JUATIKA
JURNAL AGRONOMI TANAMAN TROPIKA
VOL.
6 NO.
1 January 2024 DOI :https://doi.
org/10.
36378/juatika.
eissn 2656-1727 pissn 2684-785X Hal : 152 Ae 164 Exploration of Phosphate Soluting Bacteries Located Near Rubber Plant (Hevea brasiliensi.
Field on Different Topography Agung Rinata.
Achmad Himawan dan Elisabeth Nanik Kristalisasi INSTIPER Yogyakarta Jln.
Nangka II.
Maguwoharjo (Ringroad Utar.
Yogyakarta.
Kampus II:
Jl.
Petung No.
Papringan.
Yogyakarta.
Indonesia *E-mail: wawanhimawan2014@gmail.
ABSTRACT
The exploration of phosphate-solubilizing bacteria in the root-soil of rubber plants was conducted to determine their beneficial effects.
This research aimed to identify the sampling locations of rubber plantations where phosphate solubilizing bacteria are suspected to be present, isolate and identify colonies of these bacteria, assess their ability to break down P elements from Pikovskaya media and determine their genus.
The research took place at the INSTIPER Yogyakarta Central Laboratory.
Kec.
Maguwoharjo.
Kab.
Sleman from February to April 2022.
A descriptive method was employed, involving surveying rubber plants in different topographies, sterilizing tools and materials, collecting soil samples from rubber plants, and isolating and identifying bacteria.
The research analysis included macroscopic and microscopic observations and measuring the diameter of the transparent zone.
Phosphate solubilizing bacteria were found in the sampling locations of Karanggondang Village and Popongan Village.
Macroscopic observations revealed 17 isolates with bacterial color morphology ranging from milky white, transparent white, to yellow.
The average diameter of the transparent zone, from highest to lowest, was as follows: B5.
2A .
3 m.
A1.
1B .
3 m.
A1.
2A .
, and C1.
The phosphate solubilizing bacteria isolate A1.
2A was identified as the Escherichia Genus while isolating C1.
1 was identified as the Genus Acetobacter.
Keywords: exploration, rubber plant, topography, identification, phosphate solubilizing bacteria Copyright A 2024.
The authors.
This is an open access article under the CC BY license .
ttps://creativecommons.
org/licenses/by/4.
Rinata et.
INTRODUCTION
Rubber, scientifically known as Hevea brasiliensis, plays a significant role in Indonesia's economic activities as one Indonesia holds the title of being the world's largest producer and exporter of rubber, according to the BPS .
The root soil of rubber plants serves as a habitat for various Among these bacteria, phosphatesolubilizing bacteria are particularly beneficial for the growth of rubber plants.
Despite high phosphate concentrations in the soil, only a small fraction, 1% of total P, is available to the plants due to its poor solubility and fixation ability.
This is attributed to the interaction of phosphate with other elements in the soil, such as calcium (C.
, aluminium (A.
, and iron (F.
, resulting in the formation of calcium phosphate, aluminium phosphate, and iron phosphate.
Consequently, becomes unavailable for plant uptake.
However, phosphate-solubilizing bacteria have the ability to enhance the availability of phosphorus for plants by converting inorganic phosphate into forms that can be readily absorbed (Asril et al.
, 2.
Previous exploratory research has phosphatesolubilizing bacteria in the rhizosphere of various plants, including banana nipah (Marista et al.
, 2.
, oil palm (Nugraha et al.
, 2.
, corn (Panjaitan et al.
, 2.
and sugar palm (Syarwani et al.
, 2.
However, there has been relatively phosphatesolubilizing bacteria in the root-soil of rubber plants.
Therefore, this research aims to identify potential sampling locations within rubber plantations where phosphate-solubilizing bacteria are likely to be present.
Additionally, this study aims to assess the abundance of phosphate-solubilizing bacteria on the roots of rubber plants Juatika Vol.
