Harmonia axyridis have become extremely popular through their extensive elytra and spot variation for many years. However, phenotypic polymorphism and striking visual phenotypes are not yet to be extensively studied. With growing attention to polymorphism of H. axyridis, it is imperative to study involved in genomic research in order to elucidate the molecular mechanisms underlying the diversity of elytra variation. To figure out suitable approach for analyzing genes of H. axyidis, we assessed RNAi as a tool for functional analysis. In order to demonstrate the utility of RNAi technique on H. axyridis, we conducted cDNA library screening to affirm silencing effect. We choose random genes from cDNA clones and the random genes were synthesized into dsRNA. After the injection of dsRNA into last instar larvae, we observed morphological defects or deaths. To investigate the effectiveness of knockdown in H. axyridis, we performed quantitative real time PCR on H. axyridis that appears the most distinctive features. These results provide that RNAi is highly applicable in H. axyridis and it will be useful for the gene functional analysis.
polydnavirus, Cotesia plutellae bracovirus (CpBV), is symbiotic to an endoparasitoid wasp, C. plutellae, which specifically parasitizes young larvae of the diamondback moth, Plutella xylostella. CpBV contains some genes originated from other insect viruses. CpBV-E94k1 and CpBV-E94k2 are homologous to corresponding baculovirus gene E94k, and may play an important role in host-parasitoid interactions. This study was conducted to confirm the origin and function of CpBV-E94k by analyzing its sequence and functional assays. Our phylogenetic analysis indicates that CpBV acquires these E94k genes from baculoviruses. These two genes were expressed during entire period parasitization period. Expression of these E94ks was also tissue-specific because they were expressed in the hemocyte and fat body, but not in the other tissues. Subsequent analysis of gene function by RNA interference showed that it clearly inhibited host immune and developmental processes
Immune defense is indispensible for insect survival. However, uncontrolled and excessive immune responses would be highly detrimental and energy-consuming processes. An insect cytokine, plasmatocyte-spreading peptide (PSP), induces hemocyte-spreading behavior as well as activating phenoloxidase (PO) in the beet armyworm, Spodoptera exigua. A hemocyte transcriptome of S. exigua contains a partial sequence of a putative PSP-binding protein (SePSP-BP). SePSP-BP was expressed in all developmental stages especially in hemocytes and fat body. A quantitative RT-PCR showed that the bacterial infection significantly up-regulated the expression level of SePSP-BP. A double-stranded RNA specific to SePSP-BP (dsRNASePSP-BP) was injected and suppressed SePSP-BP expression even in response to bacterial challenge. The larvae treated with dsRNASePSP-BPsuffered high mortality to infection of nonpathogenic bacteria and prolonged high PO activity after the immune challenge. These results suggest that SePSP-BP may play a role in suppressing immune responses as a negative controller
Our previous study showed that transgenic (TG) pigs harboring human EPO (hEPO) gene have been shown to have reproductive disorders, including low pregnancy rates, irregular estrus cycle and low little size. To investigate these reasons, we assessed estrus behavior (standing response) and plasma 17B-estradiol (E2) level, which partly reflect reproductive function, during the estrus cycles after synchronization and superovulation by hormone treatments. Then, we analysed blood composition and expression of hEPO gene in TG pigs. Pigs were injected with PG600. After 10 days, pigs were fed with Regumate porcine for 6 days. Blood samples were collected from jugular vein. Analysis of blood composition and E2 level were measured by Hemavet 950 and E2 ELISA kit, respectively. And, the expression of hEPO gene in reproductive organs was quantitated by real-time RT-PCR. The percentage of estrus behavior in TG was significantly decreased. Hematocrit (HCT), hemoglobin (Hb) concentration and red blood cell (RBC) number were significantly higher in TG than wild type (WT). On the other hand, high expression of hEPO gene in TG was observed in the mammary gland as well as in the uterus. Moreover, plasma E2 level was significantly higher in TG than WT. These results suggest that nonspecific expression of hEPO gene in the other organs of TG may affect blood composition and plasma E2 level, thereby causing reproductive disorders.
