Isolation and Identification of Endophytic Fungi and Bacteria from <i>Phalaenopsis deliciosa</i>

CHEN Yaoli; YU Longchun; QIAN Yue; SONG Xiqiang; ZHANG Zhe

doi:10.15886/j.cnki.rdswxb.2019.04.011

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Volume 10 Issue 4

Jul. 2019

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Article Navigation > Journal of Tropical Biology > 2019 > 10(4): 372-379

CHEN Yaoli, YU Longchun, QIAN Yue, SONG Xiqiang, ZHANG Zhe. Isolation and Identification of Endophytic Fungi and Bacteria from Phalaenopsis deliciosa[J]. Journal of Tropical Biology, 2019, 10(4): 372-379. doi: 10.15886/j.cnki.rdswxb.2019.04.011

Citation:

CHEN Yaoli, YU Longchun, QIAN Yue, SONG Xiqiang, ZHANG Zhe. Isolation and Identification of Endophytic Fungi and Bacteria from Phalaenopsis deliciosa[J]. Journal of Tropical Biology, 2019, 10(4): 372-379. doi: 10.15886/j.cnki.rdswxb.2019.04.011

Isolation and Identification of Endophytic Fungi and Bacteria from Phalaenopsis deliciosa

doi: 10.15886/j.cnki.rdswxb.2019.04.011

1. Ministry of Education Key Laboratory of Genetics and Germplasm Innovation of Tropical Special Forest Trees and Ornamental Plants, College of Forestry, Hainan University, Haikou, Hainan 570228, China;
2. Hainan Yaode Agricultural Science and Technology Co. LTD, Haikou, Hainan 570228, China

Received Date: 2019-08-10
Rev Recd Date: 2019-09-01

Abstract

Endophytic fungi and bacteria are critical throughout the life cycle of Orchidaceae plants. Phalaenopsis deliciosa（Orchidaceae） was used as test material to isolate the endophytic fungi and bacteria from its fresh roots in routine sterile culture, and the isolates were then identified by using molecular markers to reveal their characteristics and diversity. The results showed that a total of 38 strains of endophytic fungi were isolated from the roots of P. deliciosa and identified as 5 OTUs belonging to 1 phylum, 2 classes, 4 orders, 4 families and 5 genera, and that the dominant genera were Lasiodiplodia and Aspergillus, while a total of 54 strins of endophytic bacteria were isolated and 17 OTUs were identified to belong to 3 phyla, 5 classes, 6 orders, 9 families, 12 genera, and that the dominant genera were Bacillus and Enterobacter. The results will provide reference for symbiotic germination and raising of mycorrhiza-based seedlings of P. deliciosa and for further study of the relationship between endophytic microorganisms and orchids.
- Phalaenopsis,
- endophytic fungi,
- endophytic bacteria,
- molecular identification

