The Effect of Trichoderma Biourine Application on Growth, Occurrence of Fusarium Wilt Disease and Yield of Several Shallot Varieties

Biourine is a liquid fertilizer that contains complete elements, namely nitrogen, phosphorus, and potassium in small amounts as well as zinc, iron, manganese, and copper. Biourine can provide an increase in plant yields that is almost the same as plant fertilizers, besides that it can control Fusarium wilt disease. This study aims to determine the effect of Trichoderma biourin application on plant growth, Fusarium wilt disease and onion yield. The research was conducted using an experimental method in Senteluk Village, Batu Layar District, West Lombok Regency, West Nusa Tenggara using a Split Plot Design consisting of 2 factors. As the main plot, the shallot varieties consist of three levels, namely Bali Karet, Ampenan and Keta Monca, while as a sub-plot, the Trichoderma biourin application method consists of four levels, namely: without biourine, spraying the soil surface, spraying seed tubers, and spraying on plants 21 days after planting. The treatment was a combination of shallot varieties and Trichoderma biourine application method, each of which was repeated three times, so there were 36 experimental units. The results showed that: (a) The application of liquid biourine by spraying the soil surface, spraying seed tubers and spraying on plants 21 days after planting could increase the growth and yield of shallot bulbs. (b) The Bali Karet shallots varieties are more resistant to Fusarium wilt disease when compared to Ampenan and Keta Monca varieties. Keywords— Biourine, Trichoderma, Fusarium wilt disease, shallot, Bali Karet.

Fungal attack F. oxysporum f. sp. cepae can cause shallots to wilt quickly, leaves turn yellow and twist and the base of the stem rots. Fusarium wilt disease has caused damage and reduced tuber yield by up to 50% [4]. Attacks on plants if symptoms like this are found, then the plants are removed and destroyed [5]. Fusarium wilt disease develops in shallot planting centers in NTB starting from West Lombok, East Lombok, Sumbawa, and Bima which causes damage and reduces the yield of tubers by more than 45% [6].
Fusarium wilt disease on shallots is very difficult to control, because this fungus has chlamydospores which are structures that can survive in the soil as a saprophyte for about three to four years even without a host plant [7]. Thus, it is necessary to find an alternative to control Fusarium wilt that is effective and environmentally friendly. One control technique that has good prospects is biological technology using biourine fermented with Trichoderma fungus and the use of shallots varieties that have induced resistance to Fusarium wilt disease.
Biourine is a liquid fertilizer that contains complete elements, namely nitrogen, phosphorus, and potassium in small amounts as well as zinc, iron, manganese, and copper. Biourin can provide an increase in plant yields that is almost the same as plant fertilizers [8]. One of the microbes used for biourin fermentation is Trichoderma spp. Biourine containing Trichoderma spp. able to stimulate the growth of mustard greens when compared to mustard plants without being given biourine, besides that biourine containing Trichoderma spp., has the potential to protect mustard plants from clubroot disease (Plasmodiophora brassicae), this disease is an important disease in mustard and cabbage plants [9].
Trichoderma fungi isolated from the rhizosphere of shallot plants were reported to be effective in controlling the fungus F. oxysporum f. sp. cepae in vitro with inhibition percentage of 45% [10]. The fungus T. harzianum in suppressing the growth of the fungus F. oxysporum f. sp. cepae through the mechanism of competition for space and nutrients, mycoparasites and antibiosis [11]. In a greenhouse experiment, it was reported that the fungus T. harzianum was able to inhibit the incidence of Fusarium wilt disease in shallots up to 75% [12].
Several reports explain that the Trichoderma fungi is not only used for biourine fermentation, but also for the fermentation of other materials. The fungus T. harzianum used to ferment liquid biocompost from cow dung applied to vanilla plants can control Fusarium wilt disease [10]. The fungus T. harzianum applied to soybeans could inhibit the development of Fusarium wilt disease [13]. The fungus T. harzianum can control Fusarium wilt disease on banana plants [10]. Fusarium wilt disease in maize can be inhibited by the fungus T. harzianum [14]. The fungus T. harzianum is effective in controlling Fusarium wilt disease in soybeans [15]. The fungus T. harzianum was able to suppress Fusarium wilt disease in shallots [16]. The use of the fungus T. harzianum in the form of a tablet bioactivator formulation of 15 g/pot effectively controlled the fungus F. oxysporum f.sp. cepae on shallots reached 42.26% [17] and was able to increase plant growth and yield of shallots [18].
Induced resistance is the resistance of plants to pathogen infection because plants have been infected by other microorganisms before, both of the same type or of other types. Induced resistance can also occur after plants are inoculated early with biotic elicitors (avirulent, nonpathogenic, saprophytic microorganisms) [19]. Fusarium wilt control using onion varieties that have induced resistance have good prospects. The results of the preliminary study showed that the varieties of Bali Karet, Ampenan varieties, Keta Monca varieties, Bima Brebes and Super Philip varieties induced with the fungus T. harzianum caused immunity to Fusarium wilt disease [12] and induced resistance to Fusarium wilt disease [16].
The effect of Trichoderma biourine application in increasing growth and yield of several varieties of shallots in the field has never been studied. Therefore, a research was conducted on "The Effect of Trichoderma Biourine Application on the Growth and Yield of Three Shallot Varieties".

