郭文武
从事柑橘细胞工程、分子育种等研究
个性化签名
- 姓名:郭文武
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学术头衔:
博士生导师, 教育部“新世纪优秀人才支持计划”入选者
- 职称:-
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学科领域:
园艺学
- 研究兴趣:从事柑橘细胞工程、分子育种等研究
郭文武,男,1970年9月生,湖北省郧县人,博士、教授、博士生指导教师。1998年在华中农业大学获果树学博士学位;1999、2003年,分别晋升为副教授、教授。2001年3月至2003年8月,美国佛罗里达大学柑橘研究与教育中心博士后。现为华中农业大学园艺林学学院教授,作物遗传改良国家重点实验室固定研究人员。任国际SCI刊物《Scientia Horticulturae》编委,并为《Plant Cell Reports》、《In Vitro Plant》、《J Amer Soc Hort Sci》、《J Hort Soc & Biotechnology》等SCI刊物审稿。
从事柑橘细胞工程、分子育种等研究。提出了柑橘二倍体无核育种的新思路,在获得一批异源四倍体体细胞杂种的基础上,实现了温州蜜柑胞质雄性不育特性的定向转移,创造了几例二倍体胞质杂种;优化了柑橘转基因技术体系,获得一批转绿色荧光蛋白(GFP)等基因的植株,为柑橘有关基础研究和遗传育种奠定了基础;转GFP基因植株用于细胞融合,为植物体细胞杂交过程中的再生优势现象提供了直观的时空证据,同时为进一步的相关研究提供了新的思路和理想的实验体系。
先后主持国家自然科学基金(2项)、霍英东青年教师基金、湖北省杰出青年基金、瑞典国际科学基金(3项)、武汉市晨光计划等10多个项目。获湖北省自然科学奖二等奖(2001)、中国农学会青年科技奖(2004)、国家发明专利(2004)、湖北省优秀博士学位论文(2000)等奖项。入选武汉市213人才工程培养计划(2003)和教育部新世纪优秀人才支持计划(2004)。
先后多次参加国际柑橘学大会、国际园艺学大会、国际植物组织培养与生物技术大会等会议并口头报告。在《Theor Appl Genet》、《Plant Science》、《Plant Cell Reports》、《Euphytica》、《J Hort Sci & Biotech》、《植物学报》、《园艺学报》等刊物上发表论文40余篇,其中SCI收录论文近20篇,论文被SCI他引80余次。
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【期刊论文】柑橘与枳属间体细胞杂种再生及其对脚腐病抗性的评价
郭文武, 周长河, 伊华林, 邓秀新*
植物学报,2000,42(7):668-672,-0001,():
-1年11月30日
Page橘柚(Citrus reticulata Blanco×C. paradisi Macf.)胚性细胞悬浮系原生质体与枳(Poncirus (L.) Raf.)叶肉原生质体经电场诱导融合,4-5个月再生150余棵小植株。再生植株根系发达,叶片具三叶特征。随机检查20余株再生苗根尖染色体数目,表明都为四倍体(2n=4x=36)。随机取7株进行RAPD分析,表明被检测植株 都为杂种。用引起脚腐病的寄生疫霉菌(Phytophthora prasitica Dastar)毒素接种体细胞杂种及双亲叶片,结果表明,Page橘柚中度感病,枳高抗,体细胞杂种为抗病类型。
电融合, 体细胞杂种, PAPD, 脚腐病抗性, 柑橘
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郭文武, W.W. Guo
Plant Cell Rep (2004) 22: 752-758,-0001,():
-1年11月30日
CMS (cytoplasmic male sterility) can be controlled by the mitochondrion genome in higher plants, including Satsuma mandarin. Somatic fusion experiments in citrus combining embryogenic callus protoplasts of one parent with leaf protoplasts of a second parent often produce cybrid plants of the leaf parent, a phenomenon occurring most often with interspecific fusion combinations. In an attempt to practically exploit this cybridization phenomenon, we conducted somatic fusion experiments combining embryogenic suspension-derived protoplasts of Satsuma mandarin, Citrus unshiu Marc. cv. Guoqing No.1 (G1), a male-sterile cultivar, with leaf protoplasts of other seedy types—Hirado Buntan Pink pummelo (HBP) [Citrus grandis (L.) Osbeck], Sunburst mandarin (C. reticulata Blanco), Orie Lee hybrid (C. reticulata cv. Clementine Murcott tangor), and Murcott tangor [C. reticulata C. sinensis (L.) Osbeck], respectively-in an attempt to generate seedless cybrids by the targeted transfer of CMS. The genetic identities of regenerated plants from all four parental combinations were determined by flow cytometry, SSR, CAPS (or PCR-RFLP), RFLP, and chloroplast-SSR analyses. Regenerated plants from the first three parental combinations were diploids, and the cybrid nature of G1 + HBP with the mitochondrion genome from G1 and the chloroplast genome from HBP was confirmed, whereas the cybrid nature of the remaining two combinations was difficult to confirm because of the close phylogenetic relatedness of both fusion parents, as expected. Plants from G1 + Murcott were confirmed as tetraploid somatic hybrids. This is the first report of targeted citrus cybrid production by symmetric fusion with male-sterile Satsuma as the callus parent and other seedy cultivars as the leaf parents.
