The 8th Korean-ASEAN Joint Symposium on Biomass Utilization and Renewable Energy

2014

BIODIVERSITY OF POTERAN ISLAND, MADURA, INDONESIA

Aunurohim 1)*, Nengah Dwianita Kuswytasari1), Nur Hidayatul Alami1)

Department of Biology, Institut Teknologi Sepuluh Nopember, Surabaya,

Indonesia

*corresponding author’s Tel: 81-654-40-738, E-mail: aunurohim@bio.its.ac.id

Abstract

The concept of sustainability island is currently a trend in the world overcome

the global crisis. One of the small islands in the East of the island of Madura
named Poteran have succeeded exploration in their natural resource for
further mapping. The survey shows that almost all locations on the island had
been inhabited, soil relatively fertile and planted with several types of
economic value. Marine resources as seaweed need to be handled for more
valuable commodity as carragenan. In addition, 49 bacterial, 9 yeast and 50
isolates of fungi had been obtained, as far as amylolytic, proteolytic,
cellulolytic, lignin degradation and sulfat reducing. These biodiversity indicates
that the Poteran island potentially to be used as one of sustainability island in
the future.

Keywords : sustainability island, Poteran island, biodiversity

INTRODUCTION
The small island in Indonesia still less to be considered, both in terms of its
internal requirements or its usefulness for large islands around it. Often is not
realized actually a small island that has the ability to meet its own internal needs, it
can even contribute to the larger island, because it has the potential unexploited.

The concept of Small Island Developing States (SIDS) has become one
concept that is currently supported for development, especially in
developing countries, in which one goal is to meet their own needs
independently, including energy (Weisser 2004; Weisser 2004b).

Poteran Island which is part of the archipelago in region of Sumenep, East Java,
Indonesia is recommended to be used as a sustainable island because it was
close to the island of Madura. As one of the candidates, the island Poteran must
have the potential to be the exploitation, one of the main focus is agriculture.
Hence the diversity of plants and other natural resources also need to be identified
to support the island Poteran as sustainable islands that meet the needs of
internal or external.

This study is the first step of a three-year research plan, which aims only to
explore biodiversity. The next step will be determined from the results of
this exploration.

MATERIAL AND METHODS

The first activity is to make observations Poteran island as a subject by using a

motor vehicle, to find out if the island deserves to be passed by a motorcycle or a
car.

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Then take the sampling point as a boundary mark for field sampling botany
and microbiology.

Soil sampling

Soil excavated using a trowel that has been sterilized with 96% alcohol and
taken at a depth of 0-10 cm with a sterile bottle. Soil samples were taken
from 5 different sampling points, ie P1 (with coordinates S 07 ° 03'48 .0 "E
113 ° 57'02 .8"), P4 (S 07 ° 04'21 .4 "E 114 ° 01'31 .8"), P5 (S 07 ° 05'28 .3
"E 114 ° 03'10 .6"), P7 (S 07 ° 05'17 .8 "E 113 ° 59'14 .8"), and P8 (S 07 °
05'21 .9 "E 113 ° 56 '30.3 "), with each point taken 5 different plots.

Bottles sterilized by spraying with 96% alcohol and then wiped with tissue
paper. Excavated soil around plant roots on using a crowbar and a screw that
had previously been sterilized with alcohol to a depth of 10 cm. Bottle caps
that have sterilized opened and plugged in the side of the excavation pit. Once
plugged bottles full of urine can be encouraged to get in on the ground side of
the excavation. Excavated portion of land that was attached to the urine bottle
using a screw to seal the bottle of urine on the ground. Once closed and
brought to the surface of the urine bottle again sterilized with 96% alcohol
sprayed and stored in the ice box. Upon arriving at the Laboratory of
Microbiology and Biotechnology, Department of Biology ITS, soil samples
contained in bottles of urine re-sterilized with 96% alcohol and placed in a
freezer at a temperature of -20⁰C.