6 No.
Furthermore, the research seeks to evaluate the bacteria's capability to break down phosphorus elements in Pikovskaya media and determine the genus of the phosphate-solubilizing bacteria.
MATERIAL AND METHODS
Research Time and Place The study was conducted at the Central Laboratory of the Instiper campus.
Maguwoharjo.
Sleman.
Yogyakarta.
Various topographies in the Semarang Regency area were selected as the sampling sites for rubber plant soil.
The research took place between February and April 2022.
Research Tool and Material The plastic clips, markers, knives, notes, electric stove, aluminium foil, ruler, thermohygrometer, pH meter, pH stick.
Lux meter, camera (HP), autoclave, analytical balance, measuring cup.
Laminar Air Flow (LAF) Cabinet, bunsen, erlenmeyer, petri dish, stirrer, microscope, tube needle, cover glass, slide, pipette, and refrigerator are the tools utilized.
The material employed consists of soil from the roots of rubber plants acquired from rubber plantations in various topographies within the Semarang Regency area.
Nutrient Agar and Pikovskaya media, distilled water, 70% alcohol, label paper, tissue paper, and Gram staining solution are utilized.
Research Design The research uses descriptive The illustrative method is carried out through surveys in the field and the identification of bacteria in the laboratory.
Research Implementation The research was carried out in several Sterilizating Tools and Materials Tools and materials are sterilized by wrapping them in aluminium foil and brown umbrella paper, then putting them in an autoclave at a temperature of 121oC with a pressure of 15 psi .
er square inc.
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Juatika Vol.
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for 60 minutes.
Taking soil samples at the roots of rubber plants Soil samples at the roots of rubber plants were taken at different topography.
At each location, five different plants were selected from different points.
The soil from each plant was wrapped in aluminium foil and put into a chiller box, then taken to the laboratory.
Isolating and Identifying Bacteria Isolation is carried out from the ground part of the rubber plant.
Isolation of bacteria was carried out by dilution:
weighing 5 g of rubber plant soil, putting the soil in a beaker, mixing it with 1 liter of distilled water, and then stirring it until The soil diluted to 10-5 is then poured into a cup containing Nutrient Agar media.
After that, it was incubated at 28AC for five days.
Bacteria that grew after Then, the bacteria were inoculated again on Pikovskaya media to observe the transparent zone.
After microscopic observation was carried out.
The research flow diagram can be seen Sterilizating Tools and Materials Soil sampling at the roots of rubber plants in three different locations Isolation of bacteria using nutrient agar media Transparent zone test using Pikovskaya medium Gram stain of phosphate solubilizing bacteria Identify phosphate solubilizing bacteria Figure 1.
Research flow diagram Data Analysis Data were analyzed descriptively.
Macroscopic observations include colony color, colony edge shape, colony surface type, colony elevation, colony size and transparent zone diameter.
Microscopic observations include the gram staining results and the bacteria's shape.
RESULT AND DISCUSSION
1 Sampling locations for rubber plantations where phosphate solubilizing bacteria are suspected.
Rubber locations were carried out in three different locations.
The first location is in Karanggondang Village.
Kec.
Pabelan.
Kab.
Semarang.
The second location is at SEAT (STIPER Edu Agro Touris.
in Lemahireng Village.
Kec.
Bergas.
Kab.
Rinata et.
Semarang.
The third location is in Popongan Village.
District.
Bringin.
Kab.
Semarang.
Observation data on the Juatika Vol.
6 No.
environmental conditions of rubber plantations at three different locations can be presented in Table 1.
Table 1.
Observation data on environmental conditions of rubber plantations at three different Location Height of Temp.
Moisture Lighting Soil Site .
(A C) (%) Intensity (Lu.
Karanggondang Village SEAT Popongan Village Note: the weather at the time of data collection was cloudy Determination of Bacterial Colonies Suspected Phosphate Solubilizing Bacteria The results of sampling taken from Karanggondang Village amounted to 6 Macroscopic observations from sampling Karanggondang Village can be seen in Figures 1 Ae 6.
