cDNA ASAL DAUN KARET
KESIMPULAN DAN SARAN
Kesimpulan
Berdasarkan pemaparan serangkaian hasil penelitian di atas dapat disimpulkan bahwa aplikasi cDNA-AFLP telah berhasil diterapkan untuk mengidentifikasi dan mendapatkan fragmen asal transkrip (transcript-derived fragment, TDF) dari cDNA tanaman karet. Fragmen tersebut diekspresikan secara diferensial, baik antar klon karet resisten AVROS 2037 dan PPN 2444 maupun antar waktu setelah inokulasi daun dengan C. cassiicola pada masing- masing klon. Hasil yang diperoleh dengan kuat menunjukkan bahwa beberapa TDF memiliki homologi tinggi dengan protein membran, seperti putative Ran binding protein, protein transporter, GTP-binding protein dan sugar transferase. Beberapa TDF lain memiliki homologi dengan enzim yang regulasi ekspresinya berhubungan dengan ketersediaan nitric oxide (NO) dalam sel, seperti aconitase dan arginase. Di samping itu juga diperoleh TDF yang ekspresinya meningkat dalam kondisi adanya cekaman, baik berupa toksin maupun suhu tinggi, seperti
heat shock protein (HSP).
Keterlibatan enzim yang berhubungan dengan NO yang diperoleh dalam penelitian ini mengindikasikan bahwa kemungkinan besar NO berperan penting dalam interaksi tanaman karet dengan C. cassiicola, seperti yang telah dilaporkan pada beberapa tanaman lain, misalnya kedelai, tembakau dan Arabidopsis. Dalam interaksi tersebut NO bersama dengan spesies oksigen lainnya seperti H2O2 merupakan molekul sinyal yang mengaktifkan mekanisme pertahanan melalui kontribusinya dalam respon hipersensitif penye bab kematian sel yang terlokalisir sehingga patogen tidak berkembang ke jaringan sehat. Situasi tersebut kelihatannya sejalan dengan yang terjadi pada tanaman karet, dimana perkembangan gejala pada klon resisten tidak ber kembang lebih lanjut, sehingga kematian sel hanya terbatas di sekitar daerah infeksi.
Berdasarkan hasil tersebut, kelihatannya TDF yang memiliki homologi dengan aconitase dan arginase merupakan kandidat transkrip yang berperan dalam sistem pertahanan tanaman karet terhadap serangan C. cassiicola.
Saran
Hasil penelitian ini merupakan tahap awal untuk memahami mekanisme resistensi klon karet terhadap infeksi cendawan C. cassiicola. Oleh karena itu disarankan penyempurnaannya dengan penelitian lebih lanjut, antara lain adalah sebagai berikut :
1. Menggunakan kandidat TDF sebagai pelacak untuk dihibridisasikan dengan pustaka cDNA asal daun karet klon resisten AVROS 2037 yang diberi perlakuan C. cassiicola . Hal ini merupakan salah satu cara untuk memperoleh cDNA yang lebih lengkap dan utuh dari TDF tersebut.
2. Sekuen sebagian nukleotida TDFs telah dimiliki, termasuk transkrip yang memiliki homologi tinggi dengan arginase, aconitase dan HSP. Untuk memperoleh cDNA lengkap dan utuh informasi tersebut dapat digunakan untuk mendisain primer gen spesifik untuk mengamplifikasi kedua daerah ujung 5’ dan 3’ yang saling overlap. Dalam hal ini teknik teknik RACE (Rapid Amplification of cDNA Ends) dapat diaplikasikan.
3. Kajian pola ekspresi arginase, aconitase dan HSP pada berbagai klon karet resisten dan rentan yang tersedia diperlukan untuk mengevaluasi akumulasi kandidat transkrip tersebut pada berbagai klon karet. Untuk tujuan tersebut dapat digunakan teknik RT-PCR atau hibridisasi.
4. Apabila arginase, aconitase atau HSP turut berperan dalam meka nisme resistensi klon karet terhadap C. cassiicola, maka keduanya dapat digunakan sebagai marka resistensi dalam program pemuliaan tanaman karet. Di samping itu gen penyandi protein tersebut dapat diintroduksikan ke klon karet lainnya yang memiliki sifat-sifat unggul yang diinginkan, namun rentan terhadap serangan C. cassiicola.
Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucl. Acids Res. 25: 3389–3402.
