|本期目录/Table of Contents|

[1]郭成,李金花,柴兆祥.天祝高寒草地三线镰孢的分离鉴定及β-tubulin基因序列分析[J].应用与环境生物学报,2011,17(01):51-56.[doi:10.3724/SP.J.1145.2011.00051]
 GUO Cheng,LI Jinhua & CHAI Zhaoxiang.Isolation, Identification and Analysis of β-tubulin Gene Sequence of Fusarium tricinctum at Different Altitudes in Tianzhu Alpine Grassland[J].Chinese Journal of Applied & Environmental Biology,2011,17(01):51-56.[doi:10.3724/SP.J.1145.2011.00051]
点击复制

天祝高寒草地三线镰孢的分离鉴定及β-tubulin基因序列分析()
分享到:

《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
17卷
期数:
2011年01期
页码:
51-56
栏目:
研究论文
出版日期:
2011-02-25

文章信息/Info

Title:
Isolation, Identification and Analysis of β-tubulin Gene Sequence of Fusarium tricinctum at Different Altitudes in Tianzhu Alpine Grassland
作者:
郭成李金花柴兆祥
(1甘肃农业大学草业学院 兰州 730070)
(2甘肃省作物遗传改良与种质创新重点实验室 兰州 730070)
(3草业生态系统教育部重点实验室 兰州 730070)
(4中-美草地畜牧业可持续发展研究中心 兰州 730070)
Author(s):
GUO Cheng LI Jinhua & CHAI Zhaoxiang
(1College of Grassland Sciences, Gansu Agricultural University, Lanzhou 730070, China)
(2Gansu Key Laboratory of Crop Improvement and Germplasm Enhancement, Lanzhou 730070, China)
(3Key Laboratory of Grassland Ecosystem, Ministry of Education, Lanzhou 730070, China)
(4Sino-U. S. Centers for Grazingland Ecosystem Sustainability, Lanzhou 730070, China)
关键词:
天祝高寒草地三线镰孢鉴定β-tubulin基因
Keywords:
Tianzhu Alpine Grassland Fusarium tricinctum identification β-tubulin gene
分类号:
S812.2 : S154.36
DOI:
10.3724/SP.J.1145.2011.00051
文献标志码:
A
摘要:
按30 m的梯度将海拔2 880~3 360 m分为16个采样点,在同一海拔高度按照“Z”字形取样,共取5个点,对16个采样点的土样混匀后以根残体分离法在镰刀菌选择性培养基上进行分离,对分离得到的镰刀菌菌落进行纯化和单孢分离后,以形态学为基础,参照Nelson分类系统进行鉴定. 结果表明:在分离到的212个镰刀菌菌株中,有5个菌株为三线镰孢(Fusarium tricinctum). 随机选取2株三线镰孢菌株进行β-tubulin基因序列分析,将PCR产物回收测序后在Genbank 上比对,菌株TZh-4-3-7和TZh-11-5-2与GenBank上登记的6个新西兰菌株的亲缘关系最近,同源性达96%. 利用DNAStar软件绘制其系统发育树状图,结果显示,菌株TZh-4-3-7和TZh-11-5-2与以上6个F. tricinctum菌株均位于系统发育树的同一分支,聚为一类. 本研究采用的β-tubulin基因序列分析结果验证了利用形态学鉴定镰刀菌的准确性;将β-tubulin基因序列分析应用于三线镰孢的鉴定中,拓宽了鉴定途径,提高了鉴定的准确性;并了解了三线镰孢在天祝高寒草原不同海拔高度的分布,丰富了我国草原镰刀菌资源库,为进一步探讨镰刀菌与草原退化的关系奠定了基础. 图4 参45
Abstract:
In order to know the distribution of Fusarium tricinctum in Tianzhu Alpine Grassland in Gansu, China, rhizosphere soils around gramineous grass were sampled through a zigzag way from 16 sampling sites at 30 m interval at a elevation ranging between 2 880~3 360 m, 5 sub-samples in total were picked up from each sampling site, and root debris from the soils was plated on Fusarium specific medium. The Fusarium colonies were purified and single-spored. Based on the morphological characteristics of Fusarium isolates, and according to the references related to Nelson’s Fusarium taxonomical system, 212 Fusarium isolates were obtained and 5 of them were identified belonging to F. tricinctum. Two F. tricinctum isolates of the 5 were randomly selected for sequences analysis of β-tubulin gene. The PCR product of the two isolates were collected, purified, and sequenced. The sequences were aligned in GenBank. It was showed that there was a very close relationship of isolates TZh-4-3-7 and TZh-11-5-2 with the 6 F. tricinctum isolates in New Zealand, and their max similarity was 96%. Microsoft DNAStar was used to draw the phylogenetic tree of isolates TZh-4-3-7 and TZh-11-5-2. The tree showed that both TZh-4-3-7 and TZh-11-5-2 were in the same cluster as the 6 New Zealand isolates. The sequence analysis of β-tubulin gene determined the morphological identification of the 5 F. tricinctum isolates in this study, and the analysis was used to identify F. tricinctum for the first time in China. This study would enrich the future research on grassland degeneration associated with Fusarium. Fig 4, Ref 45

