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Fungal Endophyte Diversity and Bioactivity in the Indian Medicinal Plant Ocimum sanctum Linn.

Chowdhary K, Kaushik N - PLoS ONE (2015)

Bottom Line: Hexane extract of M. phaseolina recovered from Hyderabad in first collection demonstrated highest activity against S. sclerotiorum with IC50 value of 0.38 mg/ml.Additionally, its components 2H-pyran-2-one, 5,6-dihydro-6-pentyl and palmitic acid, methyl ester as reported by GC-MS Chromatogram upon evaluation for their antiphytopathogenic activity exhibited IC50 value of 1.002 and 0.662 against respectively S. sclerotiorum indicating their significant role in antiphytopathogenic activity of hexane extract.The production of 2H-pyran-2-one, 5,6-dihydro-6-pentyl from M. phaseolina, an endophytic fungus is being reported for the first time.

View Article: PubMed Central - PubMed

Affiliation: TERI University, 10th Institutional Area, Vasant Kunj, New Delhi, India.

ABSTRACT
Endophytic mycopopulation isolated from India's Queen of herbs Tulsi (Ocimum sanctum) were explored and investigated for their diversity and antiphytopathogenic activity against widespread plant pathogens Botrytis cinerea, Sclerotinia sclerotiorum, Rhizoctonia solani and Fusarium oxysporum. 90 fungal isolates, representing 17 genera were recovered from 313 disease-free and surface sterilised plant segments (leaf and stem tissues) from three different geographic locations (Delhi, Hyderabad and Mukteshwar) during distinct sampling times in consequent years 2010 and 2011 in India. Fungal endophytes were subjected to molecular identification based on rDNA ITS sequence analysis. Plant pathogens such as F. verticillioides, B. maydis, C. coarctatum, R. bataticola, Hypoxylon sp., Diaporthe phaseolorum, Alternaria tenuissima and A. alternata have occurred as endophyte only during second sampling (second sampling in 2011) in the present study. Bi-plot generated by principal component analysis suggested tissue specificity of certain fungal endophytes. Dendrogram revealed species abundance as a function of mean temperature of the location at the time of sampling. Shannon diversity in the first collection is highest in Hyderabad leaf tissues (H' = 1.907) whereas in second collection it was highest from leaf tissues of Delhi (H' = 1.846). Mukteshwar (altitude: 7500 feet) reported least isolation rate in second collection. Nearly 23% of the total fungal isolates were considered as potent biocontrol agent. Hexane extract of M. phaseolina recovered from Hyderabad in first collection demonstrated highest activity against S. sclerotiorum with IC50 value of 0.38 mg/ml. Additionally, its components 2H-pyran-2-one, 5,6-dihydro-6-pentyl and palmitic acid, methyl ester as reported by GC-MS Chromatogram upon evaluation for their antiphytopathogenic activity exhibited IC50 value of 1.002 and 0.662 against respectively S. sclerotiorum indicating their significant role in antiphytopathogenic activity of hexane extract. The production of 2H-pyran-2-one, 5,6-dihydro-6-pentyl from M. phaseolina, an endophytic fungus is being reported for the first time.

No MeSH data available.


