三株野生重伞灵芝的鉴定及其多糖、三萜含量分析
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摘要
采用形态鉴定、ITS鉴定和ITS-RFLP分子标记方法对采集自福建的3株野生灵芝进行鉴定,并将其生物学特性及化学成分与赤芝(Ganoderma lingzhi)菌株ZK进行比较。结果表明:采集的3株野生灵芝为重伞灵芝(G.multipileum)。与赤芝菌株ZK相比,3株野生重伞灵芝菌株LQ、LJ和Y1菌丝生长速度快(P<0.01),LQ、LJ、Y1最适生长温度分别为32、32、34℃,均高于赤芝ZK;供试菌株均偏好酸性生长环境;重伞灵芝LQ和Y1的产量分别为每袋(12.1±1.3)、(11.7±0.9)g,达到ZK产量(17.6±1.8)g的66%以上。Y1与LQ的子实体多糖含量分别为(12.9±0.1)、(10.7±0.3)mg/g,高于赤芝ZK的子实体多糖含量(9.2±0.2)mg/g(P<0.01);LJ、Y1的三萜含量约为7.5mg/g,LQ的三萜含量为(6.5±0.3)mg/g,分别是ZK三萜含量的84.27%和73.03%。从HPLC图谱可知重伞灵芝有许多赤芝没有的峰,三萜酸的种类比较丰富。
Three wild Ganodermastrains were collected from Fujian province in China and were identified by morphological and molecular characterization(ITS and ITS-RFLP).Then their biological characteristics and effective components were compared with the reference Ganoderma lingzhi strain ZK.The results showed that the three wild Ganodermastrains(LQ,LJ,Y1)were all identified to be Ganoderma multipileum.The wild G.multipileum strains showed significantly faster mycelial growth than strain ZK(P<0.01).The optimum temperature for maximum growth of strain LQ,LJ and Y1 were 32,32 and 34 ℃,respectively.These optimal temperatures were higher than the optimum temperature of strain ZK.All three wild G.multipileum strains preferred acidic growth condition.Fruiting body yield of G.multipileum LQ and Y1 were(12.1±1.3)and(11.7±0.9)g per bag,reaching more than 66% of the yield of ZK [(17.6±1.8)g per bag].Polysaccharides content in fruiting body of strain Y1 and LQ were(12.9±0.1)and(10.7±0.3)mg/g respectively,which were significantly higher than that of strain ZK(9.2±0.2)mg/g(P <0.01).Triterpenoids content in fruiting body of strain LJ,Y1 and LQ were 7.5,7.5 and(6.5±0.3)mg/g,respectively,which were 84.27%,84.27% and 73.03% of that in strain ZK,respectively.HPLC analysis showed that G.multipileum strains contained some chemicals that are not otherwise found in G.lingzhi,suggesting their rich triterpenoids variety.
Three wild Ganodermastrains were collected from Fujian province in China and were identified by morphological and molecular characterization(ITS and ITS-RFLP).Then their biological characteristics and effective components were compared with the reference Ganoderma lingzhi strain ZK.The results showed that the three wild Ganodermastrains(LQ,LJ,Y1)were all identified to be Ganoderma multipileum.The wild G.multipileum strains showed significantly faster mycelial growth than strain ZK(P<0.01).The optimum temperature for maximum growth of strain LQ,LJ and Y1 were 32,32 and 34 ℃,respectively.These optimal temperatures were higher than the optimum temperature of strain ZK.All three wild G.multipileum strains preferred acidic growth condition.Fruiting body yield of G.multipileum LQ and Y1 were(12.1±1.3)and(11.7±0.9)g per bag,reaching more than 66% of the yield of ZK [(17.6±1.8)g per bag].Polysaccharides content in fruiting body of strain Y1 and LQ were(12.9±0.1)and(10.7±0.3)mg/g respectively,which were significantly higher than that of strain ZK(9.2±0.2)mg/g(P <0.01).Triterpenoids content in fruiting body of strain LJ,Y1 and LQ were 7.5,7.5 and(6.5±0.3)mg/g,respectively,which were 84.27%,84.27% and 73.03% of that in strain ZK,respectively.HPLC analysis showed that G.multipileum strains contained some chemicals that are not otherwise found in G.lingzhi,suggesting their rich triterpenoids variety.
