宁夏黄花棘豆中产苦马豆素内生真菌的分离及鉴定

Abstract

Abstract: To provide strains and a novel knowledge for the further study of the molecular regulation mechanism of endophytic fungi produced swainsonine in locoweed, this research was performed to isolate and identify swainsonine-producing fungal endophytes from Oxytropis ochrocephala Bunge in Ningxia. The fungal endophytes from the plant tissues were isolated and purified. Each strain was analyzed with gas chromatography-mass spectrum to determine the presence of swainsonine in fungal mycelia.The swainsonine-producing strain was induced to produce conidia or conidiospores in the culture condition of ultraviolet light and darkness alternation. The fungal DNA were extracted from mycelia of the swainsonine-producing fungal endophyte. The GPD, ITS and mt SSU of the strain were amplified and sequenced for the analysis of homology and genetic evolution. The taxon of the strain was determined based on the morphology and the result of genetic evolution analysis. The results indicated that five isolates were harvested from Oxytropis ochrocephala Bunge in Ningxia. Only one strain was determined to synthesize swainsonine. The strain could produce abundant conidia or conidiospores after induced culture. The morphology and genetic evolution relationship of the strain were closely related to the Undifilum. Therefore, the strain was identified as Undifilum oxytropis in this study.

Key words: Oxytropis ochrocephala Bunge; fungal endophyte; swainsonine; Undifilum oxytropis

CLC Number:

S816.79

ZHANG Lei-lei, HE Sheng-hu, YU Yong-tao, ZHAO Qing-mei. Isolation and Identification of Swainsonine-producing Fungal Endophyte from Oxytropis ochrocephala Bunge in Ningxia[J]. , 2013, 40(12): 165-172.

References

1 王凯,曹光荣,段得贤,等.黄花棘豆对山羊的毒性研究[J].畜牧兽医学报,1990,21(1):80~86.
2 王凯.青海省三种常见棘豆主要营养成分研究[J].青海大学学报,1998,16(2):5~7.
3 卢萍.内蒙古三种棘豆属植物中苦马豆素相关因子的研究[D].呼和浩特:内蒙古农业大学,2007.
4 史志诚.中国草地重要有毒植物[M].北京:中国农业出版社,1997.
5 余永涛,王建华,王妍,等.西藏3种疯草中合成苦马豆素内生真菌的鉴定[J].中国农业科学,2009a,42(10):3662~3671.
6 余永涛,王建华,赵清梅,等.甘肃棘豆中产苦马豆素内生真菌的分离与鉴定[J].西北农林科技大学学报(自然科学版),2009b,37(2):40~46.
7 余永涛.产苦马豆素疯草内生真菌的分离鉴定及其遗传多态性研究[D].杨凌:西北农林科技大学,2009.
8 李建科,杨具田,潘和平,等.家畜黄花棘豆、甘肃棘豆中毒调查研究[J].中国兽医科技,1987(5):22~23.
9 陶定章,李仲珊.宁夏南华山马场家畜黄花棘豆中毒的调查[J].中国兽医科技,1986(4):5~6.
10 曹光荣,李绍君,段得贤,等.黄花棘豆有毒成分的分离与鉴定[J].西北农业大学学报,1989,17(3):1~7.
11 曹光荣,张守信,陈义忠.宁夏黄花棘豆中毒调查报告[J].畜牧兽医杂志,1998(3):14~19.
12 Braun K,Cook T,Liddell C M. Locoism of cattle caused by a species of Alternaria[J]. Phytopathology,1997,87:S11~S12.
13 Braun K. Fungal endophyte infection and swainsonine toxicity in locoweed[D]. LasCruces,NM: New Mexico State University,1999.
14 Braun K,Romero J,Liddell C,et al. Production of swainsonine by fungal endophytes of locoweed[J]. Mycological Research,2003,107(8): 980~988.
15 Colegate S M,Dorling P R. Plant associated toxins. Agricultural,phytochemical and ecological aspects[M]. Wallingford: CAB International,1994.
16 Cook D,Gardner D R,Ralphs M H,et al. Swainsonine concentrations and endophyte amounts of Undifilum oxytropis in different plant parts of Oxytropis sericea[J]. Journal of Chemical Ecology,2009,35: 1272~1278.
17 Cook D,Gardner D R,Grum D,et al. Swainsonine and endophyte relationships in Astragalus mollissimus and Astragalus lentiginosus[J]. J Agric Food Chem,2011,59: 1281~1287.
18 Dorling P R,Huxtable C R,Colegate S M.Inhibition of lysosomal a-mannosidase by swainsonine,an indolizidine alkaloid isolated from Swainsona canecens[J].Biochem J,1980,191: 649~651.
19 Gardner D R,Molyneux R J,Ralphs M H. Analysis of swainsonine: Extraction methods,detection,and measurement in populations of locoweeds(Oxytropis spp.)[J]. J Agric Food Chem,2001,49: 4573~4580.
20 Kadivalt G V,Chapapas S D. Production,characterization and species secificity of five monoclonal antibodies to Mycobacterium tuberculosis[J]. J Clin Mucribiol,1987,25(1): 76~80.
21 Li K N,Rouse D,German T L. PCR primers that allow intergeneric differentiation of ascomycetes and their application to Verticillium spp.[J]. Appl Environ Microbiol,1994,60(12): 4324~4331.
22 Oldrup E,McLain-Romero J,Padilla A,et al. Localization of endophytic Undifilum fungi in locoweed seed and influence of environmental parameters on a locoweed in vitro culture system[J]. Botany,2009,88: 512~521.
23 Pryor B,Creamer R,Shoemaker R,et al. Undifilum,a new genus for endophytic Embellisia oxytropis and parasitic Helminthosporium bornmuelleri on legumes[J]. Botany,2009,87: 178~194.
24 Ralphs M H,Creamer R,Baucom D,et al. Relationship between the endophyte Embellisia spp. and the toxic alkaloid swainsonine in major locoweed species (Astragalus and Oxytropis)[J]. Journal of Chemical Ecology,2008,34: 32~38.
25 Ralphs M H,Cook D,Gardner D R,et al. Transmission of the locoweed endophyte to the next generation of plants[J]. Fungal Ecology,2011,4: 251~255.
26 Tulsiani D R P,Harris T M,Touster O. Swainsonine inhibits the biosynthesis of complex glycoproteins by inhibition of Golgi mannosidase Ⅱ[J]. Journal of Biological Chemistry,1982,257(14): 7936~7939.
27 Wang Q,Na Q,Na G H,et al. Embellisia oxytropis,a new species isolated from Oxytropis kansuensis in China[J]. Mycotaxon,2006,95: 255~260.
28 White T J,Bruns T,Lee S,et al. PCR protocols: A guide to methods and applications[M]. New York: Academic Press,1990.
29 Yu Y T,Zhao Q M,Wang J N,et al. Swainsonine-producing fungal endophytes from major locoweed species in China[J]. Toxicon,2010,56: 330~338.

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Isolation and Identification of Swainsonine-producing Fungal Endophyte from Oxytropis ochrocephala Bunge in Ningxia

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