References

[1]	MISAGHI I J,DONNDELINGER C R.Endophytic bacteria in symptom-free cotton plants [J].Phytopathology,1990,80(9):808-811.
[2]	严婉荣,赵廷昌,肖彤斌,等.生防细菌在植物病害防治中的应用[J].基因组学与应用生物学,2013,32(4):533-539.
[3]	MCCORMICK M K,WHIGHAM D F,CANCHANI-VIRUET A.Mycorrhizal fungi affect orchid distribution and population dynamics [J].New Phytologist,2018,219(4):1-9.
[4]	盖雪鸽,邢晓科,郭顺星.兰科菌根的生态学研究进展[J].菌物学报,2014,33(4):753-767.
[5]	SWARTS N D,DIXON K W.Terrestrial orchid conservation in the age of extinction [J].Annals of Botany,2009,104(3):543-556.
[6]	何劲.石斛内生菌及其拮抗真菌的研究[D].贵阳:贵州大学,2006.
[7]	张芳芳.五唇兰根部内生细菌筛选及其促生效应研究[D].海口:海南大学,2015.
[8]	LODEWYCKX C,VANGRONSVELD J,PORTEOUS F,et al.Endophytic bacteria and their potential applications [J].Critical Reviews in Plant Sciences,2002,21(6):583-606.
[9]	TSAVKELOVA E A,CHERDYNTSEVA T A,BOTINA S G,et al.Bacteria associated with orchid roots and microbial production of auxin [J].Microbiology Research,2007,162(1):69-76.
[10]	陈瑞蕊,林先贵,施亚琴.兰科菌根的研究进展[J].应用与环境生物学报,2003,9(1):97-100.
[11]	程萍,郑燕玲,黎永坚,等.石斛兰镰刀菌叶斑病的生物防治研究[J].中国农学通报,2008,24(9):357-361.
[12]	GUO L D,HYDE K D,LIEW E C Y.Identification of endophytic fungi from Livistona chinensis based on morphology and rDNA sequences [J].New Phytologist,2000,147(3):617-630.
[13]	CAPORASO J G,KUCZYNSKI J,STOMBAUGH J,et al.QIIME allows analysis of high-throughput community sequencing data [J].Nature Methods,2010,7:335-336.
[14]	EDGAR R C.Search and clustering orders of magnitude faster than BLAST [J].Bioinformatics,2010,26(19):2460-2461.
[15]	SáNCHEZ M S,BILLS G F,ZABALGOGEAZCOA I.Diversity and structure of the fungal endophytic assemblages from two sympatric coastal grasses [J].Fungal Diversity,2008,33:87-100.
[16]	CHEN J,HU K,HOU X,et al.Endophytic fungi assemblages from Dendrobium medicinal plants (Orchidaceae)[J].World Journal of Microbiology and Biotechnology,2011,27(5):1009-1016.
[17]	GRAHAM R R,DEARNALEY J D W.The rare Australian epiphytic orchid Sarcochilus weinthalii associates with a single species of Ceratobasidium[J].Fungal Diversity,2012,54(1):31-37.
[18]	BODDINGTON M,DEARNALEY J D W.Morphological and molecular identification of fungal endophytes from roots of Dendrobium speciosum [J].Proceedings of the Royal Society of Queensland,2008,114:13–17.
[19]	BAYMAN P,GONZáLEZ E J,FUMERO J J,et al.Are fungi necessary?How fungicides affect growth and survival of the orchid Lepanthes rupestris in the field [J].Journal of Ecology,2002,90(6):1002-1008.
[20]	TUPAC J O,ACKERMAN J D,BAYMAN P.Diversity and host specificity of endophytic Rhizoctonia-like fungi from tropical orchid [J].American Journal of Botany,2002,89(11):1852-1858.
[21]	RASMUSSEN H N.Recent developments in the study of orchid mycorrhiza [J].Plant and Soil,2002,244(1):149-163.
[22]	ESSER K,HOCK B.The Mycota(vol.9) [M].Berlin:Springer-Verlag,2012:207-230.
[23]	SCHULZ B J E,BOYLE C J C,SIEBER T N.Microbial root endophytes,soil biology(vol.9,part II)[M].Berlin:Springer-Verlag,2006:153-177.
[24]	陈娟,孟志霞,邢咏梅,等.5 种兰科药用植物可培养内生真菌的鉴定及多样性分析[J].中国药学杂志,2017,52(4):267-271.
[25]	陈娟,张丽春,邢咏梅,等.石斛属植物内生真菌及菌根真菌物种多样性研究进展[J].中国药学杂志,2013,48(19):1649-1653.
[26]	王亚妮.兰科石斛属植物根部内生真菌多样性研究及应用[D].北京:北京林业大学,2013.
[27]	柯海丽,宋希强,谭志琼,等.野生五唇兰根部内生真菌多样性研究[J].生物多样性,2007,15(5):456-462.
[28]	MOSQUERA-ESPINOSA A T,BAYMAN P,PRADO G A,et al.The double life of Ceratobasidium:orchid mycorrhizal fungi and their potential for biocontrol of Rhizoctonia solani sheath blight of rice [J].Mycologia,2013,105(1):141-150.
[29]	帅红艳.广西环江产铁皮石斛内生真菌的分离鉴定及其生物活性的研究[D].南宁:广西大学,2008.
[30]	CHEN X M,DONG H L,HU K X,et al.Diversity and antimicrobial and plant-growth-promoting activities of endophytic fungi in Dendrobium loddigesii Rolfe.[J].Journal of Plant Growth Regulation,2010,29(3):328-337.
[31]	WANG X,LI Y,SONG X,et al.Influence of host tree species on isolation and communities of mycorrhizal and endophytic fungi from roots of a tropical epiphytic orchid,Dendrobium sinense (Orchidaceae)[J].Mycorrhiza,2017,27(7):709-718.
[32]	张萍,宋希强.兰科植物内生细菌物种多样性及其促生机理研究进展[J].热带亚热带植物学报,2012,20(1):92-98.
[33]	李骜.华石斛根部可培养内生细菌分离鉴定及其促生研究[D].海口:海南大学,2015.
[34]	WILKINSON K G,SIVASITHAMPARAM K,DIXON K W,et al.Identification and characterization of bacteria associated with Western Australian orchids [J].Soil Biology and Biochemistry,1994,26(1):137-142.
[35]	TSAVKELOVA E A,CHERDYNTSEVA T A,KLIMOVA S Y,et al.Orchid-associated bacteria produce indole-3-acetic acid,promote seed germination,and increase their microbial yield in response to exogenous auxin [J].Archives of Microbiology,2007,188(6):655-664.
[36]	WILKINSON K G,DIXON K W,SIVASITHAMPARAM K,et al.Effect of IAA on symbiotic germination of an Australian orchid and its production by orchid-associated bacteria [J].Plant and Soil,1994,159(2):291-295.
[37]	TSAVKELOVA E A,CHERDYNTSEVA T A,NETRUSOV A I.Auxin production by bacteria associated with orchid roots [J].Microbiology,2005,74(1):46-53.
[38]	TSAVKELOVA E A,LOBAKOVA E S,KOLOMEITSEVA G L,et al.Associative cyanobacteria isolated from the roots of epiphytic orchids [J].Microbiology,2003,72(1):92-97.

