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The OnSPN2 from the nipa palm hispid beetle octodonta nipae is a multipurpose defense tool against proteases from different peptidase families

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INSECT SCIENCE 2025年第0期 2025 Jan 19页

作者： Zhang, Huajian Xu, Jiawei Chen, Mintao Yin, Jiawei Hou, Youming Tang, Baozhen Hainan Univ Sch Trop Agr & Forestry Sch Agr & Rural Sch Rural Revitalizat Haikou Peoples R China Hainan Univ Sanya Inst Breeding & Multiplicat Sch Breeding & Multiplicat Sanya Peoples R China Fujian Agr & Forestry Univ State Key Lab Agr & Forestry Biosecur Fuzhou Peoples R China Fujian Agr & Forestry Univ Dept Plant Protect Fujian Prov Key Lab Insect Ecol Fuzhou Peoples R China

Serpins (serine protease inhibitors) constitute a superfamily of proteins with functional diversity and unusual conformational flexibility. In insects, serpins act as multiple inhibitors, by forming inactive acyl-enzyme complexes, in regulating Sp & auml;tzles activation, phenoloxidases (POs) activity, and other cytokines. In this study, we present the cloning and characterization of octodonta nipae serpin2 (OnSPN2), a 415 residues protein homologous to Tenebrio molitor 42Dd-like. Notably, OnSPN2 features an arginine residue (R364) at the P1 position, and additional arginine residues (R362, R367) at the P3 and P3 ' positions, respectively which is crucial for protease inhibition. Immunohistochemistry (IHC) and Western blot analyses revealed that OnSPN2 is primarily synthesized in plasmatocytes and then released into the plasma to exert its function. RNA interference results indicated that OnSPN2 knockdown may depress serine protease in melanization and remarkably increase the transcript level of Attacin in hemolymph, but its messenger RNA levels were not changed upon immune induction. Reciprocal co-immunoprecipitation assay results confirmed that OnSPN2 binds to OnPPAF1 and OnSP8, indicating its role as a negative regulator in the PO and AMP pathway. Intriguingly, several cathepsin-L isoforms were identified in the OnSPN2 immunoprecipitated samples. The cathepsin-L inhibition assays and protein-protein docking results, identified cathepsin-L as a potential target of OnSPN2. These results indicate that OnSPN2 is produced as an intracellular resident and additionally is associated with the PO and AMP pathway. OnSPN2 represents a multiple defense tool that may provide multiple antiproteolytic functions.

关键词： octodonta nipae serpin serine protease cathepsin-L phenoloxidase cascade

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Role of prophenoloxidase 1 from the beetle octodonta nipae in melanized encapsulation of a wasp egg

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DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY 2024年 150卷 105082页

作者： Zhang, Xia-Fang Cui, Wen Wang, Mou-Jun Zhou, Yan Fu, Ting-Ting Jiang, Kun Hou, You -Ming Tang, Bao-Zhen Fujian Agr & Forestry Univ State Key Lab Ecol Pest Control Fujian & Taiwan Cr Fuzhou 350002 Peoples R China Fujian Agr & Forestry Univ Dept Plant Protect Fujian Prov Key Lab Insect Ecol Fuzhou 350002 Peoples R China

Exploring the function of the host immune system can help to understand the host-parasitoid interaction. Prophenoloxidase (PPO) is crucial in defensive melanization during the encapsulation of wasp eggs. However, the existence of multiple PPO sequences increases the difficulty of exploring the specific functions of individual PPOs. We previously identified three PPOs in the nipa palm hispid beetle, octodonta nipae. Our current work showed that OnPPO1 and OnPPO2 possessed the typical characteristics of the type III copper family, but OnPPO3 lacked the conserved histidine residues, and its active sites were substituted with Gln. OnPPOs showed the highest expression in hemocytes, but OnPPO3 presented extremely low abundance compared with that of OnPPO1 and OnPPO2, and only OnPPO1 showed a quick response after wasp infection. OnPPO1 knockdown decreased the encapsulation index and inhibited melanization, whereas silencing of OnPPO3 appeared to have no adverse effect on encapsulation and melanization, and silencing of OnPPO2 presented low RNAi efficiency. Moreover, the cleavage of recombinant OnPPO1 produced a 62 kDa fragment with high PO activity. OnPPO1 could be produced by oenocytoids, granulocytes and plasmatocytes, and was distributed around wasp eggs during capsule formation. Overall, our results indicate that proteolytic cleavage of OnPPO1 plays key roles in the melanized encapsulation of wasp eggs. This finding illuminates the mechanism of PPO activation in this invasive beetle and provides guidance for its biological control.