参考文献/References:

1 Xu ZX (许志信), Zhao ML (赵萌莉), Han GD (韩国栋). Eco-environmental deterioration and strategies for preventing it in Inner Mongolia. Grassl China (中国草地), 2000, 5: 59~63
2 Yao T (姚拓), Long RJ (龙瑞军). Dynamics of soil microbial population under disturbance in Tianzhu Alpine Grassland. Acta Pratacult Sin (草业学报), 2006, 15 (2): 93~99
3 Yin GT (殷桂涛). The main cause and control strategy of grassland degradation in Ta city. Acta Pratacult Sin (草业学报), 2004, 21 (7): 10~12
4 Xiao HL (肖辉林), Zheng XJ (郑习健). Effects of plant diversity on soil microbes. Soil & Environ Sci (土壤与环境), 2001, 10 (3): 238~241
5 Zhou ZB (周智彬), Li PJ (李培军). Ecological distribution of soil microorganism in artificial greenbelt in hinterland of Takilimakan Desert and their relations with soil factors. Chin J Appl Ecol (应用生态学报), 2003, 14 (8): 1246~1250
6 Xia BC (夏北成), Zhou JZ, James MT. Effect of vegetation on structure of soil microbial community. Chin J Appl Ecol (应用生态学报), 1998, 9 (3): 196~300
7 Shang ZH (尚占环), Ding LL (丁玲玲), Long RJ (龙瑞军), Ma YS (马玉寿). Relationship between soil microorganisms above-ground vegetation, and soil environment of degraded alpine meadows in the headwater areas of the Yangtze and Yellow Rivers, Qinghai-Tibetan Plateau. Acta Pratacult Sin (草业学报), 2007, 16 (1): 34~40
8 Wardle DA, Nicholson KS. Synergistic effects of grassland plant species on soil microbial biomass and activity: Implications for ecosystem-level effects of enriched plant diversity. Function Ecol, 1996, 10: 410~416
9 Liu SG (刘世贵), Ge SR (葛绍荣), Long ZF (龙章富). Studies on soil microorganism numbers and microbiota of degenerated rangelands in northwest region of Sichuan, P. R. China. Acta Pratacult Sin (草业学报), 1994, 3 (4): 70~76
10 Ma LP (马丽萍), Zhang DG(张德罡), Yao T (姚拓). Study on the dynamics of soil cellulose decomposer in alpine grasslandunder disturbance in Tianzhu. Grassl & Turf (草原与草坪), 2005, 1: 29~33
11 Zhao J (赵吉), Liao YN (廖仰南), Zhang GZ (张桂枝), Shao YQ (邵玉琴). Soil microbial ecology on the grassland ecosystem. Grassl China (中国草地), 1999, 21 (3): 57~67
12 Shao YQ (邵玉琴), Zhao J (赵吉), Yang J (杨劼). Distribution characteristics of soil microbial numbers in recovered grassland and degenerated grassland. J Desert Res (中国沙漠), 2004, 24 (2): 223~226
13 Guo JX (郭继勋), Zhu TC (祝廷成). Study on numbers and biomass of soil microorganism in an Eurolepidium Chinese grassland. Acta Ecol Sin (生态学报), 1997, 17 (1): 78~82
14 Ma S (马爽), Yang CD (杨成德), Xue L (薛莉), Chen XR (陈秀蓉). Seasonal dynamic of soil microorganism in typical grassland under different carrying capacities in Longdong of Gansu. Grassl & Turf (草原与草坪), 2004, 2: 27~30
15 Gao T (高婷), Zhang YP (张源沛). Study on the relationship between edaphon amount and distributing type of vegetation and soil in desert grassland. Pratacult Sci (草业科学), 2006, 23 (12): 22~25