Avrova AO, Venter E, Birch PRJ, Whisson SC. 2003. Profiling and quantifying differential gene transcription in Phytophthora infestans prior to and during the early stages of potato infection. Fungal Genetics and Biology
40: 4-14.
Bachem CWB, van der Hoeven RS, de Bruijn SM, Vreugdenhil D, Zabeau M, Visser RGF. 1996. Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP : Analysis of gene expression during potato tuber development. Plant J. 9: 745-753.
Bachem CWB, Oomen RJFJ, Visser RGF. 1998. Transcript imaging with cDNA-AFLP: A step by step protocol. Plant Mol. Biol. Reporter 16: 157- 173.
Baker B, Zambryski P, Staskawicz B, Kumar DSP. 1997. Signaling in plant- microbe interactions. Sciences 276: 726-733.
Baulkwill WJ. 1989. The history of natural rubber production. Di dalam: Webster CC, Baulkwill WJ, editor. Rubber. Longman Scientific and Technical. hlm 1-56.
Bent AF, Kunkel BN, Dahlbeck D, Brown KL, Schmidt R, Giraudat J, Leung J, Staskawicz BJ. 1994. RPS2 of Arabidopsis thaliana: a leucine-rich repeat class of plant disease resistance genes. Science 265: 1856-1859.
Besse P, Seguin M, Lebrun P, Chevallier MH, Nicolas D, Lanaud C. 1994. Genetic Diversity among Wild and Cultivated Populations of Hevea brasiliensis Assessed by Nuclear RFLP Analysis. Theor Appl Genet, 88: 199-207.
Breton F, d’Auzac J. 1999. Cassiicolin, a host-selective toxin produced by
Corynespora cassiicola, causal agent of Hevea leaf fall disease. International Rubber Research Development Board Meeting, Haicau, Chinese People’s Republic, 18-22 October 1999. CIRAD-CP, Hevea Programme, France. 5p.
Breton F, d’Auzac J, Garcia D, Sanier C, Esbach JM. 1996. Recent research on
Corynespora cassiicola. Di dalam: Proceeding of workshop on Corynespora leaf fall disease of Hevea rubber. Medan, December 16-17, 1996. Indonesian Rubber Research Institute, hlm 49-78.
Breton F, Sanier C, d’Auzac AJ. 2000. Role of cassiicolin, a host-selective toxin, in pathogenicity of Corynespora cassiicola, causal agent of a leaf fall disease of Hevea. J. Rubb.Res. 3(2): 115-128.
Briard M, Le Clerc V, Grzebelus D, Senalik D, Simon PW. 2000. Modified Protocols for Rapid Carrot Genomic DNA Extraction and AFLP Analysis using Silver Stain or Radioisotopes. Plant. Mol. Biol. Rep. 18: 235-241. Brosch G, Ransom R, Lechner T, Walton JD, Loidl P. 1995. Inhibition of maize
histone deacetylases by HC toxin, the host-selective toxin of Cochliobolus carbonum. Plant Cell 7: 1941-1950.
Bushnell WR, Berquist SE. 1975. Aggregation of host cytoplasm and the formation of papillae and haustoria in powdery mildew of barley.
Phytopathology 65: 310-318.
Chalhoub BA, Thibault S, Laucou V, Rameau C, Hofte H, Cousin R. 1996. Silver staining and recovery of AFLP amplification products on large denaturing polyacrylamide gels. BioTechniques 22: 216-220.
Chee KH. 1988. Studies on sporulation, pathogenicity and epidemiology of
Corynespora cassiicola on Hevea rubber. J. Nat. Rubb. Res. 3(1): 21-29. Chen H, McCaig BC, Melotto M, He SY, Howe GA. 2004. Regulation of plant
arginase by wounding, jasmonate, and the phytotoxin coronatine. The J of Biol Chem. 279: 45998-46007.
Chevallier MH. 1988. Genetic Variability of Hevea brasiliensis Germplasm Using Isozymes Markers. J. Nat. Rubb. Res., 3(1): 42-53.
Clouse SD, Gilchrist DG. 1987. Interaction of the asc locus in F8 paired lines of tomato with Alternaria alternata f. sp. Lycopersici and AAL-toxin.
Phytopathology 77: 80–82.
Collinge DB, Slusarenki AJ. 1987. Plant gene expression in response to pathogenes. Plant Mol. Biol. 8: 389-414.
