This paper addresses a scheduling problem aimed at minimizing makespan in a permutation flow shop with two machines and an inspection process that must be conducted at least once every certain number of outcomes from the first machine. A mathematical programming approach and a genetic algorithm, incorporating Johnson's rule and a specific mutation process, were developed to solve this problem. Johnson's rule was used to generate an initial population, while the mutation process ensured compliance with the inspection constraints. The results showed that within a computation time limit of 300 seconds, the mathematical programming approach often failed to provide optimal or feasible solutions, especially for larger job sets. For instance, when the process times of both machines were similar and the inspection time was longer, the mathematical programming approach failed to solve all 10 experiments with just 15 jobs and only had a 50% success rate for 100 jobs. In contrast, the proposed genetic algorithm solved all instances and delivered equal or superior results compared to the mathematical programming approach.

In insects, the glutathione S-transferase is initiated in both the detoxification process and the protection of cellular membranes against oxidative damage. In this study, we identified the open reading frame (ORF) sequence of GST-iso1 and 2 from Tenebrio molitor (TmGST-iso1 and 2). To investigate the expression patterrns of TmGST-iso1 and 2 in response to herbicide, 0.06, 0.6, and 6 ㎍/㎕ of butachlor (FarmHannong, Seoul, South Korea) was challenged into T. molitor larvae, resulting that the TmGST-iso1 were highly induced at 3 and 24 h-post injection. Whereas, the highest expression of TmGST-iso2 was detected at 24 h after treatment. This study may contribute to basic information about the detoxifying activities of T. molitor.

It is well known that the JNK pathway regulates AMP production against pathogenic infection in both vertebrates and invertebrates. Tenebrio molitor hep (Tmhep) is an homolog of MAP kinase kinase in mammals. Here, we investigate the immunological function of Tmhep in responses in microbial infection using RNA interference technology. The results showed that silencing of Tmhep increased the larval mortality against microbial challenge, as well as reduced AMP production compared to the control group (dsEGFP-treated group). Conclusively, Tmhep plays an critical role in antimicrobial defense in T. molitor larvae.

The baculovirus expression system (BES) utilize the p10 or polyhedrin promoter, a very late promoter that exhibits strong transcriptional activity primarily at the end of viral infection, to produce useful recombinant proteins. The burst sequence of the very late promoter is essential for strong transcription, and VLF-1 is a transcription factor that binds specifically to the burst sequence, and it has been reported that it can regulate the amount and timing of expression of protein by the very late promoter. Recently, a VLF-1 constitutively expressing cell line was constructed to increase the production of the target protein, but the effect was minimal. In this study, to find the optimal VLF-1 expression conditions to increase target protein production efficiency, we controlled the expression of VLF-1 through various promoters and evaluated the target protein expression efficiency by the p10 promoter accordingly.

Apolipophorin-III (apoLp-III) is a hemolymph protein whose function is to facilitate lipid transport in an aqueous medium in insects. Recently, apolipophorin-III in Galleria mellonella and Hyphantria cunea was shown to play an unexpected role in insect immune activation. We show here a novel possible function/role of the apoLp-III in insects. To investigate the genes which have a relationship with apoLp-III in fall webworm larvae, we reduced endogenous Hc apoLp-III mRNA levels in larvae via RNA interference (RNAi). The RNAi-mediated Hc apoLp-III reduction resulted in the reduction of antioxidants, like MnSOD, catalase, and glutathione S transferase as well as immune proteins. In particular, expression of MnSOD commonly decreased in fat body, midgut, and hemocytes following the knockdown of Hc apoLp-III, which induced an elevated level of superoxide anion in Hyphantria cunea larvae. The observed effect of Hc apoLp-III RNAi suggests that Hc apoLp-III is related to the action/expression of antioxidants, especially MnSOD.

A new insect member of the STAT family of transcription factors (HcSTAT) has been cloned from the lepidopteran, Hyphantria cunea. The domain involved in DNA interaction and the SH2 domain are well conserved. The gene is transcribed at a low level during all stages of development, and transcribed in hemocyte, fat body, midgut, epidermis, and Malpighian tubule. Especially, hemocyte and Malpighian tubule showed transcriptional activation of HcSTAT upon Gram-negative and -positive bacteria challenge. Gram-negative and -positive bacteria challenge specifically results in nuclear translocation of HcSTAT protein and induction of DNA-binding activity that recognizes a STAT target site in H. cunea hemocyte. In vivo treatment with sodium orthovanadatetranslocates HcSTAT to the nucleus in hemocyte cells.

