LAPORAN PRAKTIKUM GENETIKA IKAN_ kelas B Kelompok 18 (triploid,dihibrid,ginogenesis).pdf
May 7, 2019 | Author: BastianDamanik | Category: N/A
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LAPORAN PRAKTIKUM GENETIKA IKAN GINOGENESIS, GINOGENESIS, TRIPLOIDISASI DAN HIBRIDISASI HIBRIDISASI Di ajukan untuk M emenuhi menuhi Salah Satu Satu Tugas M ata Kul iah Gene Genetik tik a I kan
Disusun oleh: Kelompok 18 Perikanan B
Fakhri Fathurahman
230110130090 230110130090
Nuraya Asfariah
230110130091 230110130091
Bastian Damanik
230110130152 230110130152
UNIVERSITAS PADJADJARAN FAKULTAS PERIKANAN DAN ILMU KELAUTAN PROGRAM STUDI PERIKANAN JATINANGOR 2014
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KATA PENGANTAR
Puji syukur kehadirat Allah SWT yang telah memberikan rahmat serta hidayahnya, serta kesehatan kepada kita semua, salawat serta salam semoga terlimpah curah kepada junjungan kita Nabi Muhammad SAW. Yang telah menyampaikan wahyu Allah Allah kepada kita. Laporan akhir ini adalah tugas dari mata kuliah Genetika Perikanan yang disusun untuk memenuhi hasil pengamatan praktikan di laboratorium yang di berikan kepada kami, yang berjudul “ GINOGENESIS, TRIPLOIDISASI DAN HIBRIDISASI”.
Percobaan tersebut kami jadikan dasar pada penulisan laporan
ini. Kami menaruh perhatian yang seksama terhadap semua masukan dari berbagai pihak demi penyempurnaan penyempurnaan isi tugas ini. Oleh karena itu, kritik dan saran sangat diharapkan demi kesempurnaan laporan ini. Kami berharap semoga laporan ini dapat bermanfaat bagi civitas akademika yang membutuhkannya. Amin.
Jatinangor, 18 Desember 2014
Penulis
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DAFTAR ISI KATA PENGANTAR............................................ ................................................................... ......................... .. i DAFTAR ISI ............................................ .................................................................. ....................................... ................. ii DAFTAR GAMBAR .......................................... ............................................................... ............................ ....... iv DAFTAR LAMPIRAN ........................................... ................................................................. ........................ .. ..v I.
PENDAHULUAN 1.1 Latar Belakang Belakang .......................................... ................................................................ ................................ .......... 1.2 Identifikasi Masalah .......................................... ................................................................ ........................ .. 1.3 Tujuan......................................... ............................................................... ............................................. ......................... .. 1.4 Manfaat ........................................... .................................................................. ........................................... ....................
II.
1 2 2 2
TINJAUAN PUSTAKA 2.1 Biologi Ikan Komet ....................... ............................................... ............................................ .................... 3 2.2 Biologi ikan mas............................................ ................................................................... ............................ ..... 4 2.3 Reproduksi ikan komet.......................................... .............................................................. .................... 5 2.4 Reproduksi Ikan Mas ............................................ ................................................................ .................... 6 2.5 Spermatozoa .......................................... ................................................................. .................................... ............. 7 2.6 Pemijahan Buatan ....................... ............................................. ............................................. ......................... .. 8 2.6.1 Ginogenesis...................................... Ginogenesis................................................................. ......................................11 ...........11 2.6.2 Hibridisasi....................... Hibridisasi............................................. ............................................ .................................11 ...........11 2.6.3 Triploidisasi........................... Triploidisasi................................................ .......................................... ...........................12 ......12 2.7 Embriogenesis...................... Embriogenesis........................................... ........................................... ................................12 ..........12
III.
