禾谷镰孢菌组蛋白去乙酰化酶基因（HDACs）功能验证
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摘要
由禾谷镰孢菌(F. graminearum)引起的小麦赤霉病是一类很重要的真菌病害。自2003年禾谷镰孢菌野生型菌株PH-1的全基因组序列公布以来,其功能基因组学研究取得了重要进展。因缺乏有效抗病基因以及抵抗初侵染或对其完全免疫的种质资源,目前,关于抗赤霉病的小麦或大麦商业化研究尚未见报道。基于体内同源重组原理的基因敲除技术在禾谷镰孢菌基因功能研究方面的遗传转化体系已经非常成熟,本研究通过构建组蛋白去乙酰化酶基因(HDACs)两个家族classI (HDF4)、classII (HDF1、HDF2、HDF3)的突变体,以及hdf1hdf2、hdf1hdf3两个双突变体,研究HDACs在禾谷镰刀菌(F. graminearum)的生长、致病过程中的分子生物学机制,为更好的防治该病害提供一定的理论基础。
禾谷镰孢菌的基因组预测class I HDAC与酵母的PRD3同源,并且三个class II HDAC基因与酵母里的HOS2、HDA1、HOS3同源。在侵染小麦穗的致病性实验表明,HDF4和HDF1 (FGSG_01353)基因在致病侵染过程中起了重要作用。但是,突变体hdf4的营养生殖减慢了相当多,不产生气生菌丝,并且不产生分生孢子。而突变体hdf1只是轻微的降低了一些。在有性生殖杂交实验中,突变体hdf1是不育的,取代了有性生殖阶段的子囊壳,并且该突变体产生大量的分生孢子。在6小时的芽管里,突变体就产生了大量的油滴结构。并且,突变体hdf1接种小麦穗时,只在接种部位发病,并不扩展,而hdf4接种小麦穗完全不发病。实验结果表明,对HDF1的敲除,很有可能影响和调控了一些下游基因,这些基因包括有性生殖相关基因,产孢相关基因,油滴产生机制相关基因以及与致病相关。
Fusarium head blight (FHB), caused by Fusarium graminearum, is one of the most devastating fungal diseases in wheat. Functional genomics of F. graminearum has been significantly important since the wide-type PH-1 genome was sequenced in 2003. Shortage of effective resistance gene and sources for the prevention or immune of the fungus, no wheat cultivars resistant to WHB are commercialized. Gene knock out based on homologous recombination, along with the corresponding transformation system, has been widely used in functional genomic study of F. graminearum. In this study, we constructed the mutants of two histone deacetylase (HDAC) families class I (HDF4), class II (HDF1, HDF2, HDF3), and the double mutants hdf1hdf2 and hdf1hdf3 in F. graminearum. The molecular mechanism of HDACs genes in development and pathogenicity of F. graminearum was then studied, which will serve theoretical basis for control of the disease.
The F. graminearum genome has one predicted class I HDAC gene that is homologous to yeast RPD3. It also contains three class II HDAC genes that are homologous to HOS2, HDA1, and HOS3 in S. cerevisiae. Mutants deleted for these four HDAC genes were generated with the split-marker approach. Infection assays with flowering wheat heads indicated that the HDF4 and HDF1 (FGSG_01353) genes are important for plant infection. While the hdf4 mutant was severely reduced in vegetative growth, the hdf1 mutant had relatively normal growth rate. The latter produced fewer conidia and shorter aerial hyphae. On mating plates, the hdf1 mutant was sterile. Instead of forming protoperithecia or perithecia, it produced abundant sporodochia with massive amount of macroconidia. In 16 h germlings, the mutant accumulated numerous lipid droplets. The hdf1 mutant was normal in the initial penetration but failed to spread to the rachis of flowering wheat heads. The hdf4 did not caused disease in wheat head. These data suggested that deletion of HDF1 likely affected proper regulation of subsets of genes involved in sexual reproduction, conidiation, lipid metabolism, and plant infection.
禾谷镰孢菌的基因组预测class I HDAC与酵母的PRD3同源,并且三个class II HDAC基因与酵母里的HOS2、HDA1、HOS3同源。在侵染小麦穗的致病性实验表明,HDF4和HDF1 (FGSG_01353)基因在致病侵染过程中起了重要作用。但是,突变体hdf4的营养生殖减慢了相当多,不产生气生菌丝,并且不产生分生孢子。而突变体hdf1只是轻微的降低了一些。在有性生殖杂交实验中,突变体hdf1是不育的,取代了有性生殖阶段的子囊壳,并且该突变体产生大量的分生孢子。在6小时的芽管里,突变体就产生了大量的油滴结构。并且,突变体hdf1接种小麦穗时,只在接种部位发病,并不扩展,而hdf4接种小麦穗完全不发病。实验结果表明,对HDF1的敲除,很有可能影响和调控了一些下游基因,这些基因包括有性生殖相关基因,产孢相关基因,油滴产生机制相关基因以及与致病相关。
Fusarium head blight (FHB), caused by Fusarium graminearum, is one of the most devastating fungal diseases in wheat. Functional genomics of F. graminearum has been significantly important since the wide-type PH-1 genome was sequenced in 2003. Shortage of effective resistance gene and sources for the prevention or immune of the fungus, no wheat cultivars resistant to WHB are commercialized. Gene knock out based on homologous recombination, along with the corresponding transformation system, has been widely used in functional genomic study of F. graminearum. In this study, we constructed the mutants of two histone deacetylase (HDAC) families class I (HDF4), class II (HDF1, HDF2, HDF3), and the double mutants hdf1hdf2 and hdf1hdf3 in F. graminearum. The molecular mechanism of HDACs genes in development and pathogenicity of F. graminearum was then studied, which will serve theoretical basis for control of the disease.
The F. graminearum genome has one predicted class I HDAC gene that is homologous to yeast RPD3. It also contains three class II HDAC genes that are homologous to HOS2, HDA1, and HOS3 in S. cerevisiae. Mutants deleted for these four HDAC genes were generated with the split-marker approach. Infection assays with flowering wheat heads indicated that the HDF4 and HDF1 (FGSG_01353) genes are important for plant infection. While the hdf4 mutant was severely reduced in vegetative growth, the hdf1 mutant had relatively normal growth rate. The latter produced fewer conidia and shorter aerial hyphae. On mating plates, the hdf1 mutant was sterile. Instead of forming protoperithecia or perithecia, it produced abundant sporodochia with massive amount of macroconidia. In 16 h germlings, the mutant accumulated numerous lipid droplets. The hdf1 mutant was normal in the initial penetration but failed to spread to the rachis of flowering wheat heads. The hdf4 did not caused disease in wheat head. These data suggested that deletion of HDF1 likely affected proper regulation of subsets of genes involved in sexual reproduction, conidiation, lipid metabolism, and plant infection.
