Regenerasi Beberapa Varietas dan Overekspresi Gen

BIDANG ILMU : PERTANIAN
LAPORAN HASIL
PENELITIAN DISERTASI DOKTOR
TAHUN ANGGARAN 2010
Judul
Peneliti
: Regenerasi Beberapa Varietas dan Overekspresi
Gen soSPS1 Untuk Meningkatkan Kandungan
Tomat (Lycopersicon esculentum, Mill.)
:::: Ir. Parawati Dewanti , MP
Dibiayai Oleh Direktorat Jenderal Pendidikan Tinggi, Kementerian Pendidikan
Nasional sesuai dengan Surat Perjanjian Penugasan Dalam Rangka Pelaksanaan
Penugasan Penelitian Disertasi Doktor Tahun Anggaran 2010
Nomor : 492/SP2H/PP/DP2M/VI/2010, tanggal 11 Juni 2010.
Universitas Brawijaya
Malang
2010
Ringkasan
Tomat (Lycopersicon esculentum Mill.) merupakan komoditas sayuran buah
yang berperan penting dalam perekonomian di Indonesia. Tanaman ini merupakan
tanaman dikotil yang termasuk dalam famili Solanaceae. Dalam proses fotosintesis
tanaman tomat akan menghasilkan sukrosa dan pati. Fungsi sukrosa pada tanaman
yang sedang mengalami pertumbuhan adalah sebagai sumber energi dan sumber
kerangka karbon dalam metabolisme. Upaya untuk meningkatkan produksi sukrosa
pada tanaman tomat dapat dilakukan melalui metode transformasi genetik.
Transformasi gen sudah pernah dilakukan pada beberapa tanaman dikotil
seperti tomat, tembakau dan lainnya. Banyak macam gen yang dapat ditransformasi
ke tanaman, antara lain gen yang mengkode untuk protein SPS. Sukrosa Phophat
Synthase (SPS) merupakan enzim yang menentukan kandungan sukrosa dan
merupakan enzim kunci dalam asimilasi karbon pada tanaman. Kloning gen SPS
(Sucrose Phophat Synthase) dari tebu yang dinamakan dengan SoSPS1 sudah
berhasil dilakukan di Laboratorium Biologi Molekul Universitas Jember.
Tujuan dari penelitian ini adalah untuk mengetahui peningkatan kandungan
sukrosa pada buah tomat akibat over-ekspresi gen SPS. Hasil penelitian diharapkan
dapat diperoleh tanaman tomat Produk Rekayasa Genetik (PRG) dengan kadar
sukrosa meningkat dibandingkan tanaman kontrol.
Hasil penelitian menunjukkan bahwa : (1) Varietas Zamrud, kombinasi zat
pengatur tumbuh IAA : BAP (0,2 ppm : 2 ppm) dan eksplan kotiledon mempunyai
kemampuan regenerasi lebih tinggi dibandingkan perlakuan yang lain ; (2) Infeksi
pada eksplan kotiledon dan tunas menghasilkan tanaman putatif
transforman
sebesar 9% dan 4% dan telah diperoleh tanaman tomat putatif transforman sebanyak
8 klon ; (3) Hasil overekspresi SoSPS1 menghasilkan aktivitas enzim SPS meningkat
15%, kandungan sukrosa daun 2,16 kali dan kandungan sukrosa buah meningkat
1,99 kali.
Kata Kunci : Tomat (Lycopersicum esculentum Mill.), Sukrosa, Overekspresi, Gen
SoSPS1
SUMMARY
Tomato (Lycopersicon esculentum Mill.) is a fruit vegetables play an important
role in the economy in Indonesia. This plant is a dicotyledonous plant belonging to the
Solanaceae family. In the process of photosynthesis of tomato plants will produce
sucrose and starch. The function of sucrose in plants is emerging as an energy
source and source of carbon skeleton in metabolism. Efforts to increase the
production of sucrose in the tomato plant can be done through genetic transformation
methods.
Gene transformation has been done on some dicotyledonous plants such as
tomatoes, tobacco and others. Many kinds of genes that can be transformed into
plants, such as genes coding for proteins SPS (Phophat Sucrose Synthase). Phophat
Sucrose Synthase (SPS) is an enzyme that determines the content of sucrose and is
a key enzyme in carbon assimilation in plants. Gene cloning SPS (Sucrose Synthase
Phophat) from sugar cane called with SoSPS1 already successfully done at the
Laboratory of Molecular Biology, University of Jember.
The purpose of this study was to determine the increase of sucrose content in
tomatoes due to over-expression of SPS genes. This results can be obtained from
tomato transgenic plants with sucrose content increased compared with control
plants.
