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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

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

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

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

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

Regulation and function of endoplasmic reticulum autophagy in neurodegenerative diseases

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Neural Regeneration Research 2025年第1期20卷 6-20页

作者： Xiu-Yun zhao De-En xu Ming-Lei Wu Ji-Chuan Liu Zi-Ling shi Quan-Hong Ma Department of Neurology and Clinical Research Center of Neurological Disease the Second Affiliated Hospital of Soochow UniversitySuzhouJiangsu ProvinceChina Institute of Neuroscience&Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases Soochow UniversitySuzhouJiangsu ProvinceChina Department of Neurology Jiangnan University Medical CenterWuxiJiangsu ProvinceChina

The endoplasmic reticulum,a key cellular organelle,regulates a wide variety of cellular *** reticulum autophagy,one of the quality control systems of the endoplasmic reticulum,plays a pivotal role in maintaining endoplasmic reticulum homeostasis by controlling endoplasmic reticulum turnover,remodeling,and *** this review,we briefly describe the endoplasmic reticulum quality control system,and subsequently focus on the role of endoplasmic reticulum autophagy,emphasizing the spatial and temporal mechanisms underlying the regulation of endoplasmic reticulum autophagy according to cellular *** also summarize the evidence relating to how defective or abnormal endoplasmic reticulum autophagy contributes to the pathogenesis of neurodegenerative *** summary,this review highlights the mechanisms associated with the regulation of endoplasmic reticulum autophagy and how they influence the pathophysiology of degenerative nerve *** review would help researchers to understand the roles and regulatory mechanisms of endoplasmic reticulum-phagy in neurodegenerative disorders.

关键词： autophagy endoplasmic reticulum endoplasmic reticulum autophagy endoplasmic reticulum quality control system endoplasmic reticulum receptors endoplasmic reticulum-associated degradation neurodegeneration neurodegenerative disease selective autophagy unfolded protein response

The pivotal role of microglia in injury and the prognosis of subarachnoid hemorrhage

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Neural Regeneration Research 2025年第7期20卷 1829-1848页

作者： Wenjing Ning shi Lv Qian Wang Yuzhen xu Department of Rehabilitation The Second Affiliated Hospital of Shandong First Medical UniversityTaianShandong ProvinceChina Department of Central Laboratory The Affiliated Taian City Central Hospital of Qingdao UniversityTaianShandong ProvinceChina

Subarachnoid hemorrhage leads to a series of pathological changes,including vascular spasm,cellular apoptosis,blood–brain barrier damage,cerebral edema,and white matter ***,which are the key immune cells in the central nervous system,maintain homeostasis in the neural environment,support neurons,mediate apoptosis,participate in immune regulation,and have neuroprotective *** evidence has shown that microglia play a pivotal role in the pathogenesis of subarachnoid hemorrhage and affect the process of injury and the prognosis of subarachnoid ***,microglia play certain neuroprotective roles in the recovery phase of subarachnoid *** approaches aimed at modulating microglia function are believed to attenuate subarachnoid hemorrhage *** provides new targets and ideas for the treatment of subarachnoid ***,an in-depth and comprehensive summary of the role of microglia after subarachnoid hemorrhage is still *** review describes the activation of microglia after subarachnoid hemorrhage and their roles in the pathological processes of vasospasm,neuroinflammation,neuronal apoptosis,blood–brain barrier disruption,cerebral edema,and cerebral white matter *** also discusses the neuroprotective roles of microglia during recovery from subarachnoid hemorrhage and therapeutic advances aimed at modulating microglial function after subarachnoid ***,microglia in subarachnoid hemorrhage are targeted with TLR inhibitors,nuclear factor-κB and STAT3 pathway inhibitors,glycine/tyrosine kinases,NLRP3 signaling pathway inhibitors,Gasdermin D inhibitors,vincristine receptorαreceptor agonists,ferroptosis inhibitors,genetic modification techniques,stem cell therapies,and traditional Chinese ***,most of these are still being evaluated at the laboratory *** clinical studies and data on subarachnoid hemorrhage are required to improve the treatment of subarachnoid hemor