Isolation of Bacteria

Isolation was done by diluting rise followed by a dispersive method (spread plate)
were performed aseptically in a Laminar Air Flow. Soil samples as much as one
small scalpel inserted into a test tube containing 9 ml of sterile distilled water and
then homogenized and distilled water until evenly mixed. The tube is then called
-1 -1
with 10 dilution. Then from 10 dilution is taken as one ml and transferred to a
test tube containing 9 ml of sterile distilled water and homogenized by using a
 -2
vortex. The tube is then called with 10 dilution. Dilution continues until the dilution
-10 -7 -8 -9 -10
obtained 10 . Dilution of 10 , 10 , 10 , and 10 were taken 100 ml solution
using a micropipette then transferred into a petri dish containing medium Nutrient
Agar (NA). Then leveled inoculum over the entire surface of solid medium by using
Drygalski NA and incubated in an incubator at a temperature of 37⁰C for 48 hours.

Colonies of bacteria growing with its own characteristics and then purified in
order to obtain pure isolates. The bacterial colonies growing on medium-rise
dilution was taken and streaked with a loop using a needle to scratch the
surface of the method 16, and then incubated at 37⁰C for 24 hours. Purification
of bacterial colonies is done in stages at least 3 times the displacement.

Having obtained pure bacterial colonies by macroscopic observation, we then

observed microscopically. Microscopic observation was done by one drop of sterile
distilled water dropped on the surface of the object glass. Colonies of bacteria
were taken as a loop and streaking on glass objects evenly, then fixed over a
Bunsen flame until the distilled water evaporate. Provision has been made
preparations were then called preparations pillowcase. Mixture commentator
added with 2-3 drops of ethylene blue dye and wait 1-2 minutes. Preparations
were washed using running water to remove excess dye excess. Mixture wind
dried up to be completely dries (Harley and

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The 8th Korean-ASEAN Joint Symposium on Biomass Utilization and Renewable Energy
2014

Prescott, 2002). Preparations covered with glass cover and then sprinkled
with immersion oil.

Yeast

A total of 10 gr of soil sample was mixed with 90 ml of sterile physiological water

were homogenized and precipitated 5 minutes. The suspension was taken from
the bottle and made serial dilutions in sterile distilled water. From each dilution
series were determined, as many as one ml was taken and put into a petridish and
then added Yeast Malt Extract Agar (YMEA) warm solution containing
chloramphenicol 1%. Then incubated at 25-30⁰C for 3 days. Data on the number
of yeast colonies on plates used to calculate the amount of yeast per gram of soil
(cfu/g).

Isolation of Yeast

a total of 10 ml of the supernatant obtained from 10 g soil samples were

mixed with 90 ml of sterile physiological water put in 40 ml of multiplication
medium (enrichment) yeast. Medium used is the medium Yeast Malt Broth
(YMB). The cultures were incubated by using a shaker for 3 days. After
that, the suspension inoculated into sterile petri dishes that had been
added to 1 ml of 1% solution of chloramphenicol. Subsequently, the media
was poured into a YMEA and incubated at room temperature for 3 days.

At the end of the incubation period was observed macroscopic characteristics of yeast
isolates were grown. Colonies that have different macroscopic characteristics of the
medium YMEA subsequently inoculated slant. Then incubated at 25-30⁰C for ± 3 days. If
you've got a uniform cell shape, then obtained pure isolates were ready to test the
identification and testing of potentially screening yeast in phosphate dissolving, degrading
cellulose, lipids, and proteins. If unpurified isolates obtained, then be purified by using the
streak method, to obtain pure isolates for testing purposes and collections. Each pure
isolates were obtained subsequently given a different code.

Fungi
Solid medium used was Potato Dextrose Agar (PDA), containing 100 mg of
Chloramphenicol. Medium is made from 200 grams of potatoes, boiled in
1000 ml of distilled water for 2 hours. The resulting extract is then added 20
grams of dextrose / glucose, 15 g agar and 100 mg Chloramphenicol,
homogenized and heated with a hot plate magnetic stirrer until boiling.
Medium is sterilized by autoclaving at 121⁰C and 1,5 atm pressure for 15
minutes.

Isolation performed aseptically and made duplicate. The suspension is the result
of enrichment by 1 ml dissolved in 9 ml of sterile distilled water and then the
suspension is referred to as dilution 10-1. Subsequently 1 ml suspension of a 10-1
dilution dissolved in 9 ml of sterile distilled water and is referred to as dilution 10-2.
Dilution is done in stages until a dilution of 10-3. 0.1 ml of dilution was taken and
inoculated into the center of the Petri dish containing approximately 15 ml of PDA
medium 100 ‰ sterile Chloramphenicol is solid, with the method of spread
(spread plate). Incubation was carried out at room temperature for 3-7 days (Elias,
2009).

