Lytic polysaccharide monooxygenases (LPMOs) catalyze the oxidative cleavage of glycosidic bonds in crystalline polysaccharides including chitin and cellulose. The recent discovery of LPMO family proteins in many insect species suggests that they presumably play a role in chitin degradation in the cuticle/exoskeleton, tracheae and peritrophic matrix during insect development. Insect LPMOs belong to auxiliary activity family 15 (AA15/LPMO15) and have been classified into at least four groups based on phylogenetic analysis. In this study, we identified and investigated the physiological functions of group I LPMO15 (MaLPMO15-1 and PhLPMO15-1) in two longhorn beetle species, Monochamus alternatus and Psacothea hilaris. In both species, depletion of LPMO15-1 transcripts by RNAi resulted in a lethal pupal-adult molting defect. The insects were unable to shed their old pupal cuticle and died entrapped in their exuviae. Furthermore, TEM analysis revealed a failure of degradation of the chitinous procuticle layer of their old cuticle, retaining intact horizontal laminae and vertical pore canals containing perpendicularly oriented chitin fibers (pore canal fiber, PCF) in their core. These results indicate that MaLPMO15-1 and PhLPMO15-1 are required for turnover of the chitinous old cuticle, which is critical for insect molting.

This study investigates the impact of Bursaphelenchus xylophilus infection on the gut bacterial communities of the pine sawyer beetle, Monochamus alternatus, with a distinction between male and female individuals. Utilizing specific primers, we determined the infection status and analyzed the bacterial composition across different taxonomic levels. Regardless of infection status or sex, Proteobacteria and Firmicutes were found to dominate the phylum level, with significant contributions from Actinobacteria and Bacteroidetes. At the class level, Bacilli, Gammaproteobacteria, Alphaproteobacteria, and Actinobacteria emerged as prevalent groups. Meanwhile, the genus level was characterized by a high abundance of Lactobacillus, Rickettsia, Bacillus, and Rahnella. Our analysis of alpha diversity metrics, including Observed Species, Shannon Index, Inverse Simpson Index, and Phylogenetic Diversity, revealed no significant differences attributable to B. xylophilus infection; however, notable variations were observed between sexes. Principal Coordinates Analysis and Non-metric Multidimensional Scaling further underscored that the differences in gut bacterial communities were more pronounced between male and female beetles than between infected and uninfected individuals. These findings highlight the influence of host sex over infection status in shaping the gut microbiome of Monochamus alternatus, providing new insights into the complex interactions between host biology, gut microbiota, and pathogen infection.

The Japanese pine sawyer (JPS), Monochamus alternatus Hope, is an important vector of the pine wood nematode, Bursaphelenchus xylophilus (Steiner and Buhrer), which causes pine wilt disease. JPS selects a characteristic life cycle of a four- or five-instar pathway, which indicates the number of instars before pupation. In this study, we investigated the potential presence of a three-instar pathway and sought to determine the minimum larval age required for JPS to construct a pupal chamber. The results showed that no third instars made pupal chambers, suggesting that there is no three-instar pathway. The initiation time of digging the tunnel to make the pupal chamber ranged widely from Julian date 274 (30 September) to 332 (27 November). This timing became an average age stage of 4.8 instar, which equals an 80% completion of the fourth instar. Thus, the minimum larval age at which JPS constructs the tunnel to pupate the next year is estimated to be late in the fourth larval instar. Further, we discuss the diapause ecology in relation to the larval development of JPS in different thermal environments. Tentatively, we suggest that the regulation of diapause induction in JPS involves a dual process of reaching a critical developmental stage and stimulus (tentatively a cold temperature). This hypothesis will be helpful for future studies of diapause mechanisms and the selection of the instar-pathway in JPS and related species.

우리는 솔수염하늘소(Monochamus alternatus) 장에 존재하는 공생미생물들을 분리하였다. 그중 다양한 식물 들을 대상으로 성장 촉진 효과가 보고된 세균을 단일배양 분리하였다. 이 세균은 16S rRNA sequencing을 통하여 Enterobacter roggenkampii로 동정되었다. 우리는 분리된 E. roggenkampii에 대하여 genomic sequencing을 수행하 였고 유전학적 특성을 확인하였다. 우리는 E. roggenkampii가 식물의 성장을 촉진할 수 있는 다양한 유전자들을 가지고 있는 것을 확인하였고 그 중 IAA-Asp hydrolase 유전자를 가지고 있음을 알 수 있었다. 동시에, 분리된 E. roggenkampii와 같은 속의 세균을 대상으로 다양한 API kit와 기질 첨가 배지를 이용하여 생화학적 특성을 비교하였다. 향후 IAA-Asp 가수분해효소를 생산하는 잠재적인 식물 생장 촉진 비료 미생물로 등록하여 다양한 작물을 대상으로 생장 촉진 효과를 확인할 것이다.

