Antagonism Trichoderma harzianum Rifai in Suppresing the Intensity of Antraknosa ( Colletotrichum capcisi Sydow.) Disease

Cayenne pepper ( Capsicum frutescens L.) is a horticultural plant with considerable commercial worth and nutritional value. Cayenne pepper is one of the most important horticultural products at the national level, thus its production must be increased adequately. However, the obstacle of cayenne pepper production caused by anthracnose disease is a frequent stumbling block in the manufacturing of cayenne pepper. Anthracnose disease or fruit rot caused by the fungus Colletotrichum capcisi Sydow can result in yield losses ranging from 20-90%. Trichoderma harzianum Rifai. is a soil saprophytic fungus that naturally can be used as a biological agent, because it has antagonism against pathogens in the form of competition for space and nutrients, mycoparasites and antibiosis. The experimental results in the laboratory showed that the biological agent Trichoderma harzianum Rifai was able to suppress the development of anthracnose disease.


Introduction
Because Indonesian people are the most avid consumers of chili in the world, chili is one of the most essential components of traditional Indonesian cuisine (Paulus & Ellen, 2016). Cayenne pepper is widely used as raw material for the food industry such as sauces, chili powder, and the pharmaceutical industry (Sofiarani & Ambarwati, 2020). In addition, cayenne pepper is consumed in fresh or processed form which is generally used as an additional ingredient and flavoring in cooking to enhance the taste of food.
Because of its spicy taste and distinctive aroma, cayenne pepper is favoured with the substance capsaicin that *Corresponding Author: Yenny muliani; Email: yennymuliani62@gmail.com, Department of Agriculture, Universitas Islam Nusantara Copyright © 2022 Al-Hayat: Journal of Biology and Apllied Biology gives chili its spiciness. Capsaicin is useful for regulating blood circulation; strengthening the heart, pulse, and nerves; preventing flu and fever; raising enthusiasm in the body; and reducing rheumatism (Paulus & Ellen, 2016).
In Indonesia, the demand for cayenne pepper is rising in line with the country's population growth and the wide range of dishes and menu items that include this flavor (Irfandi, et.al, 2021). Various attempts to boost cayenne pepper production in Indonesia have been made in the past and continue to be made in the present to suit local consumption demands. Efforts to enhance cayenne pepper output often meet roadblocks, one of which is the interruption of cayenne pepper production, both in quality and quantity, caused by anthracnose disease.
Anthracnose or fruit rotis one of the important diseases in cayenne pepper plantscaused by the fungus Colletotrichum capcisi Sydow (Habibi & Wijayanto, 2019 To prevent losses, efforts must be made to control anthracnose. The most common method of control used by farmers is the application of synthetic fungicides, often to combat anthracnose. Farmers use fungicides with active ingredients propineb, mankozeb, and benomyl. However, continual and irresponsible use of synthetic fungicides will leave residues on plants, harm the environment, and be hazardous to human health (Amelia, Marsuni, & Budi, 2020). Because using synthetic fungicides gives negative impact for the environment, one way to stop plant diseases without harming the environment is to use biological agents; for instance, by using fungi that are antagonistic to Colletotrichum capsica.
Trichoderma harzianum Rifai. is an antagonist fungus that naturally can be used as a biological agent, because it has antagonism against pathogens in the form of competition for space and nutrients, mycoparasites and antibiosis. This mushroom is frequently used in the control of plant-disturbing organisms (Safitri, Martina, & Roza, 2019). In addition, this fungus also offers numerous benefits, including simple isolation, broad adaptability, easy detection in the planting area, the capacity to grow swiftly on varied substrates, a diverse spectrum of microparasitism, and the fact that it is not pathogenic to plants (Ministry of Agriculture, 2019).

Research Methods
The research was carried out in Neglasari Village, Ciherang Village, Pacet District, Cianjur Regency. The region is at an elevation of 1,080 -2,962 masl, with an average temperature ranging from 17°C to 26°C.