Experimental Design
The study used an experimental method in Senteluk Village, Batu Layar District, West Lombok Regency, West Nusa Tenggara using a Split Plot Design consisting of 2 factors. As the main plot, the shallot varieties consist of three levels, namely Bali Karet, Ampenan and Keta Monca, while as a sub-plot, the Trichoderma biourine application method consists of four levels, namely: without biourine, soil surface spraying, seed tuber spraying, and spraying on plants 21 days after planting. The treatment was a combination of shallot varieties and Trichoderma biourine application method, each of which was repeated three times, so there were 36 experimental units.

Experiment Execution
The production of Trichoderma biourine is carried out as follows: cow urine is collected in a holding tank. Furthermore, the standard solution of Trichoderma was put into a urine reservoir, then closed the fermentation container, and incubated for 4 weeks. Open the lid of the container once a week and stir for 15 minutes. After 4 weeks, the circulation was carried out using a ladder for 24 hours to remove the ammonia element which is pathogenic for plants. Trichoderma fermented biourin is ready to be applied to shallot plants.
The shallot seeds used were the Bali Karet, Keta Monca and Ampenan varieties purchased from seed breeders. Shallot seeds that are good to use are healthy and quality seeds with a shelf life of 2 months and there are visible growing points on the roots. The day before planting the seeds are cut off about part.
Tillage was carried out using a hoe to level the soil and making experimental plots with a size of 2 m × 4 m for each treatment plot. After processing the soil, basic fertilization is carried out using Phonska fertilizer of 100 kg/ha (50% of the recommendation). Basic fertilizer application was carried out by immersing it next to the planting hole, then the experimental plot was covered with plastic mulch.
The application of biourine was carried out according to the treatment, namely: by spraying the soil surface before installing plastic mulch, spraying shallot bulbs for 30 minutes, and spraying shallot plants after 21 days. Planting is done by inserting shallot seed bulbs into a hole with a depth of 2 cm and the hole is covered again with soil. Planting is done with a spacing of 20 × 20 cm.

Variable Observation
Observation of disease incidence was carried out by counting the number of wilted plants, observations were made from the age of 7 days after planting (DAP) until the shallots plants were 35 DAP. Disease incidence (%) is calculated using the following formula: where : Observations of growth components, namely plant height and number of leaves of shallot plants were carried out from the age of 7 DAP to 35 DAP. Observations of yield components were carried out at harvest, namely at the age of more than 70 DAP. Observation of harvested dry shallot bulb weight was carried out by weighing the weight of bulbs at harvest per plot and then converted to hectares. Observation of the weight of stored dry shallots was carried out by weighing all parts of the plant in a dry state or after being stored in a wind-dried state for one month.

Data Analysis
Observational data were analyzed using Diversity Analysis with a significance level of 5% and further tested using the Honest Significant Difference test or HSD at the same significant level.

III. RESULT AND DISCUSSION
The results of the analysis of variance showed that the treatment of Trichoderma biourine application and shallot varieties were significantly different, while the interactions did not show significant differences in the incidence of Fusarium wilt disease, plant height growth and the number of shallots and shallot yields, namely the number of tillers, dry shallot bulb weight. harvest and weight of dry shallot bulbs stored.

