The effect of various carrier mediums on the growth of trichoderma asperellum TR3 and Trichoderma sp.

Trichoderma sp. is the most common growth in soil, especially with high organic matter. The fungi are helpful as the growth trigger and potential biological agent in controlling plant diseases. The relationship between Trichoderma sp. and plants are mutualism, with plants taking benefit in terms of growth and disease control, while Trichoderma sp. gets nutrients produced by plants. The study aims to determine the ability growth of Trichoderma asperellum TR3 and Trichoderma sp. in the various carrier media forming pellets in vitro condition. The research was held at the Laboratory of the Faculty of Agriculture, Alkhairaat University Palu. The study used a Completely Randomized Design with four treatments, namely T1 = rice bran (RB), T2 = tofu dregs (TD), T3 = tapioca flour (TF), and T4 = talc powder+tofu dregs (TP+TD). The results showed that T4 (TP+TD) and T1 (RB) could be used as carrier medium in the pellet form for Trichoderma asperellum TR3 and Trichoderma sp.


Introduction
The antagonist fungi Trichoderma sp. is found in all soil types with diverse habitats.The fungi are helpful as the growth trigger and their potential as a biological agent in controlling plant diseases.In addition, Trichoderma sp. also has several advantages, such as being more straightforward in isolation, broad adaptability, easy to find in planting areas, proliferating on various substrates, having a wide range of microparasites, and not being pathogenic in plants.Trichoderma sp. is also used as a biopesticide which antagonistic to plant pathogens.The fungi known to have antagonistic ability became potential biological controllers in several plants.The relationship between Trichoderma sp. and plants are mutualism, with plants benefitting from growth and disease control, while Trichoderma sp.gets plant nutrients [1,2].
Trichoderma sp.positively influences plant roots, growth, and crop production.On the other hand, Trichoderma sp. has a preventive function against disease [3,4].Besides that, Trichoderma sp.affects microorganisms living in the soil.Trichoderma sp.affects soil structure, soil holding capacity, and water holding capacity, increasing nutrient availability and reducing the threat of drought.Apart from the roots, Trichoderma sp. also plays an endophyte in the stem and leaf tissues of annual and perennial plants [2,5].There are several reasons why the antagonist fungi were chosen as the biological control.The fungi have a high reproductive capacity and short life cycle, forming spores that last long under extreme conditions, are relatively safe to use, and are easy to produce.Another benefit of antagonist fungi is that they are compatible with a wide range of insecticides and impossible to develop resistance [2,6].
The great benefit of Trichoderma sp. is their propagation can work long-term using pellet media.Pellets contain carbohydrates, protein, and nutritional matter needed by Trichoderma sp., increasing their development.Rice bran is one of the materials used as a carrier medium for the growth of Trichoderma sp. in manufacturing pellets.Rice bran is a waste processing grain into the rice.It is quality varies depending on the rice variety.Rice bran is one of the results of a mill in producing rice [7].Rice bran contains various nutrients such as oil, protein, carbohydrates (mainly starch), betaglucan, and pectin.In addition, rice bran contains 15-20% oil, 12-16% protein, 34-52% available carbohydrates, 7-11% crude fiber, and 7-10% ash [8].Based on the description, the research was 1230 (2023) 012090 IOP Publishing doi:10.1088/1755-1315/1230/1/012090 2 conducted on the growth ability test of Trichoderma asperellum TR3 and Trichoderma sp. on various carrier media to determine the best carrier medium for the growth Trichoderma sp.

Methodology
The study used a modified method [9].Pellet was made from Trichoderma asperellum TR3 from Ratnawati collection (2019, unpublished) and Trichoderma sp.The research activities were conducted at the Laboratory of the Faculty of Agriculture, Alkhairaat University, Palu, Central Sulawesi.Trichoderma asperellum TR3 and Trichoderma sp.rejuvenated on potato dextrose agar medium.

Carrier Medium Composition
The material used as a carrier medium growth of Trichoderma asperellum TR3 and Trichoderma sp. is rice bran, tofu dregs, tapioca flour, talc powder mixed with garlic flour, brown sugar, and sterile water as needed (Table 1).

Cultivation of Trichoderma asperellum TR3 and Trichoderma sp.
Each portion of the carrier medium, based on Table 1, was wrapped with aluminum foil, then sterilized inside an autoclave.The antagonist fungi Trichoderma asperellum TR3 and Trichoderma sp.age used the research at 7 th days old.The fungi were made in suspension and diluted to reach a mass density of 2.57 x 10 8 .
Each treatment is mixed with Trichoderma asperellum TR3 and Trichoderma sp. according to a predetermined dosage.Sterile water was added according to the dosage, then mixed well.Each treatment is put into a plastic straw using a spatula.All treatments have been put into plastic straws, kept for 7 th days under light, and air-dried utilizing a fan.

