Identification of a new strain of Tricholoma giganteum by ITS and optimization of culture medium for mycelium
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摘要:目的
鉴定1株巨大口蘑Tricholoma giganteum新的野生菌株，并优化其菌丝的最适培养基.
方法以1株野生巨大口蘑为研究对象，采用组织分离法，从野生巨大口蘑子实体中分离纯化得到纯菌丝，利用分子生物学方法进行鉴定，通过测定菌丝生长速度、菌丝长势等指标对其进行培养条件研究，并运用均匀设计法对培养基进行优化，采用二次多项式逐步回归方法对试验结果进行分析.
结果和结论该野生菌株经鉴定为巨大口蘑，GeneBank编号为JX193694，命名为SCAU3，其菌丝生长的最适温度为30~32 ℃，最适pH为7~8，最佳碳源为麦芽糖，最佳氮源为酵母膏.经均匀设计法优化得到最适培养基组合:麦芽糖26 g·L-1，酵母膏2 g·L-1，MgSO4 2.7 g·L-1，KH2PO4 1.8 g·L-1，维生素B1 16 mg·L-1，pH7.7.
Abstract:ObjectiveA new wild strain of Tricholoma giganteum was identified and the culture medium of mycelium was optimized.
MethodThe pure mycelium of T. giganteum was obtained from the wild fruitbody by the tissue isolation and identified by molecular biological techniques. The culture condition of the mycelium was researched by measuring mycelial growth rate and growth vigor. The optimum growth medium of mycelium was determined by the uniform-design-method. The method of quadratic polynomial step regression analysis was employed for analyzing the experiment results.
Result and conclusionThe wild strain was identified as T. giganteum, named as SCAU3. The accession number of ITS sequence was JX193694 in GeneBank. The results show that the optimum carbon source and nitrogen source for mycelium growth are maltose and yeast extract respectively. The optimum temperature is 30-32 ℃, and the optimum pH in medium was 7-8. The optimum medium for mycelial growth is maltose 26 g·L-1, yeast extract 2 g·L-1, MgSO4 2.7 g·L-1, KH2PO4 1.8 g·L-1, vitamin B1 16 mg·L-1, pH7.7.
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Keywords:
- Tricholoma giganteum /
- ITS identification /
- optimum culture medium /
- uniform design /
- temperature /
- pH
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图 2 菌株SCAU3的ITS-PCR扩增产物电泳图
Figure 2. Electrophoresis of PCR products of ITS from SCAU3
M:DNA maker DL2000;1、2: SCAU3.
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图 3 新菌株SCAU3与其他巨大口蘑的进化树
Figure 3. The phylogenetic tree of strain SCAU3 and Tricholoma giganteum related species
表 1 培养温度对野生巨大口蘑菌丝生长的影响
Table 1 Effect of temperature on the mycelial growth of Tricholoma giganteum
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表 2 培养基pH对野生巨大口蘑菌丝生长的影响
Table 2 Effect of pH on the mycelial growth of Tricholoma giganteum
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表 3 培养基碳源对野生巨大口蘑菌丝生长的影响
Table 3 Effects of carbon sources on the mycelial growth of Tricholoma giganteum
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表 4 培养基氮源对野生巨大口蘑菌丝生长的影响
Table 4 Effects of nitrogen sources on the mycelial growth of Tricholoma giganteum
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表 6 各回归项的回归系数检验
Table 6 The test results of the regression coefficient of the regression items
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表 7 样本配方的观察值、拟合值和拟合误差
Table 7 The observed value, fitted value and fitted error of the samples
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表 8 不同培养基对野生巨大口蘑菌丝生长速度的影响
Table 8 Effects of the growth of Tricholoma giganteum in different culture media