Figure 2.
Isolate from Karanggondang Village, 2 samples, 1 replication, 1 colony Figure 3.
Isolate from Karanggondang Village, 2 samples, 2 replicates, 2 colony Rinata et.
Juatika Vol.
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Figure 4.
Isolate from Karanggondang Village, sample 1, replication 1, purification of Figure 5.
Isolate from Karanggondang Village, 2 samples, 4 replicates, and 2 colony purifiers.
Figure 6.
Isolate from Karanggondang Village, sample 1, replication 4, purification of Macroscopic results from a sampling taken at SEAT of 8 colonies.
Macroscopic observations from sampling at SEAT can be seen in Figures 7 Ae 14.
Figure 7.
Isolate from SEAT, sample 4, replication 4, purification of colony 1.
Rinata et.
Juatika Vol.
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Figure 8.
Isolate from SEAT, sample 4, replication 1, purification of colony 2.
Figure 9.
Isolate from SEAT, sample 4, replication 1, purification of colony 3.
Figure 10.
Isolate from SEAT, sample 5, replication 2, purification of colony 1 Figure 11.
Isolate from SEAT, sample 5, replication 2, purification of colony 1 Rinata et.
Juatika Vol.
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Figure 12.
Isolate from SEAT, sample 5, replication 4, purification of colony 1 Figure 13.
Isolate from SEAT, sample 5, replication 2, purification of colony 1 Figure 14.
Isolate from SEAT, sample 5, replication 4, purification of colony 2 The sampling results from Popongan Village amounted to 3 colonies.
Macroscopic observations from Popongan Village can be seen in Figures 15 Ae 17.
Figure 15.
Isolate from Popongan Village, sample 1, replication 3, purification of Rinata et.
Juatika Vol.
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Figure 16.
Isolate from Popongan Village, sample 1, replication 3, purification of Figure 17.
Isolate from Popongan Village, sample 5, replication 3, purification of Based on the appearance of the images above, a summary can be made, which is presented in Table 2.
Table 2.
Macroscopic morphology of bacteria suspected to be phosphatesolubilizing bacteria No.
Location Karanggondang Village SEAT Popongan Village Isolate A2.
A A2.
2 A1.
1 A2.
2 A1.
2A A1.
1B B4.
1 B4.
2 B4.
3 B5.
B5.
B5.
B5.
B5.
C1.
C1.
C5.
Table 2 shows that 17 isolates from the three locations had different morphologies of bacteria thought to be phosphate-solubilizing bacteria.
this can Colony Color White Milk Yellow White Milk Yellow White Milk Yellow White Milk Transparent White Transparent White Yellow Yellow White Milk Yellow Yellow Transparent White Transparent White White Milk be seen from the different colors of the bacterial colonies.
Determination of bacterial colonies capable of dissolving phosphate Rinata et.
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After macroscopic observations of colonies suspected of being phosphate-solubilizing bacteria were carried out, the transparent zone was continued to be observed.
After testing the transparent zone, 4 colonies of phosphate-solubilizing Observations of the transparent zone can be seen in Figures 18 Ae 21.
Figure 18.
Transparent zone of isolate A1.
1B Figure 19.
Transparent zone of isolate A1.
2A Figure 20.
Transparent zone of isolate B5.
2A Figure 21.
Transparent zone of isolate C1.
1 Rinata et.
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Bacteria will release extracellular enzymes called phosphatase enzymes.
This enzyme will dissolve the insoluble phosphate into soluble, thus forming a transparent zone around the colony.
The diameter of the transparent zone is measured using a ruler.
Observe the transparent zone of bacterial colonies in the petri dish at three different points to find the average area of the transparent zone in each petri dish.
The average measurement of the transparent zone area can be seen in Table 3.
Table 3.
Average transparent zone diameter No.
Isolate A1.
1B A1.
2A B5.
2A C1.