关键词： octodonta nipae Prophenoloxidase Parasitoid Encapsulation Melanization

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The Endosymbiotic Wolbachia and Host COI Gene Enables to Distinguish Between Two Invasive Palm Pests;Coconut Leaf Beetle, Brontispa longissima and Hispid Leaf Beetle, octodonta nipae

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JOURNAL OF ECONOMIC ENTOMOLOGY 2018年第6期111卷 2894-2902页

作者： Ali, Habib Muhammad, Abrar Bala, Nafiu Sanda Hou, Youming Fujian Agr & Forestry Univ State Key Lab Ecol Pest Control Fujian & Taiwan C Fuzhou 350002 Peoples R China Fujian Agr & Forestry Univ Fujian Prov Key Lab Insect Ecol Coll Plant Protect Fuzhou 350002 Fujian Peoples R China

To elucidate taxonomic eminence of identical pest species is essential for many ecological and conservation studies. Without proficient skills, accurate molecular identification and characterization are laborious and time-consuming. The coconut leaf beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), is biologically and morphologically identical to hispid leaf beetle, octodonta nipae (Maulik) (Coleoptera: Chrysomelidae), and is known as the most harming nuisances of palm cultivation worldwide. The present examination was to establish Wolbachia genotyping analysis along with host cytochrome oxidase subunit I (COI) gene for accurate identification between these individuals of the same family (Chrysomelidae). Here, we have cloned and sequenced a gene coding Wolbachia surface protein (wsp) and COI gene regions amplified from both species by polymerase chain reaction. The nucleotide sequences were directly determined (approximate to 600 bp for wsp and approximate to 804 bp for COI) and aligned using the multiple alignment algorithms in the ESPript3 package and the MEGA5 program. Comparative sequence analysis indicated that the representative of wsp and COI sequences from these two beetles were highly variable. To ensure this bacterial variation, multilocus sequence typing (MLST) of bacterial genes was conducted, and the results vindicated the same trend of variations. Furthermore, the phylogenetic analysis also indicates that B. longissima and O. nipae being the two different species harbors two distinct Wolbachia Hertig and Burt (Rickettsiales: Anaplamataceae) supergroups B and A, respectively. The present outcomes quickly discriminate between these two species. Considering its simplicity and cost-effectiveness, it can be used as a diagnostic tool for discriminating such invasive species particularly B. longissima and O. nipae which has overlapping morphologic characters.

关键词： Brontispa longissima octodonta nipae Wolbachia multilocus sequence type invasive species

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Entomopathogenic nematode Steinernema carpocapsae surpasses the cellular immune responses of the hispid beetle, octodonta nipae (Coleoptera: Chrysomelidae)

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MICROBIAL PATHOGENESIS 2018年第0期124卷 337-345页

作者： Sanda, Nafiu Bala Muhammad, Abrar Ali, Habib Hou, Youming Fujian Agr & Forestry Univ State Key Lab Ecol Pest Control Fujian & Taiwan C Fuzhou Fujian Peoples R China Fujian Agr & Forestry Univ Dept Plant Protect Fujian Prov Key Lab Insect Ecol Fuzhou Fujian Peoples R China