Apolipophorin-Ⅲ (apoLp-Ⅲ) is a hemolymph protein whose function is to facilitate lipid transport in an aqueous medium in insect. Recently, apolipophorin-Ⅲ in Galleria mellonella and Hyphantria cunea was shown to play an unexpected role in insect immune activation. We show here a novel possible function/role of apoLp-Ⅲ in insects. To investigate the genes which have a relationship with apoLp-Ⅲ in fall webworm larvae, we reduction of endogenous Hc apoLp-Ⅲ mRNA levels in larvae via RNA interference (RNAi). The RNAi-mediated Hc apoLp-Ⅲ reduction resulted in the reduction of antioxidants, like MnSOD, catalase, and glutathione S transferase as well as immune proteins. In particular, expression of MnSOD commonly decreased in fat body, midgut, and hemocytes following the knockdown of Hc apoLp-Ⅲ, which induced an elevated level of superoxide anion in H. cunea larvae. The observed effect of Hc apoLp-Ⅲ RNAi suggests that Hc apoLp-Ⅲ is related to the action/expression of antioxidants.

Innate immunity responses are triggered by the immune challenge and therefore involve signaling processes. The cellular response is initiated by hemocytes and mainly involves phagocytosis and encapsulation of intruders by these cells. To address whether Hc-STAT is activated upon bacterial challenge, we examined the subcellular location of STAT protein in hemocyte by immunostaining. A new insect member of the STAT family of transcription factors (Hc-STAT) has been cloned from the lepidopteran, Hyphantria cunea. The domain involved in DNA interaction and the SH2 domain are well conserved. The gene is transcribed at a low level during all stages of development, and the protein is present in hemocytes, fat body, midgut, epidermis, and Malphigian tuble (Mt). Especially, hemocytes and Mt showed transcriptional activation of Hc-STAT upon Gram (-) bacteria and fungal challenge. Gram (-) bacteria and fungal challenge specifically results in nuclear translocation of Hc-STAT protein and induction of DNA-binding activity that recognizes a STAT target site in H. cunea hemocyte. In vitro treatment with pervanadate translocates Hc-STAT to the nucleus in hemocyte cells. Here we report the first evidence for the involvement hemocyte JAK/STAT pathway upon microbial infection in lepidopteran insect.

Arabidopsis E3 SUMO ligase SIZ1 (AtSIZ1) controls vegetative growth and development including responses to nutrient deficiency and environment stresses. Here, we analyzed the effect of AtSIZ1 on the stability and amount of seed proteins. Proteomic analysis showed that the amount of three major nutrient reservoir proteins, CRUCIFERIN (CRU) 1, 2 and 3, were decreased in siz1-2 mutants. However, quantitative real-time RT-PCR showed that transcript levels of CRU1, 2 and 3 genes were rather significantly higher in siz1-2 mutants than wild-type plants. Yeast two hybrid analysis revealed that AtSIZ1 interacts with CRU1, CRU2 and CRU3, strongly suggesting that CRU1, 2 and 3 proteins are sumoylated by AtSIZ1. In addition, the analysis of amino acid composition by HPLC showed that the contents of amino acids were a bit high in siz1-2 mutants. Our data indicate that AtSIZ1 plays an important function for accumulation of seed storage proteins through its ligase activity.

The transition from vegetative growth to flowering is a major developmental switch in the plant cycle and the timing of flowering is very critical for reproduction of plant species. In transition to flowering in plants, Flowering locus C (FLC) is one of the crucial factors. Here, we showed How the stability and activity of FLC are regulated by sumoylation mechanism. By pull-down assay, we showed that FLC interact with E3 SUMO ligase in vitro and vivo. And we showed that FLC is sumoylated in vitro condition with AtSUMO1 protein. In transgenic plants with overexpression of FLC and inducible expression of AtSIZ1, sumo E3 ligase led to increase of FLC protein level and delayed the post-translation degradation of FLC indicating that Arabidopsis E3 sumo ligase AtSIZ1 stabilizes FLC. Also, the plants with overexpression of mutant FLC (K154R, a mutation of the sumoylation site on FLC) flowered considerably earlier than plants with overexpression of FLC but comparable with wild type indicating that sumoylation is a important part for function of FLC. Our data indicate that the sumoylation of FLC is critical for its role in the control of flowering time.