METODOLOGI PRAKTIKUM 3.1 Waktu dan Tempat ........................................... .................................................................. ......................... .. 3.2 Alat dan Bahan .......................................... ................................................................ ................................ .......... 3.3 Prosedur Praktikum Praktikum ........................................... ................................................................. ........................ .. 3.3.1 Persiapan Alat Alat ................................. ...................................................... ....................................... .................. 3.3.2 Pemijahan Buatan ........................................... ................................................................. ........................ .. 3.3.3 Hibridisasi .......................................... ................................................................. .................................... ............. 3.3.4 Ginogenesis Ginogenesis ............................................ .................................................................. ................................ .......... 3.3.5 Triploidisasi ............................................ .................................................................. ................................ .......... 3.3.6 Embriogenesis Embriogenesis ............................................ ................................................................... ............................ ..... 3.3.7 Pemeliharaan Larva Larva ............................................ ................................................................ ....................
13 13 13 14 14 14 14 14 15 15
3.4 Metode Praktikum ............................. ................................................... ........................................ .................. 16 3.5 Rancangan Ra ncangan Praktikum Prakti kum ......................................................... ............................................................... ...... 16 3.5.1 FR ............................................ ................................................................... ............................................. ........................ .. 16
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IV.
Hasil dan Pembahasan
V.
4.1 Pemijahan Buatan .......................................... ............................................................... ............................ ....... 4.2 Hibridisasi .......................................... ............................................................... ....................................... .................. 4.3 Ginogenesis Ginogenesis ........................................... .................................................................. .................................... ............. 4.4 Triploidisasi ........................................... .................................................................. .................................... ............. 4.5 Embriogenesis Embriogenesis ........................................... ................................................................. ................................ .......... 4.6 Keberhasilan Keberhasilan Pemijahan ........................................... ............................................................ ................. Kesimpulan dan Saran
18 18 20 21 22 22
5.1 Kesimpulan............................................ ................................................................... .................................... ............. 23 5.2 Saran ........................................... .................................................................. ............................................. ........................ .. 23 DAFTAR PUSTAKA ............................................ ................................................................... ......................... .. 24 LAMPIRAN ......................................... ............................................................... ........................................... ..................... 25
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DAFTAR GAMBAR Nomor
Judul
Halaman
1
Ikan Komet.................................. Komet............................................................. .................................................... .........................
3
2
Ikan Mas...................................... Mas........................................................... ............................................ .......................
4
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DAFTAR LAMPIRAN Nomor
Judul
Halaman
1
Dihibrid pada pengamatan 30 jam ................................................. .................................................
26
2
Ginogenesis pada pengamatan 30 jam............................... jam........................................ .........
26
3
Pengamatan dihibrid selam 26 jam................................................ jam................................................
27
4
Pengamatan Ginogenesis selama 26 jam......................................... jam.........................................
27
5
Triploid pada pengamatan p engamatan 10 jam .................................................. ..................................................
28
6
Dihibrid pada pengamatan 20 jam ................................................. .................................................
28
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BAB I PENDAHULUAN 1.1 Latar Belakang
Budidaya adalah suatu metode yang dikembangkan untuk melestarikan dan mempertahankan jumlah populasi mahluk hidup agar tidak hanya melakukan penangkapan ikan saja tetapi tahu bagaimana cara mengembang biakkannya. Dewasa ini rekayasa dalam bidang genetika semakin sering dilakukan untuk meperbanyak produksi. Hal ini terjadi karena permintaan akan ikan yang semakin tinggi dan untuk memperbaiki kualitas ikan agar didapatkan individu yang unggul dari biasanya yang berguna dalam proses budidaya. Rekayasa genetik yang umum dilakukan ada tiga yaitu Ginogenesis, Hibridisai dan Triploid. Ginogenesis adalah proses proses pemurnian dengan mengambil genetik murni yang berasal dari induk betina dengan cara merangsang terjadinya proses pembuahan tanpa adanya sumbangan bahan genetik ge netik jantan (sperma yang telah di non-aktifkan dan digunakan untuk memacu terjadinya perkembangan sel telur menjadi larva). Proses ginogenesis dapat dilakukan dengan dua cara yaitu proses mitosis dan miosis. Hibridisasi merupakan perkawinan silang antara dua spesies individu yang masih memiliki hubungan kekerabatan yang dekat (dalam hal ini adalah tingkat famili). Hibridisasi bertujuan untuk untuk mendapatkan mendapatkan benih dengan sifat lebih baik dari yang dipunyai tertuanya terutama dalam pertumbuhan, kematangan gonad, ketahanan terhadap penyakit serta lingkungan lingkungan burukdan burukdan efesiensi pemanfaatan makanan.
