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LAMPIRAN
Lampiran 1 Komposisi media media dasar Murashige-Skoog (MS) Unsur Hara Makro Bahan Kimia (mg L-1)
NH4NO3 1 650
KNO3 1 900
CaCl2.2H2O 440
MgSO4.7H2O 370
KH2PO4 170
Unsur Hara Mikro Bahan Kimia (mg L-1)
H3BO3 22.3 Na2MoO4.2H2O 8.6 KI 6.2 CoCl.6H2O 0.83 MnSO4.4H2O 0.025 ZnSO4.7H2O 0.25 CuSO4.5H2O 0.025 Na2EDTA.2H2O 37.3 FeSO4.7H2O 27.8
Lampiran 2 Komposisi vitamin Murashige-Skoog (MS)
Vitamin Bahan Kimia (mg L-1)
Glycine 0.2
Myoinositol 100
Nicotinic Acid 0.05
Pyridoxine HCl (B6) 0.05
Thiamine HCl (B1) 0.01
Lampiran 3 Komposisi vitamin Morel and Wetmore (MW)
Vitamin Bahan Kimia (mg L-1)
Calcium Panthotenat 0.5 Myoinositol 100 Nicotinic Acid 1 Pyridoxine HCl (B6) 1 Thiamine HCl (B1) 1 Biotine 0.001
39 Lampiran 4 Hasil analisis sidik ragam peubah pertambahan tinggi tunas
pucuk jeruk Siam Simadu pada umur delapan minggu setelah perlakuan kolkisin Sumber keragaman db JK KT F Pr>F Kolkisin 3 3.02 1.01 10.50 0.00 Ulangan 4 0.51 0.13 1.34 0.31 Galat 12 1.15 0.10 Total 19 4.69
Lampiran 5 Hasil analisis sidik ragam peubah jumlah buku tunas pucuk jeruk Siam Simadu pada umur delapan minggu setelah perlakuan kolkisin Sumber keragaman db JK KT F Pr>F Kolkisin 3 48.55 16.18 13.21 0.00 Ulangan 4 9.30 2.33 1.90 0.18 Galat 12 14.70 1.23 Total 19 72.55
Lampiran 5 Hasil analisis sidik ragam peubah jumlah daun tunas pucuk jeruk Siam Simadu pada umur delapan minggu setelah perlakuan kolkisin Sumber keragaman db JK KT F Pr>F Kolkisin 3 62.80 20.93 11.07 0.00 Ulangan 4 17.30 4.33 2.29 0.12 Galat 12 22.70 1.89 Total 19 102.80
Lampiran 6 Hasil analisis sidik ragam peubah panjang daun tunas pucuk jeruk Siam Simadu pada umur delapan minggu setelah perlakuan kolkisin Sumber keragaman db JK KT F Pr>F Kolkisin 3 0.17 0.06 10.19 0.00 Ulangan 3 0.03 0.01 1.89 0.20 Galat 9 0.05 0.01 Total 15 0.25
40
Lampiran 7 Hasil analisis sidik ragam peubah lebar daun tunas pucuk jeruk Siam Simadu pada umur delapan minggu setelah perlakuan kolkisin Sumber keragaman db JK KT F Pr>F Kolkisin 3 0.02 0.01 4.20 0.04 Ulangan 3 0.00 0.00 0.36 0.78 Galat 9 0.02 0.00 Total 15 0.04
Lampiran 8 Hasil analisis sidik ragam peubah tinggi tunas yang baru muncul dari tunas samping jeruk Siam Simadu pada umur delapan minggu setelah perlakuan kolkisin
Sumber keragaman db JK KT F Pr>F
Kolkisin 3 0.73 0.24 7.71 0.00
Ulangan 10 0.41 0.04 1.29 0.33
Galat 12 0.38 0.03
Total 25 1.64
Lampiran 9 Hasil analisis sidik ragam peubah jumlah daun tunas yang baru muncul dari tunas samping jeruk Siam Simadu pada umur delapan minggu setelah perlakuan kolkisin
Sumber keragaman db JK KT F Pr>F
Kolkisin 3 2.03 0.68 1.97 0.17
Ulangan 10 6.58 0.66 1.91 0.14
Galat 12 4.13 0.34
Total 25 12.15
Lampiran 10 Hasil analisis sidik ragam peubah panjang daun tunas yang baru muncul dari tunas samping jeruk Siam Simadu pada umur delapan minggu setelah perlakuan kolkisin
Sumber keragaman db JK KT F Pr>F
Kolkisin 3 0.00 0.00 0.16 0.92
Ulangan 10 0.06 0.01 0.67 0.73
Galat 12 0.11 0.01
41 Lampiran 11 Hasil analisis sidik ragam peubah lebar daun tunas yang baru
muncul dari tunas samping jeruk Siam Simadu pada umur delapan minggu setelah perlakuan kolkisin
Sumber keragaman db JK KT F Pr>F
Kolkisin 3 0.02 0.01 4.42 0.03
Ulangan 10 0.03 0.00 1.82 0.16
Galat 12 0.02 0.00
RINGKASAN
FITRI YULIANTI. Induksi Tetraploid Jeruk Siam Simadu (Citrus nobilis Lour.) Menggunakan Kolkisin secara In Vitro. Dibimbing oleh AGUS PURWITO, DINY DINARTI dan ALI HUSNI.
