Heat shock proteins (HSPs) are highly conserved cellular proteins that contribute to adaptive responses of organisms to a variety of stressors. In response to stressors, cellular levels of HSPs are increased and play critical roles in protein stability, folding and molecular trafficking. The mRNA expression pattern of two well-known heat shock protein transcripts, HSP70 and HSP90 were studied in two tissues of nerve ganglia, cerebral ganglion and pleuropedal ganglion of Pacific abalone (Haliotis discus hannai). It was observed that both HSP70 and HSP90 transcripts were upregulated under heat stress in both ganglion tissues. Expression level of HSP70 was found higher than HSP90 in both ganglia whereas cerebral ganglion showed higher expression than pleuropedal ganglion. The HSP70 and HSP90 showed higher expression at Day-1 after exposed to heat stress, later decreased at Day-3 and Day-7 onwards. The present result suggested that HSP70 and HSP90 synthesize in nerve ganglion tissues and may provide efficient protection from stress.

말에서 주요 경제형질인 운동과 관련된 연구는 중요하지만, 현재까지의 연구는 물리학적, 생리학적 연구에 치중되어 있어 분자수준의 연구는 미비한 실정이다. 이에 본 연구는 선행연구를 통하여 경주마에서 RNA-sequencing을 수행하여 운동 전·후 alternative transcript 이형에 따라 발현 양상이 상이한 유전자(DYNC1LI2, COBLL1, AXL, PLEKHG1)를 발굴하였다. 이 중, DYNC1LI2 유전자를 선택하여 분자생물학적 분석 및 운동성과의 관계에 대하여 연구를 수행하였다. 그 결과, DYNC1LI2 유전자의 2가지 전사 이형은 긴 형태의 전사체(DYNC1LI2a)와 결손이 일어나 상대적으로 짧아진 전사체(DYNC1LI2b)의 형태로 존재하는 것을 확인하였고, 두 가지 전사 이형 모두가 말의 각 조직(갑상선, 결장, 골격근, 맹장, 심장, 신장, 척수, 폐)에 존재함을 확인하였다. 또한, 운동 전과 운동 후 발현량 분석을 통해 두 가지 전사 이형이 동일하게 운동에 따라 발현이 감소하는 것을 확인하였다. 추가적으로 두 가지 전사 이형의 아미노산 비교 분석 결과, 엑손영역에 결손이 일어나는 부분은 단백질의 인산화 및 당질화와 관련이 있음을 확인하였다. 이는 DYNC1LI2a가 DYNC1LI2b에 비해 더욱 단백질의 안정화 작용을 하는 것을 의미하며, DYNC1LI2 유전자가 운동에 따라 발현이 달라짐에 따라 차후 말에서 운동관련 연구에 대한 기반 자료로써 사용될 수 있음을 시사한다.

머루(Vitis coignetiae)는 국내에 자생하는 야생종 포도의 한 종류로서, 포도 새눈무늬병에 저항성이다. 내병성 포도 육종에 활용할 유용한 육종소재를 개발하고자 머루의 잎과 과실로부터 cDNA libraray를 제작하였다. 머루의 cDNA library의 총 5,760개의 EST 클론을 분석하여 676개의 contig와 2,306개의 singleton을 분리하여 총 2,982개의 unigene을 분리하였다. NCBI 데이터베이스의 BLAST를 통한 상동성을 검색한 결과, 2,241개의 클론이 기능이 알려진 유전자이었고, 그 중에서 1,442개는 생물학적인 대사에 관여하고 836개는 세포구성물과 관련이 있었다. EST와 contig의 평균 크기는 각각 702bp와 757bp이었다. 포도새눈무늬병균에 감염된 머루 cDNA library로부터 Proline-rich cell wall protein, thaumatin-like protein, class IV chitinase, and pathogenesis-related (PR) protein 10 등의 다양한 방어관련 유전자와 광합성관련유전자 및 수분스트레스 저항성 관련유전자가 많이 검출되었다. 본 연구를 통해 얻어진 머루의 EST 자료는 포도의 새로운 유전자원에 관한 연구 및 내병성 포도 신품종 육종 프로그램에 기본자료로 유용하게 활용될 것이다.

