,other references have reported that SiNAC110 expression could be induced by abiotic ***,we here selected SiNAC110 for further characterization and functional ***,the predicted SiNAC110 protein encoded indicated SiNAC110 has a conserved NAM（no apical meristem）domain between the 11–139 amino acid *** analysis then indicated that SiNAC110 belongs to subfamily III of the NAC gene *** localization analysis revealed that the SiNAC110-GFP fusion protein was localized to the nucleus in Arabidopsis *** expression profiling analysis indicated that expression of SiNAC110 was induced by dehydration,high salinity and other abiotic *** functional analysis using SiNAC110 overexpressed Arabidopsis plants indicated that,under drought and high salt stress conditions,the seed germination rate,root length,root surface area,fresh weight,and dry weight of the SiNAC110 overexpressed lines were significantly higher than the wild type（WT）,suggesting that the SiNAC110 overexpressed lines had enhanced tolerance to drought and high salt ***,overexpression of SiN AC110 did not affect the sensitivity of SiNAC110 overexpressed lines to abscisic acid（ABA）*** analysis of genes involved in proline synthesis,Na＋/K＋transport,drought responses,and aqueous transport proteins were higher in th

关键词： foxtail millet (Setaria italica (L.) NAC-like transcription factor drought stress high salt stress ABA-independent pathway

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Analysis of Specific Binding and Subcellular Localization of Wheat ERF Transcription Factor W17

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Agricultural Sciences in China 2008年第6期7卷 647-655页

作者： ZHAO Yun-xiang liU Pei XU Zhao-shi CHEN Ming li lian-cheng CHEN Yao-feng XIONG Xiang-jin MA You-zhi Institute of Crop Science Chinese Academy of Agricultural Sciences/The National Key Facility for Crop Gene Resources and Genetic Improvement （NFCR1）/Key Laboratory of Crop Genetics and Breeding Ministry of Agriculture Beijing 100081 P.R.China Agronomy College Northwest A ＆ F University Yangling 712100 P.R.China

The study aims to detect the subcellular localization of ERF （ethylene-responsive element binding factor） transcription factor W17 protein, the interaction between W 17 and cis-acting regulatory elements GCC-box and DRE in vitro, the binding and transactivating ability in vivo, and the role of W17 in higher plant stress-signal pathway. Recombinant plasmid W17/163hGFP was introduced into onion epidermal cells by the particle bombardment method with a PDS 1000/He. Transformed cells were incubated for 24 h at 22℃ in the dark and green fluorescence was monitored under a confocal microscope. The gene W17 was fused N-terminus of GST （glutathione-S-transferase） in prokaryotic expression vector pGEX-4T-1 and then transformed into E. coli strain BL21 （DE3）. IPTG （0.5 mmol L-1） was added to induce the expression of recombinant GST/W17 for 3 h. The fused proteins were purified by GST purification columns, and then subjected to gel retardation assay with a 32p-labeled GCC or DRE sequence. The different reporter and effector plasmids were introduced into tobacco leaves through agroinfiltration, then transformed leaves stained by X-Gluc, faded with 75% alcohol and monitored under a Stereozooming microscope. The GFP fused with W17 protein was localized in the nuclei; SDS-PAGE assay demonstrated that the fused protein GST/W17 could be induced and purified with molecular weight at around 42.2 kD under the induction of IPTG. Purified fused protein was able to specifically bind to both the wild-type GCC-box and DRE element, but had no interaction with either the mutant DRE or GCC-box; W17 protein can bind to GCC-box and transactive downstream GUS gene in vivo. W17 can localize into the nuclei, and it may be involved not only in biotic stresses controlled by GCC-box, but also in abiotic stresses （e.g., salt-） induced signaling pathway.

关键词： ERF/AP2 domain ERF DRE element GCC-box subcellular localization

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G-protein β subunit AGB1 positively regulates salt stress tolerance in Arabidopsis

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Journal of Integrative Agriculture 2015年第2期14卷 314-325页

作者： MA Ya-nan CHEN Ming XU Dong-bei FANG Guang-ning WANG Er-hui GAO Shi-qing XU Zhao-shi li lian-cheng ZHANG Xiao-hong MIN Dong-hong MA You-zhi College of Agronomy Northwest A&F University/State Key Laboratory of Arid Region Crop Adversity Biology Institute of Crop Sciences Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement/Key Laboratory of Biology and Genetic Improvement of Triticeae CropsMinistry of Agriculture Beijing Hybrid Wheat Engineering and Technology Research Center

