Isolation, Screening for Bioactivities and Identification of Selected Endophyte Fungi by Sequencing of 18S RRNA/Its Genes

Endophytic fungi occur within plant tissues such as leaves and stems of healthy plants without producing any apparent infections and symptoms. Experiments were conducted to isolate endophytic fungi from healthy medicinal plants as well as detecting any reoccurrence of any particular endophytic f...

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Main Author:	Cheah, Yoke Kqueen
Format:	Thesis
Language:	English English
Published:	2001
Subjects:	Endophytic fungi Sequestration (Chemistry)
Online Access:	http://psasir.upm.edu.my/id/eprint/8437/1/FSMB_2001_10_IR.pdf
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id	my-upm-ir.8437
record_format	uketd_dc
spelling	my-upm-ir.84372024-01-23T01:31:23Z Isolation, Screening for Bioactivities and Identification of Selected Endophyte Fungi by Sequencing of 18S RRNA/Its Genes 2001-08 Cheah, Yoke Kqueen Endophytic fungi occur within plant tissues such as leaves and stems of healthy plants without producing any apparent infections and symptoms. Experiments were conducted to isolate endophytic fungi from healthy medicinal plants as well as detecting any reoccurrence of any particular endophytic fungi predominantly in selected local medicinal plants. Endophytic fungi have also been recognised as repository of novel secondary metabolites, which have beneficial biological activities, biocontrol of insects and oligosaccharides degrading enzymes. Thus, isolated endophytic fungi were screened for their bioactive properties by using Thin Layer Chromatography and Agar Diffusion Assay. Specially engineered yeast strains i.e. UCS and UCK from Kyowa Hakko Company, Japan were used to screened for anti-tumour activity. 18S ribosomal Ribonucleic Acid (18S rRNA) I Internal Transcribed Spacer (ITS) gene sequencing were conducted to identified certain isolates. Isolation of the endophytic fungi from healthy local medicinal plants showed that 61 out of 72 (84.7%) yielded endophytic fungi. Most of the endophytes were obtained from the leaves and very few from the stems. The reoccurrence rate of the endophytic fungi was 1.39% (1 in 72). Nevertheless, no predominant endophytic fungi association with types of medicinal plant was observed. All the isolated endophytic fungi were able to degrade starch, xylan, mannan and inulin. 98.33% of the endophytic tested were able to degrade sago starch and rice starch. About 96.67% of the isolates were detected producing potato starch and starch wheat unmodified degrading enzyme. However, 95% of the endophytes produced tapioca starch and com starch degrading enzyme. Among the isolated endophytic fungi, 16.9% of them were considered important with regard to the bioactive screening results. Thus, 22 isolates from 130 isolated fungi gave satisfactory results in bioactive screening. There were 16 isolates that gave positive results in bio-activity test against UCS/UCK yeast strain. This showed that 16 out of 130 isolates produced potential bioactive compound for anti-tumour. Four out of the twenty two important isolates were identified through microscopic examination and 18S IITS gene sequencing. Isolate 12L was identified as Penicillium spp. through microscopic morphology observation. Those isolates identified by 18S IITS gene sequencing included 1B, Endothia gyrosa; 19L, Colfetotrichum gloeosporioides and 22L, Botryosphaeria ribis . BLAST programme was used as a tool to determine the homology of the sequence obtained with the database sequence. Endophytic fungi Sequestration (Chemistry) 2001-08 Thesis http://psasir.upm.edu.my/id/eprint/8437/ http://psasir.upm.edu.my/id/eprint/8437/1/FSMB_2001_10_IR.pdf text en public masters Universiti Putra Malaysia Endophytic fungi Sequestration (Chemistry) Food Science and Technology Radu, Son English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
advisor	Radu, Son
topic	Endophytic fungi Sequestration (Chemistry)