The Nipa palm hispid, octodonta nipae (Maulik) is an important invasive pest of palm trees particularly in Southern China. How this beetle interacts with invading pathogens via its immune system remains to be dissected. Steinernema carpocapsae is a pathogenic nematode that attacks a number of insects of economic importance. The present study systematically investigates the cellular immune responses of O. nipae against S. carpocapsae infection using combined immunological, biochemical and transcriptomics approaches. Our data reveal that S. carpocapsae efficiently resists being encapsulated and melanized within the hosts hemolymph and most of the nematodes were observed moving freely in the hemolymph even at 24 h post incubation. Consistently, isolated cuticles from the parasite also withstand encapsulation by the O. nipae hemocytes at all-time points. However, significant encapsulation and melanization of the isolated cuticles were recorded fol- lowing heat treatment of the cuticles. The host's phenoloxidase activity was found to be slightly suppressed due to S. carpocapsae infection. Furthermore, the expression levels of some antimicrobial peptide (AMP) genes were significantly up-regulated in the S. carpocapsae-challenged O. nipae. Taken together, our data suggest that S. carpocapsae modulates and surpasses the O. nipae immune responses and hence can serve as an excellent biological control agent of the pest.

关键词： Immune response Steinernema carpocapsae Antimicrobial peptides Phenoloxidase octodonta nipae

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Functional conservation and division of two single-carbohydrate-recognition domain C-type lectins from the nipa palm hispid beetle octodonta nipae (Maulik)

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DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY 2019年 100卷 103416-000页

作者： Zhang, Hua-Jian Lin, Ya-Ping Liu, Min Liang, Xin-Yu Ji, Ya-Nan Tang, Bao-Zhen Hou, You-Ming Fujian Agr & Forestry Univ State Key Lab Ecol Pest Control Fujian & Taiwan C Fuzhou 350002 Fujian Peoples R China Fujian Agr & Forestry Univ Dept Plant Protect Fujian Prov Key Lab Insect Ecol Fuzhou Fujian Peoples R China

As an invasive pest, the complete and effective innate immune system is crucial for the nipa palm hispid beetle octodonta nipae (Maulik) to adjust to new environments. C-type lectins (CTLs) are large families of carbohydrate-binding proteins that possess one or more characteristic carbohydrate-recognition domains (CRD) and function as pattern-recognition receptors, which play important roles in mediating humoral and cellular immunity. In the present study, for the first time, we report two CTL-Ss (single-CRD CTLs) from O. nipae (Maulik) (designated OnCTL1 and OnCTL2). The two CTL-Ss share high identity at conserved amino acids associated with conserved carbohydrate binding sites Gln-Pro-Asp (QPD) motifs and clearly show a 1:1 orthologous relationship in insects, which endow them with functional conservation and diversification. mRNA abundance analysis showed that OnCTL1 was upregulated upon Staphylococcus aureus and Escherichia coil challenge at 6 and 12 h, while OnCTL2 underwent no changes upon E. coli challenge and was even downregulated after S. aureus infection. Knockdown of OnCTL1 significantly decreased the transcripts of two key serine proteases (prophenoloxidase activating factors), OnPPAF1 and OnPPAF3, followed by the reduction of haemolymph phenoloxidase activity;it also increased the expression of Defensin 2B. In contrast, silencing of OnCTL2 significantly decreased the expression of Defensin 2B and Attacin 3C, the encapsulation index, and the phagocytosis rate compared to the dsEGFP group. The spreading results showed that more irregularly shaped plasmatocytes and lower levels of aggregation were found in OnCTL2-silenced pupae than in the dsOnCTL1 and dsEGFP groups. We can infer from the results of this study that the two OnCTLs play important roles in the immune system and generate a functional division: OnCTL1 seems to function more in humoral immunity including mediating bacterial recognition and activating the phenoloxidase cascade, and OnCTL2 plays

关键词： octodonta nipae C-type lectin Phagocytosis Encapsulation Prophenoloxidase

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Identification of three prophenoloxidase-activating factors (PPAFs) from an invasive beetle octodonta nipae Maulik (Coleoptera: Chrysomelidae) and their roles in the prophenoloxidase activation

