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Study on the Molecular Variation and PCR Detection of Strawberry mottle virus

Agricultural Sciences in China 2009年第10期8卷 1203-1209页

作者： YANG hong-yi LI Li-li DAI hong-yan zhang zhi-hong College of Life Sciences Northeast Forestry University Harbin 150040 P.R. China College of Horticulture Shenyang Agricultural University Shenyang 110161 P.R. China College of Horticulture Northeast Agricultural University Harbin 150030 P.R.China

Strawberry mottle virus （SMoV） is an important viral pathogen infecting strawberry （Fragaria spp.）. The study was conducted to analyze the characterization of the molecular variation of SMoV and develop the methods for detection of SMoV by nested PCR and transcriptional enhancement techniques. The 3 non-coding region （NCR） and large coat protein （LCP） gene of SMoV genome were amplified by reverse transcription-polymerase chain reaction （RT-PCR）. The specific segments were cloned and sequenced. The characterization of the molecular variation for some isolates of SMoV and phylogenetic analysis were studied. Based on the primers located in the conserved region of genome of SMoV, SMoV could be steadily detected using semi-nested PCR and transcriptional enhancement techniques. Both semi-nested PCR and transcriptional enhancement techniques were considerably more sensitive than the standard RT-PCR. The nucleotide sequences of NCR and partial LCP gene of Chinese isolates were obtained, and sequence analysis of the partial LCP gene of various SMoV isolates showed nucleotide identities ranging from 76.8 to 99.7%. There was a slight tendency for isolates to group according to their geographical origin. All 3 Polish isolates, 4 isolates of 7 Dutch isolates, and 3 isolates of 4 Chinese isolates formed a small separate branch, respectively. Two Germanic isolates had a far relationship with other isolates, and formed a separate clade. A high level of sequence variability was found among SMoV isolates, and the Germanic isolates were likely to a special strain group.

关键词： Strawberry mottle virus RT-PCR semi-nested PCR variation phylogenetic analysis

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Quasi-Classical Trajectory Study of the Chemical Reaction Ca+CH_3I

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Chemical Research in Chinese Universities 2008年第2期24卷 223-225页

作者： zhang zhi-hong CHEN Mao-du CONG Shu-lin Department of Physics Dalian University of Technology Dalian 116024 P. R. China Department of Physics Ludong University Yantai 264025 P. R. China

The Ca＋CH3I→CaI＋CH3 reaction system has been studied with the quasi-classical trajectory method on the extended Lond-Eyring-Polanyi-Sato（LEPS） potential energy surface. At collision energy Ecol=10.78 kJ/mol, the calculated results show that the CaI vibrational population peaks are located at v=2. The calculated cross section decreases slowly with the collision energy increasing. The angle product distributions tend toward backward scattering. The calculated （P2（J^1·K）） values deviate slightly from-0.5 and decrease with increasing collision energy. The Quasiclassical trajectory calculation（QCT） results are in reasonable agreement with experimental data. Moreover, the dynamics of the reaction has been discussed.

关键词： Quasi-classical trajectory method Ca+CH3I Potential energy surface

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Isolation and characterization of the secondary wall-related SND1 gene in hawthorn

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Journal of Integrative Agriculture 2018年第9期17卷 2007-2014页

作者： CHEN Ke-qin GUO Yun-na SONG Meng-ru DAI hong-yan zhang zhi-hong Group of Molecular Biology of Fruit Trees College of Horticulture Shenyang Agricultural University Group of Fruit Germplasm Evaluation & Utilization College of Horticulture Shenyang Agricultural University

Secondary wall-associated NAC domain protein1 （SND1） is a key regulator directly regulating the expression levels of MYB46 and MYB83 in the regulation network for secondary wall synthesis, especially in plant fibres. In this study, a SND1 gene was isolated from hawthorn （Crataegus pinnatifida） and named as CpSND 1 because it has a conservative N-terminal DNA- binding domain with AtSNDI. Arabidopsis plants overexpressing CpSND1 had similar phenotypes as plants overexpressing AtSND1, including inhibited growth, upward-curling leaves, sepal dysplasia and sterility. In addition, overexpressing CpSNDI in Arabidopsis also induced the expression of downstream genes, including lignin, cellulose and xylan biosynthesis genes as well as MYB genes. Our results provided functional information of CpSND1 for future genetic engineering in hawthorn.

关键词： hawthorn transcription factor SND1 secondary cell wall transcriptional regulation

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Binding Equilibrium Studies Between Co^(2+) and HSA or BSA

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Chemical Research in Chinese Universities 2000年第3期16卷 276-279页

作者： BIAN He dong LIANG hong TU Chu qiao zhang hong zhi SHEN Pan wen Research Institute of Bioinorganic Chemistry Department of Chemistry and Chemical EngineeringGuangxi Normal UniversityGuilin 541004P.R.China Department of Chemistry Nankai UniversityTianjinP.R.China

The interaction of HSA and BSA with Co2+ and a carboxyl atom of Asp is investigated. The number of binding sites and the cooperation among the binding sites are analyzed. According to the equilibrium dialysis results and the study of competition between Co2+ and Cu2+, Ca2+ or Zn2+ to be bound to HSA or BSA, it is suggested that there are three strong binding sites of in both HSA and BSA.