spellingShingle	Endophytic fungi Sequestration (Chemistry) Cheah, Yoke Kqueen Isolation, Screening for Bioactivities and Identification of Selected Endophyte Fungi by Sequencing of 18S RRNA/Its Genes
description	Endophytic fungi occur within plant tissues such as leaves and stems of healthy plants without producing any apparent infections and symptoms. Experiments were conducted to isolate endophytic fungi from healthy medicinal plants as well as detecting any reoccurrence of any particular endophytic fungi predominantly in selected local medicinal plants. Endophytic fungi have also been recognised as repository of novel secondary metabolites, which have beneficial biological activities, biocontrol of insects and oligosaccharides degrading enzymes. Thus, isolated endophytic fungi were screened for their bioactive properties by using Thin Layer Chromatography and Agar Diffusion Assay. Specially engineered yeast strains i.e. UCS and UCK from Kyowa Hakko Company, Japan were used to screened for anti-tumour activity. 18S ribosomal Ribonucleic Acid (18S rRNA) I Internal Transcribed Spacer (ITS) gene sequencing were conducted to identified certain isolates. Isolation of the endophytic fungi from healthy local medicinal plants showed that 61 out of 72 (84.7%) yielded endophytic fungi. Most of the endophytes were obtained from the leaves and very few from the stems. The reoccurrence rate of the endophytic fungi was 1.39% (1 in 72). Nevertheless, no predominant endophytic fungi association with types of medicinal plant was observed. All the isolated endophytic fungi were able to degrade starch, xylan, mannan and inulin. 98.33% of the endophytic tested were able to degrade sago starch and rice starch. About 96.67% of the isolates were detected producing potato starch and starch wheat unmodified degrading enzyme. However, 95% of the endophytes produced tapioca starch and com starch degrading enzyme. Among the isolated endophytic fungi, 16.9% of them were considered important with regard to the bioactive screening results. Thus, 22 isolates from 130 isolated fungi gave satisfactory results in bioactive screening. There were 16 isolates that gave positive results in bio-activity test against UCS/UCK yeast strain. This showed that 16 out of 130 isolates produced potential bioactive compound for anti-tumour. Four out of the twenty two important isolates were identified through microscopic examination and 18S IITS gene sequencing. Isolate 12L was identified as Penicillium spp. through microscopic morphology observation. Those isolates identified by 18S IITS gene sequencing included 1B, Endothia gyrosa; 19L, Colfetotrichum gloeosporioides and 22L, Botryosphaeria ribis . BLAST programme was used as a tool to determine the homology of the sequence obtained with the database sequence.
format	Thesis
qualification_level	Master's degree
author	Cheah, Yoke Kqueen
author_facet	Cheah, Yoke Kqueen
author_sort	Cheah, Yoke Kqueen
title	Isolation, Screening for Bioactivities and Identification of Selected Endophyte Fungi by Sequencing of 18S RRNA/Its Genes
title_short	Isolation, Screening for Bioactivities and Identification of Selected Endophyte Fungi by Sequencing of 18S RRNA/Its Genes
title_full	Isolation, Screening for Bioactivities and Identification of Selected Endophyte Fungi by Sequencing of 18S RRNA/Its Genes
title_fullStr	Isolation, Screening for Bioactivities and Identification of Selected Endophyte Fungi by Sequencing of 18S RRNA/Its Genes
title_full_unstemmed	Isolation, Screening for Bioactivities and Identification of Selected Endophyte Fungi by Sequencing of 18S RRNA/Its Genes
title_sort	isolation, screening for bioactivities and identification of selected endophyte fungi by sequencing of 18s rrna/its genes
granting_institution	Universiti Putra Malaysia
granting_department	Food Science and Technology
publishDate	2001
url	http://psasir.upm.edu.my/id/eprint/8437/1/FSMB_2001_10_IR.pdf
_version_	1794018749561438208

Isolation, Screening for Bioactivities and Identification of Selected Endophyte Fungi by Sequencing of 18S RRNA/Its Genes

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