KARAKTER MORFOLOGI DAN MOLEKULER ISOLAT Phytophthora palmivora ASAL KELAPA DAN KAKAO

HIASINTA FJ MOTULO, MEITY S-SINAGA, GEDE SUASTIKA, HAJRIAL ASWIDINNOOR, ALEX HARTANA

Abstract


ABSTRAK
Phytophthora palmivora merupakan patogen penyebab penyakit
gugur buah pada tanaman kelapa dan busuk buah pada tanaman kakao.
Penelitian ini bertujuan untuk membedakan isolat P. palmivora asal kelapa
dan asal kakao berdasarkan karakter morfologi dan molekuler.
Pengambilan sampel penyakit gugur buah kelapa dan busuk buah kakao
dilakukan di Kabupaten Banyuwangi dan Jember, Jawa Timur, Kabupaten
Minahasa dan Bolaang Mongondow, Sulawesi Utara, dan Kabupaten
Gorontalo, Gorontalo. Analisis morfologi, ekstraksi DNA dan amplifikasi
DNA dengan PCR dilakukan di Laboratorium Mikologi dan Laboratorium
Virologi, Departemen Proteksi Tanaman, Faperta IPB. Analisis perunutan
DNA dilakukan di Laboratorium Biologi Molekuler, Balai Besar
Bioteknologi dan Sumberdaya Genetik dan Laboratorium Bioteknologi,
LIPI Serpong. Penelitian dilaksanakan pada bulan April 2005 sampai
Februari 2007. Berdasarkan karakter morfologi seperti diameter koloni,
panjang dan lebar sporangium, tipe koloni, bentuk sporangium, per-
bandingan panjang dan lebar sporangium serta runutan DNA ruas ITS
menunjukkan bahwa keduapuluh-dua koleksi isolat yang menunjukkan
gejala penyakit gugur buah kelapa dan busuk buah kakao adalah P.
palmivora. Isolat P. palmivora asal kelapa berbeda dengan isolat P.
palmivora asal kakao berdasarkan diameter koloni, panjang dan lebar
sporangium serta runutan DNA ruas ITS. Duapuluh-dua isolat P.
palmivora asal kelapa dan asal kakao mempunyai sporangium yang mudah
lepas dari sporangiospora (caducous), pedikel yang pendek dan papila
serta bervariasi dalam bentuk dan ukuran sporangium. Bentuk sporangium
terdiri dari 4 tipe yaitu ovoid, limoniform, obturbinate, dan obpyriform.
Ukuran sporangium berkisar antara 40 – 62 µm panjang dan 28 – 43 µm
lebar. Isolat P. palmivora memiliki tipe koloni rosaceous, stelate dan
cottony. Rata-rata diameter koloni isolat asal kelapa 54.8 cm lebih tinggi
dari isolat asal kakao 43,4 cm. Hasil perunutan DNA hasil PCR
menunjukkan adanya keragaman genetik antar isolat asal kelapa dan kakao
di Indonesia. Isolat asal kakao berbeda dengan isolat asal kelapa
berdasarkan perunutan DNA ruas ITS. Isolat P. palmivora asal kelapa dan
kakao dari Indonesia tidak berada dalam satu kelompok dengan isolat
yang berasal dari Thailand, Taiwan, Korea, Puerto Rico, Ghana, dan
Cameron.
Kata kunci : Kelapa, Cocos nucifera, kakao, Theobroma cacao, penyakit,
P. palmivora, morfologi, molekuler, keragaman, runutan
DNA-ITS, Jawa Timur, Sulawesi Utara, Gorontalo, Jawa
Barat
ABSTRACT
Morphology  and  molecular  characteristics  of  P.
palmivora isolates from coconut and cacao
Phytophthora palmivora, is the pathogen of coconut nutfall and
cacao black pod diseases. This study was conducted to differentiate the
isolates of P. palmivora from coconut and those from cacao fruit based on
morphology and molecular characteristics. Samples of nutfall of coconut
and black pod of cacao were collected from Banyuwangi and Jember
Districts, East Java, Minahasa and Bolaang Mongondow Districts, North
Sulawesi, and Gorontalo District, Gorontalo. Morphological analysis,
DNA extraction and amplification of PCR-DNA were conducted in
Micology Laboratorium and Virology Laboratorium, Plant Protection
Division, Faperta IPB. Sequencing DNA analysis was conducted in
Molecular Biology Laboratory, Balai Besar Bioteknologi dan Sumberdaya
Genetik and Biotechnology Laboratory LIPI Serpong. This research was
conducted from April 2005 to February 2007. Comparative morphological
evaluated i.e. diameter of colony, length and width of sporangium, l/w
ratio, type of colony and sequence Internal Transcribed Sequence (ITS)-
DNA showed that all isolates of Phytophthora isolated from coconut and
cacao  in  Indonesia  were  Phytophthora  palmivora.  Morphology
characteristics of pathogen isolates from cacao were smaller and
significantly different in length, width, length/width ratio of
sporangium and diameter of colony compared to coconut’s isolates.
Sporangia of 22 isolates were caducous with short pedicel, but were
variable in shape and size. The culture produced ovoid, limoniform,
obturbinate, dan obpyriform sporangia, average 40-62 µm in length and
28-43 µm in width. The colony types were stelate, cottony and rossaceous
with average diameter of coconut isolates 54.8 cm and cacao isolates 43.4
cm. Specific fragment of 900 bp was successfully amplify from coconut
and cacao infected by P. palmivora. The DNA sequence analysis of the
nuclear ribosomal internal transcribed spacer (ITS) region showed that the
coconut isolates were not in the same cluster with the cacao isolates. Based
on sequence analysis, the P. palmivora isolates from Indonesia showed
different cluster from those of Taiwan, Ghana, Puerto Rico and Costa Rica
isolates.
Key words :  Coconut, Cocos nucifera, cacao, Theobroma cacao,
diseases, P. palmivora, diversity, morphology, molecular,
sequencing  ITS-DNA,  East  Java,  North  Sulawesi,
Gorontalo, West Jav

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DOI: http://dx.doi.org/10.21082/jlittri.v13n3.2007.111-118

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