The Potential of Trichoderma sp . as Biological Agent to Support the Germination of Corn Seeds ( Zea Mays )

27 Abstract -This research is motivated by the fact that corn seeds in storage are often characterized with low seed viability and infected by diseases so that the seeds grow abnormally and even die. Corn seeds have thick/hard cell walls. It is acknowledged that the corn seed walls contain cellulose, which can be broken down by microorganisms. One such microorganism is the Trichoderma fungus, which is also a biological agent. This study aims to analyze the effect of the biological agents Trichoderma sp. on corn seed germination and identify the best concentration of Trichoderma sp. against corn seed germination. The corn seeds used in this study were locally grown, which had been stored for 6 months. The seed coating (bio seed coating of local corn) was tested at several concentrations of Trichoderma sp. (treatment), namely without treatment (control), 5g, 10g, 15g, and 20g. Each concentration was repeated 4 times, using several parameters to analyze normal sprouts and germination capacity. The analysis results confirmed the potential of Thrichoderma biological agents on corn seed germination. The T1 treatment with 5g Trichoderma biological agents significantly improve corn seed germination.


I. INTRODUCTION
Farmers on agricultural land in North Maluku mostly cultivate corn. Corn is a food crop that once dominated food staple in 2016-2019 in West Halmahera (Jailolo) [7]. The local government expects that farmers can innovate to increase corn productivity to reach production goal of 12 tons/ha, while farmers can only produce 5 tons/ha [1]. The need for fast planting material to meet the substantial demand will be constrained by the duration of seed germination. The use of seeds as plant propagation needs special treatment before the seeds are planted because they have a very hard skin structure. The entire seed germination takes at least three months if the seeds do not obtain special treatment. This period is called the dormancy period, which is caused by the physical structure of the seed, which eventually inhibits water imbibition [2]. Contends that physical dormancy can be distorted by microorganisms, in that the activity of these fungi can help shorten the dormancy period. Trichoderma sp. is a fungus capable of producing cellulosic enzymes. Cellulosic enzymes are able to degrade cellulose located in plant cell walls. Trichoderma sp. is also known to play the role as a biological fertilizer in decomposing organic matter in the soil as well as a biological agent in controlling soil borne diseases or root diseases [9] . In addition, according to [4], the fungus grows in the testa seed erodes and cracks the hard skin. This can reduce mechanical resistance to seed germination with physiological dormancy. One way to bring biological agents to seeds is through seed coating. This method is considered better because farmers can plant seeds without administering additional treatment [10]. According to [3], seed coating is a method to improve seed quality by adding organic matter to the coating formula. Seed coating can control and improve germination and has the potential to be used for inoculating seeds with living microorganisms, which can protect seeds from pests and plant diseases during nursery phase and outset of growing season, increase seedling vigor, and reduce the use of chemical pesticides during planting [8].

II. RESEARCH METHODOLOGY
The research was carried out in the Agrotechnology laboratory of the Faculty of Agriculture, UNKHAIR. This study employed a completely randomized design (CRD). The study involved several treatments involving Trichoderma sp. at different concentrations, ranging from 0 g, 5g, 10g, 15g, to 20g. Each treatment was repeated 4 times, so there were 20 experimental units, and one treatment involved 20 corn seeds, resulting in a total sample of 400 corn seeds. Description: T0 = control bio seed bio seed coating T1 = 5g Thrichoderma T2 = 10g Thrichoderma T3 = 15g Thrichoderma T4 = 20g Thrichoderma

A. Research Procedure The Preparation of Corn Seeds
Corn kernels used in this study were locally grown by corn farmers in Ternate city. These seeds had large size as these were

Original Paper
The Potential of Trichoderma sp. as Biological Agent to Support the Germination of Corn Seeds (Zea Mays) sorted in consideration of involving physiologically matured seeds. The seeds used in the present study had been stored 6 months.

The Production of Trichoderma sp.
Upon preparing Trichoderma sp. seed coating material, pure cultures of Trichoderma were obtained from agricultural soil cultures. For further dilution aiming at 107 to obtain pure isolates, these seeds were isolated in PDA media, taken and propagated in rice media. The finished Trichoderma sp was separated according to the treatment and processed to seed coating.

Bio Seed Coating of Thrichoderma
Bio seed coating material mixed with tapioca was then coated on the walls of corn seeds containing 14% moisture. The procedure began with coating corn seeds with Trichoderma according to treatment. These seeds were then left for ± 5 minutes and placed in a container (germinator) according to treatment. Observations were made for 14 days.

B. Observation Parameters
The observation of Trichoderma viride fungal responses toward the viability of corn seeds includes normal germination, abnormal germination, germination, maximum growth potential, growth speed, and vigor index.

Normal Sprouts (NS)
Normal sprouts show the potential to develop further into normal plants (Stefan 2013). The formula for calculating normal sprouts is as follow: Germination Rate (GR) Germination rate is determined based on the percentage of normal germination on day 5 (observation I) and day 7 (observation II) after the seeds germinated. The final germination is calculated by the formula: The criteria for selecting normally germinated seeds are those whose main structures (root system, embryonic shaft called epicotyl and hypocotyl, and cotyledons) show the ability to develop into normal plants if planted in the field in a suitable environment.

Maximum Growth Potential (MGP)
Maximum growth potential was measured by counting the number of sprouts that grew normally or abnormally at 7 DAG's (days after germination). The maximum growth potential is calculated by using the formula below: Growth Simultaneity (GS) Simultaneous growth was calculated based on the percentage of normal sprouts at 4 DAP. Observations were made on the number of normal seedlings between the first and second counts. Simultaneous growth is calculated by the formula:

Vigor Index (VI)
The observation of the vigor index was carried out by focusing on the number of normal sprouts on the first count, namely day 5 (observation I) (ISTA, 2010).