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The 8th Korean-ASEAN Joint Symposium on Biomass Utilization and Renewable Energy
2014

RESULT

Mapping was done by using the motorcycle and perform soil sampling in
accordance with prescribed sampling points are P1 to P8. Coordinate
points P1 to P8 is shown as follows:

P1 S 07 03’48.0” E 113 57’02.8”

P2 S 07 04’26.2” E 113 59’35.4”

P3 S 07 04’22.4” E 114 00’35.4”

P4 S 07 04’18.8” E 114 01’31.8”

P5 S 07 05’28.3” E 114 03’10.6”

P6 S 07 06’31.6” E 114 02’47.5”

P7 S 07 05’17.8” E 113 59’14.8”

P8 S 07 05’21.9” E 113 56’30.3”

And basic mapping Poteran Island shown below:

Figure 1. Poteran Island and the location of sampling points used. This location is a
representation of the path that can be traversed by car or motorcycle. One goal of this mapping
is to determine the existence of biodiversity that can be enhanced as good commodity
Quality of soil

From the eight points, then conducted soil sampling and obtained the following
data:

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Table 1. Data quality of the soil in several locations on the island of Poteran, Madura

Site
Corg (%)
Ntot (%)
C/N
OM (%)
P.Bray 1
K
KTK

location

(mg/kg)
(me/100gr)
me/100 gr)
P1
2,23
0,28
8
3,86
3,79
0,40
28,60

P4
0,45
0,08
6
0,78
21,31
0,09
4,90

P5
1,33
0,19
7
2,31
6,96
0,57
25,24

P6
1,39
0,20
7
2,41
2,14
0,28
25,26

P7
1,39
0,20
7
2,41
2,14
0,28
25,26

P8
4,81
0,56
9
8,32
0,53
0,29
20,93

The data above shows those in terms of soil quality on the island in
Poteran are low fertile but viable as location for cultivation.

Plants and Herbs

Plant and herbs that found at Poteran islands are as follows:

Table 2. Herbs and plants found on the island Poteran (as long a road)

No
Scientific name
Local name
Criteria

1.
Tectonia grandis
Jati
Abundant

2.
Hibiscus tiliaceus
Waru Laut
Abundant

3.
Leucaena leucocephala
Lamtoro
Frequent

4.
Leucaena glauca
Petai cina
Frequent

5.
Codiaeum variegatum
Puring
Occasion

6.
Musa paradisiaca
Pisang
Abundant

7.
Cocos nucifera
Kelapa
Frequent

8.
Manihot utilisima
Ketela pohon
Dominant

9.
Acacia ariculiformis
Akasia
Occasion

10.
Ceiba petandra
Randu
Occasion

11.
Mangifera indica
Mangga
Frequent

12.
Muntingia calabura
Kersen
Occasion

13.
Cassia siamea
unknown
Frequent

14.
Carica papaya
Pepaya
Occasion

15.
Sonneratia caseolaris
Bogem
Occasion

16.
Sesbania grandiflora
Turi
Occasion

17.
Rhizopora stylosa
Bakau
Occasion

18.
Elaeis sp
Kelapa Sawit
Occasion

19.
Livistona chinensis
Palem Kipas
Occasion

20.
Azadirachta indica
Mimbo
Frequent

21.
Lamtana camara
unknown
Abundant

22.
Borassus flabellifer
unknown
Frequent

23.
Monihot glaziovii
Ketela Hutan
Rare
24.
Bambusa sp
Bambu
Frequent

25.
Pandanus amaryllfolius
Pandan Wangi
Frequent

26.
Nicotiana tabacum
Tembakau
Abundant

27.
Pithecelobium dulce
Asem londho
Occasion

28.
Tamarindus indica
Asem Jawa
Occasion

29.
Artocarpus altilis
Sukun
Frequent
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The 8th Korean-ASEAN Joint Symposium on Biomass Utilization and Renewable Energy
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30.
Capsicum frutescens
Lombok
Dominant

31.
Terminalia catappa
Ketapang
Occasion

32.
Artocarpus heterophyllus
Nangka
Occasion

33.
Zea mays
Jagung
Occasion

34.
Calathea gigantea
unknown
Frequent

35.
Arachis hypogea
Kacang tanah
Frequent

36.
Casuarina equisetifolia
Cemara laut
Occasion

Seaweed
Euchema cottonii is seaweed commodity that is quite important in Poteran.
Seaweed farmers have relied that E.cottonii as one of the two commodities to
increase their income other than fishing. However, the basic concept of
planting is just making a sale in the form of wet biomass, instead of other uses
such as carrageenan. Prajapati et al (2014), states that the carrageenan is one
of the most important commodities from seaweed, and have high value.