Cooper B. 2001. Collateral gene expression changes induced by distinct plant vuruses during the hypersensitive resistance reaction in Chenopodium amaranticolor. Plant Journal 26: 339-349.
Cowel IG. 1998. cDNA Libraries. Di dalam: Rapley R, Walker JM, editor.
Molecular Biomethods Handbook. New Jersey: Humana Press. hlm: 131- 143.
Creelman RA, Mullet JE. 1997. Oligosaccharins, barassinolides and jasmonates: Nontraditional regulators of plant growth, development and gene expression. Plant Cell 9:1211-1223.
Das M, Harvey I, Chu LL, Sinha M, Pelletier J. 2001. Full-length cDNAs: more than just reaching the ends. Physiol Genomics 6: 57-80.
Dellagi A, Birch PRJ, Heilbronn J, Lyon GD, Toth IK. 2000. cDNA-AFLP analysis of differential gene expression in the prokaryotic plant pathogen Erwinia carotovera. Microbiology 146: 165-171.
Delledonne M, Xia Y, Dixon RA, Lamb C. 1998. Nitric oxide functions as a signal in plant disease resistance. Nature 394: 585–588.
Delledonne M, Zeier J, Marocco A, Lamb C. 2001. Signal interaction between notric oxide and reactive oxygen intermediates in the plant hypersensitive disease resistance response. Proc. Natl. Acad. Sci. 98: 13454-13459. Ditjenbun (Direktorat Jenderal Bina Produksi Perkebunan) 2004. Statistik
Perkebunan Indonesia 2001 -2003. Karet. Jakarta: Ditjenbun.
Dixon RA, Harrison MJ. 1990. Activation, structure and organisation of genes involved in microbial defence in plants. Adv. Genet. 28: 165-234.
Dong X. 1998. SA, JA, ethylene and disease resistance. Curr. Opin. Plant Biol. 1, 316–323.
Doyle JJ, Doyle JL. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull. 19: 11-15.
Durner J, Wendehenne D, Klessig DF. 1998. Defense gene induction in tobacco by nitric oxide, cyclic GMP, and cyclic ADP-ribose. Proc. Natl. Acad. Sci. 95: 10328-10333.
Durrant WE, Rowland O, Piedras P, Hammond-Kosack KE, Jones JDG. 2000. cDNA-AFLP reveals a striking overlap in race-specific resistance and wound response gene expression profiles. The Plant Cell 12: 963-977. Ellis J, Jones D. 1998. Structure and function of proteins controlling strain
spesific pathogen resistance in plants. Curr Opin Plant Biol 1: 288-293. Emmet RW, Parbery DG. 1975. Appressoria. Annu. Rev. Phytopathol. 13: 147-
167.
Feron R, Mariani C, Vriezen WH. 2004. Application of the mRNA Capture Kit in cDNA-AFLP. Biochemica 3: 23-24.
Gabriel DW, Rolfe BG. 1990. Working models of specific recognition in plant- microbe interaction. Annu. Rev. Phytopathol. 28: 365-391.
Gapkindo (Gabungan Perusahaan Karet Indonesia). 2005. Indonesian natural rubber statistic yearbook 2005. Jakarta: Gapkindo
Gilchrist DG, Grogan RG. 1976. Production and nature of a host-specific toxin from Alternaria alternata f. sp. lycopersici. Phytopathology 66: 165–171. Glick BR, Pasternak JJ. 1994. Molecular Biotechnology. Principles and
Applications of Recombinant DNA. ASM Press Washington DC.
Goldraij A, Polacco JC. 1999. Arginase is inoperative in developing soybean embryos. Plant Physiol. 119: 297-303.
Grant MR, Godiard L, Straube E, Ashfield T, Lewald J, Sattler A, Innes RW, Dangle JL. 1995. Structure of the Arabidopsis Rpm1 gene enabling dual specifcity disease resistance. Science 269: 843-846.
Gubler U, Hoffman BJ. 1983. A simple and very efficient method for generating cDNA libraries. Gene 25: 263-269.
Hadi H. 2003. Analisis genetik sifat ketahanan tanaman karet terhadap penyakit gugur daun Corynespora. Ringkasan Disertasi. Institut Pertanian Bogor. 25 hal.
Hammond-Kosack KE, Jones JDG. 1996. Resistance gene-dependent plant defense responses. Plant Cell 8: 1773-1791.
