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Identification, in silico characterization, and expression analysis of Tenebrio molitor Cecropin‐2
Ali Mohammadie Kojour Maryam,Jang Ho Am,Edosa Tariku Tesfaye,Keshavarz Maryam,Kim Bo Bae,Bae Young Min,Patnaik Bharat Bhusan,Han Yeon Soo,Jo Yong Hun 한국곤충학회 2021 Entomological Research Vol.51 No.2
Antimicrobial peptides (AMPs) are considered to be candidate effectors for eliciting humoral immune responses against infectious pathogens in the host. Cecropins are α‐ helical peptides of 30–40 amino acids, which are known to permeabilize bacterial membranes, and play authoritative roles in the innate immune system of insects. In the present study, we identified the full‐length open reading frame (ORF) encoding the Tenebrio molitor cecropin‐2 (TmCec2) gene using the Tribolium castaneum cecropin‐2 (TcCec2) gene to query a T. molitor Rnaseq database. Phylogenetic analysis revealed close identity of TmCec2 with TcCec2. TmCec2 was ubiquitously expressed in the insect during all developmental stages, with the highest expression observed in the adult. Tissue‐specific TmCec2 expression was highest in larval hemocytes and in the adult integument and hemocytes. Microbial challenge experiments revealed that TmCec2 was highly induced in response to gram‐positive and gram‐negative bacteria, and fungi. These data provide credible evidence for a putative role of TmCec2 in insect innate immunity against a plethora of pathogens.
An overview of insect innate immunity
Ali Mohammadie Kojour Maryam,Han Yeon Soo,Jo Yong Hun 한국곤충학회 2020 Entomological Research Vol.50 No.6
Due to the exposure of insects to various sources of pathogens during different stages of their life cycle and their vulnerability towards subsequent infections, moreover lack of morality issues, economical breeding and short-term-life cycle, insect’s immunity has been considered as a high-potential candidate to study underlying mechanisms of defense responses against all sort of invaders, as well as pandemic diseases. Currently, the world is enduring monumental pressure to meet global food production demands. One viable option would be to mass rear edible insects, such as coleopteran meal worm Tenebrio molitor, which is thought to be a substantial protein source. In addition, using antimicrobial peptides as an alternative for antibiotic source against multidrug-resistant pathogens, makes insects a valuable option to solve this health issue. As a consequence, sufficient knowledge of insect immunity will lead us to reach advanced diagnostic and treatment technologies. To accomplish these goals, crucial importance of identification and functional characterization of the main signaling pathways, such as Toll and immune deficiency (IMD), prompted us to review the mechanisms of the signaling pathways involved in immune response in well-known insect models.
살진균제인 캡탄 처리 후 갈색거저리의 해독효소 유전자 발현
장호암,백형선,김보배,Maryam Ali Mohammadie Kojour,Bharat Bhusan Patnaik,조용훈,한연수 한국응용곤충학회 2022 한국응용곤충학회지 Vol.61 No.1
The application of fungicides is indispensable to global food security, and their use has increased in recent times. Fungicides, directly or indirectly, have impacted insects, leading to genetic and molecular-level changes. Various detoxification mechanisms allow insects to eliminate reactive oxygen species (ROS) toxicity induced by agrochemicals including fungicides. In the present study, we analyzed the mRNA expression levels of detoxifying enzymes in Tenebrio molitor larvae following exposure to non-lethal doses (0.2, 2, and 20 μg/μL) of a fungicide captan. Transcrips of peroxidases (POXs), catalases (CATs), superoxide dismutases (SODs), and glutathione-s-transferases (GSTs) were screened from the T. molitor transcriptome database. RT-qPCR analysis showed that TmPOX5 mRNA increased significantly 24 h post-captan exposure. A similar increase was noticed for TmSOD4 mRNA 3 h post-captan exposure. Moreover, the expression of TmCAT2 mRNA increased significantly 24 h post-treatment with 2 μg/μL captan. TmGST1 and TmGST3 mRNA expression also increased noticeably after captan exposure. Taken together, these results suggest that TmPOX5 and TmSOD4 mRNA can be used as biomarkers or xenobiotics sensors for captan exposure in T. molitor, while other detoxifying enzymes showed differential expression. 최근 살진균제는 세계 식량 안보에 없어서는 안될 필수 요소이며, 그 사용량은 증가하고 있다. 살진균제는 직접적 또는 간접적으로 곤충에 영향을 미쳐 유전자 및 분자 수준의 변화를 일으킨다. 곤충은 다양한 해독 매커니즘을 통해 살진균제를 포함한 농약으로부터 유발되는 활성산소 (ROS) 독성을 제거한다. 본 연구는 살진균제 캡의 비치명적 투여량(0.2, 2, and 20 μg/μL)을 주입 후 갈색거저리의 유충에서 해독효소의 mRNA 발현량을 분석했다. 갈색거저리의 전사체 분석을 통해 해독 매커니즘 관련 유전자인 퍼옥시다제(POX), 카탈라제(CAT), 슈퍼옥사이드 디스뮤타제(SOD) 및 글루타티온-S-트랜스퍼라제(GST)를 발굴하였다. 처리 24시간 후 TmPOX5 mRNA가 유의하게 증가한 것으로 나타났다. 처리 3 시간 후 TmSOD4의 mRNA가 유사하게 증가하였다. 또한 2 μg/μL 처리 24시간 후 TmCAT2의 mRNA 가 유의하게 증가하였다. 캡탄 노출 후 TmGST1 및 TmGST3의 mRNA 발현량도 증가하였다. 결론적으로, TmPOX5 및 TmSOD4 유전자는 갈색거저리에서 캡탄 노출에 대한 바이오마커 또는 생체이물 센서로 작용할 수 있음을 시사한다.