关键词： Co 2+ serum albumin Equilibrium dialysis Coordination analysis Positive cooperative effect

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Expression and functional analysis of Fa PHO1;H9 gene of strawberry(Fragaria×ananassa)

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Journal of Integrative Agriculture 2017年第3期16卷 580-590页

作者： CAO Fei LI He WANG Shou-ming LI Xiao-ming DAI hong-yan zhang zhi-hong College of Horticulture Shenyang Agricultural University Shenyang 110866 P.R. China

Although the phosphate 1（PHO1）gene family has been implicated in inorganic phosphate transport and homeostasis,the underlying mechanism of this gene in the strawberry has not yet been *** the present study,we analyzed the expression of the PHO1;H9 gene in the strawberry（Fragaria×ananassa）,revealing the involvement of this gene in the regulation of phosphorus（P）*** coding sequence（CDS）of the PHO1;H9 gene,was isolated from the cultivated strawberry‘Sachinoka’and named as Fa PHO1;*** full-length CDS of this gene was 2 292 bp,encoding 763 amino acids,and the protein contained both SYG1/Pho81/XPR1（SPX）and ERD1/XPR1/SYG1（EXS）domains,which were involved in phosphate（Pi）***-time reverse transcription-polymerase chain reaction（RT-PCR）data suggested that the level of Fa PHO1;H9 expression was consistent with the P content in different organs,except for the ***,its expression level was also correlated with P content in fruits of different developmental *** expression of Fa PHO1;H9 was also consistent with P content in leaves under different concentrations of P fertilizer ***,transgenic Arabidopsis lines were generated,and the P content in Arabidopsis plants over-expressing Fa PHO1;H9was significantly higher than that in wild-type ***,we proposed that Fa PHO1;H9 functions in P transport.

关键词： Fragariaxananassa PHO1 H9 phosphorus expression fruits transgenic Arabidopsis

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Isolation and Characterization of Transcriptionally Active Ty1-copia Retrotransposons in Fragaria × ananassa

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Agricultural Sciences in China 2010年第3期9卷 337-345页

作者： MA Yue HE Ping SUN Hai-yue ZHAO Gui-ling DAI hong-yan zhang zhi-hong College of Horticulture Shenyang Agricultural University Shenyang 110866 P.R.China College of Forestry Shenyang Agricultural University Shenyang 110866 P.R.China

One possible mechanism suggested for somaclonal variation is the activation of transposable elements. The activation of retrotransposons by stresses and external changes is commonly observed in plants. In previous study, we isolated the reverse transcriptase （RT） gene sequences of Ty 1-copia retrotransposons from tissue culture strawberry （Fragaria x ananassa） plant, but not the transcriptionally active sequence. For further understanding the relationship between retrotransposon and somaclonal varation, in this study, we isolated the transcriptionally active RT gene sequences from strawberry plants subjected to different abiotic stresses. These retrotransposons were activated by spraying strawberry leaves with 2 mmol L^-1 salicylic acid （SA）, 50 mmol L^-1 methyl jasmonate （MeJA）, 50 mmol L^-1 abscisic acid （ABA）, 50 mmol L^-1 2,4- dichlorophenoxyacetic acid （2,4-D） or by inducing callus growth in 2 types of MS media： first medium supplemented with 0.5 mg L^-1 6-benzylaminopurine （6-BA）, 0.5 mg L^-1 gibberellic acid （GA3）, 1.0 mg L^-1 thidiazuron （TDZ）, and 0.1 mg L^-1 2,4-D, and the second medium supplemented with 0.5 mg L^-1 6-BA, 0.5 mg L^-1 GA3, 2.0 mg L^-1 TDZ, and 0.02 mg L^-1 indole butyric acid （1BA）. Analysis of gene sequences of 17 RTs revealed that none of them contained stop codons and/or indels disrupting the reading frame. These different stress-origin transcriptionally active RTs were remarkably similar to each other- FATEXP2-8 and FATEYS9-7 showed 100% sequence identity. Analysis of pylogenetic of these transcriptionally active RTs and the RT sequences from genome showed that there were close phylogenetic relationships of most of the transcriptionally active RTs. The results of this study have contributed to the background information necessary for future studies for evaluating the relationship between retrotransposons and somaclonal variation.