Based on interviews with local farmers, there is about 28 seaweed’s farmers in

Poteran. Each farmer has about 4-5 “ancak” which is one of the seaweed
cultivation techniques traditionally using bamboo. Each “ancak” within 45 days of
cultivation can produce about 800-1000 kg wet seaweed, where the price of wet
seaweed is around 14,000 IDR / kg. From these data, if we predicted cash flow of
money circulating in the community at any time of harvest is about 1.96 billion
IDR, -.

However, this condition can be improved if the main focus is no longer sold
seaweed biomass per kg, but it can be worth as carrageenan with
165,000 / kg or up to 10 x higher than the wet biomass. Therefore, the
results of these observations will be continued research to determine the
ratio of wet biomass of seaweed and carrageenan produced.

Microbial Richness

While the microbial inhabitants of the land, has successfully identified 49

bacterial isolates, 9 isolates of yeast and 50 isolates of fungi. From forty-
nine bacterial isolates will further test the potential of amylolytic, proteolytic,
cellulolytic and lipolilitik. As for yeast isolates will be tested further as a
potential phosphate solvent, cellulolytic, proteolytic and lignin degrader.
Meanwhile, for fungi isolates will be tested as sulphate reducer.

CONCLUSION

Exploration of biodiversity on the island Poteran at first year has been identified some
natural resources such as plants, seaweed, and also soil microbes.

There are 36 species of higher plants with different levels of classification

Potential seaweed Eucheuma cottonii is still focused on the wet biomass, yet on the other
potentials that have a higher economic value such as carrageenan.

There were 49 isolates of bacteria, 9 isolates of yeast and 50 isolates of fungi that
will be developed for some further potential as amylolitic, proteolytic, cellulolitic,
phosphate solvent, lignin degrader, and also sulphate reducers

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Acknowledgements

Thanks to Biology department of ITS, LPPM of ITS, DP2M, and head of village of Poteran,
and the individual personnel that can not mention one by one. This project has been
funded by BOPTN ITS with number of contract 013674.175/IT2.7/PN.08.01/2013.

REFERENCES

Alexopoulus, C. J., C. W. Mims and M. Blackwell. 1996. Introductory

th
Mycology. 4 ed. John Wiley & Sons, Inc. Canada.

Anonym. 2005. Analisa kimia tanah, tanaman, air dan pupuk. Balai
Penelitian Tanah Badan Penelitian dan Pengembangan Pertanian,
Departemen Pertanian Republik Indonesia. 143 hal.

Gandjar, I., R. A. Samson, K. van den Tweel-Vermeulen, A. Oetari dan I.

Santoso. 1999. Pengenalan Kapang Tropik Umum. Yayasan Obor Indonesia.
Jakarta.

Ilyas, Muhammad. 2005. Kelimpahan dan Keragaman Kapang pada

Sampel Tanah di Sekitar Kawasan Taman Nasional Gunung Ciremai, Jawa
Barat. Jurnal Biologi Indonesia 5 (3): 245-257.

Lee, J. Y. and B. K. Hwang. 2002. Diversity of Antifungal Actinomycetes in

Various Vegetative Soil of Korea. Journal Microbiology 48: 407-417.
Prajapati, V D. Maheriya, P M. Jani, G K. Solanki, H K. 2014. Review :
Carrageenan: A Natural Seaweed Polysaccharide and its application.
Carbohydrates Polymers 105 : 97-112.

Waluyo, L. 2004. Mikrobiologi Umum. UMM Press. Malang.

Weisser, D. (2004b). On the economics of electricity consumption in small island

developing states: a role for renewable energy technologies? Energy Policy , 127 - 140.

Weisser, D. (2004). Power sector reform in small island developing: what

role for Renewable Energy Technologies? Renewable and Sustainable
Energy Reviews , 101 - 127 .

