수국은 수국과(Hydrangeaceae) 수국속(Hydrangea)의 낙엽관목 식물로 크고 화려한 화형을 가져 절화, 분화 및 조 경수로 전세계적 인기가 있는 식물이다. 나무수국은 수국 (H. macrophylla)과 비교하여 삽목율이 낮은 것으로 알려져 있지만 나무수국의 묘목생산을 위한 삽목 연구 및 두 종간 삽목율 차이 원인 규명에 관한 연구는 미미하다. 본 연구는 IBA(Indol-3-butyric acid) 500mg·L-1 처리시 삽수의 침지 시간에 따른 삽목율 조사를 통해 적정 호르몬 처리 시간을 제 시하고 나무수국과 수국의 해부학적 구조 관찰을 통한 삽목율 차이 발생의 원인을 규명하고자 실시하였다. 나무수국의 적정 호르몬 처리 시간을 규명하기 위해 IBA 500mg·L-1을 무처 리, 30분, 2시간, 4시간 침지처리를 하였다. 종간 삽목율 차이 발생의 원인 규명을 위해 나무수국과 수국의 줄기 단면과 삽 목 후 시간 경과에 따른 발근을 해부학적으로 관찰하였다. 연 구의 결과 나무수국의 삽목시 IBA 500mg·L-1에 2시간 이상 침지처리가 다른 처리구와 비교하여 발근율이 높고 발생 뿌리 수가 가장 많았다. 또, 나무수국의 삽목율이 수국과 비교하여 낮은 것은 줄기의 세포 구조상 방사조직의 형태, 섬유세포의 밀도, 도관의 발달, 전분 함유 세포의 수 등에 차이가 관찰되 었고 이러한 세포 구조적 차이들의 영향으로 나무수국이 수국 보다 삽목 후 뿌리 조직 세포분열이 7일 늦게 시작되는 것이 확인되었다. 본 연구의 결과로 나무수국의 삽목 번식의 기초 자료로 활용되어 묘목 생산 효율 증대에 활용되길 바란다.
To elucidate the effect of cellular phone electromagnetic wave (EMW) exposure on the developing cerebellar cortex of neonatal Sprague-Dawley rats, animals were exposed to cellular phone electromagnetic waves for 1 hr per day for 3 weeks. At the end of the experimental period, animals were sacrificed by cardiac perfusion, after which histological samples were prepared and observed microscopically. In the EMW exposure group, external granule cells were remained partially in the external granular layer without migrating into the internal granular layer. In addition, dark stained shrunken Purkinje cells with pyknotic nuclei increased and the outline of cells became irregular and showed degenerative signs, such as mitochondrial swelling and disrupted cristae. Moreover, the cisternae of rough endoplasmic reticula and Golgi complex were severely swollen. Bergmann glial cells adjacent to the dark stained Purkinje cells were swollen and cytoplasmic organelles were scant. Dark stained shrunken granule cells were also observed and the outline of cells was irregular. The results of the present study suggest that cellular phone EMW exposure to neonatal Sprague-Dawley rats leads to a partial delay of early migration of cerebellar cortical cells and degenerative changes in Purkinje cells, Bergmann glial cells and granule cells.
Peptidoglycan recognition proteins (PGRPs) are family of innate immune molecules that recognize bacterial peptidoglycan. PGRP-LE, a member of the PGRP family, selectively binds to diaminopimelic acid (DAP)-type peptidoglycan to activate both the immune deficiency (IMD) and proPhenoloxidase (proPO) pathways in insects. A PGRP-LE-dependent induction of autophagy to control Listeria monocytogenes has also been reported. We identified and partially characterized a novel PGRP-LE homologue, from Tenebrio molitor and analyzed its functional role in the survival of the insect against infection by a DAP-type PGN containing intracellular pathogen, L. monocytogenes. The cDNA is comprised of an open reading frame (ORF) of 990 bp and encodes a polypeptide of 329 residues. TmPGRP-LE contains one PGRP domain, but lacks critical residues for amidase activity. Quantitative RT-PCR analysis showed a broad constitutive expression of the transcript at various stages of development spanning from larva to adult. RNAi mediated knockdown of the transcripts followed by a challenge with L. monocytogenes showed a significant reduction in survival rate of the larvae, suggesting a putative role of TmPGRP-LE in sensing and control of L. monocytogenes infections in T. molitor. These results implicate PGRP-LE as a defense protein necessary for survival of T. molitor against infection by L. monocytogenes.
CD63, a member of tetraspanin membrane protein family, plays pivotal role in cell growth, motility, signal transduction, host-pathogen interactions and cancer. In this work, the cDNA encoding CD63 homologue (TmCD63) was cloned from larvae of coleopteran beetle, Tenebrio molitor. The cDNA is comprised of an open reading frame of 705 bp, encoding putative protein of 235 amino acid residues. In silico analysis shows that the protein has four putative transmembrane domains and one large extracellular loop. The characteristic ‘Cys-Cys-Gly’ motif and ‘Cys188’ residues are highly conserved in the large extracellular loop. Phylogenetic analysis of TmCD63 revealed that they belong to the insect cluster with 50-56% identity. Analysis of spatial expression patterns demonstrated that TmCD63 mRNA is mainly expressed in gut and Malphigian tubules of larvae and the testis of the adult. Developmental expression patterns of CD63 mRNA showed that TmCD63 transcripts are detected in late larval, pupal and adult stages. Interestingly, TmCD63 transcript was upregulated the maximum 4.5 fold in response to DAP-type peptidoglycan during the first 6 h, although other immune elicitors also made significant increase in the transcript level at later time-points. These results suggest that CD63 might contribute to T. molitor immune response against various microbial pathogens.
Apolipophorin III (apoLp-III) is a well-known hemolymph protein having a functional role in lipid transport and immune response of insects. We cloned full-length cDNA encoding putative apoLp-III from larvae of the coleopteran beetle, Tenebrio molitor (TmapoLp-III), by identification of clones corresponding to the partial sequence of TmapoLp-III, subsequently followed with full length sequencing by a clone-by-clone primer walking method. The complete cDNA consists of 890 nucleotides, including an ORF encoding 196 amino acid residues. Excluding a putative signal peptide of the first 20 amino acid residues, the 176-residue mature apoLp-III has a calculated molecular mass of 19,146 Da. Genomic sequence analysis with respect to its cDNA showed that TmapoLp-III was organized into four exons interrupted by three introns. Several immune-related transcription factor binding sites were discovered in the putative 5’-flanking region. BLAST and phylogenetic analysis reveals that TmapoLp-III has high sequence identity (88%) with Tribolium castaneum apoLp-III but shares little sequence homologies (<26%) with other apoLp-IIIs. Homology modeling of Tm apoLp-III shows a bundle of five amphipathic helices, including a short helix 3’. The ‘helix-short helix-helix’ motif was predicted to be implicated in lipid binding interactions, through reversible conformational changes and accommodating the hydrophobic residues to the exterior for stability. Highest level of TmapoLp-III mRNA was detected at late pupal stages, albeit it is expressed in the larval and adult stages at lower levels. The tissue specific expression of the transcripts showed significantly higher numbers in larval fat body and adult integument. In addition, TmapoLp-III mRNA was found to be highly up-regulated in late stages of L. monocytogenes or E. coli challenge. These results indicate that TmapoLp-III may play an important role in innate immune responses against bacterial pathogens in T. molitor.