引文
白娟,刘继彦,郑玲. 2009.组蛋白去乙酰化酶抑制剂抗肿瘤的研究进展.现在肿瘤医学, 17(6): 1194-1196.
陈东亮,李纪元,范正琪,范妙华. 2010.根癌农杆菌介导真菌遗传转化的影响因素及应用.安徽农业科学, 38(7): 3317-3320.
陈坚,张晓琴,傅继梁. 2000.组蛋白脱乙酰化酶及其与基因转录的关系.生物化学与生物物理进展, 27(6): 609-612.
陈婷,陆军,孙晖,董梅,韩松岩,黄百渠. 2003.酵母组蛋白乙酰基转移酶GCN5和去乙酰化酶RPD3在大肠杆菌中的表达.遗传, 25(5): 567-572.
陈延伟,李华. 2009.肺癌患者外周血中单个核细胞HDAC活性的研究.临床医药实践, 18(6): 405-406.
戴旭明. 1997.小鼠凝血因子Ⅸ胚胎干细胞基因打靶研究. [博士学位论文].上海:第二军医大学.
邓祖喜. 1998.小麦栽培与病虫害防治.北京:科学普及出版社.
符稳群,纪春艳,杨晓琦,张敏敏,张振华. 2009.稻瘟病菌组蛋白脱乙酰化酶SIR2 HDACs的功能初探.热带亚热带植物学报, 17(4): 371-377.
符稳群,林莹,黄惠玲,张云珊,池华,杨晓琦,张敏敏,张振华. 2009.水稻组蛋白脱乙酰化酶HD2 HDACs蛋白质的生物信息学分析.漳州师范学院学报(自然科学版), 22(4): 92-98.
高力,陈飞,周立人. 2005.小麦品种望水白的抗赤霉病性遗传分析.麦类作物学报, 25(5): 5-9.
宫本贺,熊和平,马雄风,喻春明,王延周. 2010.基因枪介导法转化苎麻获得转基因植株的研究.作物杂志, 1: 87-90.
贺松,张德纯. 2009.基因敲除方法及其应用.中国微生态学杂志, 21(2): 78-93.
黄小红. 2005.四川省小麦赤霉病菌的种群结构研究. [博士学位论文]四川:四川农业大学.
康印东,王立明. 2009.组蛋白去乙酰化酶抑制剂的免疫抑制功能研究进展.第二军医大学学报, 30(1): 80-83.
康振生,黄丽丽, Buchenauer H,韩青梅,蒋选利. 2004.禾谷镰刀菌在小麦穗部侵染过程的细胞学研究.植物病理学报, 34(4): 329-335.
康振生. 1996.植物病原真菌的超微结构.北京:中国科学技术出版社.
李保华,张卫林. 2007.组蛋白乙酰化/去乙酰化与基因表达调控研究进展.安徽农业科学, 35(18): 5364-5365
李樊,刘义,何钢. 2008.基因敲除技术研究进展.生物技术通报, 80-82, 92.
李娟,杨金奎,梁连铭,张克勤. 2006.丝状真菌遗传转化系统研究进展.江西农业大学学报, 28(4): 516-520.
李瑞国,安晓荣,苟克勉,陈永福. 2002.基因打靶技术及其应用前景.生物技术通报, 5: 6-9.
卢丽丽. 2008.基因枪法导入GUS基因诱导小麦条锈菌毒性突变的研究. [硕士学位论文].杨凌:西北农林科技大学.
吕学诜,吕仁杰,吕超. 2009.基因敲除与基因沉默.黑龙江医药科学, 32(5): 40-47.
王晨芳. 2008.小麦与条锈菌互作过程中活性氧迸发的组织学和细胞化学研究. [博士学位论文].杨凌: 西北农林科技大学.
王友冰,余兴龙. 2003.基因枪技术发展综述.现代科学仪器, 3: 30-34.
王政逸,王秋华,李德葆. 2003.根癌土壤杆菌介导的丝状真菌转化.菌物系统, 22: 339-344.
吴红. 2002.基因敲除动物模型的建立.实验动物科学与管理, 19(1): 32-35.
徐雍皋,陈利锋. 1993.小麦赤霉病防治理论研究与实践.南京:江苏科学技术出版社.
薛猛. 2002.基因打靶技术的研究进展.生物学杂志, 18(1): 7-8.
姚金保,葛永福,王文书. 1997.小麦品种苏麦3号抗赤霉病基因的染色体定位研究.作物学报, 23(4): 450-453.
姚金保.陆维忠. 2000.中国小麦抗赤霉病育种研究进展.江苏农业学报, 16(4): 242-248.
殷红,陈子兴. 2003.组蛋白去乙酰化异常与恶性肿瘤.国际医学(生理、病理科学与临床分册), 23(3): 238-240.
俞大绂. 1955.中国镰刀菌属(Fusarium)菌种的初步名录.植物病理学报, 1: 12-17.
俞刚,陈利锋,姚红燕,柴一秋. 2003.脱氧雪腐镰刀菌烯醇在小麦赤霉病病程中的作用.植物病理学报, 33(1): 40-43.
袁婷露,曹秀云. 2008.禾谷镰刀菌致病力和致病基因的研究进展.安徽农业科学, 36(14): 5915-5916, 5934.
张凯鸣,马鸿翔,陆维忠,蔡志翔,陈怀谷,袁生. 2006.小麦赤霉病与DON积累的抗性及其相关SSR位点差异.作物学报, 32(12): 1788-1795.
张鹏,马鸿翔. 2009.小麦赤霉病镰孢菌毒素的生物合成及其分子调控.生物技术通报, 1: 11-15
Allemand I, Augulo J F. 1995. Transgenic and knock out models for studying DNA repair. Biochimie, 77(10): 826-832.