关键词： apoptosis blood–brain barrier brain edema microglia neuroinflammation neuron neuroprotection subarachnoid hemorrhage vasoconstriction white matter injury

Flexible molecules dedicate to release strain of inverted inorganic perovskite solar cell

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Journal of Energy Chemistry 2025年第1期100卷 87-93页

作者： Hongrui Sun Sanlong Wang Pengyang Wang Yali Liu Shanshan Qi Biao shi Ying zhao Xiaodan Zhang Institute of Photoelectronic Thin Film Devices and Technology Renewable Energy Conversion and Storage Center State Key Laboratory of Photovoltaic Materials and CellsNankai University Key Laboratory of Photoelectronic Thin Film Devices and Technology of Tianjin Haihe Laboratory of Sustainable Chemical Transformations Engineering Research Center of Thin Film Photoelectronic Technology of Ministry of Education Collaborative Innovation Center of Chemical Science and Engineering (Tianjin)

The tensile strain in inorganic perovskite films induced by thermal annealing is one of the primary factors contributing to the inefficiency and instability of inorganic perovskite solar cells （IPSCs）,which reduces the defect formation ***,a flexible molecule 5-maleimidovaleric acid （5-MVA） was introduced as a strain buffer to release the residual strain of *** anhydride and carboxyl groups in 5-MVA interact strongly with the uncoordinated Pb2+through Lewis acid-base reaction,thus tightly“pull”the perovskite *** in-between soft carbon chain increased the structural flexibility of CsPbI2.85Br0.15perovskite materials,which effectively relieved the intrinsic internal strain of CsPbI2.85Br0.15,resisted the corrosion of external strain,and also reduced the formation of defects such as VIand *** addition,the introduction of 5-MVA improved crystal quality,passivated residual defects,and narrowed energy level ***,power conversion efficiency （PCE） of NiOxbased inverted IPSCs increased from 19.25% to 20.82% with the open-circuit voltage enhanced from 1.164 V to 1.230 *** release of strain also improved the stability of CsPbI2.85Br0.15perovskite films and devices.

关键词： Inverted inorganic perovskite solar cells Flexible molecules Strain release Crystallization Energy barrier High PCE

Effect of antisense oligonucleotides of telomerase on the growth of RCZ cells derived from renal cell carcinoma in culture

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吉林大学学报(医学版) 2000年第1期26卷 32-34页

作者： xu Ning,zhao Zhongwen,shi Aiping,Wang Chunxi (Dept.of Urology,The First Clinical College,NBUMS Jilin Changchun 130021)\ \ 〔 Dept.of Urology The First Clinical CollegeNBUMS Jilin Changchun 130021

目的 :研究端粒酶反义抑制剂对肿瘤细胞的影响。方法 :以端粒酶 RNA模板区为靶点 ,序列为 TAGGGTTAGACAA的硫代磷酸修饰的反义寡核苷酸 ( PS- ASON)及 1 3个寡核苷酸的随机引物与肾癌细胞株 RCZ细胞共同孵育 ,计数 1～ 43d细胞数 ,计算... 详细信息

目的 :研究端粒酶反义抑制剂对肿瘤细胞的影响。方法 :以端粒酶 RNA模板区为靶点 ,序列为 TAGGGTTAGACAA的硫代磷酸修饰的反义寡核苷酸 ( PS- ASON)及 1 3个寡核苷酸的随机引物与肾癌细胞株 RCZ细胞共同孵育 ,计数 1～ 43d细胞数 ,计算细胞倍增时间 ,通过 MTT法测定 ASON对细胞生长的抑制率。结果 :ASON实验组与随机引物对照组比较能明显减少细胞增殖数目 ( P<0 .0 1 )、增加抑制率 ( P<0 .0 1 )及延长倍增时间 ( P<0 .0 0 5) ;并且其抑制效果显示出剂量的依赖性 ( P<0 .0 5) ,具有序列选择特异性。结论 :以端粒酶 RNA模板区为靶点的反义寡核苷酸抑制肾癌 RCZ细胞增殖 ,可能对肿瘤治疗有潜在的意义

关键词： telomerase antisense nucleotides renal cell carcinoma