The results showed that: (1) Zamrud variety, a combination of growth
regulators IAA: BAP (0.2 ppm: 2 ppm) and cotyledon explant regeneration ability is
higher than the other treatments, (2) Infection of the explant cotyledons and shoots
produce putative transformed plants by 9% and 4%, and have obtained putative
transformants of tomato plants by 8 clones, (3) The overexpression SoSPS1 produce
SPS enzyme activity increased by 15%, the leaf sucrose content increased 2.16 times
and
the
fruit
sucrose
content
increased
by
1.99
times
.
Keywords: Tomato (Lycopersicum esculentum Mill.), Sucrose, overexpression, gene
SoSPS1
DAFTAR PUSTAKA
Afroz, A., Z. Chaudhry, R. Khan, H. Rashid AND S. A. Khan. 2009. Effect of GA3 on
regeneration response of three tomato cultivars (Lycopersicon esculentum).
Pak. J. Bot., 41(1): 143-151.
Ahsan N., S.H. Lee., D.G. Lee., M. Anisuzzaman (2007). The effect of wounding
type, preculture, infection method and cocultivation temperature on the
Agrobacterium mediated gene transfer in tomatoes. Annals of Applied Biology
151 : 363-372
Balitsa.2009.
Varietas
Unggul
Harapan
Tomat
Hibrida
dari
Balitsa.
http://hortikultura.litbang.deptan.go.id.
Chaudhry, Z., A. Afroz AND H. Rashid. 2007. Effect of variety and plant growth
regulatoers on callus proliferation and regeneration response of three tomato
cultivars (Lycopersicon esculentum). Pak. J. Bot., 39(3): 857-869.
Cortina C. & F. A. Culi´añez-Maci`.2004. Tomato transformation and transgenic plant
production. Plant Cell, Tissue and Organ Culture 76: 269–275
Ellul P, Garcia-Sogo B, Pineda B, Rios G, Roig LA & Moreno V. 2003. The ploidy level
of transgenic plants in Agrobacteriu-mmediated transformation of tomato
cotyledons (Lycopersicon esculentum L. Mill.) is genotype and procedure
dependent. Theor. Appl. Gen. 106 (2): 231–238
Farid Uddin, M.D., SH MD Asraful Islam, N. Sultana and M.H. Kabir Shiragi. 2004.
Effect of variety and plant growth regulators in MS medium on callus
Proliferation from Virus infected Tomato plant. Asian network for scientific
information Biotechnology. 3(2): 181-186.
Gubis. J., Z. Lajchova, J. Farago, and Z. Jurekova. 2004. Effect of growth regulators
on shoot induction and plant regeneration in tomato (Lycopersicon esculentum
Mill.)
Gubis, J., Z. Lajchova, L. Klcova and Z. Jurecova. 2005. Influence of growth
regulators on plant regeneration in tomato. Hort. Sci. 3 : 118 – 122
Hackel A., N. Schauer, F. Carrari, A. R. Fernie, B. Grimm and C. Kuhn. 2006. Sucrose
transporter LeSUT1 and LeSUT2 inhibition affects tomato fruit development in
different ways. Plant Journal. 45 :180–192
Miswar, B. Sugiharto, T. Handoyo, Dan S. A. Made, 2007. Peranan Sucrose
Phosphate Synthase (SPS) dan Acid Invertase (AI) Internoda Tebu
(Saccharum officinarum L.) dalam Akumulasi Sukrosa. Agritrop, 26 (4) : 187 .193
Miswar. 2007. Peningkatan biosintesis sukrosa tanaman tebu (Saccharum officinarum
L.) melalui over ekspresi gen sucrose phosphate synthase (SPS). Disertasi
UGM.Yogya.
Plevnes, D.R., Danuta K., Marta G., Katarzyna K., and Jadwiga K. 2006. The Effect of
Growth Regulators and Culture Conditions on The Callus Induction in Tomato
(Lycopersicon Sp.). Acta Sci.Pol., Hortorum Cultus 5(2) : 23-34
Qiu D., G. Diretto, R. Tavarza, G. Giuliano. 2007. Improved protocol for
Agrobacterium mediated transformation of tomato and production of transgenic
plants containing carotenoid biosynthetic gene CsZCD. Scientia Horticulturae
112 : 172–175
Santoso T.J., A. Sisharmini dan M. Herman.2009. Respon Regenerasi Beberapa
Genotipe Dan Studi Transformasi Genetik Tomat (Lycopersicon esculentum Mill.)
Melalui Vektor Agrobacterium tumefaciens. Balai Besar Penelitian dan
Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian.
Sheeja T. E., A.B. Mondal and R.K. S. Rathore. 2004. Efficient Plantlet Regeneration
in Tomato (Lycopersicon esculentum Mill.). Plant Tissue Cult. 14(1) : 45-53
Wijayani, A. dan W. Widodo. 2005. Usaha meningkatkan kualitas beberapa varietas
tomat dengan sistem budidaya hidroponik. Ilmu Pertanian. 12 (1) : 77 – 83