Buah tanpa biji merupakan salah satu kriteria yang diperlukan untuk dapat meningkatkan kualitas jeruk Siam Simadu. Metode yang paling efektif untuk mendapatkan jeruk Siam Simadu tanpa biji adalah dengan cara menyilangkan antara tetua tetraploid dan diploid. Tujuan penelitian ini adalah untuk mendapatkan planlet jeruk Siam Simadu tetraploid yang akan dijadikan sebagai tetua untuk menghasilkan jeruk Siam Simadu tanpa biji. Penelitian dilaksanakan di Laboratorium Biologi Sel dan Jaringan BB-Biogen dan Laboratorium Micro Technique Departemen Agronomi dan Hortikultura IPB, Bogor pada Bulan Januari-Agustus 2013. Tunas pucuk tanpa daun, tunas samping tanpa daun dan embrio somatik fase globular jeruk Siam Simadu diberikan perlakuan empat konsentrasi kolkisin yang berbeda (0%, 0.1%, 0.2%, dan 0.3%) selama 3 jam (tunas pucuk), 6 jam (tunas samping) dan 1 jam (globular).
Hasil penelitian induksi tetraploid tunas pucuk jeruk Siam Simadu secara in vitro menunjukkan bahwa perlakuan kolkisin menurunkan pertumbuhan tunas pucuk jeruk Siam Simadu. Daun tunas pucuk yang diberikan perlakuan kolkisin lebih tebal dibandingkan dengan daun tunas pucuk kontrol. Jumlah kloroplas dari daun tunas yang dihasilkan dari kontrol (kolkisin 0%) dan perendaman kolkisin 0.1% adalah 8.67 dan 18.25 kloroplas per pasang sel penjaga. Daun dari tunas perlakuan kolkisin 0.1% memiliki ukuran stomata yang lebih besar dibandingkan dengan tunas kontrol. Daun dari tunas perlakuan kolkisin 0.1% memiliki kerapatan stomata yang lebih rendah dibandingkan dengan tunas kontrol. Tunas jeruk Siam Simadu tetraploid dihasilkan pada perlakuan kolkisin 0.1%.
Hasil penelitian induksi tetraploid tunas samping jeruk Siam Simadu secara in vitro menunjukkan bahwa tunas yang diberikan perlakuan kolkisin lebih tinggi tanamannya dari pada tunas kontrol. Tunas yang diberikan perlakuan kolkisin memiliki daun yang lebih lebar dari pada tunas kontrol. Daun tunas yang diberikan perlakuan kolkisin lebih tebal dibandingkan dengan daun tunas kontrol. Perlakuan kolkisin meningkatkan jumlah kloroplas per pasang sel penjaga. Daun dari tunas yang diberikan perlakuan kolkisin memiliki ukuran stomata yang lebih besar dibandingkan dengan tunas kontrol. Daun dari tunas yang diberikan perlakuan kolkisin memiliki kerapatan stomata yang lebih rendah dibandingkan dengan tunas kontrol.
Hasil penelitian induksi tetraploid embrio somatik jeruk Siam Simadu secara in vitro menunjukkan bahwa perlakuan kolkisin dengan konsentrasi tinggi dapat meningkatkan persentase planlet abnormal. Planlet tetraploid Jeruk Siam Simadu dihasilkan pada perlakuan kolkisin 0.2% dan 0.3%.
SUMMARY
FITRI YULIANTI. The In Vitro Tetraploid Induction of Tangerine (Citrus nobilis Lour.) Using Colchicines. Supervised by AGUS PURWITO, DINY DINARTI and ALI HUSNI.
Seedless fruit is one of the criteria necessary to improve the quality of Simadu tangerine. The most effective method to obtain seedless triploid cultivars is hybridisation between tetraploid and diploid parents. The aim of this research was to obtain tetraploid Simadu tangerine planlet which would serve as parent to produced seedless Simadu tangerine. The experiment was conducted at Biology Cell and Tissue Laboratory of BB-Biogen and Micro Technique Laboratory of Agronomy and Horticulture Department, IPB, Bogor from January to August 2013. In vitro Simadu tangerine shoot-tips without leaves, lateral shoots without leaves and somatic embryos globular phase were treated with colchicines at four different concentrations (0%, 0.1%, 0.2%, and 0.3) for 3 hours (Shoot-tips), 6 hours (lateral shoots) and 1 hour (globular).
The result of in vitro tetraploid induction of tangerine shoot-tip showed that colchicine treatments reduced growth of shoot-tip of Simadu Tangerine. The leaves of colchicines treated shoots were thicker than control. Leaves from control (0% colchicine) and 0.1% colchicine treated shoots had 8.67 and 18.25 chloroplasts per pair of guard cells. Leaves from 0.1% colchicine treated shoots had stomata sizes were larger than control shoots. Leaves from 0.1% colchicine treated shoots had stomata density was lower than control shoots. The tetraploid Simadu tangerine shoots were produced by the treatment of 0.1% colchicine.
The result of in vitro tetraploid induction of tangerine shoot-tip showed that the shoots of colchicines treated were taller than control. The shoots of colchicines treated had leaves were wider than control. The colchicines treatments increased numbers of chloroplasts per stomata guard cell. Leaves from colchicine treated shoots had stomata sizes were larger than control shoots. Leaves from colchicine treated shoots had stomata density was lower than control shoots.
The result of in vitro tetraploid induction of tangerine somatic embryos showed that colchicine treatments with high concentration increased abnormal planlet percentage. The tetraploid Simadu tangerine planlets were produced using 0.2% colchicine and 0.3% colchicine.