Although the function and utility of RNA transcripts derived from matured spermatozoa remains unclear, they might play important roles in the establishment of a paternal genome and subsequently embryo development. Herein, we investigated the expression of X-chromosome linked RNA transcripts in matured bovine spermatozoa. The total RNA was extracted from the matured spermatozoa, and then converted to cDNA. Autosomal genes (ACT-β and H-2A) and X-chromosome linked genes (ANT3, HPRT, MeCP2, RPS4X, XIAP, XIST and ZFX) were analyzed for the characterization of X-chromosome linked RNA transcripts and compared to female fibroblasts by RT-PCR. The transcripts of autosomal genes (ACT-β and H2A) and X-chromosome linked genes (ANT3, HPRT, MeCP2, RPS4X and ZFX) were not detected in spermatozoa. However, XIAP (X-linked inhibitor of apoptosis protein) and XIST (X-chromosome inactive-specific transcript, a kind of paternal imprinted gene) transcripts were detected in spermatozoa, and relative levels of XIAP and XIST transcripts were similar and 0.5-fold lower when compared to female fibroblasts, respectively. Based on the findings, it is summarized that the presence of RNA transcripts of XIAP and XIST in the isolated spermatozoa may imply their role in inhibition of apoptosis and induction of X-chromosome inactivation in embryo development.

Polydnaviruses (PDVs) are a group of insect double stranded DNA viruses and symbiotically associated with host endoparasitoid wasps. Their segmented genome is located in host chromosome(s) in a proviral form. Viral replication is initiated at the ovary during late pupal stages. Little is known about the factors involved in the viral replication. This study analyzed the ovarian transcripts of an endoparasitoid wasp, Cotesia plutellae, by 454 pyrosequencing and subsequent gene annotation. Out of 2,226 contigs and 12,457 singletons, 50 transcripts categorized in DNA replication, coat proteins, and viral origins were selected as putative viral replication factors. The selected genes were analyzed in their expressions according to host wasp development. Quantitative real-time RT-PCRs showed that some of the selected genes were expressed during the viral replication at late pupal stage. Using RNA interference, five putative genes were tested in their implication in the viral replication by analyzing viral DNA amplification, structure of ovarian calyx, and parasitism. RNA interference of contig#1004 (broad complex) or contig#174 (a viral DNA polymerase gene) significantly inhibited DNA amplification without any impairment of viral formation, and subsequently resulted in significant reduction in the wasp parasitism. This study reports that two wasp genes (or not encapsidated viral genes) are implicated in the viral DNA amplification and viral coat protein production during the polydnaviral replication.

Common wheat has complex genome composition of homoeologous hexaploid (AABBDD, 2n = 6x = 42) and each homoeologous genome has high similarity. Due to these complexity, wheat genome study is a large challenge to researchers for genomic and genetic study. We analyzed expressions of individual wheat genome and rye genome specific transcripts using custom array with 2BS.2RL wheat-rye translcoation. Genomic probes were synthesized within each diploid progenitors (AA, BB, DD, 2n = 14, respectively) of wheat, common wheat, and rye (RR, 2n = 14). Total RNA isolated from seedlings of T. urartu, Ae. speltoides, Ae. squarrosa, ‘Chinese Spring’, ‘Chaupon’, and 2BS.2RL were hybridized on arrays. Each homoeologous gene differentially expressed in hexaploid wheat and rye were identified on the custom array and the transcripts were clustered based on hybridization values. qRT-PCR was performed to verify the custom array result with a set of five genes by highly replicated experiments (three biological and three technical replications). The qRT-PCR results demonstrated genome specific expression of five genes in sympathy with array results. Here we provide information of each individual genome specific transcripts and it will we a useful data to study complex wheat genome compositions.

