Fas-associated death domain protein (FADD) functions as an apoptotic adapter in mammals, recruiting caspases for death-inducing signaling complexes, while in lower animals, it interacts with IMD and DREDD to initiate antimicrobial responses. In this study, we examined the T. molitor FADD sequence (TmFADD) using molecular informatics methods to understand its involvement in the host's immune response against microorganisms. Knocking down TmFADD transcripts resulted in increased susceptibility of T. molitor larvae to E. coli, underscoring the significance of FADD in insect defense mechanisms and providing valuable insights into insect immunity.

To understand the role of small heat shock protein (sHSPs) in rice plant response to various stresses such as the heat and oxidative stresses, a cDNA encoding a 24.1 kDa mitochondrial small HSP (Oshsp24.1) was isolated from rice by rapid amplification of cDNA ends (RACE) PCR. The deduced amino acid sequence shows very high similarity with other plant small HSPs. DNA gel blot analysis suggests that the rice genome contains more than one copy of Oshsp24.1. High level of expression of Oshsp24.1 transcript was observed in rice seedlings in response to heat, methyl viologen, hydrogen peroxide, ozone, salt and heavy metal stresses. Recombinant OsHSP24.1 protein was produced in E. coli cells for biochemical assay. The protein formed oligomeric complex when incubated with Sulfo-EGS (ethylene glycol bis (succinimidyl succinate)). Our results shows that Oshsp24.1 has an important role in abiotic stress response and have potential for developing stress-tolerant plants.

In this study, we developed 11 microsatellite markers specific to A. crataegi using NGS to investigate the genetic relationships of A. crataegi populations from South Korea to circumferential Asian countries (China, Russia, Mongolia, and Japan). Further, two mitochondrial DNA (mtDNA) gene segments (COI and CytB) were sequenced from the samples. The population- and individual-based Principal Coordinates and STRUCTURE analyses collectively suggested that the South Korean population of A. crataegi is most differentiated from the Japanese population, whereas it was closer to Mongolian and Chinese populations. These results collectively suggest that northern populations, in particular, Mongolian populations can be considered as the most genetically compatible one as donee population, when the reintroduction program is launched. †These authors contributed equally to this paper.

The black-veined white, Aporia crataegi (Lepidoptera: Pieridae), which is distributed mainly in Eastern Asia is presumed to be extinct in South Korea, only with some numbers of dried specimens left, whereas the species is found casually in circumferential countries. One of the common conservation practices for such species is to launch introduction program, but prior population genetic analysis between donor and donee populations might be essential for long-term conservation. In this study, we developed 11 microsatellite markers specific to A. crataegi using Illumina paired-end sequencing to investigate the genetic relationships of A. crataegi populations from South Korea and circumferential Asian countries (China, Russia, Mongolia, and Japan). Further, two mitochondrial DNA (mtDNA) gene segments (COI and CytB) were sequenced from the samples. The population- and individual-based Principal Coordinates and STRUCTURE analyses collectively suggested that the South Korean population of A. crataegi is most differentiated from the Japanese population, whereas it was closer to Mongolian and Chinese populations. The STRUCTURE analysis based on two concatenated mtDNA gene sequences also supported different genetic composition of Japanese population from the remaining populations including that of South Korea and rather similar genetic composition between the populations of South Korea and Mongolia. These results collectively suggest that northern populations, in particular, Mongolian populations can be considered as the most genetically compatible one as doner population, when reintroduction program is launched.

Beauveria comprises entomopathogenic fungi frequently isolated from insect cadavers, among which Beauveria bassiana is the most widely studied species of entomopathogenic fungal genus for its high potential as a biological pesticide. Even though it has been reported that B. bassiana is a heterogeneous assemblage of strains, little is known about the factors that might drive the genetic diversity among various isolates. In this work, we ought to study the gene diversity of 33 isolates in order to figure out the relationship between their gene diversity and biological features. First, we analyzed gene sequences that are involved in developmental and immunological processes of the fungal isolates and host. Now we are checking whether the gene diversity is related to the genes function, the isolates geographic localization, thermo-tolerance and virulence of the isolates. The obtained results so far showed no correlation between the genetic diversity and the fungal geographic localization. But interestingly, a positive correlation has been obtained between the virulence and the genetic diversity based on the internal transcribed spacers (ITS). Further analyses are aiming to confirm the obtained data and to analyze possible correlations between the genetic diversity and morphological and transcriptional data of the studied isolates. Based on the obtained results, we could much strongly focus on interesting genes which might be related to improving or screening highly virulent fungal agents as biological control agents.