Heath MC. 1980. Reaction of nonsuscepts to fungal pathogenes. Ann. Rev. Phytopathol. 18: 211-236.
Jayasinghe CK, Silva WPK. 1996. Current status of Corynespora leaf fall in Sri Lanka. Proc. Workshop on Corynespora leaf fall disease of Hevea rubber. Medan, 16 – 17 Desember. Indonesian Rubber Research Institute. Pp : 15– 19.
Ji JP, Loeb LA. 1992. Fidelity of HIV-1 reverse transcriptase copying RNA in vitro. Biochemistry 31: 954-958.
Johal GS, Briggs SP. 1992. Reductase activity encoded by the Hm1 disease resistance gene in maize. Science 258: 985-987.
Keen NT. 1992. The molecular biology of disease resistance. Plant Mol. Biol. 19: 109-122.
Keyer K, Imlay JA. 1996. Superoxide accelerates DNA damage by elevating free-iron levels. Proc. Natl. Acad. Sci. 93: 13635– 13640
Kim WK, Mauthe W, Hausner G, Klassen GR. 1990. Isolation of high molecular weight DNA and double -stranded RNAs from fungi. Canad. J. Bot. 68: 1898-1902.
Kleinsmith LJ, Kish VM. 1995. Principles of Cell and Molecular Biology. Second Edition. Harper Collins College Publishers, New York. 810 pp. Klessig DF, Durner J. Noad R. Navarre DA, Wendehenne D, Kumar D, Zhou JM,
Shah J, Zhang S, Kachroo P, Trifa Y, Pontier D, Lam E, Silva H. 2000. Nitric oxide and salicylic acid signaling in plant defense. Proc. Natl. Acad. Sci. 97: 8849-8855.
Kombrink E, Beerhues L, Garcia -Garcia F, Hahlbrock K, Muller M, Schroder M, Witte B, Schmelzer E. 1993. Expression patterns of defense-related genes in infected and uninfected plants. In: B. Fritig and M. Legrand (Eds.). Mechanisms of Plant Defense Responses: 236-249. Kluwer Academic Publishers. Netherlands.
Lamb CJ, Lawton MA, Dron M, Dixon RA. 1989. Signals and transduction mechanisms for activation of plant defence against microbial attack. Cell
56: 215-224.
Lawrence GJ, Finnegan EJ, Ayliffe MA, Ellis JG. 1995. The L6 gene for flax rust resistance is related to the Arabidopsis bacterial resistance gene RPS2
and the tobacco viral resistance gene N. Plant Cell 7: 1195-1206.
Lekawipat N, Teerawatanasuk K, Rodier-Goud M, Seguin M, Vanavichit A, Toojinda T, Tragoonrung S. 2003. Genetic Diversity Analysis of Wild Germplasm and Cultivated Clones of Hevea brasiliensis Muell.Arg. by using Microsatellite Markers. J. Rubb. Res. 6(1): 36-47.
Lespinasse D, Rodier-Goud M, Grivet L, Leconte A, Legnate H, Seguin M. 2000. A Saturated Genetic Linkage Map of Rubber Tree (Hevea spp.) Based on RFLP, AFLP, microsatellite, and isozyme markers. Theor Appl Genet, 100: 127-138.
Liang P, Pardee AB. 1992. Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257: 967-971.
Lin J-J, Kuo J, Ma J, Saunders JA, Beard HS, MacDonald MH. 1996. Identification of Molecular Markers in Soybean-comparing RFLP, RAPD, and AFLP DNA Mapping Techniques. Plant Mol Biol Rep. 14: 156-169. Lin J-J, Ma J, Ambrose M, Kuo J. 1997. Chemiluminescent detection of AFLP
fingerprints. Focus 19: 36-38.
Liyanage NIS, Liyanage AS. 1986. A study on the production of a toxin in
Luo H, Coppenolle BV, Seguin M, Boutry M. 1995. Mitochondrial DNA Polymorphism and Phylogenetic Relationships in Hevea brasiliensis.
Molecular Breeding 1, 51-63.
Madjid A, Saleh M, Suwitoutomo S. 1977. Report on the 1974 Exchange Clones. Workshop on International Collaboration in Hevea Breeding and The Collection and Establishment of Material from Neotropics, Kuala Lumpur, 12-16 April, 1977.
