Identification of dominant fungi in medium high tides area, Angke Kapuk protected forest

Litter that falls will experience decomposition involving the role of microorganisms such as bacteria and fungi. Therefore, by using adding fungi to the leaf clutter, can make the decomposition manner can be quicker. This study aims to identify the dominant species of fungi that play a role in the decomposition of Avicennia marina leaf litter. The methods used in this study are the washing method and the particle filtration method. The research shows that it does exist 15 types of fungi that predominate in accelerating the process of litter decomposition. From the identification results, the fungus belongs to the genus Penicillium sp., Aspergillus sp., Fusarium sp., Paecilomyces, and mycelia strerile. Overall, the known types of fungi belong to the Ascomycota division but belong to different families. The frequency of emergence of the fungus Aspergillus sp. was very dominating from the 15th to the 105th day of observation with a total of 59 isolates. Aspergillus sp. has a high ability to degrade lignin compounds so that it can help speed up the process of litter decomposition, of the 13 fungi found, Aspergillus sp. was the fungus with the highest occurrence frequency compared to other types of fungi.


Introduction
Mangroves are one of the producers of aquatic life with a very significant contribution to aquatic biota, one of which is as a provider of nutrient supply for the growth of plankton in the form of dry leaves and broken branches which then experience litter decomposition and mineralization, and produce nutrients which will later be used as material in the storage process by plankton [1].Three main factors affect the decomposition process of litter, especially leaf litter, including: (a) physico-chemical conditions of the environment, (2) litter quality, and (3) composition of the decomposer community [2].
IOP Publishing doi:10.1088/1755-1315/1315/1/012032 2 The decomposition system starts offevolved from the destruction manner accomplished by means of small insects on plants and the final lifeless organic count into smaller sizes.Then continue with the organic procedures done through bacteria and fungi to decompose organic particles.The decomposition process by microorganism and fungi as decomposers is assisted by enzymes which can decompose organic materials consisting of proteins, carbohydrates and others [3].Microbial groups play aprime role in the decomposition of plant muddle in terrestrial ecosystem.Fungi make contributions significantly to the complex phenomenon of nutrient recycling in forest ecosystems due to the fact they decompose various extracellular enzymes that deconstruct various varieties of natural compounds in clutter [4].
Fungi are taken into consideration as "key players" in leaf clutter decomposition, because of their capability to provide various extracellular enzymes.These enzymes help break down the lignocellulosic layer of litter that cannot be broken down by other organisms.In particular, the most significant enzymes involve enzymes that break down plant cell wall substances, such as cellulases, hemicellulases, pectinases, phenol oxidases and polygalacturonases.The collective action of these enzymes weakens the plant cell wall, thereby exposing hemicellulose and cellulose for further enzymatic degradation.Some enzymes, such as peptidase, urease and phosphatase are important for microbial acquisition of nitrogen and phosphorus, while others such as phenol oxidase, peroxidase and laccase help catalyze lignin degradation.Some of these enzymes are widely used in agriculture, bioremediation, the food industry, medicine, and the pharmaceutical industry [2].
The mangrove forest in the Angke Kapuk Protected Forest is one of the mangrove forest ecosystems in the Province of Jakarta.Research on decomposition in the protected mangrove forest of Angke Kapuk will be limited to Avicennia marina leaf litter in areas of medium high tides, which are inundated for 9 to 20 times a month.This observe targets to become aware of the dominant species of fungi that play a role inside the decomposition of Avicennia marina leaf clutter.

Place and time of research
This research was conducted from September 2022 to January 2023 in the Angke Kapuk Protected Forest, Jakarta.Dry weight was measured at the Forest Ecology Laboratory, Faculty of Forestry and Environment, Bogor Agricultural University.Isolation and consideration of the fungus was carried out at the Forest Influence Laboratory and the Pathology Laboratory.

Tools and materials
The tools used in this take a look at had been Petri dishes, oven, analytical stability, filters with mesh sizes of 1 mm, 250 μm, and 150 μm, pipettes, tweezers, autoclaves, Laminar Airflow Cabinets, gloves, digital cameras, Erlenmeyer flasks, measuring cups, plastic samples, sample bottles, stationery, tally sheets, glass slides, label paper, test tubes, microscopes, and permanent markers.The materials used in this study were Avicennia marina leaf litter, 5% NaOCl, 96% alcohol, 70% alcohol, Potato Dextrose Agar (PDA) media, rosebengal, chloramphenicol, plastic wrap, and aluminum foil.

Isolation of the fungus from leaf litter
The determination of the fungal population was carried out using the washing method and the particle filtration method.The weight of the decomposed leaf samples taken was 20 grams for each period of Avicennia marina leaf litter collection.