Occurrence of Fusarium Wilt Disease on Shallots
The results of further tests on the effect of Trichoderma biourine application on the incidence of Fusarium wilt disease at the age of 7 DAP to 35 DAP using the 5% BNJ test are presented in Table 1.  Notes: 1) The numbers in each column followed by the same letter are not significantly different.
In Table 1, it can be seen that the application of Trichoderma biourine significantly affected the incidence of disease in shallot plants at the age of 7 DAP to 35 DAP. Trichoderma biourine which was applied by spraying the soil surface, spraying seed tubers and spraying on plants after 21 days could reduce the incidence of Fusarium wilt disease, while in control or without Trichoderma biourine treatment the incidence of Fusarium wilt at 35 DAP reached 50%. The low incidence of Fusarium wilt disease in shallots after the application of Trichoderma biourine was due to the population of Trichoderma spp. in the rhizosphere increased markedly. The results of observations of the population of Trichoderma fungi in the rhizosphere showed an increase in the population in all biourine treatments, namely an average of 43.00 x 10 3 propagules/g soil, while in the control there was no Trichoderma spp. [20]. The fungus Trichoderma in soil is able to inhibit the development of disease-causing pathogens by means of competition both in terms of space and nutrients. The fungus Trichoderma can use various nutrient sources for growth by destroying cellulose, starch, lignin, and other soluble compounds such as protein and sugar [21]. In addition, Trichoderma can also inhibit the growth of pathogenic spores and hyphae with its ability to produce furonan group antibiotics [22]. The use of biofungicides fermented with Trichoderma spp. a minimum of 5 ml/plant can control Fusarium wilt disease through the mechanism of space competition, mocoparasites and antibiosis. [16].

The Effect of Trichoderma Biourine Application on Shallot Plant Growth
The results of further tests on the effect of Trichoderma biourine application on plant height and number of shallots at the age of 7 DAP to 35 DAP using 5% BNJ are presented in Tables 2 and 3.  Tables 2 and 3 it is known that the method of application of Trichoderma biourine significantly affected plant height and number of shallots. The plant height and the highest number of scallions began to be seen in all Trichoderma biourine applications.
From the results of this study, it can be said that all methods of biourin application can increase plant height and the number of leaves of shallot plants. The growth of plant height and number of leeks after the application of Trichoderma biourine is suspected because the Trichoderma fungus contained in the biourin has a role in stimulating ethylene in plant tissues so as to stimulate plant vegetative growth [11]. The fungus T. harzianum can stimulate seed germination and plant growth [23]. Ethylene is a hormone produced by the fungus Trichoderma spp. can stimulate plant flowering [24]. Treatment of the fungus T. harzianum on soybean plants can stimulate plant growth so as to increase plant height and number of plant leaves [21]. Trichoderma spp. able to stimulate plants to form the hormones gibberellin acid (GA3), Indolasetic acid (IAA), and benzylaminopurine (BAP) so that plant growth such as plant height and number of leaves is more and is healthy, tough and affects plant resistance to disease. Furthermore, gibberellins and auxin hormones also play a role in root and stem elongation, tuber formation and increase plant development [22].