Anand A, Zhou T, Trick H N, Gill B S, Bockus W W, Muthukrishnan S. 2003. Greenhouse and field testing of transgenic wheat plants stably expressing genes for haumatin-like protein, chitinase and glucanase against Fusarium graminearum. Journal of Experimental Botany, 54: 1101-1111.
Bai G, Shaner, G E 2004. Management and resistance in wheat and barley to fusarium head blight. Annual Review of Phytopathology, 42: 135-161.
Baidyaroy, D., Brosch, G., Ahn, J.H., Graessle, S., Wegener, S., Tonukari, N.J., Caballero, O., Loidl, P. and Walton, J.D. (2001) A gene related to yeast HOS2 histone deacetylase affects extracellular depolymerase expression and virulence in a plant pathogenic fungus. Plant Cell, 13, 1609-1624.
Bluhm B H, Zhao X, Flaherty J E, Xu J R, Dunkle1 L D. 2007. RAS2 regulates growth and pathogenesisin Fusarium graminearum. Molecular Plant-Microbe Interactions, 20(6): 627–636.
Capecchi MR. 1989. Altering the genome by homologous recombination. Science, 244: 1288-1292.
CHEN GQ, ZHU J, SHI X G, Ni J H, Zhong H J, Si G Y, Jin X L, Tang W, Li X S, Xong S M, Shen Z X, Sun G L, Ma J, Zhang P, Zhang T D, Gazin C, Naoe T, Chen S J, Wang Z Y, Chen Z. 1996. In vitro studies on cellular and molecular mechanisms of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia: As2O3 induces NB4 cell apoptosis with downregulation of Bcl-2 expression and modulation of PML-RAR alpha/PML Proteins. Blood, 88: 1052-1061.
Cheong Y H, Kim k N, Pandey G K, Guptaa R, John J, Granta J J, Sheng L. 2007. CBL1, a Calcium Sensor That Differentially Regulates Salt, Drought, and Cold Responses in Arabidopsis. The Plant Cell, 15: 1833-1845
Choi J H, Kwon H J, Yoon B I, Kim J H, Han S U, Joo H J, Kim D Y. 2001. Expression profile of histone deacetylase 1 in gastric cancer tissues. Jpn J Cancer Res, 92(12): 1300-1304.
Christian A V, Wilhelm S. 2005. A secret lipase of Fusarium graminearum is a virulence factor required forinfection of cereals. The Plant Journal, 42: 364-375.
Crazzolara R, Johrer K, Johnstone R W, Greil R, Kofler R, Meister B, Bernhard D. 2002. Histone deacetylase inhibitom potently repress CXCR4 chemokine receptor expression and function in acute lymphoblastic leukaemia.Br J Haematol, 119(4): 965-969.
Cress W D, Seto E. 2000. Histone deacetylase, transcription control, and cancer. Cell Physiol, 184: 1-16. Dahleen L S, Okubara P A, Blechl A E. 2001. Transgenic Approaches to Combat Fusarium Head Blight in Wheat and Barley. Crop Science, 41:628-637.
Dangl M, Brosch G, Haas H, Loidl P, Lusser A. 2001. Comparative analysis of HD2 type histone deacetylases in higher plants. Planta, 213(2): 280-285.
de Groot M JA, Bundock P, Hooykaas P, Beijersbergen A GM. 1998. Agrobacterium tumefaciens-mediated transformation of filamentous fungi. Nat Biotechnol, 16: 839-842.
De Nadal E, Zapater M, Alepuz P M, Sumoy L, Mas G, Posas F. The MAPK Hog1 recruits Rpd3 histone deacetylase to activate osmoresposive genes. Nature, 2004, 427: 370-374
Dean M, Carrington M, Winkler C, Huttley G A, Smith M W, Allikmets R, Goedert J J, Buchbinder S P, Vittinghoff E, Gomperts E, Donfield S, Vlahov D, Kaslow R, Saah A, Rinaldo C, Detels R, O'Brien S J. 1996. Genetic restriction of HIV-1 infection and progression to AIDS by a deletion allele of the CKR5 structural gene. Hemophilia Growth and Development Study, Multicenter AIDS Cohort Study, Multicenter Hemophilia Cohort Study, San Francisco City Cohort, ALIVE Study. Sicence, 273(5283): 1856-1862.
Ding S, Mehrabi R, Koten C, Kang Z, Wei Y, Seong K, Kistler H C, Xu J R. 2008. The transducin beta like gene FTL1 is essential for pathogenesis in Fusarium graminearum. Eucarytic Cell, 8(6): 867-876.
E. Keats Shwab, Jin Woo Bok, Martin Tribus, Johannes Galehr, Stefan Graessle, and Nancy P.Keller. 2007. Histone Deacetylase Activity Regulates Chemical Diversity in Aspergillus. Eukaryotic Cell, 1656-1664.
Gao L, Gross D S. 2008. Sir2 silences gene transcription by targeting the transition between RNA polymeraseII initiation and elongation. Mol Cell Biol, 28(12): 3979-3994.
Gerlar R. 1996. Gene-targeting studies of mammalian behavior: Is it the mutation or the background genotype? Trends in Neurosciences, 19(5): 177-181.
Goswami R S, Kistler H C. 2004. Heading for a disaster: Fusarium graminearum on cereal crops. Molecular Plant Pathology, 5: 515-525.
Goswami R S, Kistler H C. 2004. Heading for disaster: Fusarium graminearum on cereal crops. Molecular Plant Pathology, 5(6): 515-525.
Goswami R S, Xu J R, Trail F, Hilburn K, Kistler H C. 2006. Genomic analysis of host–pathogen interaction between Fusarium graminearum and wheat during early stages of disease development. Microbiology, 152: 1877-1890.
Goswami R S, Xu J R, Trail F, Hilburn K, Kistler H C. 2006. Genomic analysis of host–pathogen interaction between Fusarium graminearum and wheat during early stages of disease development. Microbiology, 152: 1877-1890.
GREENSHIELDS D L, LIU G S, FENG J, SELVARAJ G, WEI Y D. 2007. The siderophore biosynthetic gene SID1, but not the ferroxidase gene FET3, is required for full Fusarium graminearum virulence. Molecular Plant Pathology, 8(4): 411-421.
Güldener U, Seong K Y, Boddu J, Cho S, Trail F, Xu J R, Adam G, Mewes H W, Muehlbauer Gary J, Kistler H C. 2006. Development of a Fusarium graminearum Affymetrix GeneChip for profiling fungal gene expression in vitro and in planta. Fungal Genetics and Biology, 43:316-325.