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pertumbuhan ikan tidak terhambat karena energi metabolisme yang digunakan untuk
perkembangan
gonad
dapat
dimanfaatkan
untuk
meningkatkan
pertumbuhan sel-sel somatik.
.2 Identifikasi Identifikasi Masalah
Dalam melakukan rekayasa genetika banyak juga faktor-faktor yang akan mempengaruhi dari perkembangan ikan tersebut. Teknik rekayasa genetika yang umum dilakukan adalah hibridisasi,seleksi, inbreeding dan lain-lain. Dalam praktikum kali ini akan mempelajari tentang teknik rekayasa genetika menggunakan hibridisasi, ginogenesis dan triploidisasi.
1.3 Tujuan Praktikum
Tujuan dari dari praktikum kali ini adalah, 1. Mahasiswa memahami teknik rekayasa genetik yaitu ginogenesis , dihibrid dan triploid . 2. Mahasiswa melakukan teknik rekayasa genetik yaitu ginogenesis, dihibrid dan triploid dengan benar dan sesuai prosedur
1.4 Manfaat
Manfaat dari praktikum kali ini adalah , 1. Mahasiswa mampu memahami teknik ginogenesis, dihibrid dan triploid. 2. Mahasiswa mampu melakukan teknik rekayasa genetik dengan baik dan sesusai prosedur
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BAB II TINJAUAN PUSTAKA
2.1 Biologi Ikan Komet
Gambar1. Ikan Komet Kebiasaan hidup di alam Ikan Komet aslinya hidup di sungai, danau, dan lain lambat atau masih menggerakkan tubuh air di kedalaman sampai dengan 20 m. Di habitat aslinya ikan Komet tinggal di iklim subtropis dan lebih suka air tawar dengan pH 6,0-8,0, dengan kesadahan air sebesar 5,0 _ 19,0 DGH, dan rentang temperatur 32-106 F (0 – 41 C). Makanan ikan Komet terdiri dari krustasea, serangga, dan bahan tanaman. Ikan Komet bertelur pada vegetasi air. Hidup di sungai-sungai, danau, kolam dan saluran dengan air tergenang dan lambat mengalir. Pemakan termasuk tumbuhan, krustasea kecil, serangga, dan detritus. Ikan Komet hidup lebih baik dalam air dingin dan bertelur pada vegetasi terendam. Ikan Komet merupakan ikan euryhaline yang mampu hidup pada salinitas
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Genus
: Carassius
Spesies
: Carassius auratus
2.2 Biologi Ikan Mas
Gambar 2. Ikan Mas Ikan mas (C. (C. carpio) carpio) merupakan salah satu ikan yang banyak dibudidayakan oleh masyarakat, karena dagingnya enak, kenyal, dan durinya mengumpul. Daerah yang sesuai untuk mengusahakan pemeli haraan ikan ini yaitu daerah yang berada antara 150 – 1000 meter, suhu optimum 25-30
o
C, pH
perairan berkisar antara 7-8 (Rukmana, 2003). Ikan mas mempunyai daya adaptasi dan laju pertumbuhan yang tinggi dengan pemberian pakan buatan yang sesuai (Santoso, 1993). Ikan mas ( C. carpio) carpio) adalah salah satu ikan perairan tawar yang hidup di danau, sungai yang perairannya tidak dalam, tidak begitu deras, dan berair hangat. Ikan mas (termasuk dalam jenis ikan pemakan hewan dan tumbuhan (omnivora). Ikan mas bersifat pemakan jasad ja sad dasar (bottom feeders), hal ini menyebabkan air keruh dan