Washing method
The washing method is used to isolate microbes from fresh and fallen leaves and twigs in the form of litter.Repeated washing is intended to remove spores on the leaf surface.Samples of decomposed Avicennia marina leaf litter were rinsed with running water, then washed with sterile distilled water.The leaf litter was then cut into smaller sizes, namely 1x1 cm in size.Pieces of leaf litter were then rinsed in sterile distilled water, after which immersed in 1% sodium hypochlorite answer for five mins, then rinsed again using sterile distilled water and then put into 70% ethanol for about 1 minute and then rinsed aseptically using sterile distilled water 3 times.All samples were dried using a sterile tissue in a Laminar Air Flow cabinet.Three pieces of leaf muddle have been positioned on PDA media in a petri dish (9 cm in diameter) and rose bengal (25 mg.L -1 ) as fungistatic and chloramphenicol (250 mg.L -1 ) as bactericidal.As a negative manage, 1 mL of sterile distilled water from the remaining rinse of the sample was unfold over the PDA medium.The petri dishes have been then incubated at 27℃.All fungal colonies that grew had been then purified on PDA media and incubated at 27℃ for 7 days.

Particle filtration method
The particle screening method is a modification of the particle screening method used by [5].Avicennia marina leaf litter become washed extensively to remove adhering silt particles after which dried at around 25℃ for eight hours.For every particle screening used 4 grams of sample.Samples were immersed in one hundred mL of sterile distilled water, then dried under aseptic conditions.The pattern became then floor sterilized by immersing in 70% alcohol for three mins, then immersed in 1% NaOCl for five mins and rinsing with sterile distilled water 2 times.The water from the second rinse was dripped onto the media and then spread evenly with a spreader.Samples of leaf litter that had been surface sterilized were dried aseptically and then pulverized using a mortar until homogeneous.The sample was then dissolved using 36 mL of sterile distilled water in a 50 mL Falcon tube and vortexed until homogeneous.The litter solution was filtered using a filter holder.The filtered litter particles were placed into PDA media, while the filtered water was diluted to a 10-4 dilution by taking 1 mL of the filtered solution, then including 9 mL of sterile distilled water and one drop of tween 80 solution.Then the solution resulting from the dilution was vortexed to make it homogeneous, and this step was carried out until the 10-4 dilution.The filtered solution and also the results of the dilution up to 10-4 were placed on the media and then flattened using a spreader.All samples in the dish were incubated at room temperature (23℃) and checked daily for up to three weeks.All fungal colonies that grew were then purified on PDA media and incubated at 23℃ to be identified morphologically.The PDA media used previously had rose bengal (25 mg.L -1 ) as a fungistatic and chloramphenicol (250 mg.L -1 ) as a bactericidal.

Morphology identification
Pure fungal cultures have been rejuvenated on PDA media, and incubated for five until seven days at room temperature.Fungus isolates that had grown at the media were determined for their macroscopic traits, colony characteristics such as the nature of the growth of hyphae, the color and diameter of the colony and the color of the mass of spores or conidia.Fungal isolates have been also grown on slide culture, through placing 4 x 4 x 2 mm pieces of agar that had been overgrown with fungus on the slide culture, which become then protected with a cowl glass.The isolates at the item glass have been positioned in a plastic box measuring 30 x 20 x 6 cm, which have been moistened in the shape of moistcotton.The fungus isolates on the slide had been left for several days at room conditions until the fungus isolates grew sufficiently advanced.Whilst the fungus isolate has developed, get rid of the quilt glass which has overgrown with the fungus carefully to eliminate the agar portions.
Then on the cut part, drip one drop of sterile distilled water.The glass culture turned into found the usage of a mild microscope to determine the microscopic traits of the fungus, the traits of the hyphae, the presence or absence of partitions on the hyphae, the form of hyphal branching, conidiophores, conidiogenesis, in addition to the traits of the conidia or spores (shape and series) and the dimensions of the spores.The characteristics acquired had been tabulated, then matched with the identity key of the fungus.After the fungus became recognized, it become recorded, the quantity of species, populace, species variety and frequency of fungal colonization found in Avicennia marina leaf litter.This interest changed into completed whenever the clutter became gathered from the sphere at some stage in the decomposition procedure, starting from day 0 (control) up to the day whilst the leaf muddle changed into completely decomposed.

Washing method
The dominant fungi found from the isolation using the washing method were 12 species (Table 1).Based on the table above, the results of the dominant fungus found using the washing method were 57 isolates.From Table 1 it can be seen that the fungus that had the most influence in supporting the process of accelerating the decomposition rate was found on the 105th day.