Han Y K, Kim M D, Lee S H, Yun S H, Lee Y W. 2007. A novel F-box protein involved in sexual development and pathogenesis in Gibberella zeae. Mol Microbiol, 63(3): 768-779.
Hasty P, Ramirez-Solis R, KrumLauf R, Bradley A. 1991. Introduction of a subtle mutation into the Hox-2.6 locus in embryonic stem cells. Nature, 35(6315): 243-246.
HOU Z, XUE C, PENG Y, KATAN T, KISTLER H C, XU J. 2002. A map kinase gene(MGV1) in fusarium graminearum is required for female fertility, heterokaryon formation,hypal growth and plant infection. Molecular Plant-Microbe Interaction, 15: 1119-1127.
Hua-Van A, Pamphile J A, Langin T, Daboussi M J. 2001. Transposition of autonomous and engineered impala transposons in Fusarium oxysporum and a related species. Mol Gen Genet, 264: 724-731.
Insinga A, Monestiroli S, Ronzoni S, Gelmetti V, Marchesi F, Andrea Viale A, Altucci L, Nervi C, Saverio M, Pelicci P G. 2005. Inhibitors of histone deacetylases induce tumorselective apoptosis through activation of the death receptor pathway. Nat Med, 11: 71-76.
Jenczmionka N J, Maier F J, Losch A P, Schafer W. 2003. Mating, conidiation and pathogenicity of Fusarium graminearum, the main causal agent of the head-blight disease of wheat, are regulated by the MAP kinase gpmK1.Current genetics, 43(2):87-95
Joyner A L, Skarnes W C, Rossant J. 1989. Production of a mutation in mouse En-2 gene by homologous recombination in embryonic stem cells. Nature, 338: 153-156.
Kasschau K D, Carrington J C. 1998. A counterdefensive strate of plant viruses: uppression of posttranscriptional gene silencing. Cell, 95(4): 461-470.
Kilby N J, Snaith M R, Murray J A. Site-specific recombinases: tools for genome engineering. Trends Genet, 1993, 9(12): 413-421.
Koller B H, Hagemann L J, Doetschman T, Hagaman J R, Huang S, Williams P J, First N L, Maeda N, Smithies O. 1989. GermLine transmission of a planned alteration made in a hypoxanthine phosphoribosyltransferase gene by homologous recombination in embryonic stem cells. Proc Natl Acad Sci USA, 86(22): 8927-8931.
Kumar A, Seringhaus M, Biery M C, Sarnovsky R J, Umansky L, Piccirillo S, Heidtman M, Cheung K H, Dobry C J, Gerstein M B, Craig N L, Snyder M. 2004. Large-scale mutagenesis of the yeast genome using a Tn7-derived multipurpose transposon. Genome Res, 14: 1975-1986.
Lagac M, Chantha S C, Major G, Matton D P. 2003. Fertilization induces strong accumulation of a histone deacetylase (HD2) and of chromatin remodeling proteins in restricted areas of the ovules. Plant Mol Biol, 53(6): 759-769.
Lathe R. 1996. Mice, gene targeting and behaviour: more than just genetic background. Trends Neurosci, 19(5):183-186.
Li L, Xue C, Bruno K, Nishimura M, Xu J R. 2004. Two PAK kinase genes, CHM1 and MST20, have distinct functions in Magnaporthe grisea. Molecular Plant-Microbe Interactions, 17(5): 547-556.
Loprevite M, Tiseo M, Grossi F, Scolaro T, Semino C, Pandolfi A, Favoni R, Ardizzoni A. 2005. In vitro study of CI-994, a histone deacetylase inhibitor, in non-small cell lung cancer cell lines. Oncol Research, 15(1): 39-48.
Lu S W, Kroken S, Lee B N, Robbertse B, Alice C L, Churchill O, Yoder C, Turgeon B G. 2003. A novel class of gene controlling virulence in plant pathogenic ascomycete fungi. Proc Natl Acad Sci USA, 100(10): 5980–5985.
LU Y, FANG J Y. 2006. Epigenetic modification and cancer. Chinese Bulletin of Life Sciences, 18(1): 10-14.
Mesterhazy A. 1995. Types and components of resistance to Fusarium head blight. Plant Breed, 114: 377-386.
Oide S, Moeder W, Krasnoff S, Gibson D, Haas H, Yoshioka K, Turgeon B G. 2006. NPS6, Encoding a Nonribosomal Peptide Synthetase Involved in Siderophore-Mediated Iron Metabolism, Is a Conserved Virulence Determinant of Plant Pathogenic Ascomycetes. The Plant Cell, 18: 2836-2853.
Patm S K, Pawa A, Dahiya R. 2001. Histone deacetylase and DNA methyltransferase in human prostate cancer. Biochem Biophys Res Commun, 287(3):705-713.
Pfaffl M W. 2001 A new mathematical modle for relative quantification in real-time RT-PCR. Nucleic Acids Res, 29: e45.
Pugh GW, Jenann H, Dickson JG. 1963. Factors affecting infection of wheat heads by Gibberel la saubinetii. Journal of Agricultural Research, 44: 207-238.
Redner R L, Wang J, Liu J M. 1999. Chromatin remodeling and leukemia: new therapeutic paradigms. Blood, 94(2): 417-428.
Robyr D, Suka Y, Xenarios I, Kurdistani S K, Wang A, Suka N, Grunstein M. 2002. Microarray deacetylation maps determine genome-wide functions for yeast histone deacetylase. Cell, 109: 437-466.
Rosato R R, Almenara J A, Grant S. 2003. The histone deacetylase inhibitor MS-275 promotes differentiation or apoptosis in human leukemia cells through a process regulated by generation of reactive oxygen species and induction of p21cip1/waf1. Cancer Res, 63: 3637-3645.
Rundlett S E, Carmen A A, Kobayashi R, Bavykin S, Turner B M, Grunstein M. 1996.HDA1 and RPD3 are members of distinct yeast histone deacetylase complex that regulate silencing and transcription. Proc Natl Acad Sci USA, 93: 14503-14508.
Schiestl R H, Petes T D. 1991. Integration of DNA fragments by illegitimate recombination in Saccharomyces cerevisiae. Proc Natl Acad Sci USA, 88: 7585-7589.
