Agrobacterium-mediated gene transfer has recently been developed to improve rice transformation. In this study, 3 different transformation methods were tested including soaking, co-cultivation, and vacuum infiltration. Agrobacterium tumefaciens GV3101 harboring the binary vector pGreen:: LeGSNOR was used in this experiment. This study aimed to identify the most appropriate method for transferring LeGSNOR into rice. Vacuum infiltration of the embryonic calli for 5 min in Ilpum resulted in high transformation efficiency based on confirmation by PCR, RT-PCR, and qRT-PCR analyses. In conclusion, we described the development of an efficient transformation protocol for the stable integration of foreign genes into rice; furthermore, the study results confirmed that PCR is suitable for efficient detection of the integrated gene. The vacuum infiltration system is a potentially useful tool for future studies focusing on transferring important genes into rice seed calli, and may help reduce time and effort.

Running business in group, in Indonesian is Sentra Peternakan Rakyat (SPR), which are sustainably running business together is the hope for all the farmers and also the government to meet the needs of national meats. In the past, some of farmers must disband and terminate its business due to wrong strategy in their business development. The important factor to running the business is not only the quantity of animal feed but also animal health and how to control the disease itself. One of important diseases we concerned is Brucellosis, is an infectious disease caused by a type of bacteria called Brucella. This study was conduct to discover the proper strategy to develop the human resources in monitoring brucellosis diseases at Sentra Peternakan Rakyat Cinarabogo, Subang. We assumed that the knowledge of Brucellosis at the farmers level is low because they could not recognize the symptom of brucellosis, how to explain the symptom and also how to solve the problem correlated with brucellosis. The sample used is 20 people consist of 9 people is from Gugus Perwakilan Pemilik Ternak (GPPT) SPR Cinagarabogo and 11 the coordinator of incorporated SPR at Cinagarabogo. The data was gathered used questioners and focus Group Discussion (FGD) that also correlated with the purpose of this study. The result showed that the knowledge level of correspondence is good. It is also showed how the correspondence explains and also solve when the symptom is appeared. The implication from this research still needs the increment of knowledge about the handling of cattle infected brucellosis by educating and training in animal health to the farmers. Moreover, the government policy still needs encouraged to be given to farmers and to solve however the livestock must be cut forced as a result of infecting brucellosis with considering the weight of livestock and the market price.

We previously identified the rice gene, OsSAP, as an encoder of a highly conserved putative senescenceassociated protein that was shown to have anti-apoptotic activity. To confirm the role of OsSAP in inducing abiotic stress tolerance in rice, we introduced OsSAP and AtBI-1, a plant homologue of Bax inhibitor-1, under the control of the CaMV 35S promoter into the rice genome through Agrobacterium-mediated transformation. The OsSAP transformants showed a similar chlorophyll index after salinity treatments with AtBI-1. Furthermore, we compared the effects of salinity stress on leaves and roots by examining the hormone levels of abscisic acid (ABA), jasmonic acid (JA), gibberellic acid (GA3), and zeatin in transformants compared to the control. With the exception of phytohormones, stress-induced changes in hormone levels putatively related to stress tolerance have not been investigated previously. Hormonal level analysis confirmed the lower rate of stress in the transformants compared to the control. The levels of ABA and JA in OsSAP and AtBI-1 transformants were similar, where stress rates increased after one week and decreased after a two week period of drought; there was a slightly higher accumulation compared to the control. However, a similar trend was not observed for the level of zeatin, as the decrease in the level of zeatin accumulation differed in both OsSAP and AtBI-1 transformants for all genotypes during the early period of salinity stress. The GA3 level was detected under normal conditions, but not under salinity stress.

Anredera cordifolia (Ten.) Steenis, in

Guanine-quadruplex-based CpG oligodeoxynucleotides (G4 CpG ODNs) have been developed as potent immunostimulatory agents with reduced sensitivity to nucleases. We designed new monomeric G4 ODNs with an antiparallel topology using antiparallel type duplex/G4 ODNs as robust scaffolds, and we characterized their topology and effects on cytokine secretion. Based on circular dichroism analysis and quantification of mRNA levels of immunostimulatory cytokines, it was found that monomeric antiparallel G4 CpG ODNs containing two CpG motifs in the first functional loop, named G2.0.0, could maintain antiparallel topology and generate a high level of immunostimulatory cytokines in RAW264 mouse macrophage-like cell lines. We also found that the flanking sequence in the CpG motif altered the immunostimulatory effects. Gc2c.0.0 and Ga2c.0.0 are monomeric antiparallel G4 CpG ODNs with one cytosine in the 3′ terminal and one cytosine/adenine in the 5′ terminal of CpG motifs that maintained the same resistance to degradation in serum as G2.0.0 and improved interleukin-6 production in RAW264 and bone marrow-derived macrophages. The immunostimulatory activity of antiparallel G4 CpG ODNs is superior to that of linear natural CpG ODNs. These results provide insights for the rational design of highly potent CpG ODNs using antiparallel G4 as a robust scaffold.