陕西宜君核桃内生真菌多样性研究
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摘要
【目的】了解陕西宜君核桃(Juglans regia L.)内生真菌的多样性及分布特征,为活性菌株筛选、抑菌机理研究等奠定基础。【方法】采用组织分离法对不同季节、组织部位的核桃内生真菌进行分离,以形态学和ITS序列分析相结合的方法进行鉴定,通过相对多度、均匀度、多样性指数等分析核桃内生真菌的多样性。【结果】共分离得到内生真菌798株,归属于38个属,有9个属的真菌属于优势类群,相对多度较高的3个属依次为Alternaria、Anthina、Colletotrichum。不同组织部位中,茎部内生真菌多样性最为丰富,分布有32个属,占菌株总数的87.84%,其中1年生茎的相对数量和多样性指数最高,分别为41.37%和1.16,干茎分离得到的内生真菌属丰度最高,为20属;不同季节中,秋季内生真菌的相对数量及多样性指数最高,分别为31.45%和1.12,夏季中分离得到的内生真菌为19属,丰度最高。【结论】宜君核桃内生真菌具有丰富的多样性,丰富度指数、相似性系数反映出不同季节、组织部位内生真菌的分布存在差异。
[Purpose] In order to study the diversity and distribution of the Juglans regia growing in Yijun of Shanxi Province.[Method]Fungal endophytes from different seasons and parts of J. regia were isolated by tissue expand method. Strains were classified by morphology and ITS rDNA sequence analysis, diversity and preference of fungal endophytes were analyzed by relative abundance(RA), evenness, diversity index(H′).[Result]There were 798 strains of endophytic fungi were isolated, belonging to 38 genera, among them 9 genera were dominant, the 3 genera with the highest relative abundance were orderly Alternaria, Anthurium and Colletotrichum. The stems possessed the highest diversity index and was distributed in 32 genera, accounting for 87.84% of the total strains,the strains isolated from 1-year-old stem possessed the highest index of the relative abundance and diversity, respectively 41.37% and 1.16, the endophytic fungi obtained from dry stem were the highest abundance, including 20 genera. In four seasons, the highest index of relative abundance and diversity was appeared in autumn, respectively 31.45% and 1.12, the endophytic fungi isolated in summer had the highest abundance, including 19 genera.[Conclusion]The fungal endophytes from J.regia in Yijun were rich. Meanwhile, the similarity coefficient(Cs) and richness show that the distribution of the endophytic fungi from J. regia in different seasons and tissues seems comparatively different.
[Purpose] In order to study the diversity and distribution of the Juglans regia growing in Yijun of Shanxi Province.[Method]Fungal endophytes from different seasons and parts of J. regia were isolated by tissue expand method. Strains were classified by morphology and ITS rDNA sequence analysis, diversity and preference of fungal endophytes were analyzed by relative abundance(RA), evenness, diversity index(H′).[Result]There were 798 strains of endophytic fungi were isolated, belonging to 38 genera, among them 9 genera were dominant, the 3 genera with the highest relative abundance were orderly Alternaria, Anthurium and Colletotrichum. The stems possessed the highest diversity index and was distributed in 32 genera, accounting for 87.84% of the total strains,the strains isolated from 1-year-old stem possessed the highest index of the relative abundance and diversity, respectively 41.37% and 1.16, the endophytic fungi obtained from dry stem were the highest abundance, including 20 genera. In four seasons, the highest index of relative abundance and diversity was appeared in autumn, respectively 31.45% and 1.12, the endophytic fungi isolated in summer had the highest abundance, including 19 genera.[Conclusion]The fungal endophytes from J.regia in Yijun were rich. Meanwhile, the similarity coefficient(Cs) and richness show that the distribution of the endophytic fungi from J. regia in different seasons and tissues seems comparatively different.
引文
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[14]王海香,申照静,杜娟,等.核桃青皮提取物对朱砂叶螨的毒力及相关酶活性的测定[J].林业科学,2008(5):70.DOI:10.3321/j.issn:1001-7488.2008.05.014.
[15]阿依佳玛丽·依玛尔,刘泽星,毛光瑞,等.不同生境春季核桃内生真菌分离及其多样性[J].西北林学院学报2016,31(6):184.DOI:10.3969/j.issn.1001-7461.2016.0632.
[16]王美琴,卢海波,赵立娟,等.核桃内生菌HT3对番茄和黄瓜的促生作用[J].山西农业科学,2013,41(2):130.DOI:10.3969/j.issn.1002-2481.2013.02.07.
[17]刘泽星,阿依佳玛丽·依玛尔,惠建超,等.陕西核桃内生真菌代谢产物抑菌活性分析[J].西南林业大学学报(自然科学),2017,37(4):126.