Paecilomyces tenuipes (P. tenuipes) is a fungus cultivated artificially by South Korean researchers, utilizing rice bran as its substrate. The increased demand for this fungus has not been met with successful cultivation methods for fruiting body production in natural environments. Therefore, we tested the effect on the growth of P. tenuipes using a Solid media based on pests. In this results, the Solid media based on M.alternatus was effective in increasing the growth of P. tenuipes and the content of cordycepin. Moreover, we confirmed the conditions for manufacturing a Solid media based on M.alternatus for P. tenuipes growth. We suggested that the growth-promoting compounds offers valuable insights for optimizing fungal cultivation conditions, thereby enhancing productivity and contributing to a broader understanding of fungal physiology in varying nutritional environments.

Insect cuticle is an extracellular matrix formed primarily from two different biopolymers, chitin and protein. During each molt cycle, a new cuticle is deposited simultaneously with degradation of the old cuticle by molting fluid cuticle degrading-enzymes, including epidermal chitinases (CHTs). Insect CHTs, belonging to family 18 glycosylhydrolase (GH18), have been classified into at least eleven subgroups based on phylogenetic analyses, and group I (CHT5) and group II (CHT10) epidermal CHTs present in molting fluid. In this study we report the physiological function of MaCHT5 and MaCHT10 in the Japanese pine sawyer, Monochamus alternatus. RNAi for either MaCHT5 or MaCHT10 resulted in larval-pupal and pupal-adult molting defects, in which the insects were unable to shed completely their old cuticle and died entrapped in their exuviae. Furthermore, TEM analysis revealed a failure of degradation of the old cuticle in both MaCHT5- and MaCHT10-deficient pharate adults. In the old pupal cuticle, the chitinous horizontal laminar and vertical pore canal essentially remained intact in the endocuticular layer. These results indicate that both CHTs are required for turnover of the chitinous old cuticle, which is critical for completion of insect molting. We also discuss the possible function of two spliced variants of MaCHT10, MaCHT10a and MaCHT10b.

Lytic polysaccharide monooxygenases (LPMOs) catalyze the oxidative cleavage of glycosidic bonds in crystalline polysaccharides including chitin and cellulose. The recent discovery of LPMO family proteins in many insect species suggests that they presumably play a role in chitin degradation in the cuticle/exoskeleton, tracheae and peritrophic matrix during insect development. Insect LPMOs belong to auxiliary activity family 15 (AA15/LPMO15) and have been classified into at least four groups based on phylogenetic analysis. In this study, we identified, characterized and investigated the physiological functions of group I LPMO15 (MaLPMO15-1 and PhLPMO15-1) in two longhorn beetle species, Monochamus alternatus and Psacothea hilaris. In both species, depletion of LPMO15-1 transcripts in last instar larvae by RNAi had no effect on subsequent larval-pupal molting and the resulting pupae developed normally. However, adverse effects on their development were observed during the pupal-adult molting period. The pharate adults were unable to shed their old pupal cuticle and died entrapped in their exuviae probably due to a failure of degradation of the chitin in their old cuticle, which is critical for completion of the insect molting and continuous growth.

본 연구에서는 소나무재선충의 매개충인 솔수염하늘소와 북방수염하늘소에 대하여 무인항공기 (무인헬리콥터)를 이용하여 스피네토람의 약효 및 약해를 조사하였다. 하늘소를 대상으로 등록된 펜토에이트 유제, 비펜트린 액상수화제, 하늘소를 제외한 딱정벌레가 대상인 에토펜프록 스 유제, 디플루벤주론 수화제와 나방류에 방제 약제로 등록된 인독사카브 수화제, 스피네토람 액상수화제 6종을 ULV기로 살포하여 솔수염하 늘소에 대한 섭식독과 접촉독을 확인한 후 선발하였다. ULV 시험 결과, 펜토에이트, 비펜트린, 인독사카브, 스피네토람의 33배, 55배 희석배수 액은 처리후 3일차에 접촉독과 섭식독에서 100% 살충율을 보였으나, 에토펜프록스는 7일차 접촉독 살충율 88.9%(33배), 88.9%(50배), 섭식독 살충율 93.4%(33배, 50배), 디플루벤주론은 7일차 접촉독 살충율 83.3%(33배), 섭식독 80.3%, 53.9%(50배)로 조금 낮은 살충률을 보였다. ULV 시험에서 선발된 가장 적합한 스피네토람의 33배액을 무인항공기로 살포하여 솔수염하늘소와 북방수염하늘소에 대한 감수성을 평가한 결 과, 98.6% - 100%의 살충율을 보였다. 그러나, 해당 약제의 매개충 방제에 적용하기 전에 항공 살포에 의한 꿀벌에 대한 위해성 평가가 필요할 것 으로 판단되었다.