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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Isolation and Identification of Endophytic Fungi and Bacteria from Phalaenopsis deliciosa

1. Ministry of Education Key Laboratory of Genetics and Germplasm Innovation of Tropical Special Forest Trees and Ornamental Plants, College of Forestry, Hainan University, Haikou, Hainan 570228, China;

2. Hainan Yaode Agricultural Science and Technology Co. LTD, Haikou, Hainan 570228, China

Received Date: 2019-08-10

Rev Recd Date: 2019-09-01

Key words:

Abstract: Endophytic fungi and bacteria are critical throughout the life cycle of Orchidaceae plants. Phalaenopsis deliciosa（Orchidaceae） was used as test material to isolate the endophytic fungi and bacteria from its fresh roots in routine sterile culture, and the isolates were then identified by using molecular markers to reveal their characteristics and diversity. The results showed that a total of 38 strains of endophytic fungi were isolated from the roots of P. deliciosa and identified as 5 OTUs belonging to 1 phylum, 2 classes, 4 orders, 4 families and 5 genera, and that the dominant genera were Lasiodiplodia and Aspergillus, while a total of 54 strins of endophytic bacteria were isolated and 17 OTUs were identified to belong to 3 phyla, 5 classes, 6 orders, 9 families, 12 genera, and that the dominant genera were Bacillus and Enterobacter. The results will provide reference for symbiotic germination and raising of mycorrhiza-based seedlings of P. deliciosa and for further study of the relationship between endophytic microorganisms and orchids.

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Reference (38)