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sisiknya adalah sisik tipe sikloid (lingkaran) (Amri, 2002). Sirip punggung (dorsal) berukuran memanjang dengan bagian belakang berjari keras dan di bagian akhir bergerigi. Letak sirip punggung berseberangan dengan permukaan sirip perut (ventral). Sirip dubur mempunyai ciri seperti sirip punggung, yakni berjari keras dan bagian akhirnya bergerigi. Garis rusuk (Linea ( Linea lateralis atau gurat sisi) ikan mas tergolong lengkap, berada di pertengahan tubuh melintang dari tutup insang sampai ke ujung belakang pangkal ekor. Kulitnya banyak mengandung kelenjar lendir, tertutup oleh sisik, sirip dan ekor yang simetris, insang tertutup tutup insang (Amri, 2002). Klasifikasi ikan mas menurut (Susanto, 2007) adalah sebagai berikut: Filum
: Chordata
Kelas
: Pisces
Ordo
: Ostariophysi
Familia
:Cyprinidae
Genus
: Cyprinus
Spesies
: Cyprinus carpio L
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yang berbeda-beda, mulai dari Tingkat kematangan Gonad (TKG) I (belum matang) sampai Tingkat Kematangan Gonad (TKG) IV (mat ang). 2.3.1 reproduksi reproduksi ikan i kan komet
Secara alami, pemijahan terjadi pada tengah malam sampai akhir fajar. Menjelang memijah, induk-induk ikan mas aktif mencari tempat yang rimbun, seperti tanaman air atau rerumputan yang menutupi permukaan air. Substrat inilah yang nantinya akan digunakan sebagai tempat menempel telur sekaligus membantu perangsangan ketika terjadi pemijahan. (Gursina, 2008). Sifat telur ikan Komet adalah menempel pada substrat. Telur ikan Komet berbentuk bulat, berwarna bening, berdiameter 1,5-1,8 mm, dan berbobot 0,17-0,20 mg. Ukuran telur bervariasi, tergantung dari umur dan ukuran atau bobot induk. Embrio akan tumbuh di dalam telur yang telah dibuahi oleh spermatozoa. Antara 2-3 hari kemudian, telur-telur akan menetas dan tumbuh menjadi larva. Larva ikan Komet mempunyai kantong kuning telur yang berukuran relatif besar sebagai cadangan makanan bagi larva. Kantong kuning telur tersebut akan habis dalam waktu 2-4 hari.
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dilakukan pemburaian dan berwarna kuning kecoklatan. Organ-organ yang teramati yaitu gelembung renang, hati, dan lambung. Sehingga gonad ikan mas dari pengamatan yang dilakukan pada tingkat kematangan gonad (TKG) IV (Khairuman dan Amri, 2007). Seperti pada gonad betina, gonad jantan ikan mas besar dan panjang, mengisi dua pertiga rongga perut atau hamper menutupi orga-organ yang lain sebelum dilakukan pemburaian. Gonad mengembung, memanjang ke depan dan berwarna putih jernih. Sehingga Sehingga gonad ikan mas dari pengamatan yang dilakukan pada tingkat kematangan gonad (TKG) IV (Khairuman dan Amri, 2007). Ciri-ciri matang gonad jantan dan betina
Ciri-ciri ikan mas betina yang siap pijah atau matang ma tang gonad adalah: ·
Pergerakan ikan lamban
·
Pada malam hari sering meloncat-loncat
·
Perut membesar/buncit ke arah belakang dan jika diraba terasa lunak
·
Lubang anus agak membengkak/menonjol dan berwarna kemerahan
Sedangkan ciri-ciri untuk ikan mas jantan gerakan lincah dan mengeluarkan cairan berwarna putih (sperma) dari lubang kelamin bila dipijit (Aswanto, 2012).
