Background : Ixeris genus has been used in traditional medicines as stomachics, sedatives, and diuretics. Ixeris dentata var albiflora is a kind of perennial herbaceous plant and one of the plants of the genus Ixeris (Asteraceae). It is well-known for edible wild vegetable in Korea, China, Japan, and Mongolia. Specially, Korean has its root and young leaf with appetizing vegetable due to bitter taste. Methods and Results : We isolated 8 genes that are involved in carotenoid biosynthesis using the Illumina/Solexa HiSeq2000 platform. In this study, a full-length cDNA clone encoding phytoene synthase (IdPSY), phytoene desaturase (IdPDS), ξ-carotene desaturase (IdZDS), lycopene β-cyclase (IdLCYB), and zeaxanthin epoxidase (IdZEP) and partial-length cDNA clones encoding lycopene ε-cyclase (IdLCYE), ε-ring carotene hydroxylase (IdCHXE), and β-ring carotene hydroxylase (IdCHXB2) were identified in I. dentata. The theoretical molecular weight (MW) and isoelectric point values of 8 genes were investigated. Sequence analyses revealed that these proteins shared high identity and conserved domains with their orthologous genes. IdPSY, IdPDS, IdZDS, IdLCYB, IdCHXB2, and IdZEP were constitutively expressed in the roots, stems, leaves, and flowers of I. dentata. Conclusion : Our study on the biosynthesis of carotenoids in I. dentata will provide basic data for elucidating the contribution of carotenoids to the considerable medicinal properties of I. dentata.

Background : Mulberry (Morus alba L.), renowned for their medicine benefits and the leave as the sole food for silkworm (Bombyx mori). To understanding the molecular mechanism of color formation and nutritive value in different mulberry fruit varieties, we use high-throughput transcriptome sequencing technique to investigated the anthocyanin and betulinic biosynthesis pathway related functional genes. In addition, the total antosyanin and betuinic acid contend were also measured. Methods and Results : The resulting cDNA library was then sequenced using an Illumina HiSeq™ 2000 system. The clean reads were assembled using Trinity software, Then perform gene family clustering to get final unigenes. The pH differential method was used to determine the total anthocyanin content (TAC) of methanol extract from the red and white mulberry, and High-performance liquid chromatography (HPLC) analysis was used to quantify the triterpenes content. In this study, total 50,149 unigenes with an average length of 1,125 nt and N50 equaling 1,861 nt were generated. Using these transcriptome sequecing, cDNAs encoding anthocyanin biosynthetic genes and triterpene biosynthetic genes were isolated. In addition, total anthocyanins and betulinic acid content were analyzed. A great amount of total anthocyanins (59.16 mg/g) were found in fully ripe fruit of Cheongil. Accumulation of betulin and betulinic acid were also detected in all stages of Cheongil and Turkey fruits with small amount. Conclusion : The results of transcriptome sequencing provide useful information at molecular lever in mulberry research, such as interesting gene discovering, marker assisted molecular breeding, and interesting metabolic pathway investigate. The gene expression results could help us understanding of the molecular mechanisms of different fruit color determining factor.

Molecular characterization of crops improved through biotechnology has traditionally been conducted using Southern blot analysis which has been used to determine T-DNA copy number, the presence or absence of backbone (sequence outside of the T-DNA) and to demonstrate generational stability of the T-DNA insert. The advancement of high-throughput DNA sequencing (HTS) technology allows efficient characterization of the transgene incorportated into the genome of the plant by rapidly sequencing the entire plant genome. By combining NGS (Next Generation Sequencing) technologies with bioinformatic methods that identify the T-DNA insert derived from the plasmid vector and genome-T-DNA junction sequences, it has been shown that conclusions equivalent to those of a Southern blot are readily obtained. NGS is done at sufficient coverage depth (>75x) across the entire genome. By mapping the sequence reads to the plasmid vector, and identifying the number of unique junctions, we can confirm insert number, copy number, absence of backbone, across multiple generations. With the widespread availability of NGS and steadily decreasing costs it is likely that academia and industry will fully transition to NGS-based molecular characterizations in the near future.