Molecular analysis
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郭文武, WENWU GUO, YANXIN DUAN, OSCAR OLIVARES-FUSTER, ZHENCAI WU, COVADONGA R. ARIAS, JACQUELINE K. BURNS, JUDE W. GROSSER*
Plant Cell Reports(2005) DOI: 10.1007/s00299-005-0952-x,-0001,():
-1年11月30日
Valencia orange (Citrus sinensis (L.) Osbeck) is the leading commercial citrus species in the world for processed juice products; however, the presence of thermostable pectin methylesterase (TSPME) reduces its juice quality. A long-term strategy of this work is to eliminate or greatly reduce TSPME activity in Valencia orange. Previous work resulted in isolation of a putative TSPME gene, CsPME4, associated with a thermostable protein fraction of Valencia orange juice. To begin research designed to overexpress CsPME4 to verify the thermostability of the protein product and/or to downregulate the gene, a sense gene cassette containing a gene-specific sequence from a putative TSPME cDNA and the enhanced GFP as a selectable marker was constructed (M2.1). Herein, M2.1 plasmid DNA was transformed (PEG-mediated) into protoplasts isolated from an embryogenic suspension culture of Valencia somaclone line B6-68, in an effort to obtain transgenic Valencia lines. A vigorous transformed line was identified via GFP expression, physically separated from non-transformed tissue, and cultured on somatic embryogenesis induction medium. One transgenic proembryo expressing GFP was recovered and multiple shoots were regenerated. The recovery of multiple transgenic plants was expedited by in vitro grafting. PCR analysis revealed the presence of the PME gene in transgenic plants, and subsequent Southern blot analysis confirmed the presence of the eGFP gene. These transgenic plants show normal growth and minor morphological variation. The thermostability of PME in these plants will be assessed after flowering and fruit set. This is the first successful transfer of a target fruit-quality gene by protoplast transformation with recovery of transgenic plants in citrus.
Citrus,, green fluorescent protein,, juice quality improvement,, in vitro grafting
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郭文武, Wen-Wu Guo a, **, Jude W. Grosser b, *
Plant Science 168 (2005) 1541-1545,-0001,():
-1年11月30日
Previous experience with citrus somatic fusion suggests that hybrid cells are generally more vigorous and have higher capacity for embryogenesis, compared to unfused cells from the embryogenic callus parent. However, this observation has never been validated due to the unavailability of an appropriate experimental design. Citrus mesophyll protoplasts never divide and regenerate into plants. Herein, mesophyll protoplasts of transgenic Valencia orange (Citrus sinensis) which expressed the green fluorescent protein (GFP) gene were fused with protoplasts isolated from an embryogenic callus line of Shekwasha mandarin (C. reticulata) to visually screen somatic hybrids. At the multicell cluster stage, the regenerated callus lines expressing GFP were morphologically identical to the callus parent regenerate. All hybrid callus lines expressing the GFP developed into embryoids, while most callus parent regenerates still remained in a callus stage. Embryoids with strong or weak GFP expression were verified to be diploid cybrid and tetraploid somatic hybrid embryoids, espectively, by flow cytometry, simple sequence repeat (SSR) and chloroplast SSR analyses. Diploid cybrid globular embryoids appeared light green in color and had a tendency to enter subsequent developmental stages earlier than the dark green tetraploids; however, GFP plants were regenerated from both sources. Thus, expression of the GFP marker gene was effectively used to visually screen somatic hybrids, and confirmed our previous observation that somatic hybrid tissue has a regeneration advantage. The intensity of GFP fluorescence and the color difference of regenerated embryoids was also an early indicator of ploidy level (based on visual GFP observation and confirmed by flow cytometry analysis), and could serve as a tool to study the regeneration mechanism of diploid cybrids via symmetric fusion in citrus and other higher plants.
Citrus, Embryogenesis, Green fluorescent protein (, GFP), , Hybrid vigor, Molecular marker, Protoplast fusion
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郭文武, W.W. Guo
Theor Appl Genet (1999) 98: 581-585,-0001,():
-1年11月30日
Chinese wampee [Clausena lansium (Lour.) Skeels], a sexually incompatible relative of citrus, is commercially cultivated in South China. In this study, embryogenic protoplasts of 'Bonanza' navel orange [Citrus sinensis (L.) Osbeck] were electrically fused with leaf protoplasts isolated from 'Chicken Heart' Chinese wampee. After 8 months of culture, fusion products regenerated into shoots. More than 70% of the shoots unexpectedly rooted well. Chromosome counting of several shoot-and root-tips revealed that their chromosome numbers were not 2n "4x" 36 as expected, but 2n "6x" 54, suggesting that chromosome doubling occurred rather than chromosome elimination in this intertribal fusion combination. RAPD analysis of embryoids and the leaves of unrooted and rooted shoots verified their hybridity. This is the first report of hexaploid somatic hybrid plant regeneration from fusion between diploids in Aurantioideae.