8 Korea-ASEAN Simposium Bersama Biomassa Pemanfaatan dan Energi Terbarukan

2014
KEANEKARAGAMAN OF Pulau Poteran, MADURA, INDONESIA

Aunurohim 1) *, Nengah Dwianita Kuswytasari1), Nur Hidayatul Alami1)

1) Departemen Biologi, Institut Teknologi Sepuluh Nopember, Surabaya,
Indonesia
* penulis yang sesuai untuk Tel: 81-654-40-738, E-mail: aunurohim@bio.its.ac.id

Abstrak

Konsep keberlanjutan pulau saat ini menjadi trend di dunia mengatasi krisis global.
Salah satu pulau kecil di timur Pulau Madura bernama Poteran telah berhasil
eksplorasi di sumber daya alam mereka untuk pemetaan lebih lanjut. Survei
menunjukkan bahwa hampir semua lokasi di pulau itu telah dihuni, tanah yang relatif
subur dan ditanami dengan beberapa jenis nilai ekonomi. Sumber daya laut sebagai
kebutuhan rumput laut yang akan ditangani untuk komoditas yang lebih berharga
sebagai karaginan. Selain itu, 49 bakteri, ragi dan 9 50 isolat jamur telah diperoleh,
sejauh amilolitik, proteolitik, selulolitik, degradasi lignin dan sulfat mengurangi.
Keanekaragaman hayati ini menunjukkan bahwa pulau Poteran berpotensi untuk
digunakan sebagai salah satu pulau keberlanjutan di masa depan.

Kata kunci: Pulau keberlanjutan, pulau Poteran, keanekaragaman hayati

PENGANTAR

Pulau kecil di Indonesia masih kurang diperhatikan, baik dari segi persyaratan internal
atau kegunaannya untuk pulau besar di sekitarnya. Sering tidak menyadari
sebenarnya sebuah pulau kecil yang memiliki kemampuan untuk memenuhi
kebutuhan internal sendiri, bahkan dapat memberikan kontribusi ke pulau yang lebih
besar, karena memiliki potensi dieksploitasi.

Konsep Pulau Kecil Mengembangkan Amerika (SIDS) telah menjadi salah satu konsep
yang saat ini didukung untuk pengembangan, terutama di negara-negara
berkembang, di mana salah satu tujuannya adalah untuk memenuhi kebutuhan
mereka sendiri secara mandiri, termasuk energi (Weisser 2004; Weisser 2004b).

Pulau Poteran yang merupakan bagian dari kepulauan di wilayah Sumenep, Jawa
Timur, Indonesia dianjurkan untuk digunakan sebagai sebuah pulau yang
berkelanjutan karena itu dekat dengan pulau Madura. Sebagai salah satu kandidat,
pulau Poteran harus memiliki potensi untuk menjadi eksploitasi, salah satu fokus
utama adalah pertanian. Oleh karena keragaman tanaman dan sumber daya alam
lainnya juga perlu diidentifikasi untuk mendukung pulau Poteran sebagai pulau yang
berkelanjutan yang memenuhi kebutuhan internal atau eksternal.

Penelitian ini merupakan langkah pertama dari rencana penelitian tiga tahun, yang
bertujuan hanya untuk mengeksplorasi keanekaragaman hayati. Langkah selanjutnya
akan ditentukan dari hasil eksplorasi ini.

BAHAN DAN METODE

Kegiatan pertama adalah membuat pengamatan pulau Poteran sebagai subjek dengan
menggunakan kendaraan bermotor, untuk mengetahui apakah pulau layak untuk
dilewati oleh sepeda motor atau mobil.

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Kemudian mengambil titik sampling sebagai tanda batas untuk botani bidang
sampling dan mikrobiologi.

Pengambilan contoh tanah

Tanah digali dengan menggunakan sekop yang sudah disterilkan dengan alkohol 96%
dan diambil pada kedalaman 0-10 cm dengan botol steril. Sampel tanah yang diambil
dari 5 titik sampling yang berbeda, yaitu P1 (dengan koordinat S 07 ° 03'48 0,0 "E 113
° 57'02 0,8"), P4 (S 07 ° 04'21 0,4 "E 114 ° 01 '31 0,8 "), P5 (S 07 ° 05'28 0,3" E 114 °
03'10 0,6 "), P7 (S 07 ° 05'17 0,8" E 113 ° 59'14 0,8 ") , dan P8 (S 07 ° 05'21 0,9 "E 113 °
56 '30 0,3"), dengan masing-masing titik diambil 5 plot yang berbeda.