[18]魏景超.真菌鉴定手册[M].上海:上海科学技术出版社,1979.
[19]薛庆婉,牛永春,邓晖.常见瓜类植物的内生真菌多样性[J].菌物学报,2015,34(2):196.DOI:10.13346/j.mycosystema.140030.
[20]毛光瑞,翟梅枝,史冠昭,等.陕西不同生境核桃内生真菌多样性[J].微生物学通报,2016,43(6):1262.DOI:10.13344/j.microbiol.china.150368.
[21]柴新义,钟媛媛.青檀果内生真菌菌群的组成及多样性[J].应用生态学报,2017,28(2):636.DOI:10.13287/j.1001-9332.201702.016.
[22]钱一鑫,康冀川,雷帮星,等.贵州白苞蒿抗肿瘤、抗氧化内生真菌的筛选与鉴定[J].中国中药杂志,2014,39(3):438.
[23]张艳,蓝桃菊,廖仕同,等.广西北部湾红树植物内生真菌多样性[J].微生物学通报,2017,44(4):783.DOI:10.13344/j.microbiol.china.160755.
[24]张婷,左雪枝,徐艳.油茶内生真菌多样性与抑菌性研究[J].河南农业科学,2017,46(4):68.DOI:10.15933/j.cnki.1004-3268.2017.04.013.
[25]郭良栋.内生真菌研究进展[J].菌物系统,2001,20(1):148 .DOI:10.3969/j.issn.1672-6472.2001.01.029.
[26]仲军梅,刘玉梅.核桃青皮的开发利用研究进展[J].食品工业科技,2014,35(19):396.DOI:10.13386/j.issn1002-0306.2014.19.078.
[27]SEENA S,WYNBERG N,B?RLOCHER F.Fungal diversity during leaf decomposition in a stream assessed through clone libraries[J].Fungal Diversity,2008,30:1.
[28]DUONG L M,JEEWON R,LUMYONG S,et al.DGGEcoupled with ribosomal DNA gene phylogenies reveal uncharacterized fungal phylotypes[J].Fungal Diversity,2006,23:121.
[29]CURLEVSKI N J A,CHAMBERS S M,ANDERSON IC,et al.Identical genotypes of an ericoid mycorrhizaforming fungus occur in roots of Epacris pulchella(Ericaceae)and Leptospermum polygalifolium(Myrtaceae)in an Australian sclerophyll forest[J].Fems Microbiology Ecology,2009,67(3):411.DOI:10.1111/j.1574-6941.2008.00637.x.
[30]NILSSON R H,RYBERG M,ABARENKON K,et al.The ITS region as a target for characterization of fungal communities using emerging sequencing technologies[J].Fems Microbiology Letters,2009,296(1):97.DOI:10.1111/j.1574-6968.2009.01618.x.
[2]CARROLL G.Fungal endophytes in stems and leaves:from latent pathogen to mutualistic symbiont[J].Ecology,1988,69(1):2.DOI:10.2307/1943154.
[3]张海龙,李善春,卢维浩,等.银杏内生真菌多样性与产黄酮类物质真菌的分离和鉴定[J].土壤,2015,47(1):135.DOI:10.13758/j.cnki.tr.2015.01.021.
[4]唐青,周思旋,雷帮星,等.贵州西南部艾纳香内生真菌多样性研究[J].菌物学报,2017,36(11):1498.DOI:10.13346/j.mycosystema.170057.
[5]LYONS P C,EVANS J J,BACON C W.Effects of the fungal endophyte Acremonium coenophialum on nitrogen accumulation and metabolism in tall fescue[J].Plant Physiology,1990,92(3):726.DOI:10.1104/pp.92.3.726.
[6]BUSH L P,WILKINSON H H,SCHARDL C L.Bioprotective alkaloids of grass-fungal endophyte symbioses[J].Plant Physiology,1997,114(1):1.DOI:10.1104/pp.114.1.1.
[7]FUNK C R,WHITE R H,BREEN J P.Importance of Acremonium endophytes in turf-grass breeding and management[J].Agriculture Ecosystems and Environment,1993,44(1/2/3/4):215.DOI:10.1016/0167-8809(93)90048-T.
[8]RAVEL C,COURTY C,COUDRET A,et al.Beneficial effects of Neotyphodium lolii on the growth and the water status in perennial ryegrass cultivated under nitrogen deficiency or drought stress[J].Agronomie,1997,17(3):173 .DOI:10.1051/agro:19970304.
[9]徐文涛,翟梅枝,王伟,等.核桃内生菌研究Ⅱ.核桃内生菌G8发酵产物的抑菌活性研究[J].西北农业学报,2008,17(1):82.DOI:10.3969/j.issn.1004-1389.2008.01.019.
