PENENTUAN LOKUS GEN DALAM KROMOSOM TANAMAN DENGAN BANTUAN MARKA DNA

Kemajuan teknik marka molekuler memberikan kemudahan bagi pemulia tanaman dalam penentuan lokasi gen yang mengendalikan karakter yang diinginkan. Penentuan gen yang mengendalikan sejumlah karakter penting dengan menggunakan marka genetik telah berhasil dilakukan pada berbagai jenis tanaman. Sebelum pemetaan suatu marka molekuler terhadap karakter yang diinginkan, diperlukan pemetaan genetik yang dikonstruksi dari sejumlah marka molekuler. Pemetaan daerah dalam kromosom yang mengendalikan karakter kualitatif dan kuantitatif mendapat perhatian yang sangat besar dalam program pemuliaan. Penentuan gen yang mengendalikan karakter kualitatif maupun kuantitatif memerlukan populasi pemetaan. Metode umum yang digunakan dalam penentuan lokasi gen yang mengendalikan karakter kualitatif ialah bulk segregant analysis (BSA). Pendekatan tersebut terbukti mampu mempercepat penentuan lokasi gen dengan biaya yang relatif rendah. Sebaliknya, penentuan lokasi gen yang mengen-dalikan sifat kuantitatif dilakukan melalui pemetaan  quantitative trait loci (QTL). Dibandingkan penentuan lokasi gen pengendali sifat kualitatif, pemetaan QTL lebih kompleks dan membutuhkan kemampuan analisis statistik untuk menentukan daerah kromosom yang terkait dengan karakter kuantitatif tersebut. Tulisan ini membahas metode penentuan lokasi gen di dalam kromosom yang bertanggung jawab terhadap karakter penting tanaman dengan memanfaatkan marka molekuler dalam pemetaan genetik dan analisis QTL

THE POTENTIAL USE OF SSR MARKERS TO SUPPORT THE MORPHOLOGICAL IDENTIFICATION OF INDONESIAN MUNGBEAN VARIETIES

Mungbean varieties were mainly characterized based on morphological traits. Molecular genetic approach is expected to help the breeder in identification of mungbean varieties in more detail and to protect intellectual property right. This study aimed to identify Indonesian mungbean varieties based on DNA fingerprint profile using a marker set to support morphological characters. A total of 22 Indonesian mungbean accessions were characterized based on 21 morphological traits and 55 simple sequence repeats (SSRs) primers. Of the total 22 mungbean varieties used in the present study, 16 varieties were improved varieties and remaining six varieties were local varieties originated from Java, Nusa Tenggara and Sulawesi collected in GeneBank of ICABIOGRAD. The results showed that the 21 morphological characters were not sufficient to differentiate 22 mungbean varieties, while SSR analysis revealed that eight multi-alleles markers and high polymorphic information content (PIC) values have been successfully selected for varietal identification. The selected markers enabled to differentiate each mungbean variety according to their genetic marker with the lowest distance of 0.125, demonstrating the robustness of the selected marker set as a tool to identify a specific DNA fingerprint profile as a varietal identity (ID). The genetic identity of a variety was shown by digital barcoding which represented a series of alleles produced by corresponding markers. The DNA fingerprint profile of each variety would be beneficial as reference identities of a mungbean variety.

Penentuan Lokus Gen dalam Kromosom Tanaman dengan Bantuan Marka Dna

Kemajuan teknik marka molekuler memberikan kemudahan bagi pemulia tanaman dalam penentuan lokasi gen yang mengendalikan karakter yang diinginkan. Penentuan gen yang mengendalikan sejumlah karakter penting dengan menggunakan marka genetik telah berhasil dilakukan pada berbagai jenis tanaman. Sebelum pemetaan suatu marka molekuler terhadap karakter yang diinginkan, diperlukan pemetaan genetik yang dikonstruksi dari sejumlah marka molekuler. Pemetaan daerah dalam kromosom yang mengendalikan karakter kualitatif dan kuantitatif mendapat perhatian yang sangat besar dalam program pemuliaan. Penentuan gen yang mengendalikan karakter kualitatif maupun kuantitatif memerlukan populasi pemetaan. Metode umum yang digunakan dalam penentuan lokasi gen yang mengendalikan karakter kualitatif ialah bulk segregant analysis (BSA). Pendekatan tersebut terbukti mampu mempercepat penentuan lokasi gen dengan biaya yang relatif rendah. Sebaliknya, penentuan lokasi gen yang mengen-dalikan sifat kuantitatif dilakukan melalui pemetaan quantitative trait loci (QTL). Dibandingkan penentuan lokasi gen pengendali sifat kualitatif, pemetaan QTL lebih kompleks dan membutuhkan kemampuan analisis statistik untuk menentukan daerah kromosom yang terkait dengan karakter kuantitatif tersebut. Tulisan ini membahas metode penentuan lokasi gen di dalam kromosom yang bertanggung jawab terhadap karakter penting tanaman dengan memanfaatkan marka molekuler dalam pemetaan genetik dan analisis QTL