Schwartzberg P L, Robertson E J, Goff S P. 1990. Targeted gene disruption of the endogenous c-abl locus by homologous recombination with DNA encoding a selectable fusion protein. Proc Natl Acad Sci USA, 87: 3210-3214.
Seo B W, Kim H K, Lee Y W, Yun S H. 2007. Functional analysis of a histidine auxotrophic mutation in Gibberella zeae. Plant Pathol, 23:51–56.
Seong K, Hou Z M, Tracy M, Kistler H C, Xu J R. 2005. Random insertional mutagenesis identifies genes associated with virulence in the wheat scab fungus Fusarium graminearum. Phytopathology, 95:744–750.
Seong K, Li L, Hou Z, Tracy Mile, Kistler H C, Xu J R. 2006. Cryptic promoter activity in the coding region of the HMG-CoA reductase gene in Fusarium graminearum. Fungal Genetics and Biology, 43: 34-41.
Shim W B, Sagaram U S,Choi Y E, So J, Wilkinsom H H, Lee YW. 2006. FSR1 is essential for virulenceand female fertility in Fusarium verticillioides and F. graminearum. Mol Plant Microbe Ineract, 19(7):725-33.
Smithies O, Gregg RG, Boggs SS, Koralewski M A, Raju S. Kucherlapati R S. 1985. Insertion of DNA sequencs into the human chromosomal betaglobin locus by homologous recombination. Nature, 317(6034): 230-234.
Sridha S, Wu K. 2006. Identification of AtHD2C as a novel regulator of abscisic acid response in Arabidopsis. The Plant Journal, 46(1): 124-133.
Thomas K R, Capecchi M R. 1987. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell, 51: 503-512.
Thompson S, Clarke A R, Pow A M, et al . 1989. GermLine transmission and stem cells. Cell, 56: 313-321. Timothy A Bolger. Year The Function and Regulation of Histone Deacetylase-4 in Neurons. [PhD Desertation]. : Department of Pharmacology and Cancer Biology Duke University. Valancius V. Smithies O. 1991. Double-strand gap repair in a mammalian gene targeting reaction. Mol Cell Biol, 11(9): 4389-4397.
Wang A, Kurdistani S K, Grunstein M. 2002. Requirement of Hos2 histone deacetylase for gene activity in yeast. Science. 298: 1412-1414
Wang YZ. 1997. Epidemiology and management of wheat scab in China. CIMMY conference on Fusarium head scab: global status and future prospects. CIMMYT, El Batan, 97-105.
Wu H, Liu X, Janeisch R. 1994. Double replacement: Strategy for efficient introduction of subtle mutations into the murine Col1a-1 gene by homologous recombination in embryonic stem cells. Proc Natl Acad Sci USA, 91: 2819-2823.
Yamamoto T, Omoto S, Mizuguchi M, Mizukami H, Okuyama H, Okada N, Saksena N K, Brisibe E A, Otake K, Fuji Y R. 2002. Double stranded nef RNA interferes with human immuno deficiency virus type 1 replication. Micobiol Immunol, 46: 809-817.
Yang B, Britton J, Kirchmaier A L.2008. Insights into the impact of histone aeetylation and methylation on Sir protein recruitment, spreading, and silencing in Saccharomyces cerevisiae. J Mol BIOl, 381(4): 826-844.
Yu H Y, Seo J, Klm J. 2008. Functional analyses of heterotrimeric G protein Gαand Gβsubunits in Gibberella zeae. Microbiology, 154(2): 392-401.
陈东亮,李纪元,范正琪,范妙华. 2010.根癌农杆菌介导真菌遗传转化的影响因素及应用.安徽农业科学, 38(7): 3317-3320.
陈坚,张晓琴,傅继梁. 2000.组蛋白脱乙酰化酶及其与基因转录的关系.生物化学与生物物理进展, 27(6): 609-612.
陈婷,陆军,孙晖,董梅,韩松岩,黄百渠. 2003.酵母组蛋白乙酰基转移酶GCN5和去乙酰化酶RPD3在大肠杆菌中的表达.遗传, 25(5): 567-572.
陈延伟,李华. 2009.肺癌患者外周血中单个核细胞HDAC活性的研究.临床医药实践, 18(6): 405-406.
戴旭明. 1997.小鼠凝血因子Ⅸ胚胎干细胞基因打靶研究. [博士学位论文].上海:第二军医大学.
邓祖喜. 1998.小麦栽培与病虫害防治.北京:科学普及出版社.
符稳群,纪春艳,杨晓琦,张敏敏,张振华. 2009.稻瘟病菌组蛋白脱乙酰化酶SIR2 HDACs的功能初探.热带亚热带植物学报, 17(4): 371-377.
符稳群,林莹,黄惠玲,张云珊,池华,杨晓琦,张敏敏,张振华. 2009.水稻组蛋白脱乙酰化酶HD2 HDACs蛋白质的生物信息学分析.漳州师范学院学报(自然科学版), 22(4): 92-98.
高力,陈飞,周立人. 2005.小麦品种望水白的抗赤霉病性遗传分析.麦类作物学报, 25(5): 5-9.
宫本贺,熊和平,马雄风,喻春明,王延周. 2010.基因枪介导法转化苎麻获得转基因植株的研究.作物杂志, 1: 87-90.
贺松,张德纯. 2009.基因敲除方法及其应用.中国微生态学杂志, 21(2): 78-93.
黄小红. 2005.四川省小麦赤霉病菌的种群结构研究. [博士学位论文]四川:四川农业大学.
康印东,王立明. 2009.组蛋白去乙酰化酶抑制剂的免疫抑制功能研究进展.第二军医大学学报, 30(1): 80-83.
康振生,黄丽丽, Buchenauer H,韩青梅,蒋选利. 2004.禾谷镰刀菌在小麦穗部侵染过程的细胞学研究.植物病理学报, 34(4): 329-335.
康振生. 1996.植物病原真菌的超微结构.北京:中国科学技术出版社.
李保华,张卫林. 2007.组蛋白乙酰化/去乙酰化与基因表达调控研究进展.安徽农业科学, 35(18): 5364-5365
李樊,刘义,何钢. 2008.基因敲除技术研究进展.生物技术通报, 80-82, 92.
