Fakultas Pertanian dan Bisnis Universitas Kristen Satya Wacana Jl. Diponegoro 52-60 SALATIGA 50711 - Telp. 0298-321212 ext 354 email:[email protected], website: ejournal.uksw.edu/agric
Terakreditasi Kementrian Riset, Teknologi dan Pendidikan Tinggi berdasarkan SK No 200/M/KPT/2020
Received: 25 September 2021 | Accepted: 24 Mey 2022
COMPATIBILITY TEST OF VARIOUS SOURCES OF INOCULANT ARBUSCULAR MYCORRHIZAL FUNGI WITH SWEET CORN PLANTS IN PEAT MEDIA
Dwi Zulfita, Surachman, Setia Budi dan Rahmidiyani Program Studi Agroteknologi Jurusan Budidaya Pertanian
Fakultas Pertanian, Universitas Tanjungpura.
Jl. Prof. Dr. Hadari Nawawi, Pontianak 78124 e-mail: [email protected]
ABSTRACT
The purpose of this study is to determine the compatibility of AMF isolates from several host plants to the yield components of sweet corn on peatlands. The field experiments were carried out with a randomized complete block design (RCBD) with 5 treatments and 3 replications.
The treatments referred to are m0 (without AMF inoculation), m1 (AMF inoculum from the host Pueraria javanica), m2 (AMF inoculum from the soybean plant host), m3 (AMF inoculum from the corn plant host), and m4 (AMF inoculum from the sorghum plant host). Observations were made on mycorrhizal infections, absorption of N, P, K nutrients, and plant yield components, including weight per ear with weight, weight per ear without husk, ear length, ear diameter, and ear weight per plot. The data acquired from the observation were statistically analyzed using analysis of variance (F test), while further tests were done with Duncan’s multiple distance test (DMRT). The results showed that AMF inoculant derived from maize rhizosphere is more compatible with corn plants than AMF inoculant from rhizosphere sorghum, P. javanica, and soybeans without inoculation. AMF inoculant from maize rhizosphere can increase root infection, absorption of N, P, K nutrients, and the best yield components of sweet corn on peatlands
Keywords: Arbuscular mycorrhizal fungi, compatibility, peatlands
INTRODUCTION
Peatland is one of the types of land with huge potential to grow crops, including corn. West Kalimantan has 1,608,000 ha of peatland area or 10.92% of the total size of the province (Central Bureau of Statistics, 2019). Even though 4,19 million ha of the peatland area have
been reclaimed, the rest of it, which made up to 9,53 million ha, can be developed for agriculture (Policy Synthesis Team, 2008).
However, the main obstacle in developing peatland areas for plant cultivation is the lack of P, K, Ca, and Mg nutrients. The Peatland area also has low levels of several microelements
such as Cu, Zn, Al, Fe, and Mn. It is also High in C/N ratio and soil acidity (Yenni, 2012).
Corn was chosen as an experimental crop to utilize peatlands due to the government’s focus on this kind of plant. The expansion of corn’s planting has caught the government’s attention, especially in using marginalized lands like peatland areas. Despite that fact, the average corn production is still relatively low due to several problems that typically occur in peatland areas, such as low fertility and high soil acidity because of organic matters decomposition accumulated in the soil (Hakim et al., 1986).
Therefore, it is essential to improve the peatland area’s quality before using it. Inoculation of Arbuscular Mycorrhizal Fungi (AMF) can be the way to do that.
This opens up opportunities for the use of Arbuscular Mycorrhizal Fungi (AMF) in corn cultivation. AMF plays a vital role for host plants to expand the absorption area of root hairs by forming mycelium around them. The formation of mycelium around the roots will increase the expansion volume, resulting in the ability of plants to absorb water and nutrients will be better than plants that do not have AMF (Hanafiah, 2001). Arbuscular Mycorrhizal Fungi (AMF) supply organic carbon compounds from their host plants for growth and development. On the other hand, fungi help host plants absorb nutrients and water from the soil, resulting in better plants growth (Rini et al., 2020; Cavagnaro, 2008).
AMF can form a mutualistic symbiosis with plant roots, resulting in better plants growth.