16 Cai Y (蔡艳), Xue QH (薛泉宏), Chen ZQ (陈占全), Chang XB (常显波), Sun XF (孙小凤), Si MR (司美茹), Lai HX (来航线), Zhang R (张荣). Ecological distribution of soil actinomycetes in eastern part of the Qinghai-Tibet plateau. Chin J Appl Environ Biol (应用与环境生物学报), 2004, 10 (3): 378~383
17 Luo M (罗明), Qiu W (邱沃). The ecological distribution of soil microorganisms in desert saline grassland in Xinjiang. Grassl China (中国草地), 1995, 17 (5): 29~33
18 Long ZF (龙章富), Liu SG (刘世贵). Soil agrochemical properties and microbiota of deraded grasslands. Acta Pedol Sin (土壤学报), 1996, 33 (2): 192~200
19 Wang GR (王国荣), Chen XR (陈秀蓉), Han YZ (韩玉竹), Yang CD (杨成德), Xu CL (徐长林). Distributional characteristics of soil microorganism of alpine shrublands in Eastern Qilian Mountains. Grassl & Turf (草原与草坪), 2006, 3: 27~30
20 Zhang CB (张崇邦), Yang JC (杨靖春). An application of the fuzzy priority ratio in the study of variation of ecological environments and microbiological ecological relation on Leymus Chinensis grassland in the Northeast of China. Grassl China (中国草地), 1994, 4: 12~16
21 Zhao J (赵吉), Liu P (刘萍), Shao YQ (邵玉琴), Li ZW (李志伟), Liao YN (廖仰南). Effect of artificial factors on microorganism and biological activity in soil of steppe. Acta Sci Nat Univ NeiMonggol (内蒙古大学学报自然科学版), 1996, 27 (4): 56~57
22 Yao T (姚拓), Yang JX (杨俊秀), Jing Y (景耀), Wu CH (吴重华). Population and ecological characteristics of soil fungi in larch plantation on Qinling Mountains. Chin J Ecol (生态学杂志), 1998, 17 (4): 7~13
23 Baldauf SL, Palmer JD. Animals and fungi are each other’s closest relatives: congruent evidence from multiple proteins. Proc Natl Acad Sci USA, 1993, 90: 11538~11562
24 Baldauf SL, Doolittle WF. Origin and evolution of the slime molds (Mycetozoa). Proc Natl Acad Sci, USA, 1997, 94 (22): 12007~12012
25 O’Donnell KE, Cigelnik, and Nirenberg HI. Molecular systematics and phylogeography of the Gibberella fujikumi species complex. Mycologia, 1998, 90: 465~493
26 Burgess LW, Summerell BA, Bullock A, Gott KP, Backhouse D. Laboratory Mannual for Fusarium Research. 3rd ed. Sydney, Australia: The University of Sydney, 1994
27 Tio M, Burgess LW, Nelson PE. Techniques for the isolation, culture and preservation of the Fusaria. Austr Plant Pathol Soc Newslett, 1977, 6: 11~13
28 Paul B. Pythium ornacarpum: A new species with ornamented oogonia isolated from soil in France. FEMS Microb Lett, 1999, 180: 337~344
29 Paul B. A new species of Pythium with filamentous sporangia having pectinolytic activities, isolated in the Burgundy region of France. FEMS Microbiol Lett, 2001, 199: 55~59
30 Smith WJ, Brett AS, Gunn L. Molecular Techniques for the Study of Soilborn Fungal Plant Panthology. Sydney, Australia: Sydney University Press, 2003