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[1] 黄年来.适合热带地区栽培的珍稀菇:巨大口蘑[J].食用菌, 2001, 23(5): 12-13. http://d.old.wanfangdata.com.cn/Periodical/syj200105008 [2] 吴建成.水平棚架梨园栽培巨大口蘑技术[J].中国食用菌, 2007, 26(4): 60-60. doi: 10.3969/j.issn.1003-8310.2007.04.020 [3] 上官建舟, 林汝楷, 吴宝芳.巨大口蘑生物学特性的研究[J].浙江食用菌, 2007, 15(1): 16-21. http://d.old.wanfangdata.com.cn/Periodical/zgsyj200803016 [4] 卢成英, 钟以举, 张敏.吉首大白口蘑(Tricholoma giganteum)研究[J].微生物学通报, 2001, 28(4): 20-23. doi: 10.3969/j.issn.0253-2654.2001.04.006 [5] 李志生.巨大口蘑及其栽培技术[J].食用菌, 2007 (5): 59-61. doi: 10.3969/j.issn.1000-8357.2007.05.049 [6] 郭翠英, 沈育芬.大白口蘑菌株厦3的分离驯化研究[J].亚热带植物科学, 2002, 31(1): 14-16. doi: 10.3969/j.issn.1009-7791.2002.01.004 [7] 刘月廉, 谭树明, 温美英, 等.野生洛巴伊口蘑菌株的分离与鉴定[J].食用菌学报, 2001, 8(2): 19-23. doi: 10.3969/j.issn.1005-9873.2001.02.004 [8] 郑贵朝, 张善信, 胡事君.高温型洛巴伊口蘑的菌种选育及栽培技术[J].食用菌, 2005, 27(4): 16-18. doi: 10.3969/j.issn.1000-8357.2005.04.011 [9] 上官建舟, 林汝楷, 吴宝芳. 荆西口蘑的驯化研究[C] //中国菌物学会, 中国食用菌协会. 首届全国食用菌中青年专家学术交流会论文集. 武汉: 中国菌物学会, 2006: 40-46. [10] 张文隽, 吴亚召, 雷萍, 等.巨大口蘑Tsw-06栽培试验初报[J].食用菌, 2011, 33(6): 21-22. http://d.old.wanfangdata.com.cn/Periodical/syj201106010 [11] 陈丽新, 韦仕岩, 黄卓忠.温度、酸碱度和培养基含水量对金福菇Tg-505菌丝生长的影响[J].西南农业学报, 2012, 25(2): 597-600. doi: 10.3969/j.issn.1001-4829.2012.02.049 [12] 张国广, 邹金美, 郑琳.一株野生大型真菌的ITS分析及生物学特征研究[J].云南民族大学学报, 2010, 19 (6): 395-399. http://d.old.wanfangdata.com.cn/Periodical/ynmzxyxb201006002 [13] 王元忠, 汤洪敏, 虞泓, 等.巨大口蘑子实体营养成分分析[J].食用菌学报, 2005, 12(2): 24-26. doi: 10.3969/j.issn.1005-9873.2005.02.005 [14] 李涛, 王元忠, 刘鸿高.巨大口蘑中微量元素的光谱测定[J].光谱学与光谱分析, 2008, 28(2): 450-452. http://d.old.wanfangdata.com.cn/Periodical/gpxygpfx200802051 [15] MAU J L, LIN H C, SONG S F. Antioxidant properties of several specialty mushrooms[J]. Food Res Int, 2002, 35 (6): 519-526. doi: 10.1016/S0963-9969(01)00150-8
[16] MIZUNO T, KINOSHITA T, ZHUANG C, et al. Antitumor-active heteroglycans from niohshimeji mushroom, Tricholoma giganteum[J]. Biosci Biotechnol Biochem, 1995, 59(4): 568-571. doi: 10.1271/bbb.59.568
[17] LIU F, OOI V E C, LIU W K, et al. Immunomodulation and antitumor activity of polysaccharide-protein complex from the culture filtrates of a local edible mushroom, Tricholoma lobayense[J]. Gener Pharmacol: Vasc Syst, 1996, 27(4): 621-624. doi: 10.1016/0306-3623(95)02058-6
[18] HYOUNG L D, HO K J, SIK P J, et al. Isolation and characterization of a novel angiotensin I-converting enzyme inhibitory peptide derived from the edible mushroom Tricholoma giganteum[J]. Peptides, 2004, 25 (4): 621627. doi: 10.1016/j.peptides.2004.01.015
[19] LI M, ZHANG G, WANG H, et al. Purification and characterization of a laccase from the edible wild mushroom Tricholoma mongolicum[J]. J Microbiol Biotechn, 2010, 20(7): 1069-1076. doi: 10.4014/jmb
[20] 张倩勉, 莫美华.巨大口蘑挥发油化学成分及抑菌作用的研究[J].现代食品科技, 2008, 24(12): 1232-1235. http://d.old.wanfangdata.com.cn/Periodical/gzspgykj200812012 [21] 莫美华, 张倩勉.巨大口蘑子实体抽提物抑菌活性研究[J].食品工业科技, 2009, 30(5): 151-153. http://d.old.wanfangdata.com.cn/Periodical/spgykj200905046 [22] WANG H X, NG T B. Purification of a novel low-molecular-mass laccase with HIV-1 reverse transcriptase inhibitory activity from the mushroom Tricholoma giganteum[J]. Biochem Bioph Res Co, 2004, 315(2): 450-454. doi: 10.1016/j.bbrc.2004.01.064
[23] 刘鸿高, 王元忠, 张东艳, 等.大白口蘑菌丝的培养优化[J].菌物学报, 2011, 30(1): 142-146. http://d.old.wanfangdata.com.cn/Periodical/jwxt201101022 [24] 刘月廉, 陈爱珠, 谭树明, 等.野生洛巴伊口蘑菌株生物学特性的研究(英文)[J].华中农业大学学报, 2004, 23(1): 36-39. doi: 10.3321/j.issn:1000-2421.2004.01.009
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