Both parties, the AMF, and the plants, benefit from getting a carbon source from photosynthesis while plants get a supply of nutrients from AMF (Ansiga et al., 2017). Bertham (2006)
explained that plants would release root exudates in the form of flavonoid compounds to form a symbiosis with AMF. Flavonoid compounds positively affect the growth of mycorrhizae at the pre-symbiotic stage. In peatlands, the available P content is low. The available P can be increased by utilizing AMF to make phosphorus fertilization more efficient (Trisilawati and Yusron, 2008). AMF can increase dissolved P through organic acids and phosphatase enzymes produced. AMF can also fix dissolved P to enter the external hyphae of AMF. An essential part of the mycorrhizal system is the mycelium which is outside the root which plays a role in the absorption of nutrients for plants.
According to Zuhry et al. (2008), the absorption of immobilized P can be increased by roots’ expansion as close to P as possible.
The presence of AMF can increase the ability of roots to absorb nutrients and water to support plant growth and development. According to Nurhayati (2012), the primary function of hyphae is to absorb water from the soil, P that accumulates in external hyphae will immediately be converted into polyphosphate compounds in the presence of phosphatase enzymes.
Arbuscular Mycorrhizal Fungi (AMF) can be symbiotic with 97% of higher plant families (Musfal, 2010), but each AMF strain has different abilities in increasing plants’ growth (Tian et al. 2004). Thus, it is necessary to select AMF isolates compatible with the cultivated plants. AMF is a symbiosis with responsive host plants with numerous roots (Simanungkalit 2004). Seasonal crops such as corn and sorghum are highly compatible hosts with endomycorrhizal (Simanungkalit 2004, Hapsoh 2008), making these two plants are considered
suitable hosts for the propagation of endomycorrhizal spores (Widiastuti 2004).
The research conducted by Rini and Rozalinda (2010) tested the Gramineae (corn and sorghum) and legumes (Centrosema pubescens and Calopogonium mucunoides) groups as host plants showed that the Gramineae group was more suitable for mycorrhizal production.
Based on previous experiments, it was unknown whether using AMF isolates from several host plants with corn would be compatible with peatlands’ sweet corn yield component. The purpose of this research is to find the compatibility of AMF isolates from several host plants to sweet corn yield components on peatlands.
RESEARCH METHOD Time and Place
This research was conducted for ± 6 months, from May 10th, 2020 to October 15th, 2020.
The study took place at the Green House of the Faculty of Agriculture, Tanjungpura University and on farmers’ land in Rasau Jaya II Village, Rasau Jaya District, Kubu Raya Regency.
Tools and Materials
The tools used in this research included a tape measure, hoe, sickle, watering can, knife, digital scale, scissors, plastic buckets, ruler, Leaf Area Meter, SPAD 502 chlorophyll meter, oven, 1000 ml measuring cup, hand sprayer, Thermo hygrometer, polynet mesh fence, labels, digital camera, stationeries, pH meter, binocular microscope, tweezers, microscope needle, glass object, cover glass, petri dish, test tubes, test tube rack, measurement cups, 100 ml measuring cylinder, 1000 ml beaker, Erlenmeyer flask 125 ml, 1 and 10 ml volumetric
pipette, and oven. The materials used in this research consisted of sweet corn seeds of the Bonanza variety; peat soil with a hemic maturity level, Zeolite, NPK Mutiara 16: 16: 16;
dolomite lime with neutralization power of 97.77%; chicken manure fertilizer given according to needs which are 20 tons ha-1;
Fungicide Banlate M-45; paper and plastic bags; black 25 cm x 20 cm polybag; corn, soybeans, Pueraria javanica and sorghum seeds as host plants; mycorrhizae derived from the pineapple plant rhizosphere; 10% KOH;
1% HCl; root staining solution (Glycerol, Lactic acid and Trypan blue) for staining observation of arbuscular mycorrhizal root infection; and Aquades distilled water.
Research Methods
This research consists of two stages: AMF trapping out of pineapple plants’ rhizosphere with various types of host plants as stage 1, and compatibility test of various sources of AMF inoculants towards sweet corn on peatland area as stage 2.
Stage 1. Trapping AMF with various host plants with pot culture technique using various host plants. This experiment used a completely randomized design (CRD) with 4 levels of host plant species: A = Pueraria javanica, B = Soybean, C = Corn, D = Sorghum. Each treatment level was repeated 6 times and there were 4 sample plants. The number of experimental units consisted of 96 pots. As a growing medium, a sterile mixture of peat and rhizosphere soil was used for each pot (1:1).