引文
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[21]叶丽云,林强,刘梅,等.钙离子和水杨酸诱导灵芝多糖和三萜的合成[J].菌物学报,2017,36(2):220-228.
[22]ZHENG LY,JIA DH,FEI XF,et al.An assessment of the genetic diversity within Ganoderma strains with AFLP and ITS PCR-RFLP[J].Microbiological Research,2009,164(3):312-321.
[2]XU P,DING ZY,QIAN Z,et al.Improved production of mycelial biomass and ganoderic acid by submerged culture of Ganoderma lucidum SB97using complex media[J].Enzyme and Microbiol Technology,2008,42(4):325-331.
[3]ZHOU XW,SU KQ,ZHANG YM.Applied modern biotechnology for cultivation of Ganoderma and development of their products[J].Applied Microbiology and Biotechnology,2012,93(3):941-963.
[4]李平作,章克昌.灵芝胞外多糖的分离纯化及生物活性[J].微生物学报,2000,40(2):217-220.
[5]李晔,朱忠敏,姚渭溪,等.灵芝三萜类化合物的研究进展[J].中国中药学杂志,2012,37(2):165-171.
[6]李国华,李晔,梅锡玲,等.灵芝三萜类化合物研究进展[J].中草药,2015,46(12):1858-1862.
[7]吴声华.中国栽培灵芝种“赤芝”的拉丁学名探究[J].食药用菌,2014,22(4):185-187.
[8]文庭池,邓春英,吴兴亮.海南岛7种野生灵芝的形态与分子鉴定[J].贵州科学,2015(6):1-8.
[9]RNG M,HOLDERNESS M,BRIDGE PD,et al.Genetic diversity of Ganodermain oil palm plantings[J].Plant Pathology,1999,48(5):595-603.
[10]邹莉,孙婷婷,王旭彤,等.松杉灵芝分离纯化及ITS分子鉴定[J].江苏农业科学,2016,44(8):278-280.
[11]傅俊生.草菇杂交育种研究及其分子遗传标记的建立[D].福州:福建农林大学,2007.
[12]胡惠萍,刘远超,莫伟鹏,等.两株西藏白肉灵芝菌株特性初探[J].食用菌学报,2017,24(1):50-54.
[13]FELSENSTEIN J.Confidence limits on phylogenies:all approach using the bootstrap[J].Evolution,1985,39:783-791.
[14]叶丽云,鲁欣,林强,等.灵芝单、双核菌丝差异研究[J].食品工业科技,2016,37(20):211-215.
[15]李文涛,余梦瑶,罗霞,等.栽培灵芝生长周期内物质的变化规律及其机制研究[J].中草药,2014,45(4):552-557.
[16]陈凌华,程祖锌,杨志坚.灵芝多糖热水提取条件的优化试验[J].浙江食用菌,2009,17(4):21-23.
[17]马敏,刘嘉宝,陈兰兰.苯酚-硫酸法测定多糖含量显色条件的优化与改进[J].江苏农业科学,2015,43(12):323-324.
[18]黄书铭,杨新林,张自强.超声循环提取灵芝中三萜类化合物的研究[J].中草药,2004,35(5):508-510.
[19]张志军,朱越,罗莹,等.灵芝中三萜化合物提取工艺[J].食品研究与开发,2009,30(9):81-83.
[20]林树钱,林冬梅,罗虹建,等.灵芝提取物质量控制研究[J].海峡药学,2018,30(1):33-36.
[21]叶丽云,林强,刘梅,等.钙离子和水杨酸诱导灵芝多糖和三萜的合成[J].菌物学报,2017,36(2):220-228.
[22]ZHENG LY,JIA DH,FEI XF,et al.An assessment of the genetic diversity within Ganoderma strains with AFLP and ITS PCR-RFLP[J].Microbiological Research,2009,164(3):312-321.