The heterotrimeric GTP-binding proteins（G-proteins） in eukaryotes consisted of α, β and γ subunits and are important in molecular signaling by interacting with G-protein-coupled receptors（GPCR）, on which to transduce signaling into the cytoplast through appropriate downstream effectors. However, downstream effectors regulated by the G-proteins in plants are currently not well defined. In this study, the transcripts of AGB1, a G protein β subunit gene in Arabidopsis were found to be down-regulated by cold and heat, but up-regulated by high salt stress treatment. AGB1 mutant（agb1-2） was more sensitive to high salt stress than wild-type（WT）. Compared with WT, the cotyledon greening rates, fresh weight, root length, seedling germination rates and survival rates decreased more rapidly in agb1-2 along with increasing concentrations of Na Cl in normal（MS） medium. Physiological characteristic analysis showed that compared to WT, the contents of chlorophyll, relative proline accumulation and peroxidase（POD） were reduced, whereas the malonaldehyde（MDA） content and concentration ratio of Na＋/K＋ were increased in agb1-2 under salt stress condition. Further studies on the expression of several stress inducible genes associated with above physiological processes were investigated, and the results revealed that the expressions of genes related to proline biosynthesis, oxidative stress response, Na＋ homeostasis, stress- and ABAresponses were lower in agb1-2 than in WT, suggesting that those genes are possible downstream genes of AGB1 and that their changed expression plays an important role in determining phenotypic and physiologic traits in agb1-2. Taken together, these findings indicate that AGB1 positively regulates salt tolerance in Arabidopsis through its modulation of genes transcription related to proline biosynthesis, oxidative stress, ion homeostasis, stress- and ABA-responses.

关键词： Arabidopsis heterotrimeric G-protein β subunit physiological processes salt stress tolerance

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区域交通路网规模综合评价方法

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北京交通大学学报（社会科学版） 2010年第3期9卷 1-5,53页

作者：李连成北京交通大学经济管理学院北京100044

区域交通路网规模的比较评价是分析区域路网规模的相对水平,进而制定相关发展规划和政策的基础。传统的人口或面积因素分析法存在片面性。本文从运输供给和运输需求的经济特性出发,强调经济发展水平、综合交通理念、路网技术结构等在路... 详细信息

区域交通路网规模的比较评价是分析区域路网规模的相对水平,进而制定相关发展规划和政策的基础。传统的人口或面积因素分析法存在片面性。本文从运输供给和运输需求的经济特性出发,强调经济发展水平、综合交通理念、路网技术结构等在路网规模比较评价中的地位,采用多因素法构建了路网规模经济系数指标,并提出评价包含各种交通方式的不同区域综合路网规模的综合评价方法。该方法既可以比较不同区域的路网规模,也可以对同一地区不同时期的路网发展水平进行评价。

关键词：运输经济路网规模综合评价法

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平面连杆机构尺度综合专家系统研究与实现

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机械科学与技术 2005年第9期24卷 1013-1017页

作者：褚金奎李连成大连理工大学精密与特种加工教育部重点实验室大连116024

将人工智能和信息技术应用到连杆机构设计领域开发了平面连杆机构尺度综合专家系统,介绍了此专家系统的各组成部分,并详细论述了知识库、推理机、方法库和综合数据库等关键模块的创建,最后介绍了专家系统的使用,实例证明本专家系统综合... 详细信息

将人工智能和信息技术应用到连杆机构设计领域开发了平面连杆机构尺度综合专家系统,介绍了此专家系统的各组成部分,并详细论述了知识库、推理机、方法库和综合数据库等关键模块的创建,最后介绍了专家系统的使用,实例证明本专家系统综合速度快、求解精度高,具有很高的实际应用价值。

关键词：连杆机构尺度综合专家系统

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降低HLW型直线振动脱介筛振动频率的生产实践

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煤炭工程 2005年第3期37卷 43-44页

作者：郭崇涛李连成淮北矿业集团临涣选煤厂中国矿业大学化工学院江苏徐州221008 峰峰集团马头选煤厂

论文主要分析了降低振动筛振动频率对其动力学参数及产品脱介效果的影响。通过现场的实验测试发现 ,适度降低振动频率对脱介效果的影响不大 ,但有助于延长脱介用振动筛使用寿命。

关键词：直线振动筛频率脱介效果可靠性

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交通强国的内涵及评价指标体系

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北京交通大学学报（社会科学版） 2020年第2期19卷 12-19页

作者：李连成国家发展改革委综合运输研究所北京100038

交通强国建设根本上是服务国家经济社会发展的,脱离这一基本目的来建设交通强国,则会在发展导向、发展路径等方面产生根本偏差。交通强国需要二三产联动、多产业协同,促进现代化经济体系建设。对交通运输业自身而言,交通强国建设要以高... 详细信息

交通强国建设根本上是服务国家经济社会发展的,脱离这一基本目的来建设交通强国,则会在发展导向、发展路径等方面产生根本偏差。交通强国需要二三产联动、多产业协同,促进现代化经济体系建设。对交通运输业自身而言,交通强国建设要以高质量运输服务为核心,以提高交通运输服务的效率和效益为目标,以快速、经济、安全、绿色、自主五大特征为指向。运输服务供给能力和运输服务质量是评价交通强国两个基本方面:运输服务供给能力要满足经济社会对资源时空配置的要求,即交通运输供给能力与运输需求基本平衡;运输服务质量要满足人们对美好出行的基本预期,并在国际竞争中能够领跑世界并具有可持续性。我国交通基础设施和运输服务规模多项指标位居世界前列甚至第一,这仅仅是与我国人口、经济总量和国土面积的世界地位相匹配而已。根据本文构建的交通强国评价指标体系,定性与定量相结合分析,我国距离交通强国还有差距。

关键词：交通强国内涵评价指标体系

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