Botol disterilkan dengan menyemprotkan dengan 96% alkohol dan kemudian

menyeka dengan kertas tisu. Digali tanah di sekitar akar tanaman menggunakan
linggis dan sekrup yang sebelumnya telah disterilkan dengan alkohol hingga
kedalaman 10 cm. Tutup botol yang telah disterilkan dibuka dan terpasang di sisi
lubang penggalian. Setelah terpasang botol penuh urin dapat didorong untuk masuk
di sisi dasar penggalian. Digali sebagian dari tanah yang melekat pada botol urin
menggunakan sekrup untuk menutup botol urin di tanah. Setelah ditutup dan dibawa
ke permukaan botol urine lagi disterilkan dengan alkohol 96% disemprotkan dan
disimpan dalam kotak es. Setelah tiba di Laboratorium Mikrobiologi dan Bioteknologi,
Jurusan Biologi ITS, sampel tanah yang terkandung dalam botol urine ulang disterilkan
dengan alkohol 96% dan ditempatkan di dalam freezer pada suhu -20⁰C.

Isolasi Bakteri
Isolasi dilakukan dengan cara pengenceran bertingkat diikuti dengan metode (spread
plate) sebar dilakukan secara aseptik dalam Air Flow Laminar. Sampel tanah sebanyak
satu pisau kecil yang dimasukkan ke dalam tabung reaksi yang berisi 9 ml air suling
steril dan kemudian homogen dan air suling sampai tercampur rata. Tabung tersebut
kemudian disebut dengan 10-1 pengenceran. Kemudian 10-1 pengenceran diambil
sebagai salah satu ml dan dipindahkan ke tabung reaksi yang berisi 9 ml air suling
steril dan dihomogenisasi dengan menggunakan pusaran. Tabung tersebut kemudian
disebut dengan 10-2 pengenceran. Pengenceran terus sampai pengenceran yang
diperoleh 10-10. Pengenceran 10-7, 10-8, 10-9, dan 10-10 diambil 100 ml larutan
menggunakan mikropipet kemudian ditransfer ke dalam cawan petri yang berisi
media Nutrient Agar (NA). Kemudian diratakan inokulum atas seluruh permukaan
media padat dengan menggunakan Drygalski NA dan diinkubasi dalam inkubator pada
suhu 37⁰C selama 48 jam.

Koloni bakteri yang tumbuh dengan karakteristik sendiri dan kemudian dimurnikan
untuk mendapatkan isolat murni. Koloni bakteri yang tumbuh pada medium-rise
pengenceran diambil dan melesat dengan lingkaran menggunakan jarum untuk
menggores permukaan metode 16, dan kemudian diinkubasi pada 37⁰C selama 24
jam. Pemurnian koloni bakteri dilakukan secara bertahap minimal 3 kali perpindahan.

Memiliki diperoleh murni koloni bakteri dengan pengamatan makroskopis, kita

kemudian diamati dengan mikroskop. Pengamatan mikroskopis dilakukan dengan satu
tetes air suling steril dijatuhkan di permukaan kaca obyek. Koloni bakteri diambil
sebagai loop dan melesat pada objek gelas merata, kemudian tetap di atas api Bunsen
sampai menguap air suling. Kesepakatan telah dibuat persiapan kemudian disebut
persiapan sarung bantal. Komentator campuran ditambahkan dengan 2-3 tetes
etilena pewarna biru dan tunggu 1-2 menit. Persiapan dicuci menggunakan air
mengalir untuk menghilangkan kelebihan zat warna berlebih. Angin campuran kering
harus benar-benar mengering (Harley dan

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Prescott, 2002). Persiapan ditutupi dengan penutup kaca dan kemudian ditaburi
dengan minyak imersi.

Ragi

Sebanyak 10 gr sampel tanah dicampur dengan 90 ml air fisiologis steril

dihomogenisasi dan diendapkan 5 menit. Suspensi diambil dari botol dan membuat
pengenceran serial dalam air suling steril. Dari setiap seri pengenceran ditentukan,
sebanyak satu ml diambil dan dimasukkan ke dalam petridish dan kemudian
ditambahkan ragi Malt Extract Agar (YMEA) solusi hangat yang mengandung
kloramfenikol 1%. Kemudian diinkubasi pada 25-30⁰C selama 3 hari. Data jumlah
koloni ragi di piring yang digunakan untuk menghitung jumlah ragi per gram tanah
(cfu / g).