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ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY 2017年第4期96卷 2017 Dec;96(4)页

作者： Zhang, HuaJian Tang, BaoZhen Lin, YaPing Chen, ZhiMing Zhang, XiaFang Ji, TianLiang Zhang, XiaoMei Hou, YouMing Fujian Agr & Forestry Univ State Key Lab Ecol Pest Control Fujian & Taiwan C Fuzhou Fujian Peoples R China Fujian Agr & Forestry Univ Dept Plant Protect Fujian Prov Key Lab Insect Ecol Fuzhou Fujian Peoples R China Fuzhou Entry Exit Inspect & Quarantine Bur PRC Fuzhou Peoples R China

A typical characteristic of the insect innate immune system is the activation of the serine protease cascade in the hemolymph. As being the terminal component of the extracellular serine protease cascade in the prophenoloxidase (proPO) activating system, proPO-activating factors (PPAFs) activated by the upstream cascade may generate active phenoloxidase, which then induces downstream melanization. In the present study, we reported three PPAFs from the nipa palm hispid beetle octodonta nipae (Maulik) (designated as OnPPAF1, OnPPAF2, OnPPAF3). All three OnPPAFs contained a single clip domain at the amino-terminus followed by a trypsin-like serine protease domain at the carboxyl-terminus, except the Ser in the active sites of OnPPAF2 and OnPPAF3 was substituted with Gly. Transcript expression analysis revealed that all OnPPAFs were highly expressed in hemolymph, whereas OnPPAF2 showed an extremely low mRNA abundance compared with that of OnPPAF1 and OnPPAF3, and that the abundance of all three OnPPAFs was dramatically increased upon bacterial challenge. Knockdown of OnPPAF1 or OnPPAF3 resulted in a reduction of hemolymph phenoloxidase activity and an inhibition of hemolymph melanization, whereas the knockdown of OnPPAF2 did not affect the proPO cascade. Our work thus implies that the three OnPPAFs may have different functions and regulation during immune responses in O. nipae.

关键词： octodonta nipae prophenoloxidase prophenoloxidase-activating factor serine protease serine protease homolog

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Molecular cloning and expression of the vitellogenin gene and its correlation with ovarian development in an invasive pest octodonta nipae on two host plants

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BULLETIN OF ENTOMOLOGICAL RESEARCH 2016年第5期106卷 642-650页

作者： Li, Jin-Lei Tang, Bao-Zhen Hou, You-Ming Xie, Yi-Xing Fujian Agr & Forestry Univ Fujian Prov Key Lab Insect Ecol Dept Plant Protect Fuzhou Fujian Peoples R China Minist Agr Key Lab Integrated Pest Management Crops Fujian T Fuzhou Peoples R China

There is an ongoing relationship between host plants and herbivores. The nutrient substances and secondary compounds found in the host plant can not only impact the growth and development process of herbivores, but, more importantly, may also affect their survival and reproductive fitness. Vitellogenesis is the core process of reproductive regulation and is generally considered as a reliable indicator for evaluating the degree of ovarian development in females. Vitellogenin (Vg) plays a critical role in the synthesis and secretion of yolk protein. In this study, the full-length cDNA of the Vg gene in an alien invasive species, the nipa palm hispid beetle octodonta nipae Maulik (Coleoptera: Chrysomelidae) (OnVg) was cloned and, the effect of host plant on the OnVg expression level and ovarian development was investigated. The results revealed that the OnVg was highly and exclusively expressed in adult females, but barely detectable in larvae, pupae and adult males. The relative expression level of OnVg and egg hatchability were much higher in females fed on Phoenix canariensis (their preferred host) than those fed on Phoenix roebelenii. A positive correlation relationship between OnVg expression and egg hatchability was also detected. Additionally, the anatomy of the female reproductive system showed that the ovaries of individuals fed on P. canariensis were considerably more developed than in females fed on P. roebelenii. The results may be applicable to many pest management situations through reproductive disturbance by alternating host plant species or varieties or by reproductive regulation through vitellogenesis mediated by specific endocrine hormones.