31 Andersen BU, Thrane A, Svendsen A, Rasmussen IA. Associated field mycobiota on malt barley. Can J Bot, 1996, 74: 854~858
32 Chelkowski J, Manka M, Kwasna H, Visconti A, Golinski P. Fusarium sporotrichioides (Sherb.), Fusarium tricinctum (Corda) Sacc. and Fusarium poae Wollenw: Cultural characteristics, toxigenicity and pathogenicity towards cereals. J Phytopathol, 1989, 124: 155~161
33 Golinski P, Perkowski J, Kostecki M, Grabarkiewicz-Szczesna J, Chelkowski J. Fusarium species and Fusarium toxins in wheat in Poland: A comparison with neighbour countries. Sydowia, 1996, 48: 12~22
34 Kosiak B, Torp M, Skjerve E, Thrane U. The prevalence and distribution of Fusarium species in Norwegian cereals: A survey. Acta Agric Scandinavica Section B: Soil & Plant Sci, 2003, 53: 168~176
35 Marasas WFO, Nelson PE, Toussoun TA. Toxigenic Fusarium Species: Identity and Mycotoxicology. University Park, Pennsylvania, USA: The Pennsylvania State University Press, 1984
36 Thrane U. Screening for fusarin C production by European isolates of Fusarium species. Mycotoxin Res, 1988, 4: 2~10
37 Burmeister HR, Plattner RD. Enniatin production by Fusarium tricinctum and its effect on germinating wheat seeds. Phytopathology, 1987, 77: 1483~1487
38 Chelkowski J, Zawadzki M., Zajkowski P, Logrieco A, Bottalico A. Moniliformin production by Fusarium species. Mycotoxin Res, 1990, 6: 41~45
39 Visconti A, Solfrizzo M, Fruchier A, Ap-Simon JW. Acuminatopyrone: Revised structure and production by Fusarium chlamydosporum and Fusarium tricinctum. J Natl Prod, 1994, 57: 695~699
40 Takeuchi M, Nakajima M, Ogita T, Inukai M, Kodama K, Furuya K, Nagaki H, Haneishi T. Fosfonochlorin: A new antibiotic with spheroplast forming activity. J Antibiot, 1989, 42: 198~205
41 Visconti A, Solfrizzo M. Isolation, characterization, and biological activity of visoltricin, a novel metabolite of Fusarium tricinctum. J Agric & Food Chem, 1994, 42: 195~199
42 Visconti A, Solfrizzo M. Visoltricin, a novel biologically active compound produced by Fusarium tricinctum. Food Addit & Contam, 1995, 12: 515~519.
43 Richard C, Martin JG, Pouleur S. Ice nucleation activity identified in some phytopathogenic Fusarium species. Phytoprotection, 1996, 77: 83~92
44 Scheetz RW, Whelan HA, Wriston JC. Purification and properties of L-asparaginase (e.c. 3.5.1.1) from Fusarium tricinctum. Arch Biochem & Biophys, 1971, 142: 184~189
45 Wick RL, Schroeder DB. In vitro production of pectolytic and cellulolytic enzymes by Fusarium tricinctum. Mycologia, 1982, 74: 460~466

备注/Memo

备注/Memo:
甘肃省科学事业费项目(甘科计[2001]21号)、甘肃省教育厅项目(No. 032-08)和甘肃农业大学科技创新基金(No. GAU-CX0504)资助
更新日期/Last Update: 2011-02-28