BAB VI Penyediaan Inokulum Steril
Gigaspora margarita dan Acaulospora tuberculata Kultur Aksenik dengan Eksplan Tomat dan Wortel
ABSTRAK
Penyediaan inokulum CMA pada umumnya menggunakan kultur pot dalam rumah kaca, kendala yang dihadapai yaitu kultur tidak murni terkontaminasi oleh
mikroorganisme lain, dan keterbatasan lahan. Salah satu upaya menanggulangi masalah tersebut adalah pemanfaatan kultur aksenik
in vitro. Namun, belum diperoleh teknik sterilisasi CMA yang tepat. Tujuan penelitian mendapat i
metode sterilisasi spora CMA, ii media dan tanaman inang sesuai atau kompatibel untuk penyediaan inokulum CMA steril, iii perkembangan
G. magarita dan A. tuberculata in vitro. Perbanyakan tanaman wortel dengan
perkecambahan biji. Perbanyakan tanaman tomat menggunakan tunas, dan media ditambah kinetin 0,1, 0,5 dan 1,0 mgl. Bahan wortel dan tomat
in vitro diadaptasikan pada medium minimum MM dan medium Strullu dan Romand
MSR. Sumber spora G. margarita dan A. tuberculata dari kultur in vivo di
rumah kaca percobaan II, disterilisasi dengan tiga metode. Masing–masing pora steril diinokulasikan ke dalam akar plantlet wortel dan diinkubasi dalam ruang
kultur cahaya 75-100 luks 12 jamhari, suhu 26
o
C dan Rh 80 . Percobaan dilaksanakan menggunakan Ra ncangan Acak Lengkap Faktorial 2x2x2x5
. Hasil
yang diperoleh menunjukkan bahwa sterilisasi terbaik menggunakan MgSO
4
.7H
2
O 0,1, 10 Tween 20 steril, dan permukaan spora dibersihkan NaOCl
2
0,52 Bayclin10. Spora disterilkan dengan antibiotik gentamisin, streptomisin dan penisilin masing- masing 100 mgl dan tetrasiklin 2,5 mg l
. Spora
G. margarita steril berkecambah antara 11-15 hari sedang A. tuberculata 40-50 hari setelah inokulasi. Media yang sesui untuk kedua jenis CMA adalah MM dan
MSR, sedang yang terbaik adalah MM. Tanaman wortel dan tomat kompatibel, namun lebih baik wortel untuk perkembangan
G. magarita dan A. tuberculata kultur aksenik. Kedua CMA tersebut berkecambah, hifa eksternal berkembang
menginfeksi akar inang, membentuk arbuskula, ve sikula dan membentuk spora baru sporulasi. Kesimpulan bahwa kedua CMA uji dapat dikembangkan secara
in vitro, walaupun perbanyakan spora masih rendah. ----------------------------------------
Kata kunci:
G. margarita, A. tuberculata, teknologi kultur-aksenik, sterilisasi
spora, perkecambahan spora CMA, wortel, tomat
73
Sterile Inoculum Supply of AMF Gigaspora margarita and Acaulospora tuberculata
Axenic Culture with Explant of Tomato and Carrot
ABSTRACT
Generally conventional production AMF inoculum was carried out with culture pot in green house or in the field, this method faced many problems
because of contamination with other microorganisms and limited farm area
.
The weakness can be overcome by AMF in vitro culture with sterile AMF inoculum.
The objectives of the research are i to obtain AMF spore sterilization technique, ii to formulate media and host plants for supporting the sufficient sterile spore,
and iii to develop the inoculum of G. margarita and A. tuberculata by axenic culture. Plant materials such as carrots multiplied by seeds and tomato could be
also propagated by axillary buds in Murashige and Skoog medium with the addition of 0.1, 0.5 and 1.0 mgl kinetin. Furthermore, planlets were adapted in
minimum medium MM and Strullu and Romand medium MSR. Spores of G. margarita
and A. tuberculata propagated in green house from experiment II with three types of sterilants were used in this experiment. Spore from each AMF was
inoculated into planlet roots of each tested plants, and incubated in light rooms 75-100 lux for 12 hoursdays, temperature 26
o
C, Rh 80 . This study is designed
in Randomized Completed Design factorial 2x2x2x5. The results showed that sterilants consisted of MgSO
4
.7H
2
O 0.1 and 10 sterile Tween 20, while the spore surface was cleaned by NaOCl
2
0.52 Bayclin10 . The spores were sterilized by antibiotics gentamycin, streptomycin, penicillin of 100 mgl each and
tetracycline of 2.5 mgl. The results showed that sterile G. margarita germinated between 11-15 days, while A. tuberculata germinated between 40-50 days after
inoculation. The most suitable media for those AMF are MM and MSR while MM is the best. These two host plants are compatible for growing and developing of
G. margarita
and A. tuberculata in monoaxenic culture. G. margarita and A. tuberculata
were able to infect tomato and carrot in the MM and MSR forming external hyphae which infect root producing, arbuscular, vesicular and
sporulation. This concluded that the development of G. margarita and A. tuberculata
can be carried out by carrot and tomato as a host in axenic culture. Association of AMF G. margarita and A. tuberculata with tomato and carrot
plants could successfully be carried out by axenic culture, however the sporulation was low.
--------------------------------
Key words :
G. margarita , A. tuberculata, monoaxcenic culture, sterilization-spore,
germination-AMF spore, tomato, carrot.
1. PENDAHULUAN
Tanaman yang bermikorhiza dapat meningkatkan penyerapan P, Zn, N, K, dan Cu, sehingga tanaman yang bermikorhiza meningkatkan pertumbuhan, serta
Kategori