Jang Ho Am,Park Ki Beom,Kim Bo Bae,Ali Mohammadie Kojour Maryam,Bae Young Min,Baliarsingh Snigdha,Lee Yong Seok,Han Yeon Soo,Jo Yong Hun 한국곤충학회 2020 Entomological Research Vol.50 No.12
Defensins are a major family of antimicrobial peptides that serve as the innate immune defense of both vertebrates and invertebrates. Due to their antimicrobial, chemotactic, and regulatory activities, Defensins have been exploited for their therapeutic potential. Insect Defensins are cysteine‐rich and contain an N‐terminal loop, α‐helix, and antiparallel β‐sheet, forming a “cysteine‐stabilized alpha beta (CSαβ)” or “loop–helix‐sheet” structure. In this study, we identified the full‐length open reading frame (ORF) sequences of Defensin (TmDef) and Defensin‐like (TmDef‐like) genes from the mealworm beetle Tenebrio molitor using in silico methods. TmDef and TmDef‐like genes encode the peptides of 72 and 71 amino acid residues, respectively. TmDefensin is comprised of a Defensin domain and the TmDefensin‐like is comprised of a signal peptide of 21 amino acid residues. Phylogenetic analysis revealed close similarities of TmDefensin with the Defensin of Acalolepta luxuriosa of the longhorn beetle family. The expression of TmDef mRNA was found to be greater than that of TmDef‐like mRNA and was mostly expressed in the pupal and adult stages. Tissue distribution showed high expression of TmDef‐like mRNA in larval hemocytes, gut, integument, and fat body, while in adults, the expression was high in gut and hemocytes. Following bacterial and fungal stimulation in vivo, TmDef was upregulated at 24 h post‐infection in whole body, fat body, and hemocytes of the larvae. Even TmDef‐like mRNA was upregulated in the gut and hemocytes at 12 and 9 h post‐infection respectively. These results suggest that TmDef and TmDef‐like genes play important roles in protecting T. molitor from microbial contact.
Jang Ho Am,Park Ki Beom,Kim Bo Bae,Ali Mohammadie Kojour Maryam,Bae Young Min,Baliarsingh Snigdha,Lee Yong Seok,Han Yeon Soo,Jo Yong Hun 한국곤충학회 2020 Entomological Research Vol.50 No.9
Antimicrobial peptides (AMPs) are effector candidates that elicit humoral immunity in invertebrates. AMPs facilitate bacterial clearance by either physically disrupting the microbial membranes or the intracellular targets. In the Coleopteran pest, Tenebrio molitor, transcriptional regulation of AMPs has been studied in the context of innate immune signaling cascades and antimicrobial immunity. Here, we report the transcriptional response of three AMP genes, Coleoptericin A, B, and C (TmCole A, B and C) in T. molitor in response to bacterial (Escherichia coli, Staphylococcus aureus), and fungal (Candida albicans) challenges. These genes were expressed essentially in the gut and hemocytes followed by the integument tissue of the T. molitor larva. Further, these genes were highly expressed in the late-larval, pupal, and early adult stages. Furthermore, while all of these transcripts were highly upregulated in the fat body and Malpighian tubules after bacterial challenge, TmCole B and TmCole C were induced in the gut after E. coli challenge. Fungal stimulation was not required for the upregulation of the transcription of Coleoptericin genes in T. molitor.