1 Transparent zone diameter .
---------------------------------------------------------------Day-3 Day-5 Day-7 Observations of the transparent zone were carried out until the 7th day because there were no significant changes in the diameter of the transparent zone on the following day.
The isolate with the widest transparent zone was isolate B5.
2A, with an average diameter on day 7 of around 7 mm.
The isolate with the smallest transparent zone was isolate C1.
1 with an average diameter on day 7th, 12.
The wider the transparent zone, the stronger the Average 3 Determination of the Genus of Phosphate Solubilizing Bacteria After observing the transparent zone, microscopic observations of the shape of the bacteria were carried out.
Gram staining is carried out so that the shape of the bacteria can be seen.
Microscopic observations can be seen in Figures 22 Ae 25.
Figure 22.
Isolate A1.
Remark: Bacterial cell shape (Stic.
Gram (-).
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Figure 23.
Isolate A1.
Description: Bacterial cell shape (Stic.
Gram (-).
Figure 24.
Isolate B5.
Description: Bacterial cell shape (Roun.
Gram (-) Figure 25.
Isolate C1.
Description: Bacterial cell shape (Stic.
Gram (-).
Based observations, the bacteria found had a rod and round shape.
Isolate code A1.
1B is rod shaped with gram (-), isolate code A1.
2A is rod shaped with gram (-), isolate code B5.
2A is round with gram (-), isolate code C1.
)1 is a rod with grams (-).
From observations ranging from macroscopic observations, transparent zone observations, and microscopic observations, the characteristics of the phosphate solubilizing bacteria obtained can be seen.
The characteristics of the phosphate-solubilizing bacteria obtained can be seen in Table 4.
Table 4.
Characteristics of phosphate solubilizing bacteria No.
Isolate Colony Edging Elevation Colony Gram Bacteria Bacteria Code Color Shape Size Coloring Shape Name 1 A1.
1B Yellow Hilly Bumpy Medium negative Stem 2 A1.
2A While Wavy Bumpy Medium negative Stem Escherichia 3 B5.
2A Yellow Hilly Bumpy Medium negative Round 4 C1.
1 Yellow Flat Flat Medium negative Stem Acetobacter Description: No.
1 and 3 cannot yet determine the name of the bacteria because the characteristics of the bacteria found are still lacking Rinata et.
Table 4 shows the characteristics phosphate-solubilizing obtained from macroscopic observations, transparent zone tests, and microscopic The phosphate-solubilizing bacteria can be Based on Table 4, the names of bacteria that can be determined are isolated A1.
2A and C1.
1, namely Escherichia Acetobacter.
After carrying out all observations to identify bacteria from each different sampling location.
Macroscopic and microscopic characteristics of phosphatesolubilizing bacteria were obtained from the four isolates.
Isolate A1.
2A has similarities to the Escherichia genus.
This genus is gram negative and rod-shaped (Marista et al.
Based macroscopic observations, transparent zone tests, and microscopic observations of the C1.
1 isolate, this isolate has similarities to the Acetobacter genus.
This bacteria is rod-shaped, gramnegative and non-motile and has a slimy wall surface (Marista et al.
, 2.
There are 2 unknown isolate codes for the genus of phosphate solubilizing bacteria: isolates A1.
1B and B5.
From the characteristics of observations of the isolate, no genus matches the characteristics of the isolate, which is in accordance with the literature.
Therefore, it is best to continue with molecular identification.
CONCLUSION
There phosphate solubilizing bacte ria f rom the Karanggondang Villa ge Popongan Village.
The r esults of macroscopic observat ions f rom obtained 17 isolates with bacterial color morphology of milky wh ite, transparent white and yello w.
The avera ge Juatika Vol.
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transparent zone f rom highest to lo west isolates B5.
2A was 15.
7 mm.
A1.
1B was 13.
7 mm.
A1.
2A 5 mm.
C1 .
1 is 12.
9 mm.
The bacteria that can be identif ied are the Genus Escherichia .
solate A1.
2A) and Acet obacter .
solate C1.
REFERENCE