The Effect of Trichoderma Biourine Application on Shallot Yield
The results of further tests on the effect of the application of Trichoderma biourine on the number of tillers of shallots, the weight of harvested dried shallots and the weight of stored dried shallots using BNJ 5% are presented in Table 4.  Table 4 shows the number of tillers, the weight of harvested dry shallots, and the lowest weight of harvested dried shallots in the control or without using biourine. This indicates that the application of Trichoderma biourine can increase the number of tillers of shallot, the weight of harvested dry shallots, and the weight of harvested dry shallots.
The increase in the number of shallot tillers, harvested dry shallot bulb weight and stored dry shallot tuber weight after application of Trichoderma biourine was thought to be due to the role of this fungus in stimulating growth and increasing yield. It was reported that the fungus T. harzainum in the rhizosphere or plant root areas secretes ethylene which is diffused into the plant body through xylem which plays a role in promoting generative growth [11]. Treatment with conidia of T. viride and T. koningii fungi for controlling blight on strawberry plants was able to stimulate early flowering [25]. Ethylene is a hormone produced by the fungus Trichoderma spp. can stimulate flowering in plants [24]. Treatment of the fungus T.harzianum on soybean plants can stimulate plant growth so as to increase yield components [21]. Trichoderma spp. can produce certain hormones to increase the weight and number of pods in soybean plants [26]. Trichoderma spp. can stimulate plants to produce hormones gibberellin acid (GA3), Indolasetic acid (IAA), and benzylaminopurine (BAP) so that plant growth becomes optimum, and affects plant resistance. Gibberellins and auxin hormones play a role in root and stem elongation, and fruit (tuber) growth and increase plant growth [27].
Several previous researchers reported the successful use of Trichoderma fungi in various formulations on various plants. The use of biaoactivators fermented with Trichoderma spp. can increase the growth and yield of shallots in dry land [28]. The use of bioactivators fermented with Trichoderma spp. can increase the induced resistance of soybean plants to plant diseases and increase the growth and yield of soybean plants [29]. Tablet and liquid bioactivator fermented with Trichoderma spp. can increase soybean plant-induced resistance to wilt disease and increase soybean growth and yield [30]. and biocompost fermented with Trichoderma spp. can increase the weight of shallot bulbs [17]. The use of Trichoderma spp. which is formulated in the form of a stimulator biocompost can increase the growth and yield of maize in dry land [31]. The use of Trichoerma spp. in the form of a liquid bioactivator formulation as much as 5 ml/plant can increase the growth and yield of shallots [32]. Furthermore, it was also reported that the fungus Trichoderma spp. which is formulated in the form of biocompost can increase the growth and yield of soybean plants [33]. The use of Trichoderma biochar can increase soybean yield [34]. The use of Trichoderma biocompost can increase the yield of shallots [35]. The use of Trichoderma biofungicide can control Fusarium wilt disease and increase shallots yield [36]. The application of Trichoderma liquid biofungicide can increase the weight of harvested dried shallots bulbs [37]. The use of Trichoderma fungus can increase the resistance induced by banana seedlings to Fusarium wilt disease. The use of the saprophytic fungus Trichoderma antagonist causes shallots plants to become resistant to Fusarium wilt disease.

The Effect of Shallot Varieties on the Occurrence of Fusarium Wilt
The results of further tests using BNJ 5% the effect of shallot varieties on the incidence of Fusarium wilt disease at the age of 7 DAP to 35 DAP are presented in Table 5.  Notes: 1) The numbers in each column followed by the same letter are not significantly different.

The Effect of Shallot Varieties on Plant Growth
The results of further tests on the effect of shallot varieties on plant height and number of shallots at the age of 7 DAP to 35 DAP using 5% BNJ are presented in Tables 6 and 7. The occurrence of differences in plant height and number of leaves of shallot plants is thought to be due to genetic factors of each variety used, in addition to the adaptability of the variety to the environment such as sunlight, irrigation, rain intensity and soil conditions. Differences in plant growth are morphological adaptability, which in turn will affect the growth and yield of a plant [40].

Effect of Shallot Varieties on Yield
The results of further tests on the effect of shallot varieties on plant height and number of tillers, weight of harvested dried shallots and weight of stored dried shallots using HSD 5% are presented in Table 8. In Table 8 it can be seen that the shallot varieties showed different effects on the number of tillers of shallots, the weight of harvested dry shallots, and the weight of stored dry shallots. The Bali Karet variety showed that the number of tillers, harvested dry onion bulbs, and stored dry shallot bulbs were higher than the Ampenan and Keta Monca varieties.
, the weight of harvested dry shallots and the weight of stored dry shallots. The difference in the effect of shallot varieties is thought to be due to genetic factors of each variety and the influence of environmental conditions of planting. Genetic factors are one of the factors that affect plant growth and yield [42]. The average number of tillers of the Bali Karet variety was 7 -14 higher than the Ampenan variety, 7 -12 and the Keta Monca variety, 5 -9. The average yield of harvested dry shallot bulbs for the Bali Karet variety was 14-16 tons/ha, higher than the Ampenan variety, 12 tons/ha and the Keta Monca variety, 11 tons/ha [3]

IV. CONCLUSION
The results showed that: 1. The application of Trichoderma biourine by spraying the soil surface, spraying seed tubers and spraying on plants 21 days after planting can increase the growth and yield of shallot bulbs.