Martin GB, Brommonschenkel SH, Chunwongse J, Spivey R, Wu T, Earle ED, Tanksley SD. 1993. Map-based cloning of protein kinase gene conferring disease resistance in tomato. Science 262: 1432-1436.
Meeley RB, Johal GS, Briggs SP, Walton JD. 1992. A biochemical phenotype for a disease resistance gene of maize. Plant Cell 4: 71-77.
Meeley RB, Walton JD. 1991. Enzymatic detoxification of HC-toxin, the host- selective cyclic peptide from Cochliobolus carbonum. Plant Physiol. 97: 1080-1086.
Mindrinos M, Katagiri F, Yu GL, Ausubel FM. 1994. The A. thaliana disease resistence gene RPS2 encodes a protein containing a nucleotide-binding site and leucine-rich repeats. Cell 78: 1089-1099.
Murayama S, Habuchi T, Yamagishi H, Terachi T. 2002. Identification of a sequence-tagged site (STS) marker link to a restorer gene for Ogura cytoplas mic male sterility in radish (Raphanus sativus L.) by non- radioactive AFLP analysis. Euphytica 129: 61-68.
Murray MG, Thompson WF. 1980. Rapid isolation of high molecular weight
DNA. Nucl. Acids. Res. 8: 4321-4323.
Navarre DA, Wendehenne D, Durner J, Noad R, Klessig DF. 2000. Nitric oxide modulate the activity of tobacco aconitase. Plant Physiology, 122: 573- 582.
Neveau C, Jaubert S, Abad P, Castagnone-Sereno P. 2003. A set of genes differentially expressed between avirulent and virulent Meloidogyne incognita near -isogenic lines encode secreted proteins. Mol. Plant- Microbe Interact. 16: 1077-1084.
Noel L, Thieme F, Nennstiel D, Bonas U. 2001. cDNA-AFLP analysis unravels a genome -wide hrpG-regulon in the plant pathogen Xanthomonas campestris pv. vesicatoria. Molecular Microbiology 41(6): 1271-1281.
Nurhaimi-Haris, Woelan S, Darussamin A. 1998. RAPD Analysis of Genetic Variability in Plant Rubber (Hevea brasiliensis Muell. Arg.) Clones.
Menara Perkebunan 66: 9 – 19.
Okayama H, Berg P. 1982. High-efficiency cloning of full-length cDNA. Mol. Cell Biol. 2, 161-170.
Onesirosan PT, Mabuni CT, Durbin RD, Morin RB, Rich DH, Arny DC. 1975. Toxin production by Corynespora cassiicola. Physiological Plant Pathology, 5: 289-295.
Orozco-Castillo, Chalmera KJ, Waugh R, Powell W. 1994. Detection of genetic diversity and selective gene introgression in coffee using RAPD marker.
Theor. Appl. Genet. 87 : 934-038.
Parker JE, Coleman MJ, Szabo V, Frost LN, Schmidt R, van der Biezen EA, Moores T, Dean C, Daniels MJ, Jones JDG. 1997. The Arabidopsis
downy mildew resistance gene Rpp5 shares similarity to the Toll and interleukin-1 receptors with N and L6. Plant Cell 9: 879-894.
Parniske M, Hammond-Kosack KE, Golstein C, Thomas CM, Jones DA, Harrison K, Wulff BB, Jones JD. 1997. Novel disease resistance specificities result from sequence exchange between tandemly repeated genes at the Cf-4/9
locus of tomato. Cell 91: 821-832.
Pawlowski K, Kunze R, De Vries S, Bisseling T. 1994. Isolation of total, poly(A) and polysomal RNA from plant tissues. In: Plant Molecular Biology Manual (Eds. SB Gelvin and RA Shilperoort). Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 1 – 13.
Perry MD, Davey MR, Power JB, Lowe KC, Bligh HFJ, Roach PS, Jones C. 1998. DNA Isolation and AFLP Genetics Fingerprinting of Theobroma cacao (L.). Plant Mol. Biol. Rep., 16: 49 – 59.
Petters J, Gobel C, Scheel D, Rosahl S. 2002. A pathogen-responsive cDNA from potato encodes a protein with homology to a phosphate starvation- induced phosphatase. Plant Cell Physiol. 43(9): 1049-1053.
Purwantara A. 1987. Studi histologi daun karet yang terserang Corynespora cassiicola (Berk. & Curt.) Wei. Menara Perkebunan 55: 47-49.