[1]	MISAGHI I J,DONNDELINGER C R.Endophytic bacteria in symptom-free cotton plants [J].Phytopathology,1990,80(9):808-811.
[2]	严婉荣,赵廷昌,肖彤斌,等.生防细菌在植物病害防治中的应用[J].基因组学与应用生物学,2013,32(4):533-539.
[3]	MCCORMICK M K,WHIGHAM D F,CANCHANI-VIRUET A.Mycorrhizal fungi affect orchid distribution and population dynamics [J].New Phytologist,2018,219(4):1-9.
[4]	盖雪鸽,邢晓科,郭顺星.兰科菌根的生态学研究进展[J].菌物学报,2014,33(4):753-767.
[5]	SWARTS N D,DIXON K W.Terrestrial orchid conservation in the age of extinction [J].Annals of Botany,2009,104(3):543-556.
[6]	何劲.石斛内生菌及其拮抗真菌的研究[D].贵阳:贵州大学,2006.
[7]	张芳芳.五唇兰根部内生细菌筛选及其促生效应研究[D].海口:海南大学,2015.
[8]	LODEWYCKX C,VANGRONSVELD J,PORTEOUS F,et al.Endophytic bacteria and their potential applications [J].Critical Reviews in Plant Sciences,2002,21(6):583-606.
[9]	TSAVKELOVA E A,CHERDYNTSEVA T A,BOTINA S G,et al.Bacteria associated with orchid roots and microbial production of auxin [J].Microbiology Research,2007,162(1):69-76.
[10]	陈瑞蕊,林先贵,施亚琴.兰科菌根的研究进展[J].应用与环境生物学报,2003,9(1):97-100.
[11]	程萍,郑燕玲,黎永坚,等.石斛兰镰刀菌叶斑病的生物防治研究[J].中国农学通报,2008,24(9):357-361.
[12]	GUO L D,HYDE K D,LIEW E C Y.Identification of endophytic fungi from Livistona chinensis based on morphology and rDNA sequences [J].New Phytologist,2000,147(3):617-630.
[13]	CAPORASO J G,KUCZYNSKI J,STOMBAUGH J,et al.QIIME allows analysis of high-throughput community sequencing data [J].Nature Methods,2010,7:335-336.
[14]	EDGAR R C.Search and clustering orders of magnitude faster than BLAST [J].Bioinformatics,2010,26(19):2460-2461.
[15]	SáNCHEZ M S,BILLS G F,ZABALGOGEAZCOA I.Diversity and structure of the fungal endophytic assemblages from two sympatric coastal grasses [J].Fungal Diversity,2008,33:87-100.
[16]	CHEN J,HU K,HOU X,et al.Endophytic fungi assemblages from Dendrobium medicinal plants (Orchidaceae)[J].World Journal of Microbiology and Biotechnology,2011,27(5):1009-1016.
[17]	GRAHAM R R,DEARNALEY J D W.The rare Australian epiphytic orchid Sarcochilus weinthalii associates with a single species of Ceratobasidium[J].Fungal Diversity,2012,54(1):31-37.
[18]	BODDINGTON M,DEARNALEY J D W.Morphological and molecular identification of fungal endophytes from roots of Dendrobium speciosum [J].Proceedings of the Royal Society of Queensland,2008,114:13–17.
[19]	BAYMAN P,GONZáLEZ E J,FUMERO J J,et al.Are fungi necessary?How fungicides affect growth and survival of the orchid Lepanthes rupestris in the field [J].Journal of Ecology,2002,90(6):1002-1008.
[20]	TUPAC J O,ACKERMAN J D,BAYMAN P.Diversity and host specificity of endophytic Rhizoctonia-like fungi from tropical orchid [J].American Journal of Botany,2002,89(11):1852-1858.
[21]	RASMUSSEN H N.Recent developments in the study of orchid mycorrhiza [J].Plant and Soil,2002,244(1):149-163.
[22]	ESSER K,HOCK B.The Mycota(vol.9) [M].Berlin:Springer-Verlag,2012:207-230.
[23]	SCHULZ B J E,BOYLE C J C,SIEBER T N.Microbial root endophytes,soil biology(vol.9,part II)[M].Berlin:Springer-Verlag,2006:153-177.
[24]	陈娟,孟志霞,邢咏梅,等.5 种兰科药用植物可培养内生真菌的鉴定及多样性分析[J].中国药学杂志,2017,52(4):267-271.
[25]	陈娟,张丽春,邢咏梅,等.石斛属植物内生真菌及菌根真菌物种多样性研究进展[J].中国药学杂志,2013,48(19):1649-1653.
[26]	王亚妮.兰科石斛属植物根部内生真菌多样性研究及应用[D].北京:北京林业大学,2013.
[27]	柯海丽,宋希强,谭志琼,等.野生五唇兰根部内生真菌多样性研究[J].生物多样性,2007,15(5):456-462.
[28]	MOSQUERA-ESPINOSA A T,BAYMAN P,PRADO G A,et al.The double life of Ceratobasidium:orchid mycorrhizal fungi and their potential for biocontrol of Rhizoctonia solani sheath blight of rice [J].Mycologia,2013,105(1):141-150.
[29]	帅红艳.广西环江产铁皮石斛内生真菌的分离鉴定及其生物活性的研究[D].南宁:广西大学,2008.
[30]	CHEN X M,DONG H L,HU K X,et al.Diversity and antimicrobial and plant-growth-promoting activities of endophytic fungi in Dendrobium loddigesii Rolfe.[J].Journal of Plant Growth Regulation,2010,29(3):328-337.
[31]	WANG X,LI Y,SONG X,et al.Influence of host tree species on isolation and communities of mycorrhizal and endophytic fungi from roots of a tropical epiphytic orchid,Dendrobium sinense (Orchidaceae)[J].Mycorrhiza,2017,27(7):709-718.
[32]	张萍,宋希强.兰科植物内生细菌物种多样性及其促生机理研究进展[J].热带亚热带植物学报,2012,20(1):92-98.
[33]	李骜.华石斛根部可培养内生细菌分离鉴定及其促生研究[D].海口:海南大学,2015.
[34]	WILKINSON K G,SIVASITHAMPARAM K,DIXON K W,et al.Identification and characterization of bacteria associated with Western Australian orchids [J].Soil Biology and Biochemistry,1994,26(1):137-142.
[35]	TSAVKELOVA E A,CHERDYNTSEVA T A,KLIMOVA S Y,et al.Orchid-associated bacteria produce indole-3-acetic acid,promote seed germination,and increase their microbial yield in response to exogenous auxin [J].Archives of Microbiology,2007,188(6):655-664.
[36]	WILKINSON K G,DIXON K W,SIVASITHAMPARAM K,et al.Effect of IAA on symbiotic germination of an Australian orchid and its production by orchid-associated bacteria [J].Plant and Soil,1994,159(2):291-295.
[37]	TSAVKELOVA E A,CHERDYNTSEVA T A,NETRUSOV A I.Auxin production by bacteria associated with orchid roots [J].Microbiology,2005,74(1):46-53.
[38]	TSAVKELOVA E A,LOBAKOVA E S,KOLOMEITSEVA G L,et al.Associative cyanobacteria isolated from the roots of epiphytic orchids [J].Microbiology,2003,72(1):92-97.