A colony of Solenopsis invicta was first intercepted on Gamman pier, Pusan port in Korea at September, 2017 by Animal and Plant Quarantine Agency. The mitochondrial DNA (mt-DNA) of workers was analyzed and compared with vary libraries of mt-DNA haplotypes to elucidate the origin of the introduced colony with the trade pattern of the Gamman pier. The mt-DNA fragment of 768 bp, which is part of the Cytochrome oxidae I gene, was amplified and sequenced. The results showed that the mt-DNA was in the clade of haplotype 5, which is endemic in southern USA, China, Taiwan, and Australia. More than 60% of containers are imported from China into Gamman pier, it may be possible to assume that the colony was inadvertently invaded through containers from China.

A genus of entomopathogenic Beaveria bassiana has been widely used in pest management, however little studies havebeen given to its virulence-related genes. To identify the roles of virulence genes, AtMT-induced random mutants weregenerated and followed by localization study with TAIL-PCR. Two genes, Complex I intermediated-associated protein30 (CIP30) and Autophagy protein 22 (Atg22), were predicted as virulence-related genes in B.bassiana JEF-007. To validatethe a possible relationship between two genes and fungal virulence, hpRNAi was performed. A hpRNAi plasmid wasconstructed as a model system to knock down of egfp gene in egfp-expressing B. bassiana transformant. A Real-TimePCR showed the knock down of egfp gene expression via hpRNAi. The CIP30 and Atg22 genes in B. bassiana willbe validated by the established RNAi technique. This work can be a strong platform for the functional genetics in B.bassiana.

염생식물 퉁퉁마디의 종자 발아에 영향을 미치는 환경 요인을 조사하고 환경 스트레스에 의해 유도되는 2CysPrx 유전자를 클로닝한 후 스트레스 조건에 따른 2CysPrx 유전자의 발현 양상에 대하여 조사하였다. 염생식물에 대한 가장 대표적인 스트레스는 염분 스트레스로서 퉁퉁마디 발아에 중요한 요인으로 작용하고 있다. 퉁퉁마디의 발아에 대한 NaCl의 한계 농도는 7%로 나타났고, 최적의 발아 조건은 NaCl이 없는 상태로 확인되었다. 퉁퉁마디 발아에서 최적 온도는 20°C로 98%의 발아율을 보였다. 스트레스에 유도되는 유전자 후보군 중 2CysPrx 유전자의 cDNA를 클론하여 분석한 결과 275개의 아미노산으로 이루어져 있고 두 개의 시스테인 잔기를 가지고 있으며 분자량은 30.1kDa 으로 나타났다. 2CysPrx 유전자는 서던 블롯에 의해 유전체에 한 카피 존재하는 것으로 나타났고, 6개의 인트론과 7개의 엑손으로 구성되어 있다. qPCR에 의한 2CysPrx 유전자의 전사율을 분석한 결과, 3.5% NaCl과 40mM H2O2 처리 조건에서 전사율이 가장 높게 나타났고, 고온(40°C)과 75μM ABA 처리 조건에서는 처리 후 8시간에 최고의 전사율을 보였으며, 저온(4°C)에서는 유전자 발현이 일어나지 않는 것으로 나타났다. 우리는 여러 환경 스트레스에 의해 유도되는 다른 유전자의 클로닝을 시도하고 있다.

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.

The aim of this study is to analyze the functional activity of an endo-β-1, 4-glucanase from the wood dwelling lower termite Coptotermes gestroi. Full length cDNA sequences of the endo-β-1,4-glucanase were obtained by primer walking in conjunction with Rapid Amplification cDNA Ends. With the obtained full length sequences, primers for amplifying open reading frame (ORF) excluding the signal peptide and glycophosphatidylinositol anchor were designed. Amplified endo-β-1,4-glucanase fragment was cloned and expressed using pET30(+) expression vector in BL21 E.coli strain. Expression of endo-β-1,4-glucanase was confirmed by Western blotting and the result revealed that only full ORF was expressed. The cellulase activity of protein preparations from the induced and non-induced cells was analyzed with Congo Red assay with the cellulase from Aspergillus niger (Sigma Aldrich) as a positive control. The activity of C. gestroi endo-β-1,4-glucanase was significantly higher than those observed in the positive control and the enzyme preparation from non-induced cells. Therefore, this study confirmed that C. gestroi endo-β-1,4-glucanase had a function of cellulose hydrolysis.