Related in: MedlinePlus

A: GC-Chromatogram of Macrophomina phaseolina recovered from leaf tissue (OSHL-2.1)- major peaks: 30.156 (2H-Pyran-2-one,5,6-dihydro-6-pentyl), 53.017(Hexadecanoic acid), 64.986 (Linoleic acid), 65.265(10-Octadeconic acid)B: Macrophomina phaseolina recovered from stem tissue-(OSHS-1.3) major peaks: 61.108 (9,12-Octadecadieonic acid, methyl ester), 55.970(Trifluoroacetoxy hexadecane). C:-Chaetomium coarctatum (OSDSS-2.5)- major peaks: 38.644 (2-Fluorobenzoic acid), 38.444 (2,5-Difluorobenzoic acid), 64.900 (Linoleic acid, ethyl ester), D: Alternaria sp (OSHL-5.2).-major peaks: 52.990(Hexadecanoic acid), 61.036 (9,15- Octadecadienoic acid, methyl ester) E-Fusarium proliferatum(OSHSS-1.3) major peaks: 52.992(Hexadecanoic acid), 64.902(Linoleic acid), 65.954(Oleic acid); F-Colletotrichum sp.-(OSML-5.4) major peaks: 61.372(7-Hydroxy3-(1,1-dimethylprop-2-enyl) coumarin), 61.052(9,12-Octadecadieonic acid).
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pone.0141444.g005: A: GC-Chromatogram of Macrophomina phaseolina recovered from leaf tissue (OSHL-2.1)- major peaks: 30.156 (2H-Pyran-2-one,5,6-dihydro-6-pentyl), 53.017(Hexadecanoic acid), 64.986 (Linoleic acid), 65.265(10-Octadeconic acid)B: Macrophomina phaseolina recovered from stem tissue-(OSHS-1.3) major peaks: 61.108 (9,12-Octadecadieonic acid, methyl ester), 55.970(Trifluoroacetoxy hexadecane). C:-Chaetomium coarctatum (OSDSS-2.5)- major peaks: 38.644 (2-Fluorobenzoic acid), 38.444 (2,5-Difluorobenzoic acid), 64.900 (Linoleic acid, ethyl ester), D: Alternaria sp (OSHL-5.2).-major peaks: 52.990(Hexadecanoic acid), 61.036 (9,15- Octadecadienoic acid, methyl ester) E-Fusarium proliferatum(OSHSS-1.3) major peaks: 52.992(Hexadecanoic acid), 64.902(Linoleic acid), 65.954(Oleic acid); F-Colletotrichum sp.-(OSML-5.4) major peaks: 61.372(7-Hydroxy3-(1,1-dimethylprop-2-enyl) coumarin), 61.052(9,12-Octadecadieonic acid).

Mentions: Fungal crude extracts of OSHL-2.1(M. phaseolina), OSHS-3.1 (M.phaseolina), OSDSS-2.5 (Chaetomium coarctatum), OSHL-5.2 (Alternaria sp.), OSML-5.4 (Colletotrichum sp.) and OSHSS-1.3 (F. proliferatum) were found positive for terpenoids in preliminary phytochemical screening. GC-MS chromatography of hexane extracts of these selected endophytic fungal isolates revealed presence of volatile compounds, fatty acids, and aliphatic constituents. GC-MS chromatogram of M. phaseolina (OSHL-2.1) showed presence of 2H-Pyran-2-one, 5, 6-dihydro-6-pentyl at RT 30.156, hexadecanoic acid at RT 53.017, linoleic acid at RT 64.986 (Fig 5A) while hexane extract of another strain of M. phaseolina (OSHS-3.1) showed presence 9,12-octadecadieonic acid, methyl ester at RT 61.108, trifluoroacetoxy hexadecane at RT 55.970 as major peaks reflecting strain variability (Fig 5B). In GC-MS chromatogram of hexane extract of Chaetomium coarctatum (OSDSS-2.5) 2-fluorobenzoic acid at RT 38.644, 2,5-difluorobenzoic acid at RT 38.444, linoleic acid, ethyl ester at RT 64.900 were observed as major peaks (Fig 5C). In Alternaria sp. (OSHL-5.2) hexane extract, hexadecanoic acid at RT 52.990, 9,15- octadecadienoic acid, methyl ester at RT 61.036 were recorded as major peaks (Fig 5D) while that of Fusarium proliferatum (OSHSS-1.3)-major peaks were hexadecanoic acid at RT 52.992, linoleic acid at RT 64.902, oleic acid at RT 65.954 (Fig 5E). Colletotrichum sp. (OSML-5.4) major peaks recorded were 7-hydroxy3-(1, 1-dimethylprop-2-enyl) coumarin at RT 61.372, 9, 12-octadecadieonic acid at RT 61.052 (Fig 5F).