李娟,杨金奎,梁连铭,张克勤. 2006.丝状真菌遗传转化系统研究进展.江西农业大学学报, 28(4): 516-520.
李瑞国,安晓荣,苟克勉,陈永福. 2002.基因打靶技术及其应用前景.生物技术通报, 5: 6-9.
卢丽丽. 2008.基因枪法导入GUS基因诱导小麦条锈菌毒性突变的研究. [硕士学位论文].杨凌:西北农林科技大学.
吕学诜,吕仁杰,吕超. 2009.基因敲除与基因沉默.黑龙江医药科学, 32(5): 40-47.
王晨芳. 2008.小麦与条锈菌互作过程中活性氧迸发的组织学和细胞化学研究. [博士学位论文].杨凌: 西北农林科技大学.
王友冰,余兴龙. 2003.基因枪技术发展综述.现代科学仪器, 3: 30-34.
王政逸,王秋华,李德葆. 2003.根癌土壤杆菌介导的丝状真菌转化.菌物系统, 22: 339-344.
吴红. 2002.基因敲除动物模型的建立.实验动物科学与管理, 19(1): 32-35.
徐雍皋,陈利锋. 1993.小麦赤霉病防治理论研究与实践.南京:江苏科学技术出版社.
薛猛. 2002.基因打靶技术的研究进展.生物学杂志, 18(1): 7-8.
姚金保,葛永福,王文书. 1997.小麦品种苏麦3号抗赤霉病基因的染色体定位研究.作物学报, 23(4): 450-453.
姚金保.陆维忠. 2000.中国小麦抗赤霉病育种研究进展.江苏农业学报, 16(4): 242-248.
殷红,陈子兴. 2003.组蛋白去乙酰化异常与恶性肿瘤.国际医学(生理、病理科学与临床分册), 23(3): 238-240.
俞大绂. 1955.中国镰刀菌属(Fusarium)菌种的初步名录.植物病理学报, 1: 12-17.
俞刚,陈利锋,姚红燕,柴一秋. 2003.脱氧雪腐镰刀菌烯醇在小麦赤霉病病程中的作用.植物病理学报, 33(1): 40-43.
袁婷露,曹秀云. 2008.禾谷镰刀菌致病力和致病基因的研究进展.安徽农业科学, 36(14): 5915-5916, 5934.
张凯鸣,马鸿翔,陆维忠,蔡志翔,陈怀谷,袁生. 2006.小麦赤霉病与DON积累的抗性及其相关SSR位点差异.作物学报, 32(12): 1788-1795.
张鹏,马鸿翔. 2009.小麦赤霉病镰孢菌毒素的生物合成及其分子调控.生物技术通报, 1: 11-15
Allemand I, Augulo J F. 1995. Transgenic and knock out models for studying DNA repair. Biochimie, 77(10): 826-832.
Anand A, Zhou T, Trick H N, Gill B S, Bockus W W, Muthukrishnan S. 2003. Greenhouse and field testing of transgenic wheat plants stably expressing genes for haumatin-like protein, chitinase and glucanase against Fusarium graminearum. Journal of Experimental Botany, 54: 1101-1111.
Bai G, Shaner, G E 2004. Management and resistance in wheat and barley to fusarium head blight. Annual Review of Phytopathology, 42: 135-161.
Baidyaroy, D., Brosch, G., Ahn, J.H., Graessle, S., Wegener, S., Tonukari, N.J., Caballero, O., Loidl, P. and Walton, J.D. (2001) A gene related to yeast HOS2 histone deacetylase affects extracellular depolymerase expression and virulence in a plant pathogenic fungus. Plant Cell, 13, 1609-1624.
Bluhm B H, Zhao X, Flaherty J E, Xu J R, Dunkle1 L D. 2007. RAS2 regulates growth and pathogenesisin Fusarium graminearum. Molecular Plant-Microbe Interactions, 20(6): 627–636.
Capecchi MR. 1989. Altering the genome by homologous recombination. Science, 244: 1288-1292.
CHEN GQ, ZHU J, SHI X G, Ni J H, Zhong H J, Si G Y, Jin X L, Tang W, Li X S, Xong S M, Shen Z X, Sun G L, Ma J, Zhang P, Zhang T D, Gazin C, Naoe T, Chen S J, Wang Z Y, Chen Z. 1996. In vitro studies on cellular and molecular mechanisms of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia: As2O3 induces NB4 cell apoptosis with downregulation of Bcl-2 expression and modulation of PML-RAR alpha/PML Proteins. Blood, 88: 1052-1061.
Cheong Y H, Kim k N, Pandey G K, Guptaa R, John J, Granta J J, Sheng L. 2007. CBL1, a Calcium Sensor That Differentially Regulates Salt, Drought, and Cold Responses in Arabidopsis. The Plant Cell, 15: 1833-1845
Choi J H, Kwon H J, Yoon B I, Kim J H, Han S U, Joo H J, Kim D Y. 2001. Expression profile of histone deacetylase 1 in gastric cancer tissues. Jpn J Cancer Res, 92(12): 1300-1304.
Christian A V, Wilhelm S. 2005. A secret lipase of Fusarium graminearum is a virulence factor required forinfection of cereals. The Plant Journal, 42: 364-375.
Crazzolara R, Johrer K, Johnstone R W, Greil R, Kofler R, Meister B, Bernhard D. 2002. Histone deacetylase inhibitom potently repress CXCR4 chemokine receptor expression and function in acute lymphoblastic leukaemia.Br J Haematol, 119(4): 965-969.
Cress W D, Seto E. 2000. Histone deacetylase, transcription control, and cancer. Cell Physiol, 184: 1-16. Dahleen L S, Okubara P A, Blechl A E. 2001. Transgenic Approaches to Combat Fusarium Head Blight in Wheat and Barley. Crop Science, 41:628-637.
Dangl M, Brosch G, Haas H, Loidl P, Lusser A. 2001. Comparative analysis of HD2 type histone deacetylases in higher plants. Planta, 213(2): 280-285.
de Groot M JA, Bundock P, Hooykaas P, Beijersbergen A GM. 1998. Agrobacterium tumefaciens-mediated transformation of filamentous fungi. Nat Biotechnol, 16: 839-842.
