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Winarso Drajad Widodo

Kandungan Hormon Endogen dan Status Nutrisi dalam Buah Tanpa Biji Muda Hasil Induksi Antibiotik pada Anggur Muscat of Alexandria. Winarso Drajad Widodo. Streptomisin (SM); Spektinomisin (SE); Aquadest. Developed. Endosperm. PTT. Embrio. Pt. Degenerated. ?. SM. SE. -3. -6. +5 HSA.

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Winarso Drajad Widodo

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  1. Kandungan Hormon Endogen dan Status Nutrisi dalam Buah Tanpa Biji Muda Hasil Induksi Antibiotik pada Anggur Muscat of Alexandria Winarso Drajad Widodo

  2. Streptomisin (SM); Spektinomisin (SE); Aquadest Developed Endosperm PTT Embrio Pt Degenerated ?

  3. SM SE -3 -6 +5 HSA -3 -6 +5 HSA control Gambar 1. Pengaruh antibiotika terhadap diameter buah

  4. seeded seedless Pertumbuhan awal buah pra fertilisasi: Dipicu oleh hormon yang diproduksi oleh pollen Pertumbuhan buah pasca fertilisasi: Diatur/distimulir oleh hormon-2 yang disintesis oleh biji muda yang ditranslokasikan ke dinding buah; jaringan nourishing adalah endosperm tanpa endosperm embrio gagal tumbuh dan sebaliknya, tanpa embrio, endosperm tidak berkembang Kandungan hormon endogen dan status nutrisi dalam buah muda

  5. 4 tahun (30-40) cm x 50 cm x 20 cm Celup SM 200 mg/l SE 200 mg/l - 3 HSB

  6. Kandungan ABA

  7. Rf Gambar 2. Aktivitas sitokinin

  8. Gambar 3. Aktivitas auksin (IAA)

  9. Gambar 4. Aktivitas GA

  10. Nutrisi: Sukrosa dan Fruktosa (gula aktif dan reaktif) rendah pada buah dari tandan yang diberi perlakuan antibiotika Nitrogen: Kandungan dalam buah tanpa biji lebih rendah dibandingkan buah berbiji Asam amino: glutamin, sistein, asparagin, tirosin, isoleusin, leusin, asam aspartat, asam glutamat dan alanin lebih rendah pada buah tanpa biji dibanding buah berbiji GA merupakan hormon terpenting dalam pertumbuhan tahap awal buah!

  11. Application of Cytokinin and Gibberellin to Promote Enlargement of Muscat of Alexandria Grape Seedless Berries induced by Streptomycin1) Winarso Drajad Widodo2 DEPARTMENT OF AGRONOMY FACULTY OF AGRICULTURE BOGOR AGRICULTURAL UNIVERSITY 1)A part of the final experiments of : Widodo, W. D. 2000. Seedlessness Induction by Antibiotics and Its Mechanism in Grapes. The Graduate School of Natural Science and Technology, Okayama University - Japan (Doctor Thesis). 2)Secretary of Crop Production Laboratory, Department of Agronomy, Faculty of Agriculture - IPB.

  12. Background • Seedlessness: the important characteristic • for table grapes • Natural seedless cultivars are very few • Breeding for producing seedless cultivars: • - seedless inheritance is unclear • - time consuming performance How to produce seedless berries artificially?

  13. Fertilized Ovules Abortion:Stenospermy Stenospermocarpic formation (pseudo seedless) ‘remaining seed’ can not be detected organolep- tically --> stenospermic seedless berries • Stenospermic seedless berries: • can be induced in seeded cultivars by Gibberellin (GA) • simple method • free from genetic factors and barriers

  14. The Problems: • Certain vinifera grapes - included MOA - are severely induced lignification by GA; berries can not develop normally • Seedless berries can not be produced artificially by GA on seeded Muscat of Alexandria, an excellent cultivar of Japan grapes

  15. Change An antibiotic, streptomycin (SM) could induce seedlessness in several seeded cultivars, evidenced by: 1. Ogasawara (1986) on Muscat Bailey-A grape 2. Ishikawa et al. (1996-1998) on Kyoho and Fujiminori grapes 3. Pommer et al. (1996) on Italia (a vinifera) grape The mechanism of seedless berry information was not sufficiently studied yet

  16. The whole research objectives: 1. to investigate seedlessness induction capability of various antibiotics 2. to investigate the process of seedless berry formation induced by antibiotics 3. to study the cause of insufficient growth of antibiotic induced seedless berries by anatomical and hormonal analysis 4. to seek the method of seedless berry enlargement, especially Muscat of Alexandria

  17. 1. Seedlessness Induction Capability of Bactericidal and Bacteriostatical Antibiotics Plant materials: Muscat of Alexandria (MOA), Kyoho and Pione Antibiotics: Bactricide group: Streptomycin and Gentamicin Bacteriostatic group: Tetracycline and Specti- mycin Treatments: 1. Streptomycin: 200 mg/l 2. Tetracycline: 80 mg/l 3. Gentamicin: 60 mg/l 4. Spectinomycin: 200 mg/l vs. Control (distilled water) Application: Cluster dipping 3 days before full bloom

  18. Seedlessness induction capability of antibiotics

  19. Seedlessness induction capability of antibiotics

  20. Seedlessness induction capability of antibiotics

  21. 2. Anatomical analysis Materials: young berries of Muscat of Alexandria collected 0, 3, 5, 7, 9, 11 and 14 days after anthesis (dAA) Method: paraffin-sectioned pistils, observed un- der light microscope to determine ovule width and endosperm nuclei number

  22. Effect of antibiotics on ovule width

  23. Integument Nucellus Endosperm Embryo a. normal c. late degenerated b. early degenerated Ovule at 7 days after anthesis (Widodo, W. D. 2000. Seedlessness Induction by Antibiotics and Its Mechanism in Grapes. Doctoral Thesis, The Graduate School of Natural Science and Technology, Okayama University, Japan.)

  24. 3. The small berry formation 3.1. Cell size and cell number observation Plant material: Muscat of Alexandria Antibiotics: Streptomycin (SM) and Spectinomycin (SE) at the concentration of 200 mg/l applied 3 days before full bloom

  25. Effect of Streptomycin and Spectinomycin on cell width of ovule wall

  26. Effect of antibiotic treatment on cell numbers along pericarp area

  27. 3.2. Hormone analysis 1. ABA: Gas Chromatography  not significant 2. Auxins: Avena Coleoptile Straight-growth Test  Not significant 3. Cytokinins: Soybean Hypocotile Culture Test  significant 4. Gibberellins: Barley Endosperm Amylase Activity Assay  significant

  28. 4. Berry enlargement experiments • GAs is the important hormones for berry • enlargement • GA (as GA3) was tested in low level con- • centration to enlarge MOA-seedless • berries induced by Streptomycin; as • well as Forchlor-fenthuron (CPPU) a • synthetic cytokinin

  29. Effect of CPPU on berry growth of seedless berry induced by streptomycin

  30. Effect of GA3 on berry growth of seedless berry induced by streptomycin

  31. GA CPPU CPPU GA GA: A= 5 ppm, 10 dAA D= 5ppm , 15 dAA B= 10 ppm, 5 dAA E= 10 ppm, 10 dAA C= 1 ppm, 10 dAA CPPU: A= 10 ppm, 5 dAA D= 10 ppm, 10 dAA B= 10 ppm, 15 dAA E= 15 ppm, 10 dAA C= 5 ppm, 10 dAA

  32. Conclusion CPPU or GA treatments as the second treatments could enlarge the small-sized seedless berries induced by Streptomycin

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