Fungal Endophyte Diversity and Bioactivity in the Indian Medicinal Plant Ocimum sanctum Linn.

Chowdhary K, Kaushik N - PLoS ONE (2015)

A: GC-Chromatogram of Macrophomina phaseolina recovered from leaf tissue (OSHL-2.1)- major peaks: 30.156 (2H-Pyran-2-one,5,6-dihydro-6-pentyl), 53.017(Hexadecanoic acid), 64.986 (Linoleic acid), 65.265(10-Octadeconic acid)B: Macrophomina phaseolina recovered from stem tissue-(OSHS-1.3) major peaks: 61.108 (9,12-Octadecadieonic acid, methyl ester), 55.970(Trifluoroacetoxy hexadecane). C:-Chaetomium coarctatum (OSDSS-2.5)- major peaks: 38.644 (2-Fluorobenzoic acid), 38.444 (2,5-Difluorobenzoic acid), 64.900 (Linoleic acid, ethyl ester), D: Alternaria sp (OSHL-5.2).-major peaks: 52.990(Hexadecanoic acid), 61.036 (9,15- Octadecadienoic acid, methyl ester) E-Fusarium proliferatum(OSHSS-1.3) major peaks: 52.992(Hexadecanoic acid), 64.902(Linoleic acid), 65.954(Oleic acid); F-Colletotrichum sp.-(OSML-5.4) major peaks: 61.372(7-Hydroxy3-(1,1-dimethylprop-2-enyl) coumarin), 61.052(9,12-Octadecadieonic acid).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4631451&req=5

pone.0141444.g005: A: GC-Chromatogram of Macrophomina phaseolina recovered from leaf tissue (OSHL-2.1)- major peaks: 30.156 (2H-Pyran-2-one,5,6-dihydro-6-pentyl), 53.017(Hexadecanoic acid), 64.986 (Linoleic acid), 65.265(10-Octadeconic acid)B: Macrophomina phaseolina recovered from stem tissue-(OSHS-1.3) major peaks: 61.108 (9,12-Octadecadieonic acid, methyl ester), 55.970(Trifluoroacetoxy hexadecane). C:-Chaetomium coarctatum (OSDSS-2.5)- major peaks: 38.644 (2-Fluorobenzoic acid), 38.444 (2,5-Difluorobenzoic acid), 64.900 (Linoleic acid, ethyl ester), D: Alternaria sp (OSHL-5.2).-major peaks: 52.990(Hexadecanoic acid), 61.036 (9,15- Octadecadienoic acid, methyl ester) E-Fusarium proliferatum(OSHSS-1.3) major peaks: 52.992(Hexadecanoic acid), 64.902(Linoleic acid), 65.954(Oleic acid); F-Colletotrichum sp.-(OSML-5.4) major peaks: 61.372(7-Hydroxy3-(1,1-dimethylprop-2-enyl) coumarin), 61.052(9,12-Octadecadieonic acid).
Mentions: Fungal crude extracts of OSHL-2.1(M. phaseolina), OSHS-3.1 (M.phaseolina), OSDSS-2.5 (Chaetomium coarctatum), OSHL-5.2 (Alternaria sp.), OSML-5.4 (Colletotrichum sp.) and OSHSS-1.3 (F. proliferatum) were found positive for terpenoids in preliminary phytochemical screening. GC-MS chromatography of hexane extracts of these selected endophytic fungal isolates revealed presence of volatile compounds, fatty acids, and aliphatic constituents. GC-MS chromatogram of M. phaseolina (OSHL-2.1) showed presence of 2H-Pyran-2-one, 5, 6-dihydro-6-pentyl at RT 30.156, hexadecanoic acid at RT 53.017, linoleic acid at RT 64.986 (Fig 5A) while hexane extract of another strain of M. phaseolina (OSHS-3.1) showed presence 9,12-octadecadieonic acid, methyl ester at RT 61.108, trifluoroacetoxy hexadecane at RT 55.970 as major peaks reflecting strain variability (Fig 5B). In GC-MS chromatogram of hexane extract of Chaetomium coarctatum (OSDSS-2.5) 2-fluorobenzoic acid at RT 38.644, 2,5-difluorobenzoic acid at RT 38.444, linoleic acid, ethyl ester at RT 64.900 were observed as major peaks (Fig 5C). In Alternaria sp. (OSHL-5.2) hexane extract, hexadecanoic acid at RT 52.990, 9,15- octadecadienoic acid, methyl ester at RT 61.036 were recorded as major peaks (Fig 5D) while that of Fusarium proliferatum (OSHSS-1.3)-major peaks were hexadecanoic acid at RT 52.992, linoleic acid at RT 64.902, oleic acid at RT 65.954 (Fig 5E). Colletotrichum sp. (OSML-5.4) major peaks recorded were 7-hydroxy3-(1, 1-dimethylprop-2-enyl) coumarin at RT 61.372, 9, 12-octadecadieonic acid at RT 61.052 (Fig 5F).

