Induksi dan Regenerasi Kalus Jagung yang Ditransformasi dengan Gen CsNitr1-L melalui Penembakan Partikel

Abstract

The success in development of transgenic plants is influenced by the regeneration system. The objective of the study was to assess the response of maize genotypes to regeneration system of organogenesis and embryogenesis, after transformed with CsNitr1-L gene through particle bombardment. Induction and callus regeneration of maize immature embryos of inbred lines Ult:cm.1#, ARC 178-123-112-XB3, and AZ2 were conducted through organogenesis, whereas those inbred lines AZ1, AZ2, P4G19(S)C2.59.3.3.1.3 and P4S3.29.4.4.1 were conducted through embryogenesis somatic. Transformation of CsNitr1-L gene was done with the distance of bombardment of 7 cm and 9 cm and calli were then selected using 10 mg/l hygromycin. All explants (100%) of inbred lines Ult:cm.1# formed organogenic callus, while callus formation of ARC 178-123-112-XB3 was 94.3% and AZ2 was 60.5%. Ult:cm.1# was the most responsive line to the regeneration of organogenesis and produced 24 green shoots, compared with ARC 178-123-112-XB3 which produced one green shoot and AZ2 that did not produce green shoots. The highest percentage of embryogenic calli formed through somatic embryogenesis was obtained on inbred lines AZ1 (85.4%) and the lowest was on P4S3.29.4.4.1 (18.9%). Inbred lines AZ1 had the highest percentage of regeneration (50.7%) and produced 62 plants, followed by P4G19(S)C2.59.3.3.1.3 that produced 17 plants (2.8%) and P4S3.29.4.4.1 which produced two plants. Preliminary identification on 31 putative transgenic plants through PCR analysis produced 22 plants (70.96%) that contained nptII gene. Keywords:Zea mays, organogenesis, somatic embryogenesis, transformant identification. ABSTRAKKeberhasilan perakitan tanaman transgenik dipengaruhi oleh proses regenerasi. Tujuan penelitian adalah mengetahui respons genotipe jagung terhadap sistem regenerasi organogenesis dan embriogenesis somatik setelah ditransformasi dengan gen CsNitr1-L melalui penembakan partikel. Induksi dan regenerasi kalus dari eksplan embrio muda jagung galur inbred Ult:cm.1#, ARC 178-123-112-XB3, dan AZ2 dilakukan secara organogenesis, sedangkan galur inbred AZ1, AZ2, P4G19(S)C2.59.3.3.1.3, dan P4S3.29.4.4.1 secara embriogenesis somatik. Transformasi gen CsNitr1-L dilakukan dengan jarak penembakan 7 cm dan 9 cm, kemudian kalus diseleksi dengan higromisin 10 mg/l. Semua (100%) eksplan galur Ult:cm.1# dapat membentuk kalus organogenik, sedangkan galur ARC 178-123-112-XB3 dan AZ2 masing-masing dapat membentuk kalus sebesar 94,3% dan 60,5%. Galur Ult:cm.1# paling responsif terhadap regenerasi secara organogenesis dan menghasilkan 24 tunas hijau dibanding dengan ARC 178-123-112-XB3 yang menghasilkan satu tunas hijau dan AZ2 yang tidak menghasilkan tunas hijau. Persentase terbentuknya kalus embriogenik yang tertinggi diperoleh pada galur AZ1 (85,4%) dan paling rendah pada galur P4S3.29.4.4.1 (18,9%). Galur AZ1 mempunyai daya regenerasi yang tertinggi (50,7%) dan menghasilkan 62 tanaman, diikuti galur P4G19(S)C2.59.3.3.1.3 yang menghasilkan 17 tanaman (2,8%) dan galur P4S3.29.4.4.1 yan...