Teknik PCR Kualitatif untuk Deteksi Produk Rekayasa Genetika Jagung Event BT11 dan GA21

In some countries, including Indonesia, labelling of GMO products is mandatory for giving consumers the right to choosebetween GMOs and conventional products. Therefore, development of methodology that can detect a specific geneticallymodified (GM) crops and to verify the absence or presence of GM material in a product including raw materials (e.g. grains)and/or their derivatives is needed. The objectives of this study were to find the most efficient screening methods to detectwhether or not a product is GM material and to develop a specific detection method to identify GM product BT11 and GA21. Inaddition, present study was also aimed to obtain a duplex detection method for both GM products. Two GM-maize, including theBT11 and GA21 lines of maize (Zea mays L.), and one plant, namely NK11 as the nontransgenic control, were used as plantgenetic materials in the event-specific detection of maize. The target gene from each sample was amplified in different reaction(simplex) using both the event specific primer and the endogenous maize reference, Zein, as internal control. Furthermore, induplex PCR, two targets were simultaneously amplified in the same reaction. The results showed that detection method of theGM product obtained from present study enabled us to screen the GM products and specifically the event of BT11 and GA21using simplex and duplex methods. The duplex method is more efficient because it can detect two GM crops in one timecompared to simplex method that only can detect GM crop one by one

Comparison of detergent and CTAB method for isolation of DNA from Salak ( Salacca zalacca (Gaert.) Voss. ‘Pondoh’)

This study conducted in Laboratory of Molecular Biology, Balai Besar Bioteknologi dan Sumberdaya Genetik Pertanian (BB- Biogen) Bogor. The aims of this study are to determine and comparing the quantity,  quality and the efficiency of DNA isolation result using detergent method and CTAB method.  The parameters observed in this study are the value of DNA concentration, purity, and visualization result using gel electrophoresis. The samples are the leaves of Salak ‘Pondoh’ (Salacca zalacca (Gaert.) Voss.). Detergent method is a method which was developed by Faculty of Biology UGM, it has simple method and relatively affordable cost. Meanwhile, CTAB method is one of the commonly used methods of DNA isolation protocol with relatively expensive cost.  Detergent method used detergent in the cell wall separation and protein removal in the sample. The CTAB method used Cetyltrimethyl ammonium bromide (CTAB) for cell membrane separation in the sample. The research methods included DNA isolation with detergent and CTAB methods, PCR analysis and electrophoresis. Data analysis was done quantitatively  using spectrophotometric method and qualitative used electrophoresis method. The result of the study  showed that DNA isolation using  CTAB method showed higher purity compared with detergent method with the purity values ranging from 1,3- 1,4 . Meanwhile, the concentration of DNA in the detergent method was higher than that of CTAB with the highest concentration of 1730 µg/ml. There is no difference between the  quality of genomic DNA isolated by CTAB and detergent methods

The Potential Use of Ssr Markers to Support the Morphological Identification of Indonesian Mungbean Varieties

Mungbean varieties were mainly characterized based on morphological traits. Molecular genetic approach is expected to help the breeder in identification of mungbean varieties in more detail and to protect intellectual property right. This study aimed to identify Indonesian mungbean varieties based on DNA fingerprint profile using a marker set to support morphological characters. A total of 22 Indonesian mungbean accessions were characterized based on 21 morphological traits and 55 simple sequence repeats (SSRs) primers. Of the total 22 mungbean varieties used in the present study, 16 varieties were improved varieties and remaining six varieties were local varieties originated from Java, Nusa Tenggara and Sulawesi collected in GeneBank of ICABIOGRAD. The results showed that the 21 morphological characters were not sufficient to differentiate 22 mungbean varieties, while SSR analysis revealed that eight multi-alleles markers and high polymorphic information content (PIC) values have been successfully selected for varietal identification. The selected markers enabled to differentiate each mungbean variety according to their genetic marker with the lowest distance of 0.125, demonstrating the robustness of the selected marker set as a tool to identify a specific DNA fingerprint profile as a varietal identity (ID). The genetic identity of a variety was shown by digital barcoding which represented a series of alleles produced by corresponding markers. The DNA fingerprint profile of each variety would be beneficial as reference identities of a mungbean variety