The technique of filling the media is that the pot is filled with sterile peat soil 1/3 part, then 1/3 part is filled with sterile rhizosphere soil and the last 1/3 is covered with sterile peat soil, so that the planting medium is composed of sterile
peat-sterile rhizosphere soil-sterile peat. Plastic cups used as pots were first sterilized with chlorax solution. The seeds of the host plants were sown on sterile peat media in a nursery, which had previously been soaked in warm water to break any dormancy that might have occurred.
After the host plant seeds turned 1 week old, they were transferred to pots that had been provided, then the spore suspension was inserted around their roots (Pueraria javanica, soybean, corn and sorghum). The plants were fertilized using red Hyponex fertilizer with a concentration of 100 mg/l for 96 pots. The fertilizer was given 3 times a week until the plants were 1 month old, then 2 times a week until the plants were 2 months old and 1 time a week until the plants were 3 months old. Plant maintenance includes watering plants 2 times a day in the morning and evening. The plants were watered until they were 3 months old, then they were put in drought stress conditions until the plants were dry. After that, the mycorrhizae are harvested and ready for observation.
Observations were made on mycorrhizal infection, spore density and AMF spore diversity.
Stage 2. Compatibility test of various sources of AMF inoculum on sweet corn on peatland.
Field experiments were conducted using a Randomized Block Design (RCBD) with 5 treatments and 5 replications. The treatments in question were m0 (without AMF inoculation), m1 (AMF inoculum from the host plant Pueraria javanica), m2 (AMF inoculum from the host soybean plant), m3 (AMF inoculum from the host corn plant) and m4 (AMF inoculum from the host sorghum plant). There were 5 treatments with 5 replications so that there were twenty-five experimental units and four sub-
samples. Observations were made on root infection and yield components, which included the weight of cobs without husk, the weight of cobs with husk, the weight of cobs per plot, length of cobs and diameter of cobs.
Data Analysis
Data accumulated from observations were analyzed statistically with analysis of variance (F test). If the F test showed a significant effect of each treatment from various AMF inoculum sources, the data will then be analyzed with Duncan Multiple Range Test (DMRT) at 5%
level. The calculations were performed using the Statistical Analysis System (SAS) program.
RESULT AND DISCUSSION
The results of AMF trapping with various host plants using pot culture technique using various host plants on % root colonization, spore density and spore diversity is shown in Table 1.
The percentage of root colonization by AMF from pineapple rhizosphere was 57.78%. Table 1 shows that the highest percentage of root colonization by AMF after trapping was shown in the corn host plant, which was 95.33%.
Meanwhile, the lowest percentage of root colonization by AMF was shown in the soybean host plant, which was 77.89%. The criteria for the percentage of root colonization by AMF on various types of host plants were classified as high to very high. According to Setiadi et.al.
(1992) that the percentage of root colonization ranging from 0 – 25% is low, 26 – 50% is moderate, 51 – 71% is high and 76 – 100% is very high.
The observations’ results after trapping culture (Table 1) showed that the number of spores had increased from 42 spores per 50 g of soil before trapping (derived from pineapple
Host Plant
Root Colonization
(%)
Spore density/
50 g soil Spore diversity
P. javanica 82,11 61 2 (Glomus (3 tipe), Acaulospora
(1 tipe))
Soya bean 77,89 60 2 (Glomus (3 tipe), Gigaspora
(1 tipe))
Corn 95,33 167 3 (Glomus (5 tipe), Gigaspora
(1 tipe), Acaulospora (2 tipe))
Sorghum 80,21 114 3 (Glomus 2 tipe), Gigaspora (1 tipe),
Acaulospora (1 tipe))
Table 1 % Root Colonization, Spore Density and Spore Diversity on Various Types of Host Plants in Arbuscular Mycorrhizal Fungi Trapping
rhizosphere) to 61 -167 spores per 50 g of soil. According to Suharno et al. (2015), the increase in the number of trapping spores was supported by controlled and stable screen house conditions, thus providing an opportunity for spores isolated from the field that had not germinated to undergo the germination process and then form new spores.
In this research, each treatment showed a varying number of spores. However, there were certain treatment combinations that tended to produce a higher number of spores compared to other treatment combinations, such as the combination-treated with corn as host plant produced 167 spores per 50 g of soil sample.
