In Vitro Culture Application in the form of Somaclonal Variation Combined with Mutagen Introduction for Plant Improvement. Fatmawati is a new type of rice potentially to be developed. The development of this new type of rice in various places of West Java, Central Java and Lampung is often hampered by the blast disease causing the empty grain resulted in the harvest failure. Hence, from January to December 2007. The Indonesia Research Institute for rice in cooperation with Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development conducted research aimed at improving the quality of Fatmawati type of rice through somaclonal variation by mutative induction. In this research, the calli were treated with 1-50 gy gamma ray prior to its regeneration. The shoots produced by this regeneration were then acclimatized in the green house until the production stage. All 342 somaclone lines were sub-sequently tested on its endurance against leaf blast disease using three races of blast isolate namely 001, 033, and 173. The research yielded 21 somaclone lines which were absolutely tolerant to blast disease. These new somaclones were then planted in the green house for further morphological and agronomical observation.
Regeneration and Growth Some Varieties of Sugarcane (Saccharum officinarum L.) through In Vitro Culture. Deden Sukmadjaja and Ade Mulyana. The research was conducted at the Laboratory of Tissue Culture The Biology of Cell and Tissue Researcher Group ICABIOGRAD, Bogor from June to November 2009 to studied growth and regenerations response some varieties of sugarcane through in vitro culture. The research activities have been carried out in three steps, i.e., callus formation, regeneration of shoots and roots regeneration. The type of explants used in the study was in vitro planlet explants of both sugarcane varieties. Seven media formulations were used for the callus induction and regeneration of shoots, while five media formulations were used for the roots regeneration. The results showed that the highest respond for calluses induction was Bulu Lawang varieties at media formulation MS + 2.4-D 2 mg.l -1 + BAP 0.4 mg.l -1 + CH 2000 mg.l -1 and PS 951 varieties at media formulation MS + 2.4-D 1 mg.l -1 + BAP 0.4 mg.l -1 . While the highest respond for regeneration of shoots was Bulu Lawang varieties at media formulation MS0 (control MS) dan PS 951 varieties at media formulation MS + BAP 1 mg.l -1 + kinetin 1 mg.l -1 + NAA 0.5 mg.l -1 + GA 3 0.5 mg.l -1 . The highest respond of roots regeneration was Bulu Lawang and PS 951 varieties at media formulation MS + IBA 1 mg.l -1 . Acclimatization of plantlets produced were grew successfully about 90-100% in greenhouse.
ABSTRAKTebu (Saccharum officinarum L.) merupakan tanaman yang diperbanyak secara vegetatif sehingga berisiko besar akan terjadinya akumulasi virus di dalam jaringan tanaman. Kultur meristem merupakan salah satu teknik eliminasi virus yang umum digunakan, namun seringkali meristem memiliki daya hidup dan daya regenerasi yang rendah. Salah satu penyebabnya adalah karena akumulasi senyawa fenol. Akumulasi senyawa tersebut dapat direduksi melalui penggunaan senyawa adsorben dan antioksidan. Penelitian ini bertujuan untuk mengetahui pengaruh polyvinylpyrrolidone (PVP) dan diethyldithiocarbamate sodium (DIECA) terhadap regenerasi meristem tebu. Bahan tanaman yang digunakan adalah tebu PS864. Eksplan yang digunakan adalah meristem dengan 1-2 primordia daun yang diisolasi di bawah mikroskop. Perlakuan meliputi PVP (100 dan 300 mg/l) dan DIECA (0 dan 20 mg/l) serta kombinasi antara kedua zat tersebut, dengan 3 ulangan (botol) dan setiap botol terdiri atas 3 meristem. Hasil penelitian menunjukkan bahwa peningkatan konsentrasi PVP atau kombinasi perlakuan PVP dan DIECA dapat meningkatkan persentase eksplan hidup, daya regenerasi, dan jumlah tunas. Kombinasi perlakuan PVP 300 mg/l dan DIECA 20 mg/l merupakan perlakuan terbaik karena persentase hidup dan daya regenerasi eksplan yang paling tinggi (100%) dengan jumlah tunas 3,8 tunas/eksplan. Kata kunci: Tebu, Saccharum officinarum L., kultur meristem, PVP, DIECA ABSTRACTBeing vegetatively propagated, sugar cane faces a high risk of virus accumulation. Meristem culture is one method that usually applied for virus elimination. However, it often has low survival and regeneration rate due to an accumulation of phenolic compounds. Accumulation of those compounds can be reduced by apply adsorbent antioxidant. This research aimed at evaluating the effect of PVP and DIECA on the regeneration capacity of meristem. The plant material was sugar cane PS864. Meristems with 1─2 primoridial leaves were used as the explants and isolated under microscope. The PVP (100−300 mg/l) and DIECA (0− 20 mg/l), or combined treatment of both antioxidants were used as treatments. Each treatment was replicated 3 times (bottles), and each bottle contained 3 meristems. The result showed that the higher concentration of PVP or combined treatment of PVP and DIECA could increase the percentage of survival, regeneration rate, and number of shoot. The combined treatment of 300 mg/l PVP, and 20 mg/l DIECA produced the highest level of survival rate (100%) which yielded 3.8 shoots/explants.