Planting Trichoderma asperellum TR3 and Trichoderma sp.
Pellet planting was carried out in a cup containing potato dextrose agar with four treatments; T1 (RB), T2 (TD), T3 (TF), and T4 (TP+TD).A pellet was planted in the potato dextrose agar.Each treatment was repeated five times.The planting process was carried out in the laminar airflow using bunsen in the aseptic condition-the observation of fungi by measuring the diameter of the colony every day in 7 th days.

Result
The growth average of Trichoderma asperellum TR3 and Trichoderma sp.colony was shown in Table 2. Based on Table 2, the antagonist fungi Trichoderma asperellum TR3 and Trichoderma sp. were stored on the 7th day and had different average colonies growth in each treatment.The treatments stored in 7 days showed the development of colonies of Trichoderma asperellum TR3, and Trichoderma sp. from T4 (TP+TD) was able to cover the entire surface of the petri dish.The T4 had the highest average value on day 7 th .It declared the carrier medium better for Trichoderma asperellum TR3 and Trichoderma sp growth.Besides T4, The best carrier medium for the growth of Trichoderma asperellum TR3 and Trichoderma sp. is T1 (RB), T3 (TF), and T2 (TD), respectively.

Discussion
Trichoderma sp. is a biological agent that has multi-functional responsibilities.Fungi are commonly used in plant protection, biological fertilizer, and disease prevention.Unfortunately, the fungus is less practicable to utilize in the field because it demands a lot of containers and labor.Trichoderma asperellum TR3 and Trichoderma sp. have been produced more efficiently in the pellet form from various acceptable carrier media, enhancing the fungi's growth capacity.
Based on Duncan, Multiple test results showed the treatment for growth Trichoderma asperellum TR3 and Trichoderma sp.stored for 7 th days significantly different.The fact can be seen from the effects of observation parameters in Table 2. On the 7 th -day consecutive observation growth of Trichoderma asperellum TR3 and Trichoderma sp. in T4 (TP+TD) containing: talc powder, tofu dregs, garlic flour, brown sugar, and sterile water showed better results for all observation parameters.The nutrient combination in T4 (TP+TD) is suitable for Trichoderma asperellum TR3 and Trichoderma sp growth.The colonies grew in the potato dextrose agar until the 7 th day of observation.This showed better results compared to other treatments.Talc powder is the hydro magnesium silicate formed from the natural minerals magnesium, silicon, oxygen, and hydrogen [10].Tofu dregs contain more than 20.13% protein, 57.26% carbohydrates, and 19.8% crude fiber [11][12][13].It means they can be used as a carrier medium for the growth of Trichoderma.Flour made from tofu dregs has an excellent nutritional composition with a water content of around 10.43%, protein of 23.25%, fat of 5.87%, carbohydrates of 26.92%, 17.3% ash, and 16.53% crude fiber [14,15].Based on the information, talc powder and tofu dregs are very suitable as the carrier medium for growing Trichoderma asperellum TR3 and Trichoderma sp.
In observing the colonies growth of Trichoderma asperellum TR3 and Trichoderma sp.carried out for 7 th days, all treatments still grow daily.The T3 (TF) has an average of 1 st day (2.54 cm), 2 nd days (4.74 cm), 3 rd days (5.3 cm), 4 th days (6.84 cm), and 5 th days (6.92 cm) higher than T1 (RB), T2 (TD) and T4 (TP+TD).However, on the 6 th day, T3 (TF) did not increase in the size of colony growth, which remained at an average of 6.92 cm, while the other treatments continued to experience colony growth.This is presumably due to contamination in the T3 (TF), resulting in the growth of Trichoderma asperellum TR3 and Trichoderma sp.colonies becoming unable to increase.The growth of T3 (TR) stopped on the 5th day of observation.Contamination is the common condition where other elements contaminate microbes in the laboratory and then break the data observation.
All treatments experienced contamination on the 3 rd day of observation.However, the treatment grew the colonies well in T4 (TP+TD).The colonies continued to spread and filled the entire surface of the potato dextrose agar until the 7 th day of observation.It could be stated as a treatment that tended better because the fungi were not affected by contamination.

Conclusion
T4 (Talc powder + Tofu dregs) and T1 (rice bran) showed the best treatments.The fungi covered the entire surface of the potato dextrose agar on the 7 th day's observation.Also, the treatment had the highest average colony growth value on the 7 th day (8.4 cm) and was unaffected by contamination.The treatment T4 and T1 can be used as carrier medium in the form of pellets for antagonist fungi Trichoderma asperellum TR3 and Trichoderma sp.

Table 1 .
Carrier medium composition of antagonist fungi.Experimental Design The research was conducted in vitro, containing four treatments with five replications.The observed variables were the growth of Trichoderma asperellum TR3 and Trichoderma sp. on pellets incubated in Petri dishes.Data about the growth of Trichoderma asperellum TR3 and Trichoderma sp. was analyzed by Completely Randomized Design and meant comparison with Duncan's Multiple Range Test at α = 5%.

Table 2 .
Colonies growth average of Trichoderma asperellum TR3 and Trichoderma sp. on the various carrier medium (cm).Numbers in the same column followed by the same letter are not significantly different according to Duncan's Multiple Range Test α = 5%.