关键词： egg hatchability expression level host plant octodonta nipae ovarian development vitellogenin

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Study on the dynamic change of cuticular hydrocarbons in different gender and eclosion age of octodonta nipae

Study on the dynamic change of cuticular hydrocarbons in dif...

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作者： Hai-Jun Xiang Bin Huang You-Ming Hou State Key Laboratory of Ecological Pest Control for Fujian-Taiwan Crops Fujian Agriculture and Forestry University Fujian Provincial Key Laboratory of Insect Ecology Department of Plant Protection Fujian Agricultureand Forestry University

octodonta nipae（Maulik）(Coleoptera: Chrysomelidae) is one of the most damaging pest of palms, which poses a high-risk threat to palm plant industry, economic palm planting, city landscaping and ecological safety. In addition, O. nipae has a short generation period but long life time, with the bionomics of severe overlapping generation, high degree of aggregation and almost lifetime breeding, which advantage for multiple mating and polyandry. Cuticular hydrocarbons（CHCs）, one of the important external chemical characteristic, has been proved to be reliable signal for intraspecific discrimination of many insects mainly through differing in compounds compositions, relative contents and absolute contents among individuals. In order to investigate whether CHCs can also play an important role in intraspecific discrimination of ***, this study used chemical immersion method and gas chromatography-mass spectrometry（GC-MS） to analysis the dynamic change of CHCs in different gender and 1 d, 5 d, 10 d, 15 d（sexual maturity） eclosion age of O. nipae. The results as follows: 1. There was no type difference between male and female O. nipae cuticular hydrocarbons. 2. There are 28 kinds of cuticular hydrocarbons in both male and female O. nipae, as follows: n-C, n-C, nC, n-C, n-C, n-C, n-C, n-C, n-C, n-C, n-C, n-C, n-C,（3 Z）-C ene, 1-C ene, 1-C ene,（1 Z）-C ene,（10 E）-C ene,（1 Z）-C ene, 5-Me-（5 Z）-C ene, 8-Me-C ane, 3-Me-C ane, 2-Me-C ane, 10-Me-C ane, 2,6,10,15-Tetramethyl-C ane, cyclotetracosane, cyclooctacosane, and cyclotriacontane. 3. With emergence days increasing, average absolute contents of male O. nipae cuticular hydrocarbons decreased firstly and then increased when it becomes sexually mature individual. 4. Average absolute contents of each cuticular hydrocarbon in male O. nipae evidently more than female O. nipae in 15 emergence days. Conclusions: The difference between absolute content and relative content of male and female O. nipae cuticular hydrocarbons can act a

关键词： octodonta nipae cuticular hydrocarbons intraspecific discrimination gas chromatography-mass spectrometry reproductive investment

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Altered immune function of octodonta nipae (Maulik) to its pupal endoparasitoid, Tetrastichus brontispae Ferriere

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COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY 2016年 198卷 100-109页

作者： Meng, E. Tang, Baozhen Hou, Youming Chen, Xinxin Chen, Jiantu Yu, Xiao-Qiang Fujian Agr & Forestry Univ Dept Plant Protect Fujian Prov Key Lab Insect Ecol 15 Shangxiadian Rd Fuzhou 350002 Fujian Peoples R China Minist Agr Key Lab Integrated Pest Management Crops Fujian T Fuzhou 350002 Fujian Peoples R China Univ Missouri Sch Biol Sci Div Cell Biol & Biophys Kansas City MO 64110 USA