In Vitro Selection and Evaluation of Banana MutantsVariety Ambon Kuning to Fusarium Wilt Disease. Deden Sukmadjaja, Ragapadmi Purnamaningsih, and Tri P. Priyatno. Fusarium wilt of banana (Musa spp.) caused by Fusarium oxysporum f. sp. cubense (Foc) is the most serious problem faced in banana cultivation in terms of plant productivity and fruit quality. Mutation breeding is one of the alternative method that can be applied in producing new banana cultivar. Mutants can be induced by chemical mutagen such as ethyl methane sulfonate (EMS) followed by in vitro selection and then evaluation of the mutants to fusarium wilt disease in glasshouse and Foc infected field. The aim of this research was obtained EMS induced and in vitro selected mutants of banana var. Ambon Kuning and evaluated Foc disease resistant clones in glasshouse and Foc infected field. The first step to obtain the explants for this research was initiation and formation of multiple bud clumps (MBC) using MS basal media supplemented with 5, 10, and 20 mg/l of benzyladenin. Plant regeneration of MBC was also studied by using MS media containing 0, 0.2, and 1 mg/l of benzyladenin. To induce mutagenesis, MBC was soaked in 0.1, 0.3, and 0.5% (v/v) EMS for 1, 2, and 3 hours. The assesment of resistant MBC mutants to Fusarium phytotoxin was conducted by using fusaric acid (FA) as selection agent in concentration of 30, 45, and 60 ppm. Putative mutant plants produced by in vitro selection were further tested using spore solution of Foc race 4 in glasshouse. Meanwhile, Foc resistance assesment in the infected field was conducted in Pasirkuda Experimental Station, Bogor Agricultural University. The results showed that MBC can be formed in MS basal media supplemented with 10 or 20 mg/l benzyladenin. The EMS played a role in obtaining mutants by producing 68 MBC putative mutants tolerant to Foc based on FA selection. Further evaluation in the glasshouse was obtained 64 Foc resistant plants from 391 putative mutants produced by in vitro selection. Evaluation in the Foc infected field showed six clones survived until generative phase (12 month of age). Keywords
<p>Protoplast<br />fusion or somatic hybridization technology is an alternative<br />technology for production hybrids of plants that are difficult<br />to be produced by conventional methods due to their sexual<br />incompatibility. An experiment was conducted to develop<br />techniques for isolation, purification, and culture of rice<br />protoplasts of cultivar IR64 and a wild rice species (Oryza<br />officinalis). Optimization of protoplast isolation and purification<br />methods from both rice genotypes were successfully<br />done. The highest protoplast density was obtained by<br />digesting embryonic callus or stems of young seedling in an<br />enzyme solution containing of 2% cellulose, 0.1% pectolyase,<br />0.5% macerozyme, 0.5% driselase, 5 mM ES, and 13% mannitol<br />in CPW solution. The protoplast digestion was done for<br />three hours by soaking in the enzyme solution followed by<br />shaking at 50 rpm under a room temperature. Purification of<br />the protoplasts were done by separating them from plant<br />debris using a 25% sucrose solution. Protoplast regeneration<br />was not successful using although different media compositions<br />and conditions. Growth process from cell division to<br />cell aggregate was only successful on IR64 protoplast culture<br />on a medium that contained AgNO3.</p>
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