研究領域
One goal of research in my lab is to understand the genetic and molecular basis of cotton fiber development. Cotton is truly a miracle crop in the world, providing fibers for textile manufacturing and affording oil and protein for human food and animal feed production. Cotton fibers are single cells resulted from elongated cells of the ovule epidermis and undergo rapid and synchronous elongation and are considered as an ideal system for studying cell elongation and cell wall development. Fiber number and fiber length, fiber fineness and fiber strength are important agronomy traits for cotton breeding, which are greatly determined by fiber cell initiation, elongation and secondary cell wall accumulation. However, so far, knowledge of the regulatory mechanism controlling cotton fiber initiation and elongation is still mostly unknown. Currently, we are trying to elucidate how plant hormones and their interactions regulate cotton fiber cell development and how to manipulate the expression of the plant hormone biosynthesis and signaling genes to improve cotton fiber quality and yield. We found that auxin, a key phytohormone involved in controlling many aspects of plant development, accumulates highly and specifically in the initiating fiber cells. Based on this observation, we hypothesized that enhancing IAA level in the ovule epidermis at the time of fiber cell initiation by the tissue-specific expression of an IAA biosynthetic gene would promote the initiation of fiber cells and thereby increase fiber yield. Through this way, we generated transgenic cottons which produced higher yield and better quality fibers. The results had been published in “Nature Biotechnology”, a high level journal, and a comment in the same volume gave high words to our work “this study illustrates the promise of the next generation of transgenic crops.”
As transgenic crops are rapidly becoming a staple of modern agriculture in the world, the food safety and gene flow problems resulted from genetically modified crops have been a great concern to both the public and the scientific community. To address this problem, we developed a new recombination system characterized with highly efficient in deleting all functional transgenes from the pollen, seed or both of tobacco plants. This system can be applicable for vegetatively propagated plants. We are now engaged in establishing a new two-component transcript activation system for gene deletion in hybrid crops, such as rice, maize and canola.
Another group in my laboratory is interested in molecular basis of insect fungal diseases. Biological control agents are being considered as supplements or alternatives to synthetic chemical insecticides that are known to have toxic effects on nontarget organisms, including animals and humans. Entomopathogenic fungi are key regulatory factors of insect populations in nature and considered as the only practical microbial control for insects that feed by sucking plant and animal juice and for the many acridid and coleopteran pests which have no known viral or bacterial pathogens. We had cloned several cuticle-degrading enzyme genes, e.g., proteinase or chitinase genes, from Beauveria bassiana, a widely used entomopathogenic fungi in insect pest control. Then, we utilized a chitin-binding domain from an insect, the silkworm moth (B. mori), to produce an engineered chitinase and proteinase in the insect pathogen B. bassiana. The engineered chitinase and proteinase showed increased kinetic properties in substrate hydrolysis than those of the wild-type chitinase and proteinase. Furthermore, when the engineered chitinase and proteinase were overexpressed in B. bassiana, respectively, the genetically modified fungus showed an increase in virulence over a strain with a constitutively expressed native chitinase or proteinase gene. We have demonstrated a strategy where genetic components of the host insect can be incorporated into the fungal pathogen in order to increase host cuticle penetration ability. To isolate new genes involved in infection, we have set up a T-DNA insertion mutant pool of B. bassiana. Some interesting genes had been cloned and characterized from this pool.