Somatic hybridization
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郭文武, W.W. Guo
Plant Cell Rep (2002) 20: 829-834,-0001,():
-1年11月30日
Citrus exocortis viroid (CEV) is widespread in citrus production areas where trifoliate orange [Poncirus trifoliata (L.) Raf.] is used as rootstock. Citrus reticulata Blanco cv. Red tangerine, a different rootstock, is tolerant to CEV. Embryogenic protoplasts of C. reticulata cv. Red tangerine were electrically fused with mesophyll protoplasts from P. trifoliata, and five embryoids were regenerated after 40 days of culture. The embryoids were cut into several pieces and subcultured on shoot induction medium. After 5 months and several subcultures, shoots initially regenerated. The plants grew vigorously with well-developed root systems and exhibited the trifoliate leaf character of P. trifoliata. Chromosome counts on four randomly selected root tips revealed them to be tetraploids (2n=4x=36). RAPD analysis of four randomly selected plants verified their hybridity. This hybridity was further confirmed by AFLP analysis using four primer pairs, from which a total of 65 specific bands were detected. Cytoplasmic genome analysis using universal primers revealed that their chloroplast DNA banding pattern was identical to that of trifoliate orange, while the banding pattern of mitochondrial DNA was identical to that of Red tangerine. The potential of this somatic hybrid as a means to control tree size and provide multi-resistance is discussed.
Citrus
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郭文武, W.W. Guo, X.X. Deng & H.L. Yi
Euphytica 116: 281-285, 2000.,-0001,():
-1年11月30日
Protoplasts from cell suspension cultures of 'Bonnaza' navel orange (Citrus sinensis (L.) Osbeck) were electrically fused with mesophyll protoplasts isolated from seedless 'Red Blush' grapefruit (Citrus paradisi). After 6 months of culture, a total of 20 plants were regenerated. Root tip chromosome counting revealed that 4 of them were tetraploids (2n=4x=36) and the rest were diploids (2n=2x=18) morphologically resembling the mesophyll parent. After 6 months of transplantation into the greenhouse, 4 of the diploid mesophyll regenerants unexpectedly flowered, but this phenomenon disappeared in the next year. This is the first report of precocious flowering in citrus via protoplast fusion. RAPD analysis further confirmed that the tetraploid regenerants were somatic hybrids while the diploid regenerants were mesophyll parent type. This somatic hybrid will be utilized as a possible pollen parent for improving the seedy pummelo cultivars in China by producing triploid seedless pummelo hybrid. The mechanism of early flowering was also discussed.
Citrus,, early flowering,, electrofusion,, RAPD,, somatic hybrids
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郭文武, W.W. Guo
Plant Cell Reports (1998) 18: 297-300,-0001,():
-1年11月30日
Abstract Protoplasts isolated from 'Page' tangelo (Minneola tangelo
Somatic hybridization
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【期刊论文】Citrus somatic hybridization with potential for direct tetraploid scion cultivar development
郭文武, BV W.W GUO, D. PRASAD, P. SERRANO, E.G. GMITTER, JR. and J.W. GROSSER*
Journal of Horticultural Science & Biotechnology (2004) 79 (3) 400-405,-0001,():
-1年11月30日
Protoplast fusion has been an effective tool with several applications for citrus cultivar improvement. The evaluation of fruit characteristics of certain previously produced citrus somatic hybrids with 'Page' tangelo or 'Murcott' tangor as parents, has shown that some allotetraploids combining these parents with other high quality scions were produced as follows: 'Murcott' tangor + 'Dancy' tangerine, 'Murcott' + LB8-8 ('Clementine'
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郭文武, W.W. Guo & X.X. Deng*
Euphytica 118: 175-183, 2001.,-0001,():
-1年11月30日
Citrus wild relatives are an untapped germplasm reservoir, which possesses many elite resistance traits. Genetic introgression into Citrus by conventional methods has been greatly hindered by polyembryony, pollen/ovule sterility, sexual/graft incompatibility, long juvenility etc. Somatic hybridization via protoplast fusion may make it possible to transfer genes from wild relatives to Citrus. To date, more than sixty sexually compatible or incompatible intergeneric somatic hybrids between Citrus and its various related wild genera have been produced by PEG-or electrically-induced fusion. These wide somatic hybrids were identified by morphology, cytology, isozyme, RAPD and RFLP analyses. Genetic variation or recombination was also revealed in some of them. Several sexually compatible combinations have flowered and set fruits. The agronomic performance of these wide somatic hybrids is diverse, and the current results are not conclusive. Somatic hybrids are being tested as rootstocks. The prospects of wide somatic hybridization of Citrus with its wild relatives are discussed.
Aurantioideae,, Citrus,, genetic improvement,, protoplast fusion,, sexually compatible/, incompatible intergeneric somatic hybrids
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