Expression profiling was conducted with the Oryza sativa alternative splicing detecting microarray v.4 (OsASDM). Probe features are designed based on rice genome IRGSP_1.0 (http://rapdb.dna.affrc.go.jp/ ). The genome contains 37,868 genes. Among these 5,254 genes have alternative spliced sites, 11,938 transcripts. In the microarray, a total of 41,953 transcripts are covered from all the loci and 9112 alternative spliced transcripts. Four 60-nt long probes were designed from each transcript starting 60 bp ahead the end of stop codon and with shifting 30 bp so 4 probes cover 150 bp in the 3’ region of the gene. Genes from chloroplast (123) and mitochondria (74) and selection markers such as gfp, gus, hyg, bar, and kan are included. In total, he 125,956 probes were designed. To find organ specific transcripts RNA was prepared from leaf, root, panicle at 1 cm (P1cm). The signal intensity files were analyzed with limma package. Background correction and normalization were performed with libraries in the package. 13,486 genes are organ specific and 1,856 transcripts are alternatively spliced. Transcripts that specifically alternatively spliced in leaf are Os02t0197600-02_UE; Chlorophyll a-b binding protein 8, Os11t0707000-01_UE; Ribulose bisphosphate carboxylase/oxygenase, Os12t0291100-01_UE; ribulose 1,5-bisphosphate carboxylase small subunit. Transcripts that specifically alternatively spliced in root are Os03t0669100-02_UE; Deoxyuridine 5’-triphosphate nucleotidohydrolase, Transcripts that specifically alternatively spliced in tissues at P1cm are Os11t0210300-02_UE; Alcohol dehydrogenase 1, Os04t0631200-02_UE; Xyloglucan endotransglycosylase. Os03t0669100-02_UE ; Deoxyuridine 5’-triphosphate nucleotidohydrolase, Os11t0210300-02_UE ; Alcohol dehydrogenase 1, Os04t0631200-02_UE; Xyloglucan endotransglycosylase. These results show that OsASDM could be used to find alternatively spliced gene at ease.

Abiotic environmental stresses cause serious economic losses in agriculture. These stresses include temperature extremes, high salinity and drought. We identified several drought stress-related novel/function unknown coding transcripts (transcription factors and functional genes) and non-coding transcripts (small noncoding transcripts such as microRNA and long noncoding transcripts) using the next generation sequencing method from rice (Oryza sativa L.), and have constructed databases of drought stress-related coding and noncoding transcripts. We used novel gene prediction programs for the selections. The expression level of the each gene was analyzed by real-time PCR. The results ended up the selection of 29 transcription factors, 6 microRNAs and 10 long noncoding RNAs. Currently, we are further characterizing these transcripts. We expect that this study could provide functional information of the drought stress-regulated novel genes, and relationships among novel coding and noncoding transcripts.

Recent global warming and climate change has presented greater challenge to the global agriculture of having to cope with more severe adversaries from various abiotic stress conditions including drought, cold, and heat. As a preliminary step towards developing a heat-tolerant japonica rice variety through molecular breeding, we examined and compared expression of several genes that have been reported being expressed specifically during rice panicle development in different rice varieties after subjecting them heat stress. Although the induction of these transcripts upon heat treatment was invariably observed in all rice varieties tested, the magnitude and kinetics of the induction were found to be different among these varieties, suggesting possible functional implication of these genes in conferring heat tolerant phenotype during reproductive organ development of these plants. General protein synthesis activity as well as pollen viability incurred by the heat stress treatment were also monitored in these plants and the result showed a close correlation overall with the induction dynamic of these transcripts under heat stress. Therefore, these genes, together with the ones involved in the regulatory network for the expression of them, could serve as candidates for useful markers with which molecular breeding of heat tolerant japonica rice can be facilitated.

Methyl jasmonate(MeJA) treatment promotes the plant secondary, especially triterpene saponin which is main compounds in Platycodon grandiflorum. We were performed 5,760 expressed sequence tags from MeJA treated hairy root. Total number of ESTs are 2,536 (811 contigs and 1,725 singletons) among them 2,299 are pass the annotation. The most abundant transcript were encoding fatty acid desaturase (FAD) and lots of secondary metabolite transcript were observed. Four MVA pathway genes (HMGS, HMGR, MK, MDD) and one MEP pathway gene (IDI) are identified. The unigene were classified into three major functional categories by standard GO (Gene Ontology) - cellular component, molecular function and biological process. In this study we were cloned four genes from ESTs : HMGS (1401bp ORF), MK (1167bp ORF), MDD (1254bp ORF), IDI (909bp ORF)