The number of spores is related to the increased chance of the spores to infect the roots. The research results from Nurbaity et al. (2011) stated that spore inoculation with the number of spores (0, 50, 100 and 150 spores) on potato plants showed the percentage of infected roots was different (0%, 31%, 37% and 52%) and it seems that the treatment with the highest number of spores (150 spores) gave the highest percentage of infected roots. Other than that, the infectivity of AMF originating from the pineapple rhizosphere with the host plant type of corn was found compatible. According to Santoso (1994), the compatibility of AMF with
host plants varies greatly depending on AMF species, host plant species and environmental conditions..
Through identifications, it was found that there were 3 AMF genera found in the trapping soil, which are Glomus, Gigaspora, and Acaulospora.
The identification was carried out by looking at differences in characteristics, morphological characteristics (shape, cell wall thickness, and surface ornamentation) and the reaction of spores to Melzer. Glomus was found in all host plant rhizospheres, while Acaulospora and Gigaspora were found only in some host plant rhizospheres.
The spores trapping’s diversity from pineapple rhizosphere was found to be 6 types of Glomus, 1 type of Gigaspora and 1 type of Acaulospora.
Table 1 shows that after AMF trapping there was an increase in spore diversity to 13 types of Glomus, 3 types of Gigaspora and 4 types of Acaulospora. Glomus always dominates the diversity of spore types after trapping culture.
According to Wanda et.al (2015) spores of the genus Glomus are the most dominant types of spores found in several ecosystem conditions because this AMF species has a wide host range. According to Johnson-Green et.al.
(1995) AMF activity and diversity were associated with seasonal changes in addition
to host growth activity. The peak of AMF spore formation in each host occurred at different times, as well as differences in the AMF species symbiotic with each host.
The results of variance on root infection, N, P and K nutrient uptake showed that inoculation various inoculum sources affected root infection, N, P and K nutrient absorption (Table 2).
Table 2 shows that the treatment of various sources of AMF inoculants was significantly different to root infection, and the absorption of N, P, and K as well. Although there were significantly different variables, the highest value was in the treatment of AMF inoculants from corn rhizosphere and significantly different from other rhizosphere AMF inoculant treatments (without inoculation, P.javanica, soybean and sorghum) on the variables of N, P, and K absorption. However, as on root infection variables, corn AMF inoculants were not significantly different from AMF inoculants from P. javanica, sorghum rhizosphere. This means that AMF inoculants from corn, sorghum, and P. javanica rhizosphere were more responsive to root infection. The highest root infection occurred in AMF inoculants from corn rhizosphere, despite the fact that it was not significantly different from the ones in AMF inoculants from sorghum and P. javanica rhizosphere. It is also significantly different from
Source of FMA Inoculants Root Infection (%)
N Nutrient Uptake (g)
P Nutrient Uptake (g)
K Nutrient Uptake (g) No Inoculation 38.00 ± 3.22b 0.40 ± 0.17d 40.44 ± 1.25e 45.30 ± 5.00e Pueraria javanica 64.30 ± 2.79a 0.78 ± 0.09c 73.10 ± 0.76c 78.63 ± 6.31c
Soya bean 39.70 ± 1.68b 0.63 ± 0.05c 63.54 ± 3.32d 63.26 ± 2.11d
Corn 68.20 ± 4.43a 3.53 ± 0.31a 100.31 ± 1.41a 147.25 ± 7.98a
Sorghum 63.70 ± 4.13a 1.36 ± 0.11b 90.23 ± 7.50b 99.57 ± 2.41b
KK (%) 6.10 11.87 5.09 6.68
Table 2 Root infection, N nutrient uptake, P nutrient uptake and K nutrient uptake in maize with various sources of Arbuscular Mycorrhizal Fungi inoculants
Note: All the results on the table is the average value ± standard of deviation. The value inside the column has superscript that followed with the same letter does not have any difference according to Duncan’s multiple-distance test at 5% level.
AMF root infections from soybean rhizosphere and without AMF inoculation.
The root infection variable in the AMF inoculant treatment from corn rhizosphere was 68.20%
which was considered high based on the criteria of Rajapakse and Miller (1992). Root infection in plants is an evidence of the presence of AMF.