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2.5 Pemijahan buatan
Sebenarnya pemijahan ikan komet dapat terjadi sepanjang tahun dan tidak tergantung pada musim. musim. Namun, di habitat habitat aslinya, ikan ini memijah pada awal musim hujan, karena adanya rangsangan dari aroma tanah kering yang tergenang air. Secara alami, pemijahan terjadi pada tengah malam sampai akhir fajar. Menjelang memijah, induk-induk ikan komet aktif mencari tempat yang rimbun, seperti tanaman air atau rerumputan yang menutupi permukaan air. Substrat inilah yang nantinya akan digunakan sebagai tempat menempel telur sekaligus membantu perangsangan ketika terjadi pemijahan (Anonim, pemijahan (Anonim, 2011). 2.5.1 Ginogenesis
Ginogenesis adalah proses terbentuknya zigot dari gamet betina tanpa kontribusi dari gamet jantan. Dalam ginogenesis gamet jantan hanya berfungsi untuk merangsang perkembangan telur dan sifat-sifat genetisnya tidak diturunkan. Ginogenesis dapat terjadi secara alami dan buatan. Nagy et al,. 1978, menyebutkan ginogenesis adalah terbentuknya zigot 2n (diploid) tanpa peranan genetik gamet jantan. Jadi gamet jantan hanya berfungsi secara fisik saja, sehingga prosesnya hanya merupakan perkembangan pathenogenetis betina
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Perlakuan Ginogenesis
Untuk mendapatkan benih ikan yang monosex secara ginogenesis ada beberapa perlakuan yang dapat dapat dilakukan yakni antara lain: 1.
Penyinaran sperma dengan sinar ultraviolet
Sebelum sperma dicampur dengan sel telur (pemijahan buatan) sperma tersebut diberi perlakuan penyinaran dengan sinar UV. Hal ini dilakukan untuk merusak bahan
genetik
sperma.
Komposisi
kimiawi
sperma
pada
plasma
inti
(nukleoplasma) diantaranya adalah DNA, Protamine, Non Basik Protein. Sedangkan seminal plasma mengandung protein, potassium, sodium, calsium, magnesium, posfat, klarida. Sedangkan komposisi kimia ekor sperma adalah protein, lecithin dan cholesterol (Gusrina, 2008). Sinar ultraviolet dengan panjang gelombang di bawah 300 nm dapat diserap secara kuat oleh bahan biologi tertentu, terutama asam nukleat, protein, dan koenzim. Tetapi sinar ini tidak sampai mengionisasi atom-atom dan molekulnya disamping itu kemampuan sinar ultraviolet untuk menembus bahan sangat terbatas. Walaupun sinar ultraviolet yang dapat masuk ke bahan biologi tersebut sedikit, tetapi hampir semua diserap. Hal ini berarti efisiensi penyerapan
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penggunaannya penggunaannya sehingga bisa diterapkan pada jumlah yang banyak. Kejut suhu dimaksudkan untuk pencegahan keluarnya polar body II telur pada saat terjadi pembelahan miosis kedua atau pencegahan pembelahan sel setelah duplikasi kromosom pada saat terjadi pembelahan mitosis pertama sehingga jumlah kromosom telur mengganda lagi pada awal perkembangan zigot (Nagy et al:, 1978). Kejut suhu disini berupa kejutan panas dan kejutan dingin. Pemberian kejutan panas lebih singkat periodeny peri odenyaa dibandingkan dengan kejut dingin. Pada saat oogenesis (proses pembentukan sel telur hingga siap untuk ovulasi), sel telur belumlah dalam keadaan 2N 2N melainkan 4N. Saat pembelahan sel miosis I terjadi,saat itu dikatakan sel telur telah matang. Saat itulah ada "loncatan" polar body I (2N), sehingga sel telur yang awalnya 4N menjadi 2N.
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memiliki tiga set kromosom (3N). Karena materi genetic sperma telah rusak maka yang akan berkembang dan mengalami pembelahan hanya pada set kromosom telur dari induk betina. Oleh karena itu ginogenesis hanya akan menghasilkan anakan yang sama dengan sifat induknya jika metode ini berhasil. Ginogenesis dapat digunakan untuk pemurnian ikan menggantikan teknik perkawinan sekerabat. Menurut Rohadi, D. S, (1996) dengan ginogenesis buatan dapat menghasilkan ikan bergalur murni dengan sifat homozigositas. Hasil pemurnian ikan dengan metode ginogenesis selama satu generasi sama dengan hasil
tujuh
sampai
delapan
generasi
perkawinan
sekerabat
sedangkan
homozogositassatu generasi ikan ginogenesis sama dengan homozigositas tiga generasi ikan hasil perkawinan sekerabat. Keberhasilan dari metode ini ditentukan
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dapat
diaplikasikan
pada
hibridisasi
ikan
komet
( Carassius
auratus), auratus),
mengingatikan mas memiliki pertumbuhan cepat, dan memiliki kekerabatan yang cukup
dekat dengan ikan komet. Hibridisasi yang dilakukan diharapkan
dapat
menghasilkan ikan komet Hibrid komet Hibrid yang memiliki mutu genetis yang lebih baik.