Glycyrrhiza ura lensis (Leguminosae) has long been known as an ant i-i nflammatory agent for gastric 띠cers , arthri t is, and rheumaLism. 1'he f1avonoid glycyr ol isolated from Glycyrrhiza uralensis dramatically inhibits phorbol ester (PMA)-induced NF-kB-dependent transcriptional activity, as determined by luciferase activity in HEK293T cells To in vestigate the global gene expression profiling by glycyrol, we performed high-density oligonucleotide microarrays, Our microarray analysis showed that gl ycY I 이 inhi bited phorbol ester-induced NF-kB-dependent transcriptional activity in inflarnmatory-related gene expression, RT-PCR analysis based on microarry data showed that NF-kB-dependent genes such as CCL2‘ CCL7. CD44, and HSPB8 in addition to NF-kB itself were significantly down-regulated by glycyrol Treatment with glycyrol inhibited ]-kB degradation induced by PMA, suggesting that glycyrol has a potential anti-in flammatory activity by modulating NF-kB-dependent pathways. Also, the microarray data showed that glycyr이 appears to induce gene expression related to p53-dependent apoptosis t hrough ENDO G instead of CAD/DFF and AlF/PDCD8 as down stream apoptosis factor in HEK293T tumor cell s and to induce an added function as oncogenes in connection with apoptoslS
Transcription factors are essential for the regulation of gene expression in plant. They are binding to either enhancer or promoter region of DNA adjacent to the gene and are related to basal transcription regulation, differential enhancement of transcription, development, response to intercellular signals or environment, and cell cycle control. The mechanism in controlling gene expression of transcription can be understood through the assessment of the complete sequence for the maize genome. It is possible that the maize genome encodes 4,000 or more transcription factors because it has undergone whole duplication in the past. Previously, several transcription factors of maize have been characterized. In this review article, the transcription factors were selected using Pfam database, including many family members in comparison with other family and listed as follows: ABI3 / VP1, AP2/EREBP, ARF, ARID, AS2, Aux/IAA, BES1, bHLH, bZIP, C2C2-CO-like, C2C2-Dof, C2C2-GATA, C2C2- YABBY, C2H2, E2F/DP, FHA, GARP-ARR-B, GeBP, GRAS, HMG, HSF, MADS, MYB, MYB-related, NAC, PHD, and WRKY family. For analyzing motifs, each amino acid sequence has been aligned with ClustalW and the conserved sequence was shown by sequence logo. This review article will contribute to further study of molecular biological analysis and breeding using the transcription factor of maize as a strategy for selecting target gene.
Maize is one of the most important food and feed crops in the world including Southeast Asia. In spite of numberous efforts with conventional breeding, the maize productions remain low and the loss of yields by drought and downy mildew are still severe in Asia. Genetic improvement of maize has been performed with molecular marker and genetic engineering. Because maize is one of the most widely studied crop for its own genome and has tremendous diversity and variant, maize is considered as a forefront crop in development and estimation of molecular markers for agricultural useful trait in genetics and breeding. Using QTL (Quantitative Trait Loci) and MAS (Marker Assisted Breeding), molecular breeders are able to accelerate the development of drought tolerance or downy mildew resistance maize genotype. The present paper overviews QTL/MAS approaches towards improvement of maize production against drought and downy mildew. We also discuss here the trends and importance of molecular marker and mapping population in maize breeding.
All aspects of plant life are controlled by the regulated synthesis of new proteins and the precise degradation of preexisting proteins, predicting up to 50% of total plants protein is replaced every week. The ubiquitin/26S proteosome pathway is known to be one of mechanisms to regulate signal pathways, developmental process and abiotic/biotic stress responses via protein degradations. In the previous study, we have identified a large number of the RING ubiquitin ligase proteins whose functions have been clarified in the protein degradation pathway. Curiously, one RING-H2 finger protein gene evidenced striking differences in expression patterns in response to salt and dehydration stress between leaf and culm-node tissues. Characterization of the gene evidenced its function as E3 ubiquitin ligase activity by using an in vitro ubiquitination assay. We have constructed a library with rice culm-node tissues under salt stress for Yeast two hybrid assay and performed primary yeast two-hybrid screening with the gene as a bait. A total of 13 candidate genes were isolated as positive interacting partners. Gene ontology of most candidate genes appears to be related with various abiotic stresses. Therefore, the RING-H2 finger protein genes might function to regulate plant abiotic stress responses via protein degradation pathways.
We carried out to study the function of ArgE in transgenic rice plants, which were confirmed by PCR analysis and hygromycin selection. Transgenic rice plants were with selectable marker gene(HPT) inserted in genome of the rice. Southern analysis with hpt probe confirmed by two restriction enzymes that copy numbers of the selectable gene was introduced into the plant genome. We displayed that the relationship between drought stress and ArgE gene with the overexpressing rice plants. From this result, we observed that the degree of leaves damage has no difference in control and transgenic lines. The total RNAs were extracted from 6 weeks-seedling in normal condition in order to examine their expression levels with ArgE-overexpressed transgenic rice. In particular, expression patterns of genes encoding enzymes involved in abiotic stress, including drought and salt stresses. OsGF14a and OsSalt were investigated by reverse transcription-PCR(RT-PCR). Expression levels of the OsSalt gene decreased significantly in transgenic rice plants compared to control plant. However, ion leakage measurement did not demonstrate any leaves damage change between control and ArgE transgenic plants exposure to mannitol treatment. These results suggest that expression of the ArgE is not involved in tolerance for drought stress in rice but may playa role of signaling networks for salt-induced genes.