Musfal (2010) stated that AMF is helpful to increase nutrient absorption, especially phosphate. This is because the external hyphae network of AMF is able to expand the absorption field. AMF produces phosphatase enzymes that can release P nutrients bound by organic acids in peat soils, and Ca in calcareous soils so that nutrients are more available to plants. This condition was also supported by the abundance of spores and the diversity of AMF species (G. clarum and A. tuberculata) obtained by corn hosts in the AMF trapping experiment and used as inoculants in this experiment (Zulfita et al. 2020).
The increase in root infection in AMF inoculants from corn rhizosphere was followed by an increase in the absorption of N, P, and K. The increase in P absorption encourages the growth of corn plants. This is indicated by the growth and yield of corn. The AMF inoculants treatment of corn rhizosphere increased because there was an increase in the absorption of N, P, and K as well. The corn that inoculated
with AMF inoculants from the corn rhizosphere was more responsive to absorb N, P and K than those inoculated with AMF from other hosts or without inoculation. Plants with mycorrhiza have a more excellent water absorption that brings easily soluble nutrients such as N and K.
The value of corn plant variables in the treatment of AMF inoculant derived from corn rhizosphere is constantly increasing compared to the others from other rhizosphere, due to a higher number of spores on it than AMF inoculants from other host plants. This is based on the AMF trapping process in the previous stage of research showing that the number of AMF spores from the corn rhizosphere was higher and more diverse.
The main reason of that is because corn has longer roots than the other host plants, so the number of spores and hyphae produced are higher. According to Muis et al. (2016), AMF is symbiotic with responsive host plants with many roots with an extensive root system. The relationship between root infection and nutrient absorption of N, P, and K are presented in Figure 1.
Figure 1. The figure explains that there is a solid relationship with a moderate to high degree of
C
Figure 1 The relationship between root infection and nutrient uptake (A) nutrient uptake of N, (B) P nutrient uptake, (C). K nutrient uptake
association between root infection and nutrient absorption N (r = 0.679), nutrient absorption P (r = 0.882) and nutrient absorption K (r = 0.825). The higher the root infection, the higher the N, P and K nutrient absorption. The increased N, P, K nutrient absorption of plants affected plant metabolism which resulted in an increase in dry weight of corn plants.
The results of the variance of the yield components showed that inoculation with various sources of inoculum had an effect on the weight of the cobs with the husk, without the husk, the length and the diameter of the cobs.
On the other hand, it had no effect on the weight of the cobs per plot (Table 3).
Table 3. As seen in table 3, it is apparent that corn plants treated with AMF inoculants from corn and sorghum rhizosphere produced the same weight per cob. However, it weighed differently from AMF inoculations from P.
javanica rhizosphere, soybean and plants without AMF inoculation. Corn plants treated with AMF inoculants from corn rhizosphere produced the heaviest cobs compared to the others. Corn plants with various treatments of AMF inoculant sources produced weight per cob without husk which was not different but different from the treatment without AMF
inoculation (Table 3). It was also seen that corn plants treated with various sources of AMF inoculants and without AMF inoculation produced no different cob weight per plot, although visually corn plants treated with AMF inoculants from corn had the heaviest weight of cobs with or without husk and cob per plot.
When there is more P available in the soil will increase the P absorption of plants, allowing the process of photosynthesis. The high rate of photosynthesis will produce large photosynthate which is then distributed to other plant parts, including the yielding organs.
There was a correlation between P absorption, ILD and plant’s dry weight with cob weight
Source of FMA Inoculants
Weight per cob (g)
Weight per cob without husks (g)
Cob weight/plot
(kg)
Cob length (cm)
Cob diameter (cm) No Inoculation 411.55 ± 37.10c 279.90 ± 39.13b 9.72 ± 0.41a 19.40 ± 0.88c 3.02 ± 0.30c Pueraria javanica 472.60 ± 24.86b 318.35 ± 12.04a 10.04 ± 0.52a 20.95 ± 0.42ab 4.43 ± 0.51ab Soya bean 464.10 ± 38.63b 317.80 ± 23.40a 9.94 ± 0.08a 20.35 ± 0.57b 3.91 ± 1.14b Corn 506.85 ± 13.75a 345.25 ± 16.44a 10.18 ± 0.24a 21.59 ± 0.37a 5.06 ± 0.08a Sorghum 493.25 ± 25.36ab 342.95 ± 13.15a 10.14 ± 0.21a 21.12 ± 0.61a 4.51 ± 0.48ab
KK (%) 5.10 5.93 3.37 2.55 14.16
Table 3 Weight per cob with cob, weight per cob without cob, weight on cob/plot, cob length and cob diameter of corn plants treated with various sources of inoculant Mycorrhizal Fungi Arbuscula
Note: All the results on the table is the average value ± standard of deviation. The value inside the column has superscript that followed with the same letter does not have any difference according to Duncan’s multiple- distance test at 5% level.
per plot (r = 0.98*, r = 0.94* and r = 0.93*) meaning that the increase in P absorption, ILD and plant dry weight consistently increased followed by the addition of cob weight per plot.