近五年主持課題
國家重點基礎研究發展規劃(973)項目:
球孢白僵菌侵染與抗逆相關功能基因的研究(2003CB114203)
“農業生物藥物”重大項目:細菌、真菌類生物殺蟲劑研究與創製 (2006AA10A212)
農業部“公益性行業(農業)科研專項經費”棉花黃萎病減災技術體系研究,(nyhyzx07-052)
國家轉基因專項項目:“轉基因優質纖維棉花新品種培育”(2008ZX08003-003)
國家轉基因專項項目:“高衣分轉基因棉花的創建與新品種選育”(2009ZX08005-023B)
國家高技術研究發展計畫(863)計畫:殺蟲真菌侵染相關基因的克隆與球孢白僵菌高效殺蟲重組菌株的研究 (2001AA214051)
國家高技術研究發展計畫(863)計畫:新型高效殺蟲重組菌株構建(2004AA214121滾動項目)
國家高技術研究發展計畫(863)計畫:細菌、真菌類生物殺蟲劑研究與創製(2006AA10A212)
國家自然科學基金重點項目:利用激素合成與傳導基因改良棉花的纖維品質與產量,(30530490)
國家自然科學基金:用DNA Shuffling技術改造殺蟲真菌分解寄主外殼的幾丁酶,(30080001)
國家自然科學基金:高效雜交幾丁質酶的創建及其在殺蟲真菌菌改造中的套用(30470059)
國家轉基因植物研究與產業化專項課題:速生、抗逆、優質轉基因楊樹桉樹研究與產業化 (JY03-B-25-02)
國家教育部高等學校博士學科點專項科研基金 GhDET2和GhGA20基因在棉花纖維發育中的功能研究( 20040625010)
國家教育部重點項目:球孢白僵菌BbCycl-1基因與菌株毒力的關係
已發表論文(部分)
Agrobacterium tumefaciens-mediated transformation of Beauveria bassiana using an herbicide resistance gene as a selection marker. Weiguo Fang, Yongjun Zhang, Xingyong Yang, Xuelian Zheng, Hui Duan, Yi Li, and Yan Pei*. Journal of Invertebrate Pathology 2004, 85:18–24
Cloning of Beauveria bassiana Chitinase Gene Bbchit1 and Its Application To Improve Fungal Strain Virulence. Weiguo Fang, Bo Leng, Yuehua Xiao, Kai Jin, Jincheng Ma, Yanhua Fan,Jing Feng, Xingyong Yang, Yongjun Zhang, Yan Pei*. Applied and Environmental Microbiology 2005, 71(1):363–370
Construction of a genetic linkage map and QTL analysis of fiber-related traits in upland cotton (Gossypium hirsutum L.). Zheng-Sheng Zhang, Yue-Hua Xiao, Ming Luo, Xian-Bi Li, Xiao-Ying Luo, Lei Hou, De-Mou Li, Yan Pei*. Euphytica 2005, 144: 91–99
The maize Knotted1 gene is an effective positive selectable marker gene for Agrobacterium-mediated tobacco transformation. Keming Luo, Xuelian Zheng, Yongqin Chen, Yuehua Xiao, Degang Zhao, Richard McAvoy, Yan Pei*, Yi Li. Plant Cell Rep 2006,25:403-409
Leaf senescence is delayed in tobacco plants expressing the maize knotted1 gene under the control of a wound-inducible promoter. Keming Luo, Wei Deng, Yuehua Xiao, Xuelian Zheng, Yi Li, Yan Pei*. Plant Cell Rep 2006, 25: 1246–1254
Psc-AFP, an antifungal protein with trypsin inhibitor activity from Psoralea corylifolia seeds. Xingyong Yang, Jun Li, Xiaowen Wanga, Weiguo Fang, Michael J. Bidochka, Rong She, Yuehua Xiao, Yan Pei*. Peptides 2006, 27(7):1726-1731
Isolation and characterization of a novel thermostable non-specific lipid transfer protein-like antimicrobial protein from motherwort (Leonurus japonicus Houtt) seeds. Xingyong Yang, Jun Li, Xianbi Li, Rong She, Yan Pei*. Peptides 2006, 27: 3122-3128
Transformation of Metarizium anisopliae mediated by Agrobacterium tumefaciens. Weiguo fang, Yan Pei and Michael Bidochka. Can. J. Microbiol. 2006, 52: 623-626
Direct amplification of intron-containing hairpin RNA construct from genomic DNA. Yue-Hua Xiao, Meng-Hui Yin, Lei Hou, Yan Pei*. BioTechniques 2006, 41:548-552
Functional Expression of the Cotton Gibberellic Acid Oxidase Homologous Gene GhGA20ox1 in Tobacco. XIAO Yue-Hua, YE Ying-Fu, FENG Yi, LI Xian-Bi LOU Ming, HOU Lei, LUO Xiao-Ying, LI De-Mou, PEI Yan*. Journal of Physiology and Molecular Biology 2006, 32(5): 563-569