UDP-glucose 4-epimerase (UGE; EC 5.1.3.2) is an enzyme that plays an essential role in the interconverts UDP-D-glucose (UDP-Glc) and UDP-Dgalactose (UDP-Gal). Five members of the Chinese cabbage (Brassica rapa) UDP-glucose 4-epimerase gene family, designated BrUGE1 to BrUGE5, have been cloned and characterized. Quantitative PCR shows that the BrUGE1and BrUGE4 mRNA are most abundant among other BrUGE genes, accounting for more than 55% of total BrUGE transcripts in most of the tissues examined. All genes showed organ specific expression pattern, two of which (BrUGE1 and 4) actively responded after Pectobacterium carotovorum subsp. carotovorum infection, while four genes (BrUGE-1, -3, -4 and -5)were shown to respond considerably against salt, drought and abscisic acid (ABA) treatments. To better understand the function of the UGE gene, we constructed a recombinant pART vector carrying the BrUGE1 gene under the control of the CaMV 35S promoter and nos terminator and transformed using Agrobacterium tumefaciens. We then investigated BrUGE1 overexpressing rice lines at the physiological and molecular levels under biotic and abiotic stress conditions. Bioassay of T3 progeny lines of the transgenic plants in Yoshida solution containing 120 mM Nacl for 2 weeks, confirmed that the BrUGE1 enhances salt tolerance to transgenic rice plants. Also T3 progeny lines of the transgenic plants, when exposed to infection caused by Xanthomonas oryzae pv oryzae, showed tolerance to bacterial blight. These results showed that BrUGE1 can be used as potential genetic resource for engineering Brassica with multiple stress resistance.

Heat shock transcription factors(HSFs) are the major heat shock factors regulating the heat stress response. They participate in regulating the expression of heat shock proteins (HSPs), which are critical in the protection against stress damage and many other important biological processes. In this study, a genome-wide analysis was carried out to identify all HSFs soybean genes. Twenty six nonredundant HSF genes(GmHsf) were identified in the latest soybean genome sequence. Chromosomal location, protein domain and motif organization of GmHsfs were analyzed in soybean genome. The phylogenetic relationships, gene duplications and expression profiles of GmHsf genes were also presented in this study. According to their structural features, the predicted members were divided into the previously defined classes A–C, as described in Arabidopsis. Using RT-PCR, the expression patterns of 26 GmHsf genes were investigated under heat stress. The data revealed that these genes presented different expression levels in response to heat stress conditions. Real-time (q)RT-PCR was performed to investigate transcript levels of five GmHsfs in response to multiple abiotic stresses. Differential expression of five GmHsfs implies their role during abiotic stresses. Subcellular localization using GFP-fusion protein demonstrated that GmHsf12 and GmHsf34 were restricted to the nucleus and GmHsf28 was localized in the nucleus and cytoplasm in plant. The results provide a fundamental clue for understanding of the complexity of the soybean HSF gene family and cloning specific function genes in further studies and applications.

The plant-specific NAC (NAM, ATAF, and CUC)-domain proteins play important roles in plant development and stress responses. Comparative time-course expression analyses were carried out to analyze the expression levels of 62 soybean NAC genes during drought stress in order to search for the stress-inducible NAC genes. Ten GmSNAC (Glycine max stress-inducible NAC) genes having the significant differential expression in response to the drought stress and abscisic acid (ABA) hormone application were further investigated for their expression profiles with various stresses such as drought, high salinity, cold and with ABA treatments by the quantitative real-time PCR analyses. In this research, the full-length cDNAs of eight GmSNAC were isolated for the further studies. Eight GmSNAC proteins were tested for their transcription activation in the yeast assay system. Two GmSNAC proteins showed the very high transcriptional activities and the other two GmSNAC proteins displayed moderate levels of transactivation while the remaining four GmSNAC proteins lacked transactivation in yeast. Subcellular localization of eight GmSNAC proteins was analyzed via the green fluorescent protein-GmSNAC fusion protein in tobacco plant cell. Three GmSNAC proteins with the C-terminal transmembrane domain were localized to the nucleus and cytoplasmic fractions. The other five GmSNAC proteins were targeted to the nucleus. The function of GmSNAC49 gene was further investigated using the overexpression transgenic Arabidopsis. Germination rate in transgenic plants over-expressing GmSNAC49 was delayed in the media supplemented with mannitol or ABA compared with that of wild-type (WT) plants. The 35S:GmSNAC49 transgenic Arabidopsis displayed improved tolerance to drought stress compared to the WT. The results of this systematic analysis of the GmSNAC family responsive to abiotic stress will provide novel tools and resources for the development of improved drought tolerant transgenic soybean cultivars

Two carotenoid biosynthetic genes, phytoene synthase (Psy) and carotene desaturase (CrtI) linked via synthetic 2A sequence under control of CaMV 35S promoter (two T0 plants 5 and 6) or β- conglycinin promoter (three T0 plants 7, 13 and 16) were transformed into soybean variety Kwangan. After agronomic and phenotypic selection at early generations, T5 progeny of PAC soybean were analyzed by Southern blot to confirm T-DNA copy numbers. A total of 27 homologous lines derived from one of three T0 plants (line 7 under the control of β- conglycinin promoter) with one copy T-DNA insertion, were separated and planted into greenhouse. Flanking sequence analysis was carried out on one of homologous line 6-2-3 and results indicated the T-DNA was intergenic inserted into chromosome 14 from 10,873,131 to 10,872,998 base of soybean chromosome. T-DNA insertion structure, flanking sequence and inserted gene expressions need to be analyzed in the further study.

