Kepel (Stelechocarpus burahol) is an original Indonesian fruit plant. Kepel is currently hard to find and cultivate conventionally. In vitro culture is a method for kepel propagation. The in vitro culture requires an established protocol for explant sterilization to be successful. For the present, there is no appropriate kepel sterilization method that has been known. This study aimed to obtain a sterilization method for kepel embryo explants appropriate for embryo culture. This research was conducted in September 2018 - January 2019 at the In Vitro Culture Laboratory, Faculty of Agriculture, Universitas Muhammadiyah Yogyakarta. This study used an experimental method arranged in a single factor. Completely Randomized Design with the treatment of various immersion methods of kepel embryo explants divided into 8 levels. They are Sodium hypochlorite (NaOCl) (5% : 5', 5% : 10', 10% : 5', 10% : 10'), Hydrogen peroxide (H2O2) (2% : 10', 2% : 15', 3% : 10', 3% : 15') and plant growth regulators 2 ppm 2,4-D + 0,5 ppm BAP, 2 ppm NAA + 0,5 ppm BAP. The results showed that 5% NaOCl for 5 minutes treatment was the best method for sterilization with a percentage of viable explant 88.89%. Then, the percentage of browning 11.11% and 2 ppm 2,4-D + 0,5 ppm BAP was effective in the organogenesis of callus of kepel embryos.
Abstract. Irsyadi MB, Sawitri WD, Purwantoro A. 2022. Molecular and phenotypic characteristics of T1 transgenic yellow cosmos (Cosmos sulphureus) carrying neomycin phosphotransferase II gene. Biodiversitas 23: 6097-6105. Yellow cosmos (Cosmos sulphureus) is a tropical ornamental flower that contains secondary metabolites useful fornatural pesticide application. Plant genetic transformation is one genetic engineering method used to increase secondary metabolite accumulation. However, published reports on this issue have yet to be made available. This is the first report of genetic transformation in transgenic yellow cosmos using neomycin phosphotransferase II (nptII) encoding gene. This study aimed to determine the efficiency transformation and phenotypic character of the T1 yellow cosmos transgenic nptII. This genetic transformation was carried out by the floral dip method mediated with Agrobacterium tumefaciens and characterized based on quantitative and qualitative observations, as well as a cluster analysis of transgenic plants. The T1 transgenic yellow cosmos was confirmed using PCR with a transformation efficiency of 73.33% based on the total plants resistant to Kanamycin and 10.57% based on the total seeds transformed. The presence of the nptII encoding gene was shown in transgenic plant samples with a size of 550 bp. In general, the introduction of the nptII gene generated no novel traits except the resistance to the kanamycin antibiotic. However, there was a decrease and increase in the number of stomata and the size of stomata in transgenic plants, respectively. In addition, there was a change in the type of ray floret to a mixture of ligulate and tubular, indicating that the nptII gene affected the phenotype of yellow cosmos. The result revealed that the nptII gene was inserted fairly randomly into the plant genome.
Kepel (Stelechocarpus burahol [Bl] Hook F. & Th.) merupakan buah asli Indonesia berbiji banyak dengan ukuran yang besar. Bagian buah yang dapat dikonsumsi hanya 49% dengan bagian lain berupa biji. Perbanyakan kepel secara konvensional masih sulit dilakukan dengan hasil yang rendah. Kultur endosperma secara in vitro adalah metode perbanyakan yang tepat untuk memperoleh tanaman triploid dengan buah tanpa biji. Sterilisasi merupakan tahap awal yang menjadi kunci keberhasilan kultur in vitro. Hingga kini belum dilaporkan metode sterilisasi endosperma kepel secara in vitro yang tepat. Penelitian ini bertujuan untuk memperoleh metode sterilisasi eksplan yang tepat untuk kultur endosperma kepel. Penelitian ini telah dilaksanakan pada bulan September 2018 – Januari 2019 di Laboratorium Kultur In Vitro, Fakultas Pertanian, Universitas Muhammadiyah Yogyakarta. Penelitian ini menggunakan metode eksperimen disusun dalam Rancangan Acak Lengkap (RAL) faktor tunggal dengan perlakuan berbagai konsentrasi bahan sterilan yang terbagi 8 aras: H2O2 (3%10’, 3%15’, 5%10’, 5%15’) NaOCl (5%5’, 5%10’, 10%5’, 10%10’) dengan 3 kali ulangan dan 3 sempel. Parameter yang diamati yaitu: persentase kontaminasi, browning, hidup, vitrifikasi, jenis kontaminasi, waktu kontaminasi dan waktu browning. Hasil penelitian diperoleh bahwa perlakuan NaOCl 10% selama 10 menit merupakan metode sterilisasi paling tepat dengan presentase ekplan hidup 44,44%, persentase eksplan vitrifikasi 66,66%, serta tidak terjadi kontaminasi dan browning
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