Bottom Line: Hexane extract of M. phaseolina recovered from Hyderabad in first collection demonstrated highest activity against S. sclerotiorum with IC50 value of 0.38 mg/ml.Additionally, its components 2H-pyran-2-one, 5,6-dihydro-6-pentyl and palmitic acid, methyl ester as reported by GC-MS Chromatogram upon evaluation for their antiphytopathogenic activity exhibited IC50 value of 1.002 and 0.662 against respectively S. sclerotiorum indicating their significant role in antiphytopathogenic activity of hexane extract.The production of 2H-pyran-2-one, 5,6-dihydro-6-pentyl from M. phaseolina, an endophytic fungus is being reported for the first time.

View Article: PubMed Central - PubMed

Affiliation: TERI University, 10th Institutional Area, Vasant Kunj, New Delhi, India.

ABSTRACT
Endophytic mycopopulation isolated from India's Queen of herbs Tulsi (Ocimum sanctum) were explored and investigated for their diversity and antiphytopathogenic activity against widespread plant pathogens Botrytis cinerea, Sclerotinia sclerotiorum, Rhizoctonia solani and Fusarium oxysporum. 90 fungal isolates, representing 17 genera were recovered from 313 disease-free and surface sterilised plant segments (leaf and stem tissues) from three different geographic locations (Delhi, Hyderabad and Mukteshwar) during distinct sampling times in consequent years 2010 and 2011 in India. Fungal endophytes were subjected to molecular identification based on rDNA ITS sequence analysis. Plant pathogens such as F. verticillioides, B. maydis, C. coarctatum, R. bataticola, Hypoxylon sp., Diaporthe phaseolorum, Alternaria tenuissima and A. alternata have occurred as endophyte only during second sampling (second sampling in 2011) in the present study. Bi-plot generated by principal component analysis suggested tissue specificity of certain fungal endophytes. Dendrogram revealed species abundance as a function of mean temperature of the location at the time of sampling. Shannon diversity in the first collection is highest in Hyderabad leaf tissues (H' = 1.907) whereas in second collection it was highest from leaf tissues of Delhi (H' = 1.846). Mukteshwar (altitude: 7500 feet) reported least isolation rate in second collection. Nearly 23% of the total fungal isolates were considered as potent biocontrol agent. Hexane extract of M. phaseolina recovered from Hyderabad in first collection demonstrated highest activity against S. sclerotiorum with IC50 value of 0.38 mg/ml. Additionally, its components 2H-pyran-2-one, 5,6-dihydro-6-pentyl and palmitic acid, methyl ester as reported by GC-MS Chromatogram upon evaluation for their antiphytopathogenic activity exhibited IC50 value of 1.002 and 0.662 against respectively S. sclerotiorum indicating their significant role in antiphytopathogenic activity of hexane extract. The production of 2H-pyran-2-one, 5,6-dihydro-6-pentyl from M. phaseolina, an endophytic fungus is being reported for the first time.

No MeSH data available.


Related in: MedlinePlus