De Nadal E, Zapater M, Alepuz P M, Sumoy L, Mas G, Posas F. The MAPK Hog1 recruits Rpd3 histone deacetylase to activate osmoresposive genes. Nature, 2004, 427: 370-374
Dean M, Carrington M, Winkler C, Huttley G A, Smith M W, Allikmets R, Goedert J J, Buchbinder S P, Vittinghoff E, Gomperts E, Donfield S, Vlahov D, Kaslow R, Saah A, Rinaldo C, Detels R, O'Brien S J. 1996. Genetic restriction of HIV-1 infection and progression to AIDS by a deletion allele of the CKR5 structural gene. Hemophilia Growth and Development Study, Multicenter AIDS Cohort Study, Multicenter Hemophilia Cohort Study, San Francisco City Cohort, ALIVE Study. Sicence, 273(5283): 1856-1862.
Ding S, Mehrabi R, Koten C, Kang Z, Wei Y, Seong K, Kistler H C, Xu J R. 2008. The transducin beta like gene FTL1 is essential for pathogenesis in Fusarium graminearum. Eucarytic Cell, 8(6): 867-876.
E. Keats Shwab, Jin Woo Bok, Martin Tribus, Johannes Galehr, Stefan Graessle, and Nancy P.Keller. 2007. Histone Deacetylase Activity Regulates Chemical Diversity in Aspergillus. Eukaryotic Cell, 1656-1664.
Gao L, Gross D S. 2008. Sir2 silences gene transcription by targeting the transition between RNA polymeraseII initiation and elongation. Mol Cell Biol, 28(12): 3979-3994.
Gerlar R. 1996. Gene-targeting studies of mammalian behavior: Is it the mutation or the background genotype? Trends in Neurosciences, 19(5): 177-181.
Goswami R S, Kistler H C. 2004. Heading for a disaster: Fusarium graminearum on cereal crops. Molecular Plant Pathology, 5: 515-525.
Goswami R S, Kistler H C. 2004. Heading for disaster: Fusarium graminearum on cereal crops. Molecular Plant Pathology, 5(6): 515-525.
Goswami R S, Xu J R, Trail F, Hilburn K, Kistler H C. 2006. Genomic analysis of host–pathogen interaction between Fusarium graminearum and wheat during early stages of disease development. Microbiology, 152: 1877-1890.
Goswami R S, Xu J R, Trail F, Hilburn K, Kistler H C. 2006. Genomic analysis of host–pathogen interaction between Fusarium graminearum and wheat during early stages of disease development. Microbiology, 152: 1877-1890.
GREENSHIELDS D L, LIU G S, FENG J, SELVARAJ G, WEI Y D. 2007. The siderophore biosynthetic gene SID1, but not the ferroxidase gene FET3, is required for full Fusarium graminearum virulence. Molecular Plant Pathology, 8(4): 411-421.
Güldener U, Seong K Y, Boddu J, Cho S, Trail F, Xu J R, Adam G, Mewes H W, Muehlbauer Gary J, Kistler H C. 2006. Development of a Fusarium graminearum Affymetrix GeneChip for profiling fungal gene expression in vitro and in planta. Fungal Genetics and Biology, 43:316-325.
Han Y K, Kim M D, Lee S H, Yun S H, Lee Y W. 2007. A novel F-box protein involved in sexual development and pathogenesis in Gibberella zeae. Mol Microbiol, 63(3): 768-779.
Hasty P, Ramirez-Solis R, KrumLauf R, Bradley A. 1991. Introduction of a subtle mutation into the Hox-2.6 locus in embryonic stem cells. Nature, 35(6315): 243-246.
HOU Z, XUE C, PENG Y, KATAN T, KISTLER H C, XU J. 2002. A map kinase gene(MGV1) in fusarium graminearum is required for female fertility, heterokaryon formation,hypal growth and plant infection. Molecular Plant-Microbe Interaction, 15: 1119-1127.
Hua-Van A, Pamphile J A, Langin T, Daboussi M J. 2001. Transposition of autonomous and engineered impala transposons in Fusarium oxysporum and a related species. Mol Gen Genet, 264: 724-731.
Insinga A, Monestiroli S, Ronzoni S, Gelmetti V, Marchesi F, Andrea Viale A, Altucci L, Nervi C, Saverio M, Pelicci P G. 2005. Inhibitors of histone deacetylases induce tumorselective apoptosis through activation of the death receptor pathway. Nat Med, 11: 71-76.
Jenczmionka N J, Maier F J, Losch A P, Schafer W. 2003. Mating, conidiation and pathogenicity of Fusarium graminearum, the main causal agent of the head-blight disease of wheat, are regulated by the MAP kinase gpmK1.Current genetics, 43(2):87-95
Joyner A L, Skarnes W C, Rossant J. 1989. Production of a mutation in mouse En-2 gene by homologous recombination in embryonic stem cells. Nature, 338: 153-156.
Kasschau K D, Carrington J C. 1998. A counterdefensive strate of plant viruses: uppression of posttranscriptional gene silencing. Cell, 95(4): 461-470.
Kilby N J, Snaith M R, Murray J A. Site-specific recombinases: tools for genome engineering. Trends Genet, 1993, 9(12): 413-421.
Koller B H, Hagemann L J, Doetschman T, Hagaman J R, Huang S, Williams P J, First N L, Maeda N, Smithies O. 1989. GermLine transmission of a planned alteration made in a hypoxanthine phosphoribosyltransferase gene by homologous recombination in embryonic stem cells. Proc Natl Acad Sci USA, 86(22): 8927-8931.
Kumar A, Seringhaus M, Biery M C, Sarnovsky R J, Umansky L, Piccirillo S, Heidtman M, Cheung K H, Dobry C J, Gerstein M B, Craig N L, Snyder M. 2004. Large-scale mutagenesis of the yeast genome using a Tn7-derived multipurpose transposon. Genome Res, 14: 1975-1986.
Lagac M, Chantha S C, Major G, Matton D P. 2003. Fertilization induces strong accumulation of a histone deacetylase (HD2) and of chromatin remodeling proteins in restricted areas of the ovules. Plant Mol Biol, 53(6): 759-769.
Lathe R. 1996. Mice, gene targeting and behaviour: more than just genetic background. Trends Neurosci, 19(5):183-186.
Li L, Xue C, Bruno K, Nishimura M, Xu J R. 2004. Two PAK kinase genes, CHM1 and MST20, have distinct functions in Magnaporthe grisea. Molecular Plant-Microbe Interactions, 17(5): 547-556.