Pusat Penelitian Karet. 1992. Hasil rumusan Lokakarya Nasional Pemuliaan Karet. Di dalam: Prosiding Lokakarya Nasional Pemuliaan Tanaman Karet. Medan, 7-9 Desember 1992. Pusat Penelitian Karet.
Richter TE, Ronald PC. 2000. The evolution of disease resistance genes. Plant Molecular Biology 42: 195-204.
Ride JP, Pearce RB. 1979. Lignification and papilla formation at sites of attempted penetration of wheat leaves by non-pathogenic fungi. Physiol. Plant Pathol. 15: 79-92.
Roberts JD, Preston BD, Johnston LA, Soni A, Loeb LA, Kunkel TA. 1989. Fidelity of two retroviral reverse transcriptase during DNA-dependent DNA synthesis in vitro. Mol. Cell Biol. 9, 469-476.
Rogers SO, Bendich AJ. 1994. Extraction of total cellular DNA from plants, algae and fungi. In : Plant Molecular Biology Manual. Second Edition. S.B. Gelvin & R. A. Schilperoort (Eds.). Section D1.
Rohlf, JR. 1993. NTSYS-pc. Numerical Taxonomy and Multivariate Analysis System. Version 1.80. Exeter Software. New York.
Roslim DI, Hartana A, Suharsono 2003. Kemir ipan Genetika Tiga Populasi Kelapa Tipe Dalam Berdasarkan Tiga Metode Analisis Data Penanda RAPD. Hayati 10: 12 – 18.
Rouli Y. 1987. Studi jaringan terserang cendawan Corynespora cassiicola (Berk & Curt) Wei pada daun tanaman karet Hevea brasiliensis Muell. & Arg. klon PPN 2444. Laporan masalah khusus, 31 hal. Jurusan Hama dan Penyakit tumbuhan, Fakultas Pertanian, Institut Pertanian Bogor.
Sambrook J, Fritsch EF, Maniatis T. 1989. Molecular Cloning. A laboratory manual. Second edition. Cold Spr ing Harbor Laboratory Press.
Santaella M, Suarez E, Lopez C, Gonzalez C, Mosquera G, Restrepo S, Tohme J, Badillo A, Verdier V. 2004. Identification of genes in cassava that are differentially expressed during infection with Xanthomonas axonopodis
pv. manihotis. Molecular Plant Pathology 5(6): 549-558.
Schena M, Shalon D, Davis RW, Brown PO. 1995. Quantitative monitoring of gene expression patterns with a complementary DNA microarray. Science
270: 467-470.
Seguin M, Besse P, Lebrun P, Chevallier MH. 1995. Hevea Germplasm Characterization using Isozymes and RFLP Markers. In: Population Genetics and Genetic Conservation of Forest Trees. Baradat WT Adams & Muller-Starck (eds). 129-133.
Shalev G, Levy AA. 1997. The transposable elements Ac induces recombination between the donor site and a homologous ectopic sequence. Genetics 146: 1143-1151.
Shukor SKA, Hidir SM. 1996. Current status of Corynespora leaf fall in Malaysia. Proc. Workshop on Corynespora leaf fall disease of Hevea rubber. Medan, 16 – 17 Desember. Pp: 21 – 28.
Simmonds NW. 1989. Rubber Breeding. In: Webster CC, Baulkwill WJ (ed).
Rubber. Longman Scientific & Technical. John Wiley & Sons. New York. hlm: 85 – 124.
Sinulingga W, Suwarto, Soepena H. 1996. Current status of Corynespora leaf fall in Indonesia. Proc. Workshop on Corynespora leaf fall disease of Hevea rubber. Medan, 16 – 17 Desember. Pp: 29 – 36.
Situmorang A. 2002. Sebaran Penyakit Gugur Daun, Virulensi dan Genetika
Corynespora cassiicola Asal Sentra Pe rkebunan Karet Indonesia. Disertasi, Program Pascasarjana IPB, 115 hal.
Situmorang A, Budiman A. 1984. Corynespora cassiicola (Berk. & Curt.) Wei penyebab penyakit gugur daun pada karet. Kumpulan makalah Lokakarya Karet 1984 PN/PT Perkebunan Wilayah I dan P4TM. Medan. 10 hal. Situmorang A, Budiman A, Pawirosoemardjo S, Lasminingsih M. 1996.