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Isolation and Identification of Endophytic Fungi and Bacteria from Phalaenopsis deliciosa

doi: 10.15886/j.cnki.rdswxb.2019.04.011

Abstract

References

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通讯作者: 陈斌, bchen63@163.com

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Isolation and Identification of Endophytic Fungi and Bacteria from Phalaenopsis deliciosa

doi: 10.15886/j.cnki.rdswxb.2019.04.011

1. Ministry of Education Key Laboratory of Genetics and Germplasm Innovation of Tropical Special Forest Trees and Ornamental Plants, College of Forestry, Hainan University, Haikou, Hainan 570228, China;

2. Hainan Yaode Agricultural Science and Technology Co. LTD, Haikou, Hainan 570228, China

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Isolation and Identification of Endophytic Fungi and Bacteria from Phalaenopsis deliciosa

doi: 10.15886/j.cnki.rdswxb.2019.04.011

Abstract

References

Proportional views

通讯作者: 陈斌, bchen63@163.com

Proportional views

Related

Isolation and Identification of Endophytic Fungi and Bacteria from Phalaenopsis deliciosa

doi: 10.15886/j.cnki.rdswxb.2019.04.011

1. Ministry of Education Key Laboratory of Genetics and Germplasm Innovation of Tropical Special Forest Trees and Ornamental Plants, College of Forestry, Hainan University, Haikou, Hainan 570228, China; 2. Hainan Yaode Agricultural Science and Technology Co. LTD, Haikou, Hainan 570228, China

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1. Ministry of Education Key Laboratory of Genetics and Germplasm Innovation of Tropical Special Forest Trees and Ornamental Plants, College of Forestry, Hainan University, Haikou, Hainan 570228, China;

2. Hainan Yaode Agricultural Science and Technology Co. LTD, Haikou, Hainan 570228, China