Loprevite M, Tiseo M, Grossi F, Scolaro T, Semino C, Pandolfi A, Favoni R, Ardizzoni A. 2005. In vitro study of CI-994, a histone deacetylase inhibitor, in non-small cell lung cancer cell lines. Oncol Research, 15(1): 39-48.
Lu S W, Kroken S, Lee B N, Robbertse B, Alice C L, Churchill O, Yoder C, Turgeon B G. 2003. A novel class of gene controlling virulence in plant pathogenic ascomycete fungi. Proc Natl Acad Sci USA, 100(10): 5980–5985.
LU Y, FANG J Y. 2006. Epigenetic modification and cancer. Chinese Bulletin of Life Sciences, 18(1): 10-14.
Mesterhazy A. 1995. Types and components of resistance to Fusarium head blight. Plant Breed, 114: 377-386.
Oide S, Moeder W, Krasnoff S, Gibson D, Haas H, Yoshioka K, Turgeon B G. 2006. NPS6, Encoding a Nonribosomal Peptide Synthetase Involved in Siderophore-Mediated Iron Metabolism, Is a Conserved Virulence Determinant of Plant Pathogenic Ascomycetes. The Plant Cell, 18: 2836-2853.
Patm S K, Pawa A, Dahiya R. 2001. Histone deacetylase and DNA methyltransferase in human prostate cancer. Biochem Biophys Res Commun, 287(3):705-713.
Pfaffl M W. 2001 A new mathematical modle for relative quantification in real-time RT-PCR. Nucleic Acids Res, 29: e45.
Pugh GW, Jenann H, Dickson JG. 1963. Factors affecting infection of wheat heads by Gibberel la saubinetii. Journal of Agricultural Research, 44: 207-238.
Redner R L, Wang J, Liu J M. 1999. Chromatin remodeling and leukemia: new therapeutic paradigms. Blood, 94(2): 417-428.
Robyr D, Suka Y, Xenarios I, Kurdistani S K, Wang A, Suka N, Grunstein M. 2002. Microarray deacetylation maps determine genome-wide functions for yeast histone deacetylase. Cell, 109: 437-466.
Rosato R R, Almenara J A, Grant S. 2003. The histone deacetylase inhibitor MS-275 promotes differentiation or apoptosis in human leukemia cells through a process regulated by generation of reactive oxygen species and induction of p21cip1/waf1. Cancer Res, 63: 3637-3645.
Rundlett S E, Carmen A A, Kobayashi R, Bavykin S, Turner B M, Grunstein M. 1996.HDA1 and RPD3 are members of distinct yeast histone deacetylase complex that regulate silencing and transcription. Proc Natl Acad Sci USA, 93: 14503-14508.
Schiestl R H, Petes T D. 1991. Integration of DNA fragments by illegitimate recombination in Saccharomyces cerevisiae. Proc Natl Acad Sci USA, 88: 7585-7589.
Schwartzberg P L, Robertson E J, Goff S P. 1990. Targeted gene disruption of the endogenous c-abl locus by homologous recombination with DNA encoding a selectable fusion protein. Proc Natl Acad Sci USA, 87: 3210-3214.
Seo B W, Kim H K, Lee Y W, Yun S H. 2007. Functional analysis of a histidine auxotrophic mutation in Gibberella zeae. Plant Pathol, 23:51–56.
Seong K, Hou Z M, Tracy M, Kistler H C, Xu J R. 2005. Random insertional mutagenesis identifies genes associated with virulence in the wheat scab fungus Fusarium graminearum. Phytopathology, 95:744–750.
Seong K, Li L, Hou Z, Tracy Mile, Kistler H C, Xu J R. 2006. Cryptic promoter activity in the coding region of the HMG-CoA reductase gene in Fusarium graminearum. Fungal Genetics and Biology, 43: 34-41.
Shim W B, Sagaram U S,Choi Y E, So J, Wilkinsom H H, Lee YW. 2006. FSR1 is essential for virulenceand female fertility in Fusarium verticillioides and F. graminearum. Mol Plant Microbe Ineract, 19(7):725-33.
Smithies O, Gregg RG, Boggs SS, Koralewski M A, Raju S. Kucherlapati R S. 1985. Insertion of DNA sequencs into the human chromosomal betaglobin locus by homologous recombination. Nature, 317(6034): 230-234.
Sridha S, Wu K. 2006. Identification of AtHD2C as a novel regulator of abscisic acid response in Arabidopsis. The Plant Journal, 46(1): 124-133.
Thomas K R, Capecchi M R. 1987. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell, 51: 503-512.
Thompson S, Clarke A R, Pow A M, et al . 1989. GermLine transmission and stem cells. Cell, 56: 313-321. Timothy A Bolger. Year The Function and Regulation of Histone Deacetylase-4 in Neurons. [PhD Desertation]. : Department of Pharmacology and Cancer Biology Duke University. Valancius V. Smithies O. 1991. Double-strand gap repair in a mammalian gene targeting reaction. Mol Cell Biol, 11(9): 4389-4397.
Wang A, Kurdistani S K, Grunstein M. 2002. Requirement of Hos2 histone deacetylase for gene activity in yeast. Science. 298: 1412-1414
Wang YZ. 1997. Epidemiology and management of wheat scab in China. CIMMY conference on Fusarium head scab: global status and future prospects. CIMMYT, El Batan, 97-105.
Wu H, Liu X, Janeisch R. 1994. Double replacement: Strategy for efficient introduction of subtle mutations into the murine Col1a-1 gene by homologous recombination in embryonic stem cells. Proc Natl Acad Sci USA, 91: 2819-2823.
Yamamoto T, Omoto S, Mizuguchi M, Mizukami H, Okuyama H, Okada N, Saksena N K, Brisibe E A, Otake K, Fuji Y R. 2002. Double stranded nef RNA interferes with human immuno deficiency virus type 1 replication. Micobiol Immunol, 46: 809-817.
Yang B, Britton J, Kirchmaier A L.2008. Insights into the impact of histone aeetylation and methylation on Sir protein recruitment, spreading, and silencing in Saccharomyces cerevisiae. J Mol BIOl, 381(4): 826-844.
Yu H Y, Seo J, Klm J. 2008. Functional analyses of heterotrimeric G protein Gαand Gβsubunits in Gibberella zeae. Microbiology, 154(2): 392-401.