A. The Potential of Trichoderma sp. as Natural Agent toward Normal Sprouts of Corn Seeds
The results of the analysis showed that Trichoderma bio seed coating generated satisfactory effect on the maximum growth potential. The maximum growth potential analysis data is presented in Figure 1: The highest number of normal sprouts was found in T1 with 5 gr Trichoderma, which was evident from the 4 to day 10 with a value of 77% -98%. This presence of normal sprouts resulted from the germination assisted by Trichoderma. Trichoderma is a biological agent that can help seed germination because this fungus contains cellulose enzymes that can erode the cell walls of the seed coat to speed up the plant germination [11,12]. Trichoderma is a mold or a type of fungus capable of producing cellulosic enzymes. Cellulosic enzymes are able to degrade cellulose, which is located in plant cell walls [5]. According [4], fungus grows in the testa seed erodes and cracks the hard skin, thereby potentially reducing mechanical resistance to seed germination with physiological dormancy.

B. The Potential of Trichoderma sp. as Natural Agent toward
Germination Rate of Corn Seeds The analysis results of the germination showed that Trichoderma generated a fine effect. The data backgrounding this finding is shown in Figure 2.  Figure 2 shows the average germination of corn seeds in treatment T0, which is identified as the lowest average, namely 47.5%. This is because T0 involved only some germinated seeds, so that the germination rate only reached 47.5%. The seeds in the T0 treatment did not grow. Due to the damage to the seeds, there were even undamaged seeds which could not germinate in the near future. Seeds of this nature are commonly referred to as fresh seeds, which do not grow. These seeds are usually still in dormancy. Usually the dormancy period is caused by several factors, one of which is a resistant or impermeable seed coat, for that an enzyme is needed that is able to erode the seed coat so that it can accelerate seed germination.

C. The Potential of Trichoderma sp. as Natural Agent toward Maximum Growth Potential of Corn Seeds
The results of the analysis showed that Trichoderma bio seed coating generated satisfactory effect on the maximum growth potential. The maximum growth potential analysis data is presented in Figure 3: The average germination of corn seeds in treatment T0 was the lowest, namely 47.5%. This was because T0 only showed some seeds germinated, which only reached 47.5%. The seeds in T0 treatment did not grow. Because of the damage to the seeds, there were even undamaged seeds, which could not germinate in the near future. These seeds are commonly referred to as fresh seeds which do not grow. These seeds are usually still in dormancy. The dormancy period is generally driven by several factors, one of which is a resistant or impermeable seed coat. To that end, an enzyme is needed to erode the seed coat so that it can accelerate seed germination. Trichoderma is a fungus that has cellulosic enzymes that are able to degrade the seed coat to accelerate germination. According to [5], Trichoderma is a mold or a type of fungus that is capable of producing cellulosic enzymes. Cellulosic enzymes are enzymes which are able to degrade cellulose located in plant cell walls.
In T1, T2, T3 and T4 treatments, the germination rate was not significantly different because these also involved Trichoderma to assist the germination. T1 with 5g Trichoderma had the highest percentage of germination rate. By implication, this concentration involving Trichoderma has the potential to increase the germination rate of corn seeds. This is because during germination the plumules are not inhibited by the Trichoderma layer , which covers the corn seeds.

D. The Potential of Trichoderma sp. as Natural Agent toward Growth Simultaneity of Corn Seeds
The analysis results showed that the effect of the Trichoderma bio seed coating on the simultaneous growth was significant.
Simultaneous growth data is presented in Figure 4 below.  Figure 4 shows the test results of T0 having the lowest level of simultaneous growth with a percentage of 46.25%, this shows that in the treatment the percentage of simultaneous growth of corn seeds is very low. There is a low percentage because many seeds do not grow at the same time, seeds which grow at the same time because the seeds have low viability. Seeds with low viability usually have been stored for a long time and will regress. Seed deterioration is a decline in the physiological quality of seeds, which can cause overall changes in the seeds, both physical, physiological, and chemical which results in a decrease in seed viability [6].

E. The Potential of Trichoderma sp. as Natural Agent toward
Vigor Index of Corn Seeds The analysis results showed that the effect of Trichoderma bio seed coating on the vigor index was markedly significant. Vigor index is presented in Figure 5.  Shows T0 (without Trichoderma) has a very low effect and is very significantly different from all treatments with an average of 42.5% while T1 (5g Trichoderma) has the highest value with an actual vigor index average of 96.25% of all treatments. The vigor index is a comparison between the number of normal sprouts on the first count and the total number of seeds germinated. The results of the t-test test showed that the lowest percentage of vigor index was in T0 (without Trichoderma) with a percentage of 42.5%, which was significantly different from the other treatments with Trichoderma at 90%. This was because the T0 treatment did not contain the required enzymes to stimulate the sprout growth, so the percentage of normal sprouts was very low.

IV. CONCLUSION
The present study has acknowledged the potential of Trichoderma sp. as biological agent on the germination of corn seeds. The findings have demonstrated very significant effect on corn seed viability particularly in terms of normal and abnormal germination, growth potential, level of uniformity, and vigor index, each of which has been confirmed in treatment involving Trichoderma sp. Seeds stored for 6 months can still be used providing that these are treated using Trichoderma with the seed coating method at 5g concentration, which has been found significantly better than other concentrations. Giving Trichoderma can also keep the seeds free from disease during storage.