关键词： transcription activated callus culture Ty1-copia retrotransposons strawberry stress

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Isolation and Identification of Virus dsRNA from Strawberry Plants

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Agricultural Sciences in China 2007年第1期6卷 86-93页

作者： LI He DAI hong-yan zhang zhi-hong GAO Xiu-yan DU Guo-dong zhang Xin-yu College of Horticulture Shenyang Agricultural University Shenyang 110161 P.R.China

The analysis of virus genome is based on nucleic acid isolation. The aims of this study were to develop a method for isolation and identification of virus double-stranded ribonucleic acid （dsRNA） and to elucidate the nucleotide sequences of strawberry virus. Using the modified method, virus dsRNA was extracted from strawberry virus indicator plants and cultivated strawberry plants and detected using agarose gel electrophoresis with ethidium bromide staining and reverse transcription-polymerase chain reaction （RT-PCR）. The quantity of virus dsRNA varied among strawberry cultivars. The quantity of dsRNA from in vitro plantlets was higher than that from the young leaves of field plants. For the field-grown plants, there was more dsRNA in the young leaves. Virus dsRNA extracted from strawberry plants was resistant to deoxyribonuclease Ⅰ （DNase Ⅰ）, but evidently, it became resistant to ribonuclease A （RNase A） only in the presence of 0.5 M NaCl. Its bands in agarose gel could be readily recycled using an agarose gel DNA purification kit. With RT-PCR, the segments of both strawberry mottle virus and Strawberry mild yellow edge virus genomes were amplified by using the virus dsRNA recycled from gel or treated with DNase Ⅰ /RNase A as templates. The system developed for dsRNA isolation and identification in strawberry plants laid a sound foundation for the work on genome analysis of strawberry virus isolates in China.

关键词： dsRNA virus strawberry isolation RT-PCR

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Fingerprinting Analysis of Rhizoma Chuanxiong of Commercial Types using ~1H Nuclear Magnetic Resonance Spectroscopy and High Performance Liquid Chromatography Method

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Journal of Integrative Plant Biology 2009年第6期51卷 537-544页

作者： Hai-Lin Qin An-Jun Deng Guan-Hua Du Peng Wang Jin-Lan zhang zhi-hong Li Institute of Materia Medica Chinese Academy of Medical Sciences Peking Union Medical College Key Laboratory of Bioactive Substances and Resources of Chinese Herbal Medicine Ministry of Education Beijing 100050 China

The ^1 H nuclear magnetic resonance （^1 H NMR） fingerprints of fractionated non-polar extracts （control substance for a plant drug （CSPD） A） from Rhizoma chuanxiong, the rhizomes of Ligusticum chuanxiong Hort., of seven specimens from different sources were measured on Fourier Transform （FT）-NMR spectrometer and assigned by comparing them with the ^1 H NMR spectra of the isolated pure compounds. The ^1 H NMR fingerprints showed exclusively characteristic resonance signals of the major special constituents of the plant. Although the differences in the relative intensity of the ^1H NMR signals due to a discrepancy in the ratio of the major constituents among these samples could be confirmed by high performance liquid chromatography analysis, the general features of the ^1H NMR fingerprint established for an authentic sample of the rhizomes of L. chuanxiong e. hibited exclusive data from those special compounds and can be used for authenticating L. Chuanxiong species.

关键词： control substance for a plant drug A ^1 H nuclear magnetic resonance fingerprint high performance liquid chromatography Ligusticumchuanxiong phenylpropenoid phthalides.

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Comparative transcriptome analysis of shortened fruit mutant in woodland strawberry (Fragaria vesca) using RNA-Seq

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Journal of Integrative Agriculture 2017年第4期16卷 828-844页

作者： WANG Shou-ming LI Wei-jia LIU Yue-xue LI He MA Yue zhang zhi-hong College of Horticulture Shenyang Agricultural University Shenyang 110866 P.R. China

Genes controlling fruit appearance determine fruit shape and size. In ethylmethane sulfonate （EMS）-mutagenized lines of Fragaria vesca accession Yellow Wonder （YW）, two fruit shapes are observed： wild-type long fruit and mutated shortened fruit （sf）. In this study, we first characterized sf based on morphology, histology, cytology and physiology. The sf was identified as a gibberellin （GA）-deficient mutant, and four complementary DNA （cDNA） libraries separately constructed from flower buds and small green fruits of YW and sfwere sequenced to comparatively analyze transcriptome differences. A total of 29 differentially expressed GA pathway genes were identified by comparisons between YWl and sfl, and 28 differentially expressed GA pathway genes were identified between YW2 and sf2. In addition, the expression patterns of 45 differentially expressed genes were validated by quantificational real-time PCR （qRT-PCR）, and the results were highly concordant with the RNA-Seq results.＇ This transcriptome analysis provides valuable information for understanding the molecular mechanisms of fruit development of strawberry.

关键词： strawberry fruit shape transcriptome RNA-Seq differentially expressed gene

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Correlation between Programmed Death-1 Ligand-1 and p53 in Patients with Lung Adenocarcinoma

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Chinese Medical Journal 2018年第8期131卷 990-993页

作者： Cheng Xu zhi-hong zhang Department of Pathology The First Affiliated Hospital of Nanjing Medical University Nanjing Jiangsu 210029 China

Cancer immunotherapy is causing a paradigm shift in tile way cancer researchers think about treating people with the disease. Unlike traditional therapies, immunotherapy harnesses the immune system to fight cancer instead of targeting the ttlmor itself＂ Lung adenocarcinoma is frequently diagnosed in advanced stages with poor prognosis and has become the most common tumor treated by immunotherapy.

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