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<p>Callus Induction and In Vitro Plant Regeneration of<br />Wheat Genotypes (Triticum aestivum L.). Atmitri<br />Sisharmini, Aniversari Apriana, and Sustiprijatno. Development<br />of a reliable in vitro plant regeneration procedure for<br />wheat is a prerequisite for its improvement by genetic transformation.<br />The purpose of this study was to obtain methods<br />of callus induction and regeneration of wheat genotypes.<br />This experiment was conducted at ICABIOGRAD. Immature<br />embryos from four wheat genotypes, ie Perdix, Naxos Wew,<br />Combi and Fasan were used to induce callus formation and<br />regeneration rate of callus. For the preparation of callus<br />induction medium, MS-L7 basal medium was supplemented<br />with combination of growth regulators 2,4 dichlorophenoxy<br />acetic acid (2,4-D) and 4-amino-3,5,6-trichloropicolinic acid<br />(picloram). While, plant regeneration medium was prepared<br />using MS basal medium supplemented with combination of<br />three growth regulators i.e. IAA, BAP and kinetin. The results<br />showed that genotype, in vitro culture medium and growth<br />regulators played a dominant role in callus induction and<br />plantlet regeneration. All the 4 genotypes responded positively<br />to callus induction, however, variability was observed<br />not only among the genotypes but also within callus<br />induction medium used. The best induction medium was<br />the MS-L7 basal medium supplemented with combination of<br />phytohormon 4 mg/l 2,4-D + 2 mg/l picloram (GIK-3) which<br />showed 100% callus induction frequency. Whereas, the best<br />regeneration medium was shown by MS basal medium with<br />combination of phytohormon 1.5 mg/l BAP dan 0.5 mg/l<br />kinetin (RG3). Regarding plant regeneration, Perdix was the<br />most responsive genotype to be regenerated with regeneration<br />frequency of 57.33%. The successfully acclimatized<br />planlets in greenhouse were obtained from Perdix and<br />Naxos Wew genotypes. These results will potentially facilitate<br />genetic transformation research of wheat in Indonesia.</p>

Maize (Zea mays L.) productivity in Indonesia is challenged to be increased using genetic engineering. Recent advances in Agrobacterium tumefaciens-mediated in-planta transformation makes it possible to transform maize with low cost, and simple method. This study aimed to confirm pIG121Hm-Cs plasmid in A. tumefaciens, and to estimate the efficiency level of A. tumefaciens-mediated in-planta transformation of Indonesian maize by using pIG121Hm-Cs plasmid containing nptII and hpt genes. A series of studies were conducted including confirmation of gene construct of pIG121Hm-Cs plasmid in A. tumefaciens, transformation of four maize lines through A. tumefaciens-mediated in-planta technique, acclimatization of transformant plants and molecular analysis of selected plants using polymerase chain reaction (PCR). The pIG121Hm-Cs plasmid was confirmed via PCR analysis using specific primers of nptII and hpt genes and resulted 700 bp and 500 bp for fragments of nptII and hpt, respectively. After selection, acclimatization and molecular analysis steps, the efficiency levels of transformation of four maize lines were low, ranging from 3.8% to 12.8%. The level of transformation efficiency of ST-27 line was the highest accounting for 12.8% of 45 planted embryos on selection medium based on PCR analysis using specific primer for nptII gene. Overall, A. tumefaciensmediated in planta transformation on maize floral pistil in this study proved to be successful and rapid. Therefore, this enhanced transformation method will be beneficial for Indonesian maize genetic engineering.

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The E5 protein is the smallest known oncoprotein linked to HPV 16 cancer development. In this study, we determined the potential of asarinin and thiazolo as an inhibitor of the E5 protein through molecular dynamics. The results showed that the binding site is unstable because of its hydrophobic nature and small size, causing considerable changes in the binding site for each of the 3 drugs examined. Except for asarinin, which still interacts with the first hydrophobic domain, they preserved their capacity to prevent endosomal acidification, hyper amplification of the EGFR pathway and contact with BAP31. It may inhibit E5-MHC I interaction. Thiazolo[3,2-a]benzymidazole-3(2H)-one,2-(2-fluorobenzylideno)-7,8-dimethyl (thiazolo) is expected to form more stable protein-ligand complexes than the other 2. However, the SASA, hydrogen bond and DCCM plots show that both compounds are equivalent to HPV 16 E5 protein. HIGHLIGHTS The smallest known oncoprotein associated with cancer development as a result of HPV 16 infection is the E5 protein The SASA, hydrogen bond, and DCCM plots, on the other hand, show that asarinin and thiazolo are equivalent to the HPV 16 E5 protein Thiazolo is predicted to produce more stable protein-ligand complexes than the others because of its unique molecular dynamic properties GRAPHICAL ABSTRACT

The sustainability and increasing the national rice production require the readiness of food and agriculture sector cope with the impacts of climate change, land degradation, drought area, sloping production and the raising of population growth. Adaptation plays an important role in ensuring the sustainability of food security. This research aimed to develop drought-tolerant variety of Inpari 30 (submergence tolerance variety) and Situ Bagendit through marker-assisted backcrossing-through pyramiding gene of identified QTLs for foreground selection and to explore SSRs and 6K SNPs for background selection distributed in 12 rice chromosome of drought tolerant donor (Cabacu) and recipient rice (Inpari 30 and Situ Bagendit). The foreground selection revealed that flanking SSRs of each QTLs (qRPF2.1, qGPP2.1, qSPP4.1 and Sub1) was less than 2 cM. The background selection through polymorphic survey of Rice 6K SNP primers revealed 2457 (53,3%) polymorphic SNPs on Inpari 30 vs Cabacu and 2563 (55,6%) polymorphic SNPs on Situ Bagendit vs Cabacu with the average distance about 0.74 cM/chromosome. The genotypic selection of F1 Inpari 30/Cabacu and F1 Situ Bagendit/Cabacu have already in heterozygote condition for these 4 QTLs target. These lines was continued for backcross breeding to develop BC1F1 Inpari 30/Cabacu and BC1F1 Situ Bagendit/Cabacu generation.

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Genetic Transformation of Tobacco with Cold ShockProtein Gene mediated by Agrobacterium tumefaciens. Seagames Waluyo, Sustiprijatno, and Suharsono. Cold shock protein (Csp) essential for organisms to survive in abiotic stress condition. CspB gene has been fused to ubiquitin promoter in the T-DNA region of pCambia 1300int, and introduced into Agrobacterium tumefaciens LBA4404. This research had an objective to transform genetically Nicotiana tabacum cv. Samsun by CspB gene under the control of Ubiquitin promoter and NOS terminator mediated by A. tumefaciens. Leaf discs were co-cultivated with A. tumefaciens LBA 4404. Based on the number of hygromycinresistant calli, the efficiency of transformation was 57.5%. In the selective medium containing 50 µg/l hygromycin, the efficiency of regeneration of transgenic shoots was 82.6%. Based on PCR analysis using primers corresponding to ubiquitin promoter and CspB gene, 18 putative tobacco transgenic containing CspB gene under the control of ubiquitin promoter.