Most studies on the contribution of the altered immune response by endoparasitoid have been restricted to the interactions between Ichneumonoidea and their hosts, while effects of parasitism by Chalcidoidea on the hosts have rarely been characterized except some wasps such as Pteromalidae. Endoparasitoid Tetrastichus brontispae Ferriere, belonging to Eulophidae (Hymenoptera), has a great potential to control some Coleopteran beetles such as octodonta nipae, one invasive species in southern China. However, the physiological mechanism underlying the escape from the melanotic encapsulation in O. nipae pupae has not been demonstrated. In the present study, effects of parasitism on the immune function of its pupal host O. nipae were investigated. The combining results that granulocytes and plasmatocytes could phagocytize bacteria from 2 to 48 h and granulocytes, plasmatocytes and oenocytoids were prophenoloxidase/phenoloxidase positive hemocytes indicated that granulocytes, plasmatocytes and oenocytoids were the main immunocompetent hemocytes in O. nipae pupae. Parasitism by T. brontispae resulted in a significant increase in the percentage of hemocytes viability and spreading at 96 h, growing percentage of granulocytes at 24 h but no effects on the total hemocyte counts, and an enhanced phenoloxidase activity only at 12 and 72 h while a significantly longer melanization time of the hemolymph at 96 h following parasitism. These results indicate that mixtures of systemic active and local active regulation are used for T brontispae to escape host encapsulation in O. nipae pupae. The present study contributes to the understanding of the diversity of virulence strategies used by parasitoids. (C) 2016 Elsevier Inc All rights reserved.

关键词： Hemocyte types Hemocyte loads Melanization octodonta nipae Parasitism Phenoloxidase Tetrastichus brontispae

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Biocontrol potential of entomopathogenic nematodes Steinernema carpocapsae and Heterorhabditis bacteriophora on an invasive pest nipa palm hispid octodonta nipae(Maulik)

Biocontrol potential of entomopathogenic nematodes Steinerne...

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作者： Sanda Nafiu Bala You-Ming Hou State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops Fujian Agriculture and Forestry University Fujian Province Key Laboratory of Insect Ecology Department of Plant Protection Fujian Agriculture and Forestry University

The destructive pest of palm trees such as coconut palm(Cocos spp.), Washingtonia filifera, Phoenix canariensis and Phoenix hanceana called Nipa palm hispid, octodonta nipae(Maulik);is an invasive species to China, which is believed to originate from Malaysia and distributed to other parts of Southeast Asia. This hispid beetle was first reported in China Hainan province in 2001 and Fujian province in 2007. The damages are caused by adults and larvae which live together within the tight leaves of the palm, unreachable to insecticidal spray. Laboratory experiments was conducted to test the efficacy of entomopathogenic nematodes Steinernema carpocapsae and Heterorhabditis bacteriophora against third instar larvae O. nipae due their abilities to locate and infects host in cryptic environment like leaf frond of palm trees. Lower concentrations of 25, 50 and 100 Ijs/larvae were used for the plate bioassay, and higher concentrations of 250, 500 and 100 Ijs/larvae were applied in the leaf bioassay experiment. The results show that, as early as 24 hours after treatment, S. carpocapsae caused 77.95% ± 5.761 highest cumulative mortalities, comparatively only 34.11% ± 4.55 were caused by H. bacteriophora at 48 HAT. Similarly, S. carpocapsae was more pathogenic to H. bacteriophora and was able to locate and caused 57.84% ± 2.56 larval mortalities in leaf bioassay at 48 HAT. Both nematodes penetrated O. nipae larvae at all concentrations. The mean penetration potential is higher for S. carpocapsae(48.80 ± 4.00 Ijs/larva) at 100 Ijs than H. bacteriophora(22.33 Ijs/larva). Unlike mortality, the results show that H. bacteriophora produced more Ijs than S. carpocapsae. Comparatively, the mean Ijs for H. bacteriophora in O. nipae larvae was 23,136.67 ± 1, 821.13 Ijs/larva and 252,852.45 ± 4, 6164 Ijs/larva in Galleria mellonella larvae. Our results revealed that S. carpocapsae can best be used for biocontrol of O. nipae, due to its ability to locate and caused higher mortality in the

关键词： Biocontrol Invasive species Infectivity Steinernema carpocapsae Heterorhabditis bacteriophora octodonta nipae

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