Particle filtration method
The number of fungi obtained from the particle filtration method was 12 different types (Table 2).Based on Table 2, the results of the dominant fungus found using the particle filtration method were 156 isolates.From these results it can be seen that the frequency of the emergence of fungi which play a role in helping the process of accelerating the rate of decomposition is higher than using the washing method.The most dominating fungus was on the 75th day with a total of 32 isolates.
The rate of litter decomposition from day 15 to day 120 decreased due to the presence of fungi which act as decomposers.The frequency of occurrence of the most dominant fungus was found on the 75th day with 44 isolates.Comparison between the rate of decomposition and the total isolate of the dominant fungi (Figure 1).3.3.2Fusarium sp.Mycelium large and cottony, frequently with some tinge red, crimson or yellow, within the mycelium or medium; conidiophores variable, slender and modest, or stout, short, irregularly branched or phialid circular, singly or grouped into sporodochia; conidia (phialospores) hyaline, variable, basically consists of two types, frequently held in small wet head; the multiple-celled macroconidia are barely curved or bent at the pointed tip, usually canoe-fashioned; microconidia one celled, ovoid or oblong, borne individually or in chains; a few conidia intermediate, two or three celled, rectangular or barely curved.To determine the type of fungus that has been found from the results of isolation, it is necessary to identify the fungus morphologically both macroscopic and microscopic (Table 3).3, the dominant fungi found during the litter decomposition period from day 15 to day 120 were Penicillium, Aspergillus sp., Fusarium sp., Paecilomyces sp., and some mycelia sterile.

Discussion
The dominant fungi found from the results of research using the particle filtration method were more than the washing method.This is because particle filtration methods are a number of the handiest techniques for culturing fungal strains from clutter and soil, due to the fact they emphasize vegetative CFUs rather than numerically dominant spore CFUs [6].Many types of fungi are found in areas of medium high tides with a salinity of <10 ppm.This occurs because those conditions are nearly similar to clean situation which are precise enough for the boom and improvement of numerous styles of fungi in comparison to conditions at a better stage of salinity.The higher the extent of environmental salinity, the greater decreased the populace of fungi.This takes place due to the fact the elements wanted for the boom and development of diverse varieties of fungi are greater considerable within the leaf clutter of Avicennia marina which undergoes a decomposition manner at a salinity level that is nearly near clean, i.e. <10 ppt [7].The elements that affect populace density and species variety of soil organisms are oxygen supply, moisture, soil temperature, nutrient content and the quantity of soil organic count number.
The fungi that dominate in assisting the litter decomposition process are Aspergillus sp., Fusarium sp., Penicillium sp., and Paecilomyces.This group of fungi is often found and isolated with high abundance in litter samples even though prior to the isolation process the sample was washed [8].When the leaf litter falls to the ground, early colonizing fungi are fungi that are able to utilize only simple sugars, and these fungi are often found on fresh leaves.After that there will be rapid colonization by soil fungi belonging to the genera Penicillium, Fusarium, and from other genera.The soil fungi that colonize the litter are called autochthonous species.Most of the autochthonous fungi are able to hydrolyze polysaccharides, even certain species such as Penicillium are able to use tannins as a carbon source, but other fungi that do not produce more complex substrate hydrolyzing enzymes will be reduced.Furthermore, colonization will be carried out by survivors, namely fungi that are able to hydrolyze complex compounds such as cellulose and even lignin from the litter substrate as their main energy source Basidiomycetes and Ascomycetes [9].
The highest number of dominant fungi was found on day 60, day 75 and day 105.This happened because the humidity level was high enough so that the frequency of the appearance of the fungus was more numerous and dominated.Fungal colonization begins with weak parasites invading old tissue.Second, there are primary sugar fungi that utilize simple carbon compounds after the litter is deposited in the soil.In stage 2, secondary saprophytic sugar-associated cellulose decomposing fungi act on dead leaves.It comprises most of the asexual forms of ascomycetes.Finally at step 3, the basidiomycetes act on the substrate as lignin degraders [2].There are three stages of fungal succession, namely pioneer community, mature community, impoverished community [10].
The dominant fungi obtained from the research results are fungi belonging to the phylum Ascomycota.The Ascomycota phylum is an important group in the early stages of the decomposition process and is one of the dominant fungal phyla in mangrove forests (Avicennia marina).This phylum generally decomposes cellulose selectively towards lignin and dominates the fungal community and is very active (production of extracellular enzymes) in the early stages of litter decomposition.Aquatic ascomycetes play an important role in the degradation of wood and leaf litter and are more abundant, whereas Basidiomycetes are found in lower numbers.Ascomycetes are better able to withstand these conditions that prevail in aquatic habitats so they are much more numerous than Basidiomycetes [11].

Conclusion
Identifying microfungal diversity in species-wealthy habitats inclusive of soil and leaf litter clutter stays a mission.However, particle filtration will help speedy accumulate simple facts approximately microfungi in leaf litter and other finely divisible natural substrates.Further studies are wanted at the factors influencing fungal range estimates, inclusive of the results of isolation media and storage temperature, and the constancy of morpho-taxa.

Figure 1 .
Figure 1.Graph of decomposition rate and total isolates 3.3 Morphological identification 3.3.1 Aspergillus sp.Erect conidiophores, modest, ends with a rounded or clavate swelling, carrying phialides at the top or radiating from across the floor; conidia (phialospores), one celled, circular, often varying in color, in dry basipetal chains.

Table 1 .
The dominant number of fungi from the washing method

Table 2 .
The dominant number of fungi from the particle filtration method

Table 3 .
Identification of the dominant fungus