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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

Phylogenetic tree based on neighbor-joining analysis of the rDNA ITS sequences of the endophytic fungal isolates obtained from various tissues of Ocimum sanctum.The bootstrap consensus tree inferred from 1,000 replicates.
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pone.0141444.g001: Phylogenetic tree based on neighbor-joining analysis of the rDNA ITS sequences of the endophytic fungal isolates obtained from various tissues of Ocimum sanctum.The bootstrap consensus tree inferred from 1,000 replicates.

Mentions: 90 endophytic fungi were isolated based on culture dependent technique. Fungal isolates were grouped into 43 different morphospecies according to their morphological characteristics: colony colour and texture, border type, and radial growth rate on MA media. All isolates of each morphospecies group were submitted to molecular identification based on rDNA ITS sequence analysis. Based on the molecular identification these morphospecies were grouped into 23 species. The detailed description of 23 different endophytic fungal isolates with respective identification, accession number from GenBank, % similarity, % query coverage and number of base pairs sequence analysed are summarised in Table 2. The evolutionary history was inferred using the Neighbor-Joining method (Fig 1). The optimal tree with the sum of branch length = 72.08408877 is shown. The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (1000 replicates) are shown next to the branches. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the Maximum composite likelihood method and are in the units of the number of base substitutions per site. All the fungal endophytes recovered in the present study belonged to phylum Ascomycota besides Rhizopus oryzae (4.4%: Mucormycotina) that belongs to phylum Zygomycota. Within Ascomycota, endophytic fungal isolates obtained are in six families of class Sordariomycetes (40%: Hypocreaceae, Glomerellaceae, Xylariaceae, Nectriaceae, Diaporthaceae and Chaetomiaceae), two families of class Dothideomycetes (33.3%: Pleosporaceae, Botryosphaeriaceae), two families of class Saccharomycetes (4.4%: Debaryomycetaceae, Trichomonascaceae) and single family of class Eurotiomycetes (16.6%:Trichocomaceae) (Fig 1). Aspergillus niger was observed as the most dominant species with 10% relative abundance (RA), followed by D. phaseolorum, Hypocrea sp. and M. phaseolina (RA = 7.86%) (Table 2). Only 3 endophytic fungal isolates namely, Bipolaris maydis with relative abundance (RA) of 2.22%, Meyerozyma guilliermondii (RA = 2.2%) and Fusarium verticillioides (RA = 1.11%) were exclusively isolated from stem tissues and rest were present in both the tissues (S1 Table and Table 2)


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

Chowdhary K, Kaushik N - PLoS ONE (2015)

Phylogenetic tree based on neighbor-joining analysis of the rDNA ITS sequences of the endophytic fungal isolates obtained from various tissues of Ocimum sanctum.The bootstrap consensus tree inferred from 1,000 replicates.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0141444.g001: Phylogenetic tree based on neighbor-joining analysis of the rDNA ITS sequences of the endophytic fungal isolates obtained from various tissues of Ocimum sanctum.The bootstrap consensus tree inferred from 1,000 replicates.
Mentions: 90 endophytic fungi were isolated based on culture dependent technique. Fungal isolates were grouped into 43 different morphospecies according to their morphological characteristics: colony colour and texture, border type, and radial growth rate on MA media. All isolates of each morphospecies group were submitted to molecular identification based on rDNA ITS sequence analysis. Based on the molecular identification these morphospecies were grouped into 23 species. The detailed description of 23 different endophytic fungal isolates with respective identification, accession number from GenBank, % similarity, % query coverage and number of base pairs sequence analysed are summarised in Table 2. The evolutionary history was inferred using the Neighbor-Joining method (Fig 1). The optimal tree with the sum of branch length = 72.08408877 is shown. The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (1000 replicates) are shown next to the branches. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the Maximum composite likelihood method and are in the units of the number of base substitutions per site. All the fungal endophytes recovered in the present study belonged to phylum Ascomycota besides Rhizopus oryzae (4.4%: Mucormycotina) that belongs to phylum Zygomycota. Within Ascomycota, endophytic fungal isolates obtained are in six families of class Sordariomycetes (40%: Hypocreaceae, Glomerellaceae, Xylariaceae, Nectriaceae, Diaporthaceae and Chaetomiaceae), two families of class Dothideomycetes (33.3%: Pleosporaceae, Botryosphaeriaceae), two families of class Saccharomycetes (4.4%: Debaryomycetaceae, Trichomonascaceae) and single family of class Eurotiomycetes (16.6%:Trichocomaceae) (Fig 1). Aspergillus niger was observed as the most dominant species with 10% relative abundance (RA), followed by D. phaseolorum, Hypocrea sp. and M. phaseolina (RA = 7.86%) (Table 2). Only 3 endophytic fungal isolates namely, Bipolaris maydis with relative abundance (RA) of 2.22%, Meyerozyma guilliermondii (RA = 2.2%) and Fusarium verticillioides (RA = 1.11%) were exclusively isolated from stem tissues and rest were present in both the tissues (S1 Table and Table 2)

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