Figure 1
Making PSA (Potato Sucrose Agar) Media

Figure 3
Propagation of Trichoderma harzianum Rifai. on Rice Media Calculating Spore Density

Trichoderma harzianum Rifai Antagonist Test
Antagonism test against Colletotrichum capsici was carried out in vitro by placing the two fungal colonies facing each other with a distance of 3 cm as shown in Figure 4 below.
Subsequently, it was incubated at room temperature and observed for its inhibitory power for ±7 x 24 hours.

Multiple Culture Test on Antagonist Test of Trichoderma harzianum Rifai
Antagonism Trichoderma harzianum….

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After inoculation, observations were taken daily by measuring the radius of the fungal colonies from the first day to the fifth day. Measurements were made using a millimeter ruler. The percentage growth inhibition data is calculated based on the following formula (Ningsih, Hastusi, & Listyorini, 2016) :

Concentration Test
The concentration test was carried out to determine the range value of several concentrations of Trichoderma harzianum Rifai applications. against the intensity of the attack of the pathogen Colletotrichum capsici in vitro and will be used as a treatment for the application of Trichoderma harzianum Rifai on cayenne pepper plants.
In vivo, the concentrations of Trichoderma harzianum Rifai tested were 10 g/l, 15 g/l, 20g/l and 25 g/l. The concentration test is carried out with the following working steps: 1. Measure each petri dish with a distance of 2.25 cm from each boundary for Trichoderma harzianum Rifai solution. and Colletotrichum capsici isolates and controls. Then mark the measured cup using a marker and ruler.

Figure 5
Trichoderma harzianum Rifai . 6. Incubate at room temperature (± 25ºC -30ºC), the inhibition was observed every day for 7 x 24 hours. observe and calculate the effectiveness of the inhibition level by measuring the diameter of the mycelia growth.

Trichoderma harzianum Rifai Antagonist Test
The results of the Antagonist Trichoderma harzianum Rifai against Colletotrichum capcisi Sydow disease using the dual culture method shows that the biological agent Trichoderma harzianum Rifai. effect in inhibiting the growth of Colletotrichum capcisi Sydow up to 81% on day 5 after incubation. This result can be shown in the The inhibition of the antagonist fungus Trichoderma harzianum Rifai against the fungus Colletotrichum capcisi occurs by several mechanisms, including competition for space and nutrients between pathogens where Trichoderma harzianum Rifai can suppress the development of pathogens in their life cycle, therefore the nutrients needed by pathogens to thrive are not met. Furthermore, the mechanism of parasitism is able to wrap around the hyphae of pathogenic fungi. Together with the coiling of the hyphae, enzymes are released that are able to remodel the cell walls of the hyphae of pathogenic fungi. The next mechanism is Antibiosis, in which it not only produces fungal cell wall enzymes, but also produces antibiotic compounds belonging to the furanon group which can inhibit the growth of spores and hyphae of pathogenic fungi (Ningsih, Hastusi, & Listyorini, 2016) (Suwahyono & Wahyudi, 2004.
The results of an in vitro concentration test with Trichoderma harzianum Rifai at 4 different concentrations are shown in table  2   Table 2.

Concentration Test Results of Trichoderma harzianum Rifai.
The results of the determined concentration test showed that at 1 DAI to 6 DAI the inhibition of Trichoderma harzianum Rifai was observed. In the four tests, namely 10 grams; 15 grams; 20 grams; 25 grams, the inhibitory power is not much different, but 25 grams/liter of water is a high concentration of inhibition compared to a concentration of 10 grams; 15 grams; 20 grams; 25 grams with the growth of hyphae diameter reaching 9 cm.

Conclusions
The biological agent Trichoderma harzianum Rifai is able to inhibit the growth of Colletotrichum capcisi Sydow up to 81% on day 5 after incubation. The results of the concentration test showed that on the fourth day, the highest concentration was 25 grams with the growth of hyphae diameter reaching 9 cm.