2.5.3. Triploidisasi
Triploidisasi merupakan salah satu metode untuk menghasilkan ikan yang steril adalah dengan manipulasi kromosom. Benih triploid diperoleh dengan memberi kejutan suhu atau tekanan setelah pembuahan dengan menggunakan spermatozoa aktif. Selain itu triploidi dapat juaga dihasilkan dengan membuahi telur ikan tetraploid dengan sperma ikan diploid normal
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BAB III METODOLOGI PRAKTIKUM
3.1 Waktu dan Tempat
Praktikum
genetika
ikan
mengenai
hibridisasi,
triploidisasi
dan
ginogenesis dilaksanakan pada hari Kamis,Jum’at,Sabtu dan Minggu, tanggal 27 – 30 November 2014. Praktikum ini dilakukan di Laboratorium Akuakultur Fakultas Perikanan Perikanan dan Ilmu Kelautan Universitas Universitas Padjadjaran.
3.2 Alat dan Bahan 3.2.1 Alat
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3.2.2 Bahan
1. Ikan Komet betina berfungsi sebagai sampel ikan yang akan diuji 2. Ikan Mas jantan berfungsi sebagai sampel ikan yang akan diuji 3. NaCl fisiologis 0,9 berfungsi berfungsi sebagai cairan untuk mengencerkan sperma 4. Hormon ovaprim
berfungsi untuk merangsang terjadinya ovulasi ovulasi telur
oleh indukan yang dipijahkan dan untuk indukan jantan berfungsi untuk meningkatkan produksi sperma yang akan dikeluarkan
3.3 Prosedur 3.3.1 Persiapan Alat
1. Mencuci akuarium hingga bersih
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5. Melakukan pengamatan.
3.3.4 Ginogenesis
1. Mengencerkan
sperma yang telah dihasilkan dihasilkan oleh indukan jantan
menggunakan larutan NaCl.
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terjadi untuk pada sel telur. Pengamatan dilakukan setiap 15 menit sekali selama 8 jam.
3.3.7 Pemeliharaan Larva
Sel telur mengalami kematian dan tidak berkembang menjadi larva setelah
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pembuahan telur. tel ur. Effendie (1979) menyebutkan bahwa untuk untu k mengetahui me ngetahui derajat penetasan telur ikan dapat menggunakan menggunakan rumus sebagai berikut :
Keterangan :
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BAB IV HASIL DAN PEMBAHASAN
4.1 Pemijahan Buatan
Pemijahan
buatan
yang
dilakukan
oleh
penulis
adalah
dengan
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genus dalam ras satu famili atau berbeda famili. Tujuan hibridisasi mendapatkan benih dengan sifat lebih baik dari yang dipunyai tertuanya terutama dalam pertumbuhan, kematangan gonad, ketahanan terhadap penyakit serta lingkungan buruk, dan efesiensi pemanfaatan makanan. Data hasil pengamatan praktikum didapatkan hasil derajat pembuahan pembuahan 44,8% dan derajat penetasan telur 4,56%.
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Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
The world's largest digital library
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
The world's largest digital library
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
The world's largest digital library
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
The world's largest digital library
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
The world's largest digital library
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
The world's largest digital library
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
The world's largest digital library
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
The world's largest digital library
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
The world's largest digital library
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
The world's largest digital library
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
The world's largest digital library
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
The world's largest digital library
Try Scribd FREE for 30 days to access over 125 million titles without ads or interruptions! Start Free Trial Cancel Anytime.
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