Isolasi Ragi

total 10 ml supernatan yang diperoleh dari sampel tanah 10 g dicampur dengan 90 ml

air fisiologis steril dimasukkan ke dalam 40 ml media multiplikasi (pengayaan) ragi.
Media yang digunakan adalah media Yeast Malt Broth (YMB). Kultur diinkubasi
dengan menggunakan shaker selama 3 hari. Setelah itu, suspensi diinokulasi ke dalam
cawan petri steril yang telah ditambahkan ke 1 ml larutan 1% kloramfenikol.
Selanjutnya, media dituangkan ke YMEA dan diinkubasi pada suhu kamar selama 3
hari.

Pada akhir masa inkubasi diamati karakteristik makroskopik isolat khamir ditanam.
Koloni yang memiliki karakteristik makroskopik yang berbeda dari media YMEA
kemudian diinokulasi miring. Kemudian diinkubasi pada 25-30⁰C selama ± 3 hari. Jika
Anda punya bentuk sel yang seragam, maka diperoleh isolat murni siap untuk menguji
identifikasi dan pengujian berpotensi skrining ragi dalam melarutkan fosfat,
merendahkan selulosa, lipid, dan protein. Jika isolat unpurified diperoleh, kemudian
dimurnikan dengan menggunakan metode beruntun, untuk mendapatkan isolat
murni untuk tujuan pengujian dan koleksi. Setiap isolat murni diperoleh kemudian
diberi kode yang berbeda.

Jamur

Medium padat yang digunakan adalah Potato Dextrose Agar (PDA), mengandung 100
mg Kloramfenikol. Sedang dibuat dari 200 gram kentang, direbus dalam 1000 ml air
suling selama 2 jam. Ekstrak yang dihasilkan kemudian ditambahkan 20 gram
dekstrosa / glukosa, 15 g agar dan 100 mg Kloramfenikol, homogen dan dipanaskan
dengan pengaduk magnet piring panas sampai mendidih. Medium disterilkan dengan
autoclave pada 121⁰C dan 1,5 tekanan atm selama 15 menit.

Isolasi dilakukan secara aseptik dan membuat duplikat. Suspensi merupakan hasil
pengayaan oleh 1 ml dilarutkan dalam 9 ml air suling steril dan kemudian suspensi
disebut sebagai pengenceran 10-1. Selanjutnya 1 ml suspensi dari 10-1 pengenceran
dilarutkan dalam 9 ml air suling steril dan disebut sebagai pengenceran 10-2.
Pengenceran dilakukan secara bertahap sampai pengenceran 10-3. 0,1 ml
pengenceran diambil dan diinokulasi ke pusat cawan Petri yang berisi sekitar 15 ml
media PDA 100 ‰ Kloramfenikol steril solid, dengan metode penyebaran (spread
plate). Inkubasi dilakukan pada suhu kamar selama 3-7 hari (Elias, 2009).
19-20 Agustus 2014, Universitas Korea, Seoul, Korea Page 3

8 Korea-ASEAN Simposium Bersama Biomassa Pemanfaatan dan Energi Terbarukan

2014

HASIL

Pemetaan dilakukan dengan menggunakan sepeda motor dan melakukan

pengambilan contoh tanah sesuai dengan poin yang ditentukan sampling P1 untuk P8.
Titik koordinat P1 ke P8 ditunjukkan sebagai berikut:

P1 S 07 03'48.0 "E 113 57'02.8"

P2 S 07 04'26.2 "E 113 59'35.4"

P3 S 07 04'22.4 "E 114 00'35.4"

P4 S 07 04'18.8 "E 114 01'31.8"

P5 S 07 05'28.3 "E 114 03'10.6"

P6 S 07 06'31.6 "E 114 02'47.5"

P7 S 07 05'17.8 "E 113 59'14.8"

P8 S 07 05'21.9 "E 113 56'30.3"

Dan pemetaan dasar Pulau Poteran ditunjukkan di bawah:

19-20 August 2014, Korea University, Seoul, Korea Page 7