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Lampiran 1. Deskripsi tetua berulang Nama Varietas : Ciherang Tahun dilepas : 2000
Tetua : IR 18349-53-1-3-1-3/IRI 19661-131-3-1///IR 64////IR 64 Rata-rata hasil : 6,0 t/ha GKG
Potensi hasil : 8,5 t/ha GKG
Pemulia : Tarjat T., Z. A. Simanulang, E. Sumadi, Aan A. Daradjat Nomor pedigri : S3383-1d-Pn-41-3-1
Golongan : Cere
Umur tanaman : 116-125 hari Bentuk tanaman : Tegak Tinggi tanaman : 107-115 cm Anakan produktif : 14-17 batang Warna kaki daun : Hijau
Warna batang : Hijau
Warna telinga daun : Putih Warna lidah daun : Putih
Warna daun : Hijau
Permukaan daun : Kasar pada sebelah bawah
Posisi daun : Tegak
Daun bendera : Tegak
Tipe malai : Intermediate
Leher malai : Terbuka
Bentuk gabah : Panjang ramping Warna gabah : Kuning bersih
Kerontokan : Sedang
Kerebahan : Sedang
Tekstur nasi : Pulen
Bobot 1000 butir : 27-28 gram
Kadar amilosa : 23 %
Ketahanan terhadap hama : Tahan terhadap wereng coklat biotipe 2, agak tahan biotipe 3.
Ketahanan terhadap penyakit
: Tahan terhadap bakteri hawar daun (HDB) strain III dan IV
Anjuran tanam : Cocok di tanam pada musim hujan dan kemarau dengan ketinggian dibawah 500 m dpl
Lampiran 2. Pembuatan larutan stok A.Isolasi DNA
1. 0,5 EDTA pH 8,0
a. Bahan-bahan yang digunakan adalah 93,05 g NaEDTA dan 10 g NaOH. b. Aquades ±400 mL disiapkan di beakerglass yang sudah diberi stirer. c. Semua bahan dilarutkan kedalam aquades.
d. pH ditera dengan pH meter (NaOH ditambahkan jika pH terlalu rendah dan HCl ditambahkan jika pH terlalu tinggi) sampai pH 8,0.
e. Aquades ditambahkan samapi volume 500 mL. 2. TBE 10 x
a. Bahan-bahan yang akan digunakan disiapkan yaitu 108 g Tris base, 55 g Boric acid, dan 40 mL 0,5 EDTA pH 8,0.
b. Akuades ±500 mL disiapkan di beakerglass yang sudah diberi stirer. c. Tris base dan Boric acid dimasukkan kedalam beakerglass untuk
dilarutkan.
d. Jika semua bahan sudah tercampur secara homogen, 0,5 EDTA pH 8,0 ditambahkan sesuai dengan ketentuan.
e. Aquades ditambahkan sampai volume 1L.
f. Larutan dimasukkan kedalam tabung dan diautoclave selama 20 menit. 3. NaCl 5 M
a. Bahan yang akan digunakan adalah 292,2 g NaCl. b. NaCl dilarutkan kedalam 800 mL aquades.
c. Jika sudah larut, tambakan aquades sampai volume 1 L. d. Larutan disaring dengan kertas saring Whatman.
4. CTAB (10% dari NaCl 0,7 M)
a. Bahan yang akan digunakan adalah 100 mL NaCl 0,7 M dan 10 g CTAB. b. CTAB dilarutkan dalam 100 mL NaCl.
c. Pelarutan dilakukan pada incubator 65°C.
d. Larutan disaring dengan milipore dan alat suntik didalam laminar air flow.
5. SDS 20%
a. Bahan yang digunakan adalah 50 g SDS. b. SDS dilarutkan dalam 200 mL aquades. 6. Buffer Extraction without SDS
a. Bahan-bahan yang digunakan adalah 50 mL Tris 1 M pH 8,0, 60 mL NaCl 5 M, dan 50 mL EDTA 0,5 M.
b. Semua bahan dicampurkan sampai homogen. c. Aquades ditambahkan sampai volume 1 L. 7. Ethanol 70%
a. Bahan yang digunakan adalah 365 mL ethanol 96%. b. Bahan dilarutkan dalam 135 mL aquades.
B.Elektroforesis 1. TBE 1
a. Bahan yang digunakan adalah 100 mL TBE 10. b. TBE 10 dilarutkan kedalam 900 mL aquades. 2. Acrylamide solution 40%
b. Acrylamide dan bis dilarutkan dalam 200 mL Aquades dengan bantuan stirer.
c. Aquades ditambahkan sampai volume 500 mL. d. Larutan disaring dengan kertas saring Whatman.
e. disimpan dalam botol yang dilapisi dengan alumunium foil. 3. Premix
a. Bahan yang digunakan adalah 100 mL Acrylamide solution 40%, 50 mL TBE 10 x, dan 345 mL aquades.
b. Semua bahan dicampurkan sampai homogen. 4. APS 10%
a. Bahan yang digunakan adalah 1 g APS. b. APS dilarutkan dalam 10 mL aquades.
c. Larutan dimasukkan kedalam corning yang sudah dilapisi alumunium foil.
Lampiran 3. Peta genetik 200 individu padi populasi BC3F2 di kromosom 12.
Peta genetik individu no. 133 - 172
Peta genetik individu no. 213 - 252
Lampiran 4. Penampilan benih padi pada tetua dan beberapa galur populasi BC3F3.
Keterangan:
Galur BIOR1-85-193-38 memiliki bobot 1000 butir terkecil sebesar 2.28 g; galur BIOR1-85-193-94 dan BIOR1-85-302-1133 memiliki bobot 1000 butir rata-rata yaitu 2.58 g; BIOR1-85-302-1167 memiliki bobot 1000 butir terbesar pada populasi BC3F3 yaitu 3.39 g. Rata-rata bobot 1000 butir tetua adalah Ciherang 24.70 g dan B11143D 30.40 g.
Lampiran 5. Peta genetik 300 individu padi terpilih populasi BC3F3 di kromosom 12.
Peta genetik individu no. 1 - 257
Peta genetik individu no. 514 - 699
Peta genetik individu no. 705 - 885
Keterangan: Individu no. 1 – 600 berasal dari galur BIOR1-85-193; individu no. 601 – 1200 berasal dari galur BIOR1-85-302.
Lampiran 6. Kegiatan seleksi tanaman padi di lapang pada populasi BC3F3.
Lampiran 7. Kegiatan pengamatan molekuler.
Lampiran 8. Kegiatan pengamatan fenotipe dan panen tanaman padi.
Lampiran 9. Populasi padi BC3F3 di rumah kaca. Isolasi DNA Amplifikasi DNA
DNA
Elektroforesis gel DNA
Pengukuran tinggi tanaman panen Penghitungan jumlah gabah