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Molecular characterization of an endophytic Phomopsis liquidambaris CBR-15 from Cryptolepis buchanani Roem. and impact of culture media on biosynthesis of antimicrobial metabolites

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ABSTRACT

An endophytic fungus Phomopsis liquidambaris CBR-15, was isolated from Cryptolepis buchanani Roem. (Asclepiadaceae) and identified by its characteristic culture morphology and molecular analysis of the ITS region of rDNA and intervening 5.8S rRNA gene. The impact of different culture media on biosynthesis of antimicrobial metabolites was tested by disc diffusion assay. Polyketide synthase gene (PKS) of the endophytic fungus was investigated using three pairs of degenerate primers LC1–LC2c, LC3–LC5c and KS3–KS4c by PCR. TLC-bioautography method was employed to detect the antimicrobial metabolites. Antimicrobial metabolites fractionated with ethyl acetate extract showed significant antimicrobial activity against the test bacteria and fungi. Biosynthesis of antimicrobial metabolites was optimum as depicted by zone of inhibition from ethyl acetate extract cultured in potato dextrose broth. Strain CBR-15 was identified as Phomopsisliquidambaris and PKS genes of the fungus were amplified with LC3–LC5c and KS3–KS4c sets of degenerate primers. These findings suggest that endophytic P.liquidambaris CBR-15 harbor iterative type I fungal PKS gene domain which indicates the biosynthetic potential of endophytic fungi as producers of natural antimicrobial metabolites. The study also demonstrates the utilization and optimization of different culture media which best supports for the biosynthesis of the antimicrobial metabolites from P.liquidambaris.

Electronic supplementary material: The online version of this article (doi:10.1007/s13205-014-0204-2) contains supplementary material, which is available to authorized users.

No MeSH data available.


a Colony morphology of P.liquidambaris CBR-15 on PDA and b microscopic features at ×40 magnification showing alpha and beta conidia
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Fig1: a Colony morphology of P.liquidambaris CBR-15 on PDA and b microscopic features at ×40 magnification showing alpha and beta conidia

Mentions: The endophytic fungus P.liquidambaris CBR-15 which is used in the present study was isolated from the leaf tissue of C. buchanani. Colony morphology on PDA, after 5 days at 25 °C, initially cottony, white to olive gray later on turns to gray to light brown and a radiating growth pattern, margin regular. Conidiomata eustromatic, black, spherical to irregular on the upper regions; Conidiophores: short to elongated, aseptate to septate and branched, Conidia are of two types: (a) alpha conidia hyaline, straight, aseptate, forming white to yellowish cirrhi; (b) beta conidia hyaline, filiform, straight or curved, aseptae morphology characters observed, were closely related with the genus Phomopsis, belongs to class coelomycetes (Fig. 1).Fig. 1


Molecular characterization of an endophytic Phomopsis liquidambaris CBR-15 from Cryptolepis buchanani Roem. and impact of culture media on biosynthesis of antimicrobial metabolites
a Colony morphology of P.liquidambaris CBR-15 on PDA and b microscopic features at ×40 magnification showing alpha and beta conidia
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4362740&req=5

Fig1: a Colony morphology of P.liquidambaris CBR-15 on PDA and b microscopic features at ×40 magnification showing alpha and beta conidia
Mentions: The endophytic fungus P.liquidambaris CBR-15 which is used in the present study was isolated from the leaf tissue of C. buchanani. Colony morphology on PDA, after 5 days at 25 °C, initially cottony, white to olive gray later on turns to gray to light brown and a radiating growth pattern, margin regular. Conidiomata eustromatic, black, spherical to irregular on the upper regions; Conidiophores: short to elongated, aseptate to septate and branched, Conidia are of two types: (a) alpha conidia hyaline, straight, aseptate, forming white to yellowish cirrhi; (b) beta conidia hyaline, filiform, straight or curved, aseptae morphology characters observed, were closely related with the genus Phomopsis, belongs to class coelomycetes (Fig. 1).Fig. 1

View Article: PubMed Central - PubMed

ABSTRACT

An endophytic fungus Phomopsis liquidambaris CBR-15, was isolated from Cryptolepis buchanani Roem. (Asclepiadaceae) and identified by its characteristic culture morphology and molecular analysis of the ITS region of rDNA and intervening 5.8S rRNA gene. The impact of different culture media on biosynthesis of antimicrobial metabolites was tested by disc diffusion assay. Polyketide synthase gene (PKS) of the endophytic fungus was investigated using three pairs of degenerate primers LC1–LC2c, LC3–LC5c and KS3–KS4c by PCR. TLC-bioautography method was employed to detect the antimicrobial metabolites. Antimicrobial metabolites fractionated with ethyl acetate extract showed significant antimicrobial activity against the test bacteria and fungi. Biosynthesis of antimicrobial metabolites was optimum as depicted by zone of inhibition from ethyl acetate extract cultured in potato dextrose broth. Strain CBR-15 was identified as Phomopsisliquidambaris and PKS genes of the fungus were amplified with LC3–LC5c and KS3–KS4c sets of degenerate primers. These findings suggest that endophytic P.liquidambaris CBR-15 harbor iterative type I fungal PKS gene domain which indicates the biosynthetic potential of endophytic fungi as producers of natural antimicrobial metabolites. The study also demonstrates the utilization and optimization of different culture media which best supports for the biosynthesis of the antimicrobial metabolites from P.liquidambaris.

Electronic supplementary material: The online version of this article (doi:10.1007/s13205-014-0204-2) contains supplementary material, which is available to authorized users.

No MeSH data available.