EFFECTS OF TRICHODERMA HARZIANUM AND FUSARIUM SOLANI INOCULANT ON THE RESIN CONTENT OF AGARWOOD

This study aimed to determine which fungal inoculants, Trichoderma harzianum or Fusarium solani in a solid or liquid form, produced the highest resin content in agarwood trees. This experimental study used a randomized completely block design (RCBD) with seven treatments in triplicates: solid Trichoderma harzianum inoculants in pellets(A) and capsules (B), liquid T. harzianum inoculants (C), solid Fusarium solani inoculants in pellets (D) and capsules (E), liquid F. solani inoculants (F), and control or without inoculants (G). Parameters observed were the color and aroma of the infected agarwood and the resin content of the harvested sapwood. Quantitative data obtained from observations/measurements were analyzed statistically with analysis of variance and a subsequent LSD test at a 5% significance level following a significant result. The results showed that treatments using different fungal inoculants had no significant effects on the resin content of agarwood. However, inoculants in pellets and capsules tended to increase the resin content. The resin content of agarwood ranged from 15.97% to 21.53%, and the highest level was obtained from treatment A (solid T. harzianum inoculants in a pellet form).


INTRODUCTION
Resin is a secondary metabolite produced by agarwood trees due to injury or microbial infection.The agarwood resin produced from the inoculation with

Materials and tools
The materials used in this study were agarwood (Gyrinops versteegii) from The parameters observed were the color, aroma, and resin content of the sapwood.
The quantitative data obtained from observations were analyzed statistically using analysis of variance (variance fingerprint) and, if a significant result was obtained, followed by the LSD test at a 5% significance level.

Research Implementation
The research was conducted in four stages: preparation, inoculation, observation and analysis, and resin content calculation.At the preparation stage, the research tools and materials were prepared.
First, the fungal inoculants were prepared at the Soil Biology Laboratory, Faculty of Agriculture, Udayana University, Denpasar.Each type of fungal isolate was cultured on PDA and inoculant media (e.g., sawdust, rice bran).Second, agarwood trees aged 3-4 years with the same diameter of 10-15 cm were selected from a field of
Third, the tools for fungal inoculation, including wood drill, inoculation needle, spatula, plasticine, and alcohol, were prepared.
Then, at the next stage, the fungal inoculant was inoculated into selected agarwood trees to stimulate sapwood formation.The inoculated trees had to be 3-4 years old or had a stem diameter of more than 3-6 cm.
Several holes measuring 0.6 cm in diameter and 2 cm in depth were made on the stems using a wood drill.These holes were drilled at least 20 cm from the ground and 10 cm apart in a circular direction at an angle of

Agarwood color
Figure 1 and Table 1 describe

Agarwood fragrance
The organoleptic measurements of the infected stems detected different fragrance levels, from slightly-fragrant to fragrant (Table 2).The fragrant aroma was obtained in treatments A (solid T. harzianum inoculants in a pellet form) and E (solid F. solani inoculants in a capsule form).The slightly fragrant to fragrant aroma was produced in treatments B, C, D, and E, while the slightly fragrant aroma was in the control (G).

Resin Content
The statistical analysis showed that fungal inoculants had no significant effects on the resin content, although resin levels tended to increase in the treatment groups  The agarwood trees respond to these pathogenic infections by producing secondary metabolites or resin compounds that release a fragrant aroma when burned (Sitepu, et al., 2011).Resin is a phytoalexin or a defense compound against pathogenic fragrance is linked to sesquiterpene compounds.A higher fragrance level is probably caused by a higher concentration of sesquiterpene compounds, while its decrease is due to the loss of these compounds, which are very volatile (Rahayu, 2009).