Overexpression of an Arabidopsis magnesium transport gene, AtMGT1, in Nicotiana benthamiana confers Al tolerance. Wei Deng, Keming Luo Demou Li, Xuelian Zheng, Xiaoyang Wei, William Smith, Chandra Thammina, Litang Lu, Yi Li, Yan Pei*. Journal of Experimental Botany 2006, 13: 4235-4243
In vitro regeneration and Agrobacterium-mediated genetic transformation of Euonymus alatus. Yougqing Chen, Litang Lu, Wei Deng, Xingyu Yang, Richard MeAvoy, Degang Zhao, Yan Pei, Keming Luo,Hui Duan, William Smith, Chandra Thammina, Xuelian Zheng, Donna Ellis, Yi Li. Plant Cell Rep 2006, 25:1043-1051
Increased Insect Virulence in Beauveria bassiana Strains Overexpressing an Engineered Chitinase. Yanhua Fan, Weiguo Fang, Shujuan Guo, ,Xiaoqiong Pei, Yongjun Zhang, Yuehua Xiao, Demou Li, Kai Jin, Michael J. Bidochka, Yan Pei*. Applied and Environmental Microbiology 2007,73(1): 295–302
‘GM-gene-deletor’: fused loxP-FRT recognition sequences dramatically improve the efficiency of FLP or CRE recombinase on transgene excision from pollen and seed of tobacco plants. Keming Luo, Hui Duan, Degang Zhao, Xuelian Zheng, Wei Deng, Yongqin Chen, Neal Stewart Jr, Richard McAvoy, Xiangning Jiang, Yanhong Wu, Aigong He, Yan Pei, Yi Li. Plant Biotechnology Journal 2007,5(2): 263-374
A regulator of a G protein signalling (RGS) gene, cag8, from the insect-pathogenic fungus Metarhizium anisopliae is involved in conidiation, virulence and hydrophobin synthesis. Weiguo Fang, Yan Pei, Michael J. Bidochka. Microbiology 2007, 153: 1017–1025
Asymmetric overlap extension PCR method bypassing intermediate purification and the amplification of wild-type template in site-directed mutagenesis. Yue-Hua Xiao, Meng-Hui Yin, Lei Hou, Ming Luo, Yan Pei*. Biotechnology Letters 2007 29: 925–930
Cotton flavonoid structural genes related to the pigmentation in brown fibers. Yue-Hua Xiao, Zheng-Sheng Zhang, Meng-Hui Yin, Ming Luo, Xian-Bi Li, Lei Hou, Yan Pei*. Biochemical and Biophysical Research Communications 2007, 358: 73–78
The cauliflower mosaic virus (CaMV) 35S promoter sequence alters the level and patterns of activity of adjacent tissue- and organ-specific gene promoters. Xuelian Zheng, Wei Deng, Keming Luo, Hui Duan, Yongqin Chen, Richard McAvoy, Shuiqing Song, Yan Pei*, Yi Li. Plant Cell Rep 2007, 26: 1195–1203
Directed evolution for increased chitinase activity. Yanhua Fan, Weiguo Fang, Yuehua Xiao, Xingyong Yang, Yongjun Zhang, Michael J. Bidochka, Yan Pei*. Appl Microbiol Biotechnol 2007, 76: 135-139
GhDET2, a steroid 5a-reductase, plays an important role in cotton fiber cell initiation and elongation. Ming Luo, Yuehua Xiao, Xianbi Li, Xiaofeng Lu, Wei Deng, Demou Li, Lei Hou, Mingyu Hu, Yi Li, Yan Pei*. The Plant Journal 2007, 51: 419–430
Sequential Amplification of Flanking Sequences by Y-shaped Adaptor Dependent Extension Using Multiple Templates. XIAO Yue-Hua, PENG Yi, LUO Ming, LI De-Mou, HOU Lei, PEI Yan*. Journal of Plant Physiology and Molecular Biology 2007, 33(1): 85-90
Expression of a Beauveria bassiana chitinase (Bbchit1) in Escherichia coli and Pichia pastoris. Yanhua Fan, Yongjun Zhang, Xingyong Yang, Xiaoqiong Pei, Shujuan Guo, Yan Pei*. Protein Expression and Purification 2007, 56: 93–99