Flowering time is a important agronomic trait for grain production in rice. So the control of flowering time is a critical step. In Arabidopsis, expression of certain key flowering gene such as FLOWERING LOCUS C (FLC) is known to be epigenetically regulated by chromatin modification through Enhancer of Zeste[E(z)], a histone methyltransferase, that core component of repressive complex, polycomb repressive complex2(PRC2). However, the chromatin mechanism involved in the regulation of rice flowering genes is presently not well known. Here we show that predict coding region of a intronic LncRNA[termed rice COLDAIR(OsCOLDAIR)], which is expected to associate with a component of PRC2, is predicted at rice FLC gene. And additionally we suggest interaction of histone methyltransferase and E3 SUMO ligase that indicate possibility of interaction with rice E(z) gene and rice E3 SUMO ligase. Our study contribute to control of rice flowering time by observing two factor that can regulate expression of related of rice FLC gene.

Drought tolerance is the ability of a plant to live, grow, and reproduce properly with limited water supply or under periodic conditions of water deficit. However, the climate changes and worldwide water shortages would result in the loss of applied water to irrigated land, increasing soil water deficit. To control the situation, we have carried out the international joint research project for the aim of developing that drought tolerance common wheat and durum wheat in Korea and Tunisia. Total 79 (41 common wheat, 39 durum wheat) Tunisian lines and 33 Korean wheat cultivars were incorporated in this study. Drought stress was applied for 25 days of stopping irrigation from the 3-leaf stage followed by re-watering for restoration in greenhouse. We selected top 13 (5 Korean line, 8 Tunisian line) tolerant lines and 11 (5 Korean, 6 Tunisian) susceptible lines based on growth parameter analysis. Primers (Operon primers and wheat Dreb1 gene) that have been known to be related drought resistance were applied to explain selected population. The correlation between PCR-based length polymorphism of selected lines and their resistance were evaluated. The obtained primer information will aid selection for drought tolerance durum as well as hexaploid common wheat.

Mutant analysis is one of most optimized genome-wide approach towards acquiring utile phenotypes and defining related genes. Gamma-irradiation, an acknowledged way of mutant-generating method, was applied to gain sets of mutant line in Brachypodium distachyon. B. distachyon is a model plant, commonly used in genus of Gramineae for the research of structure genomics and functional genomics. B. distachyon contribute to rapid and easy analysis because of its small size and quick growth. Mutant population was generated by different doses of gamma-irradiation (0, 50, 100, 150, 200, 250 Gy) in the gamma field phytotron. Distance from the source gives same irradiation duration for each plant. Plant growth parameters such as plant height, tiller number, leaf length & width, internode number & diameter, maturity and yield components (ear number biomass) were scored on M0 plants. Plant responses to different doses of radiation are evaluated and the effective radiation dosages to generate mutant using gamma-phytotron are suggested. Chronic irradiation using gamma-phytotron is useful tool to generate mutants for genomic variations such as SNP or INDEL as well as suitable for functional study of genes in Gramineae.

Rye has important genes for biotic and abiotic stress resistance. Introduction of these genes to wheat by breeding wheat-rye translocation have been intensively used in wheat breeding program. Rye chromatin 1RS and/or 2RL show superior performance in unfavorable environments. In order to develop high yielding wheat, we applied various molecular breeding strategies. To develop EST-derived 1RS specific markers, we used comparative genomics with public sequence databases of Poaceae family. Putative rye chromatin specific sequences were used to design 1RS specific markers. To identify genes related to water deficiency, cDNA AFLP analysis was used in PEG treated seedlings of 1RS RILs. For functional analysis of identified genes and markers, we used Brachypodium distachyon, as a new model plant of temperate grasses. B. distachyon were recently applied for transformation and we constructed Agrobacterium-mediated transformation system. Integration of those strategies and conventional breeding method would enhance the usefulness of rye chromatins for wheat improvement.