농작물은 다양한 외부 환경스트레스에 노출되어 있다. 환경스트레스는 작물의 성장에 영향을 주어 세계 각 지역의 농업 생산량을 심각하게 감소시키고 있다. 따라서 작물의 생산성을 높이기 위해서 다양한 환경스트레스에 내성이 강한 새로운 품종의 개발이 요구된다. 최근의 연구 동향은 환경스트레스 저항성 유전자를 작물에 도입시켜 환경 변화에 대한 저항성이 강한 작물을 개발하는 것이다. 본 연구에서는 배추의 저온, 고농도의 염과 건조 등의 환경스트레스에 대한 저항성 유전자로 추정되는 BrTSR53의 염기서열을 분석하였다. BrTSR53의 유전자의 총 길이는 481 bp이며 이중에서 ORF 부위는 234 bp이었다. 이 ORF의 염기서열 상동성을 분석한 결과 Arabidopsis에서 보고된 유전자와 유사한 것으로 나타났다. BrTSR53의 발현을 분석하기 위하여 quantitative real-time PCR을 실시하였다. 그 결과 배추를 고염 처리, 저온 처리하고 3시간 후에 가장 높은 mRNA 양을 보였으며, 건조 처리에서는 36시간 후에 발현량이 최대치를 보였다. 따라서 이 ORF는 환경스트레스에 대한 배추의 저항성 유전자임을 확인하였다. 그리고 BrTSR53 유전자를 효모발현 벡터인 pYES-DEST52에 삽입하고 western blot 분석법을 통해 효모에서 분자량이 약 13 kDa인 저항성 단백질의 발현을 확인하였다. 또한 BrTSR53 형질전환 효모는 염분 스트레스에 대한 저항성이 증가한 것으로 나타났다. 따라서 BrTSR53 유전자는 농작물의 환경스트레스 저항성을 높여줄 수 있는 주요한 유전자원으로 이용될 수 있다고 사료된다.

The inflammatory response to infections, such as bacteria and viruses is mediated by multiple host factors. The tumor related-genes are the important cytokines in mammals. However, a number of tumor related-genes are not identified in the rock bream. Here, we have reported the identification and molecular characterization of the tumor related genes. The LPS-induced TNF-α factor 1 and 2 (LITAF1, LITAF2), tumor necrosis factor superfamily member 14 (TNFSF14), tumor necrosis factor receptor superfamily member 14 (TNFRSF14) and translationally controlled tumor protein (TCTP), programmed cell death 10 (PCD10) from rock bream are used for the under investigations. The LITAF1 and LITAF2 consist of 138 and 163 amino acids with a conserved LITAF domain. TNFSF14 and TNFRSF14 comprise 266 and 181 amino acid, respectively. TCTP encompasses of 170 amino acid containing two conserved TCTP signatures. Furthermore PCD10 consists of 210 amino acids. Using quantitative real-time PCR, we have obtained expression analysis results of LITAF1 and 2, TNFSF14, TNFRSF14, TCTP, PCD10 in the various tissue. Compared to the control, the tumor related genes mRNA is detected at a high levels in gill (LITAF1, TCTP), intestine (LITAF2), liver (PCD10), spleen (TNFSF14) and RBC (TNFRSF14). We have also performed gene expression analysis in the kidney, spleen, liver and gill after challenging with Streptococcus iniae, Edwardsiella tarda and Red seabream iridovirus. We have acquired the dynamic regulated mRNA expression to each of pathogen according to the tissue. Expression of tumor related-genes mRNA are significantly increased by infected with pathogens in most of the tissue. But oddly, PCD10 mRNA is expressed significantly decreased by S. iniae infection in all of tissues. Our results reveal that rock bream tumor relatived-genes may be involved in rock bream immune responses to pathogen infections, as well as, they also act like potential biomarkers for innate immunity.

We used a microarray dataset that is deposited in the public database to evaluate plant responses to heat stress and selected two genes, OsSHSP1 (Os03g16030) and OsSHSP2 (Os01g04380), that are highly expressed under heat stress in rice. OsSHSP1 and OsSHSP2 gene transcripts were highly induced in response to salt and drought. In addition, OsSHSP1 and OsSHSP2 gene transcripts were induced under ABA and SA. Subcellular localization of proteins of 35S::OsSHSP1 were associated with the cytosol, whereas those of and 35S::OsSHSP2 were associated with the cytosol and nucleus. Heterogeneous overexpression of both genes exhibited higher germination rates than those of wild-type plants under the salt treatment, but not under heat or drought stress. The network of both genes harboring 9 sHSPs as well as at least 13 other chaperone genes might support the idea of a role for sHSPs in the chaperone network. Our findings might provide clues to shed light on the molecular functions of OsSHSP1 and OsSHSP2 in response to abiotic stresses, especially heat stress.