Epidemic of Corynespora leaf fall disease and its preventive methods on
Hevea rubber. Di dalam: Proc. Workshop on Corynespora leaf fall disease of Hevea rubber. Medan, 16 – 17 Desember, hlm: 111-132.
Soepena H. 1983. Gugur daun Corynespora pada tanaman karet di Sumatera Utara. Kongres Nasional ke VII dan Seminar Perhimpunan Fitopatologi Indonesia, Medan 21-23 September 1983. 7 hlm.
Soepena H, Suwarto, Sinulingga W. 1996. Pengendalian penyakit gugur daun
Corynespora secara kimiawi. Di dalam: Proc. Workshop on Corynespora leaf fall disease of Hevea rubber. Medan, 16 – 17 Desember, hlm: 215- 224.
Speirs J, Longhurst T. 1993. RNA extraction and fractination. Meth. Plant Biochem. 10: 1-30.
Strommer J, Gregerson R, Vayda M. 1993. Isolation and characterization of plant mRNA. Di dalam: Glick BR, Thompson JE (eds.). Methods in plant molecular biology and biotechnology. CRC Press. hlm: 49-65.
Sugui JA, Deising HB. 2002. Isolation of infection-specific sequence tags expressed during early stages of maize anthracnose disease development.
Mol. Plant Pathol. 3: 197-203.
Suwarto 1989. Patogenisitas dan pembentukan toksin isolat Corynespora cassiicola dari tanaman karet (Hevea brasiliensis) dan kedelai. Risalah Seminar Latihan/Magang Penelitian Pertanian dan Bioteknologi III, Sukamandi, 13-14 Desember 1989. Halaman 55-59.
Suwarto 2003. Produksi dan inaktivasi in vitro toksin isolat Corynespora cassiicola (Berk. & Curt.) Wei asal daun karet. Ringkasan Disertasi, Program Pascasarjana Institut Pertanian Bogor, 37 hal.
Suwarto, Pawirosoemardjo S, Sinulingga W. 1996. Responses of recommended
Hevea rubber clones to Corynespora leaf fall in Indonesia. Di dalam:
Proc. Workshop on Corynespora leaf fall disease of Hevea rubber. Medan, 16 – 17 Desember, hlm: 149-162.
Takken FLW, Joosten MHAJ. 2000. Plant resistance genes: Their structure, function and evolution. European Journal of Plant Pathology 106: 699- 713.
Tan AM, Loo TP, Vadivel G, Bachik MR, Yoon KF. 1992. Survey of major leaf diseases of rubber in Peninsular malaysia. Plrs’Bull. Rubb. Res. Inst. Malaysia. No.211: 51-62.
Tan H, Tan AM. 1996. Genetic studies of leaf disease resistance in Hevea. J. Nat. Rub. Res. 11(2): 108-114.
Tesniere C, Vayda ME. 1991. Method for the isolation of high-quality RNA from grape berry tissues without contaminating tannins and carbohydrates.
Plant Mol. Bio. Reporter 9: 242-251.
Thomas MKG, Raghotha ma, Jacob J. 2002. A simple and efficient method for the isolation of total RNA and mRNA from mature leaves of Hevea brasiliensis. Indian Journal of Nat. Rubb. Res. 15: 93-95.
Thomas CM, Vos P, Zabeau M, Jones DA, Norcott KA, Chadwick BP, Jones JAG. 1995. Plant Journal 8: 785-794.
Toruan-Mathius N, Hutabarat T. 1996. Penanda RAPD dan polimorfisme genetik tanaman kopi robusta (Coffea canephora) toleran terhadap cekaman air.
Menara Perkebunan, 64: 45 – 55.
Toruan-Mathius N, Hutabarat T, Suhendi D. 1997. The use of RAPD to evaluate genetic variability of hybrid parent in Theobroma cacao L. plants.
Menara Perkebunan, 65: 53 – 63.
Unterstenhover G. 1963. The basic principles of crop protection field trial. Pflanzenschulz -Nachrichten Bayer AG. Laverkusen p. 169-170.
Vance CP, Sherwood RT. 1976. Regulation of lignin formation in reed canarygrass in relation to disease resistance. Plant Physiol. 57: 915-919. Van der Biezen EA, Hedyani J, Parker JE, Jones JDG. 2000. cDNA-AFLP
display for the isolation of Peronospora parasitica genes expressed during