[10]陆俊,赵安琪,成策,等.核桃营养成分与生理活性及开发利用[J].食品与机械,2014,30(6):238.DOI:10.13652/j.issn.1003-5788.2014.06.057.
[11]JOSE S,GILLESPIE A R.Allelopathy in black walnut(Juglans nigra L.)alley cropping.II.Effects of juglone on hydroponically grown corn(Zea mays L.)and soybean(Glycine max L.Merr.)growth and physiology[J].Plant and Soil,1998,203(2):199.DOI:10.1023/A:1004353326835.
[12]JOSE S,GILLESPIE A R.Allelopathy in black walnut(Juglans nigra L.)alley cropping.I.Spatio-temporal variation in soil juglone in a black walnut-corn(Zea mays L.)alley cropping system in the midwestern USA[J].Plant and Soil,1998,203(2):191.DOI:10.1023/A:1004301309997.
[13]翟梅枝,王磊,何文君,等.核桃青皮乙醇提取物抑菌活性研究[J].西北植物学报,2009,29(12):2542.DOI:10.3321/j.issn:1000-4025.2009.12.025.
[14]王海香,申照静,杜娟,等.核桃青皮提取物对朱砂叶螨的毒力及相关酶活性的测定[J].林业科学,2008(5):70.DOI:10.3321/j.issn:1001-7488.2008.05.014.
[15]阿依佳玛丽·依玛尔,刘泽星,毛光瑞,等.不同生境春季核桃内生真菌分离及其多样性[J].西北林学院学报2016,31(6):184.DOI:10.3969/j.issn.1001-7461.2016.0632.
[16]王美琴,卢海波,赵立娟,等.核桃内生菌HT3对番茄和黄瓜的促生作用[J].山西农业科学,2013,41(2):130.DOI:10.3969/j.issn.1002-2481.2013.02.07.
[17]刘泽星,阿依佳玛丽·依玛尔,惠建超,等.陕西核桃内生真菌代谢产物抑菌活性分析[J].西南林业大学学报(自然科学),2017,37(4):126.
[18]魏景超.真菌鉴定手册[M].上海:上海科学技术出版社,1979.
[19]薛庆婉,牛永春,邓晖.常见瓜类植物的内生真菌多样性[J].菌物学报,2015,34(2):196.DOI:10.13346/j.mycosystema.140030.
[20]毛光瑞,翟梅枝,史冠昭,等.陕西不同生境核桃内生真菌多样性[J].微生物学通报,2016,43(6):1262.DOI:10.13344/j.microbiol.china.150368.
[21]柴新义,钟媛媛.青檀果内生真菌菌群的组成及多样性[J].应用生态学报,2017,28(2):636.DOI:10.13287/j.1001-9332.201702.016.
[22]钱一鑫,康冀川,雷帮星,等.贵州白苞蒿抗肿瘤、抗氧化内生真菌的筛选与鉴定[J].中国中药杂志,2014,39(3):438.
[23]张艳,蓝桃菊,廖仕同,等.广西北部湾红树植物内生真菌多样性[J].微生物学通报,2017,44(4):783.DOI:10.13344/j.microbiol.china.160755.
[24]张婷,左雪枝,徐艳.油茶内生真菌多样性与抑菌性研究[J].河南农业科学,2017,46(4):68.DOI:10.15933/j.cnki.1004-3268.2017.04.013.
[25]郭良栋.内生真菌研究进展[J].菌物系统,2001,20(1):148 .DOI:10.3969/j.issn.1672-6472.2001.01.029.
[26]仲军梅,刘玉梅.核桃青皮的开发利用研究进展[J].食品工业科技,2014,35(19):396.DOI:10.13386/j.issn1002-0306.2014.19.078.
[27]SEENA S,WYNBERG N,B?RLOCHER F.Fungal diversity during leaf decomposition in a stream assessed through clone libraries[J].Fungal Diversity,2008,30:1.
[28]DUONG L M,JEEWON R,LUMYONG S,et al.DGGEcoupled with ribosomal DNA gene phylogenies reveal uncharacterized fungal phylotypes[J].Fungal Diversity,2006,23:121.
[29]CURLEVSKI N J A,CHAMBERS S M,ANDERSON IC,et al.Identical genotypes of an ericoid mycorrhizaforming fungus occur in roots of Epacris pulchella(Ericaceae)and Leptospermum polygalifolium(Myrtaceae)in an Australian sclerophyll forest[J].Fems Microbiology Ecology,2009,67(3):411.DOI:10.1111/j.1574-6941.2008.00637.x.
[30]NILSSON R H,RYBERG M,ABARENKON K,et al.The ITS region as a target for characterization of fungal communities using emerging sequencing technologies[J].Fems Microbiology Letters,2009,296(1):97.DOI:10.1111/j.1574-6968.2009.01618.x.