CONCLUSION
First, the form of fungal inoculation has no significant effect on the sapwood's resin content, but the latter tends to be higher in the treatment groups than the control group.
the agarwood farms in Marga Dauhpuri Village (Marga, Tabanan, Bali Province), two isolates of Fusarium and Trichoderma, Potato Dextrose (PD) media, solid media (rice bran, corn bran, sawdust), distilled water, methanol, and capsules.The tools included a bottle clock, measuring cup, petri dish, autoclave, laminar air flow, wood drill, digital scale, bath, oven, and color book.
30 o .Afterward, 0.8 cm 3 of the fungal inoculant was placed into each hole using a sleeve (the funnel), and the hole was covered with plasticine to avoid contamination by other materials or organisms.At the third research stage, observations and analyses were conducted after eight weeks of the inoculation.Sapwood, the brown (infected) parts of the agarwood trees, was harvested by prying using a chisel, stored in a plastic bag, and then chemically analyzed.Part of the plants (wood/ agarwood) was dried for analysis in the laboratory to determine the resin content.Finally, at the last stage, each sapwood sample was extracted with 150 ml of methanol for 3 hours.The resin yield (%) was calculated by dividing the weight of the extracted resin by the weight of the extracted sapwood times 100(Pasaribu et al., 2013).The color of the infected agarwood was determined using the Munsell color chart, and the aroma was determined organoleptically.
Figure 1 and Table 1 describe the color of the infected agarwood stem with different inoculants or treatments.The color varied from light brown (whitish-brown) to brown.The brown infected stems were obtained in treatments A (solid Trichoderma harzianum inoculants in a pellet form) and E (solid Fusarium solani inoculants in a capsule form).The lightbrown to brown infected stems were produced in treatments B, C, D, and E, while the light brown stems were found in the control (G).

Figure 2 .
Figure 2. The relationship between the resin content of the sapwood and the preselected fungal inoculants Harzianum and Fusarium Solani Inoculant on The Resin Content of Agarwood (Gyrinops Versteegii (Gilg.)Domke) Mega, I.M., Kartini, N.L.,Innosensia, N.L.P.C. attacks.The accumulation of resin compounds on the infected plant stems will form sapwood with various color and aroma.Based on the data obtained in this study, it turns out that there is a relationship between the color, aroma, and resin content of agarwood.A darker or browner agarwood has a stronger aroma (fragrance) and a higher resin content.It is evident from the color, aroma, and resin content of the control, treatment A, and treatment E (light brown-brown-brown, less fragrant, and 15.97% -21.19% -21.53%).The results correspond to Mega and Nuarsa (2019), which stated that the darker or the more darkish-brown the agarwood stem, the stronger the aroma (fragrance) and the higher the resin content.These assessments are relative to the control (without inoculants), JF (Fusarium solani inoculation), and JT (Trichoderma harzianum inoculation), which each produced light brown, brown, and blackishbrown sapwood with less fragrant and 5.25, 5.31, and 7.92% resin content.On the other hand, an increase in the fragrance level is not necessarily in line with an increase in the sapwood's color intensity because Second, treatments A (Trichoderma harzianum pellet) and D (Fusarium solani pellet) increase the resin content by 34.82 and 32.69 percentage points relative to the control (G, without fungal inoculation).Despite the less significant effect on resin contents, using fungal inoculants in the form of pellets is still highly suggested so as to produce the most resin from agarwood trees.The similar results between different forms of fungal inoculants mean that pellets are not the only option to obtain the most favorable sapwood color and aroma and the highest resin levels.It is necessary to test these forms in different locations.REFERENCES Budi, R.S.W., Santoso, E. and Wahyudi, A. 2010.Identifikasi Jenis-jenis Fungi yang Potensial terhadap Pembentukan Gaharu dari Batang Aquilaria spp.Jurnal Silvikultur Tropika, 1(1), 1 -5 Mega, I M., Suanda, D.K., Kasniari, D.N. and Suena, I.W.. (2013).Formulasi Inokulan Jamur Pembentuk Gubal

Table 2 .
Aroma of the infected agarwood stems by treatment and repetition