Cloning and Characterization of a Balsam Pear Class I Chitinase gene (Mcchit1) and Its Ectopic Expression Enhances Fungal resistance in Transgenic Plants. Yue-Hua Xiao, Xian-Bi Li, Xingyong Yang, Ming Luo, Lei Hou, Shao-Hui Guo, Xiao-Ying Luo, and Yan Pei*. Biosci Biotechnol Biochem 2007, 71(5): 1211 -1219
A cuticle-degrading protease (CDEP-1) of Beauveria bassiana enhances virulence. Yong-Jun Zhang, Ming-Guang Fang, Yan-Hua Fan, Zhi-Bing Luo, Xing-Yong Yang, Di Wu and Yan Pei*. Biocontrol Science and Technology 2008, 18: 543-555
Implication of a regulator of Gprotein signaling (BbRGS1) in Conidiation and conidial thermotolerance of the insect pathogenic Fungus Beauveria bassiana. Weiguo Fang, Lisa R. Scully, Lei Zhang, Yan Pei and Michael J. Bidochka. FEMS Microbiol Lett 2008,279: 146–156
Functional characterization of a cotton late embryogenesis-abundant D113 gene promoter in transgenic tobacco. Keming Luo, Guofang Zhang, Wei Deng, Fengtao Luo, Kun Qiu, Yan Pei*. Plant Cell Rep 2008, 27: 707-717
Characterization of a Highly Active Promoter, PBbgpd, in Beauveria bassiana. Xing-gang Liao, Wei-guo Fang, Yong-jun Zhang, Yan-hua Fan, Xing-wei Wu, Qun Zhou, Yan Pei*. Current Microbiology 2008, 57: 121-126
An improved method for Beauveria bassiana transformation using phosphinothricin acetlytransferase and green fluorescent protein fusion gene as a selectable and visible marker. Kai Jin, Yongjun Zhang, Zhibing Luo, Yuehua Xiao, Yanhua Fan, Di Wu, Yan Pei*. Biotechnol Lett 2008, 30: 1379–1383
SCFP, a novel fiber-specific promoter in cotton. HOU Lei, LIU Hao, LI JiaBao, YANG Xia, XIAO YueHua, LUO Ming, SONG ShuiQing, YANG GuangWei and PEI Yan*. Chinese Science Bulletin 2008, 53(17): 2639-2645
Functional analysis of the Arabidopsis thaliana poly(A) binding protein PAB5 gene promoter in Nicotiana tabacum. Keming Luo, Wei Deng, Shan Xu, Yan Pei*. Plant Cell Rep 2008 27: 1811-1819
Light stimulates conidiation of the entomopathogenic fungus Beauveria bassiana. Yong-Jun Zhang, Zun-Hua Li, Zhi-Bing Luo, Jian-Qing Zhang, Yan-Hua Fan, Yan Pei*. Biocontrol Science and Technology 2009,19(1): 91-101
The size and ratio of homologous sequence to non-homologous sequence in gene disruption cassette influences the gene targeting efficiency in Beauveria bassiana. Jin-Cheng Ma, Qun Zhou, Yong-Hong Zhou, Xing-Gang Liao, Yong-Jun Zhang, Dan Jin and Yan Pei*. Appl Microbiol Biotechnol 2009, 82: 891–898
Mitogen-Activated Protein Kinase hog1 in the Entomopathogenic Fungus Beauveria bassiana Regulates Environmental Stress Responses and Virulence to Insects. Yong-Jun Zhang, Jianhua Zhao, Weiguo Fang, Jian-Qing Zhang, Zhi-Bing Luo, Mi Zhang, Yan-Hua Fan and Yan Pei*. Appl. Envir. Microbiol. 2009, 75: 3787-3795
註:加*的為通訊作者
專利申請
裴炎,
范艷華,張永軍,方衛國. 重組
絲氨酸蛋白酶及含有其的殺蟲真菌劑(專利號:ZL200510114696.1)
裴炎,周國慶,
侯磊,
羅小英,楊光偉,楊星勇,
李德謀,
肖月華,李名揚. 幾丁質酶和抗菌肽基因雙價抗病載體pChima(專利號:ZL99119241.9)
裴炎,侯磊,李德謀,宋水清,
李先碧,羅明,肖月華,
鄭雪蓮,曾其偉,張覓,邱坤,羅鳳濤,表達生長素合成相關基因的植物表達載體及其在棉花纖維性狀改良的套用(專利申請號:200810142518.3)
張永軍,裴炎,楊星勇,方衛國. 一種殺蟲真菌分生孢子的液體保存方法(專利號:ZL01124159.4)
張永軍,
羅志兵,
裴炎. 一種真菌孢子可濕性粉劑(專利號:ZL200510080659.3)
張永軍,裴炎,彭榮. 通過降低培養基的水活度生產殺蟲真菌液體種子和分生孢子的方法(專利:ZL200510085036.5)