Characterization of Trichoderma isolated and their lignocellulolytic activities

• Main Author: Azriah Asis
• Format: Thesis
• Language: English; English
• Published: 2017
• Subjects: QK504-(638) Cryptogams
• Online Access: https://eprints.ums.edu.my/id/eprint/38583/1/24%20PAGES.pdf; https://eprints.ums.edu.my/id/eprint/38583/2/FULLTEXT.pdf; https://eprints.ums.edu.my/id/eprint/38583/
• Institution: Universiti Malaysia Sabah

Trichoderma is a cosmopolitan fungus that prevalent in the soil and other diverse habitats. It has gained vast economical importance's because industrial enzymes production, antifungal, antibiotics, biocontrol agents and plant growth promoter. Trichoderma species produced the lignocellulolytic enzymes activities that assisted in the degradation of woody lignocellulose materials for industrial applications. Therefore, an accurately identification of Trichoderma isolates at the species level is highly desirable. In this study, the main aim to accurately identify of Trichoderma isolates at the species level based on the morphological characteristics, lignocellulolytic enzyme activities and multilocus gene sequencing based on the Internal Transcribed Spacers 1 and 2 (ITS1 and ITS2) regions of the rDNA, a partial sequence of the Translation Elongation Factor 1-alpha ( tefl) and calmodulin (ea/). A total of 53 isolates of Trichoderma were isolated from wet paddy field in Tuaran, 5abah, Malaysia. On the combination of morphological characteristics and multilocus gene sequencing analysis were positively identified three Trichoderma species, namely T. asperellum, T. harzianum and T. reesei. The phylogenetic relationships were constructed based on the Internal Transcribed Spacers 1 and 2 (ITS1 and ITS2) regions of the rDNA gene, a partial sequence of the Translation Elongation Factor 1-alpha (tefl) and calmodulin gene (ea/) by using UPGMA method and found three sections such as T. asperellum in the "Trichoderma'' section, T. harzianum in the "Pachybasium'' section, and T. reesei in the "Long1brachiatuni' section. Mycelial growth and biomass yield among three species isolates were examined on Potato Dextrose Agar and Potato Dextrose Broth, respectively, with different temperatures; 20, 25, 30, 35 and 40 °C. T. asperellum isolates were grown very well of mycelia growth and produced the highest biomass yield at 30 °c, followed by T. harzianum and T. reesei. The lignocellulolytic activities were assayed based on their ability to develop dark brown pigments, yellow halo zone, and clear white zone on tannic acid media (TAM) for lignin, Jensen Media (JM) for cellulose and modified Melin-Nokrans media (MMNM) for starch. The diameters of halo zones were measured for the analysis of their ability in degrading lignin, cellulose, and starch. The best seven Trichoderma isolates [51(9)10-1(3), E3(6)10-1(2), W2(2)10-1(2), 53(1)10-1(1 ), N2( 4)10-2(3), N2(2)10-1(2) dan 53(6)10-1(2)] were found the potential lignocellulolytic agents based on the diameter of dark brown pigments and halo zones formed. Trichoderma species are found to synthesize polyphenol oxidase, endoglucanases, and are able to hydrolyze starch to glucose in the three different media. Thus, the potential Trichoderma isolates can be further applied as biocontrol agents in controlling disease and increasing yield of agricultural crops.