It is because a more developed leaf organs will support the plant’s ability to absorb nutrients and sunlight that’s needed for a increased rate of photosynthesis process. The higher the rate of photosynthesis process, the higher the production of dry matter on the leaves, and it helps to increase the assimilate supply to the other plant organs as well including the cobs that become the plant’s sinks. The relationship between P nutrient absorption and weight per cob with grain, weight per cob without grain, and cob weight/plot are presented in Figure 2.
Figure 2 The relationship between P nutrient uptake and corn yield components (A) Weight per cob with grain, (B) Weight per cob without grain, (C). Cob weight/plot
Figure 2 explains that there is a solid relationship with a moderate to high degree of association between P nutrient absorption and weight per cob with husk (r = 0,679), weight per cob without grain (r = 0,882) and weight per cob (r
= 0,825). The higher the P nutrient absorption, the heavier the weight per ear with corn husks, the weight per ear without corn kernels and the weight of the cob/plot.
Table 3 also shows that corn plants treated with AMF inoculants from corn rhizosphere, sorghum and P. javanica produced the same cob length, but different from AMF inoculation treatment from soybean rhizosphere and without AMF inoculation. The corn plants treated with AMF inoculants from corn rhizosphere produced corncobs that were longer and larger than those treated with other AMF inoculants and without AMF inoculation.
This is because AMF from the rhizosphere of corn, sorghum, and P. javanica is better for increasing crop yields in terms of increasing the length and diameter of the cob in marginal soil conditions. This is in line with the results of field research showing the success of inoculation.
According to Lukiwati (2017) AMF depends on indigenous AMF species and the potential of the inoculant itself. It was further stated that the effectiveness of indigenous AMF populations was related to soil nutrient status factors, host plants, propagule density, and competition between AMF and other soil microorganisms. According to Susila et.al.
(2016), each plant has different functional compatibility with AMF inoculants due to plant preferences for specific AMF inoculants.
There is a relationship with root infection that occurs which also affects the absorption of P and other nutrients. There is a positive and
significant correlation between root infection and ear length (r = 0.92*) and ear diameter (r = 0.98*). In addition, there is also a correlation between the nutrient absorption of P with the length of the ear (r = 0.94*) and the diameter of the ear (r = 0.94*). This indicated that the increase in the length and diameter of the cob was a response to root infection by AMF and nutrient uptake of P.
Increased absorption of P and other nutrients by AMF hyphae allows the photosynthesis process to be more optimal and produce more assimilation. The assimilation translocated to the sink is also used to increase the length and the diameter of the ear.
CONCLUSIONS
The results of observations on the number of spores showed an increase from 42 spores per 50g of soil before trapping (derived from pineapple rhizosphere) to 60 - 167 spores per 50g of soil. The diversity of spore types (pineapple rhizosphere) such as 6 Glomus types, 1 Gigaspora type and 1 Acaulospora type were changed to 13 Glomus types, 3 Gigaspora types and 4 Acaulospora types. The percentage of root colonization before trapping (pineapple rhizosphere) was 57.78% and after trapping ranged from 77.89% - 95.33%.
AMF inoculants derived from the rhizosphere of maize and sorghum were as compatible with corn as compared to AMF inoculants from the rhizosphere of P. javanica, soybean and without inoculation. AMF inoculants from corn and sorghum rhizosphere can increase root infection, nutrient absorption of N, P, and K and also the best yield of sweet corn on peat media.
ACKNOWLEDGEMENT
The authors of this research would like to thank the Chancellor of Universitas Tanjungpura and the Dean of Faculty of Agriculture of Universitas Tanjungpura who supported the funds for this research through Universitas Tanjungpura DANA DIPA 2020.
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