Lethal and sub-lethal effects of transgenic rice containing cry1Ac and CpTI genes on the pink stem borer, Sesamia inferens, were studied to collect information for ecological risk assessment on insect-resistance of tr...Lethal and sub-lethal effects of transgenic rice containing cry1Ac and CpTI genes on the pink stem borer, Sesamia inferens, were studied to collect information for ecological risk assessment on insect-resistance of transgenic rice. In vitro insect-feeding bioassays were conducted to evaluate the lethal and sub-lethal effects of transgenic rice lines （II YouKF6 and KF6） containing cry1Ac＋CpTI genes on S. inferens at four different growth stages, viz., seedling, tillering and elongation, booting, and milk and maturing. Transgenic rice at seedling stage showed significantly high lethal effect on S. inferens with the shortest lethal duration for 50 and 100% individuals and the highest corrected mortalities after feeding on transgenic lines at this stage for 3 and 6 d. Followed by tillering and elongation stage, 50 and 100% S. inferens were dead after feeding on transgenic lines at this stage for 4 and 10 d, respectively. Moreover, corrected mortalities for 6 d feeding on transgenic lines at this stage were significantly higher than that at booting, and milk and maturing stages. Lethal effect of KF6 on S. inferens decreased significantly at booting stage. Lethal duration for 50% S. inferens significantly extended and its corrected mortalities for 6 d feeding also declined remarkably. However, lethal effect of II YouKF6 on S. inferens did not decrease significantly at this stage. Transgenic rice at booting, and milk and maturing stages did not show significant lethal effect to S. inferens and it showed the longest lethal duration for 50% individuals and the lowest corrected mortalities for 3 and 6 d feeding. A few larvae of S. inferens could survive, pupate and emerge on these two transgenic lines at booting, and milk and maturing stages. Sub-lethal effect of two transgenic lines on S. inferens also differed significantly between different developmental stages. Continuously feeding on transgenic rice lines at seedling, and tillering and elongation stages delayed the development of larvae and pupae and decreased pupation rate, but no effect was observed on eclosion rate. Larval development was significantly inhibited after feeding on transgenic rice at booting stage, but no obvious effect was observed in pupal stage, pupation and eclosion rate. There were no significant differences for larval and pupal development, pupation, and eclosion rates between feeding on transgenic and control rice lines at milk and maturing stage. Larval and pupal weights significantly declined, but no influence was observed on fecundity when S. inferens infested on transgenic rice at any stage. These showed that there were significant differences in lethal and sub-lethal effects of transgenic rice on S. inferens among developmental stages, and the effects gradually decreased with the increase of growth stages of rice plant.展开更多
[ Objective] The aim of this study was to provide the reference for the artificial breeding of aquatic animals. [ Method] Amoeba discoides and Trichodina were treated by neutral red solution with different concentrati...[ Objective] The aim of this study was to provide the reference for the artificial breeding of aquatic animals. [ Method] Amoeba discoides and Trichodina were treated by neutral red solution with different concentrations to observe the physiological activities of organelles in their death processes. Effects of neutral red solution on the growth of common aquatic animals such as Paramecium caudatum, Euglena viridis and Brachionus plicatilis were analyzed, and the specific lethal mechanism of trace neutral red to A. discoides and Trichodina was also preliminarily studied. [ Re- suit] The neutral red solution at a concentration of 0.5 mg/L damaged the physiological function of contractile vacuole in Trichodina and also had the specific lethal effect on Tdchodina, but it had no effect on the growth and reproduction of non-parasitic protozoa and B. p/icatilis. Neutral red so- lution with certain concentration led to disorder of the physiological functions of A. discoides, such as assimilation and rejection, which was a main factor that caused the death of A. discoides. [ Conclusion] With the advantages such as targeting, safety and easiness to be oxidized and decom- posed, neutral red is an ideal drug for treating the diseases caused by A. discoides and Trichodina, and its suitable concentration is 0. 5 mg/L.展开更多
BACKGROUND： Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human gr...BACKGROUND： Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human granulocyte-macrophage colony stimulating factor （rhG-MCSF） and recombinant human interleukin-4 （rhlL-4） can generate a great many dendritic cells and promote the lethal effect of T cells on human neuroblastoma, but it is unclear that whether the lethal effect is associated with the most proper concentration of dendritic cells. OBJEETIVE： To investigate the lethal effect of human umbilical cord blood mononuclear cells induced in vitro by cytokines differentiating into dendritic cells on human neuroblastoma, and its best concentration range. DESIGN ： Open experiment SEI-FING： Department of Pediatrics, the Medical School Hospital of Qingdao University MATERIALS ： The study was carried out in the Shandong Provincial Key Laboratory （Laboratory for the Department of Pediatrics of the Medical School Hospital of Qingdao University） during September 2005 to May 2006. Human umbilical cord blood samples were taken from the healthy newborn infants of full-term normal delivery during October to November 2005 in the Medical School Hospital of Qingdao University, and were voluntarily donated by the puerperas. Main instruments： type 3111 CO2 incubator （Forma Scientific, USA）, type 550 ELISA Reader （Bio-Rad, USA）. Main reagents： neuroblastoma cell line SK-N-SH （Shanghai Institute of Life Science, Chinese Academy of Sciences）, RPMI-1640 culture fluid and fetal bovine serum （Hyclone）, rhlL-4 （Promega, USA）, rhG-MCSF （Harbin Pharmaceutic Group Bioengineering Co.Ltd）, rat anti-human CDla monoclonal antibody and FITC-labeled rabbit anti-rat IgG （Xiehe Stem cell Gene Engineering Co.Ltd）. METHODS： ① Human umbilical cord blood mononuclear cells obtained with attachment methods differentiated into human umbilical cord blood dendritic cells, presenting typical morphology of dendritic cells after in vitro induction by rhG-MCSF and rhlL-4. ② Different concentrations of dendritic cells[ dendritic cells： neuroblastoma cells=20：1,50：1,100：1 （2×10^8 L^-1,5×10^8 L^-1,1×10^9 L^-1）], 1×10^9 L^-1 T cells and 1×10^7 L^-1 neuroblastoma cells were added in the experimental group. 1 ×10^9 L^-1 T cells and 1 ×10^7 L^-1 neuroblastoma cells were added in the control group. ③ Main surface marker CDla molecules of dendritic cells were detected with indirect immunofluorescence, and the percent rate of dendritic cells was counted with ultraviolet light and expressed as the expression rate of CD1a^＋ cells. ④Single effector cells and target cells were respectively set in the experimental group and control group to obtain the lethal effect. The lethal effect of dendritic cells on neuroblastoma cells was indirectly evaluated by detecting cellular survival with MTT assay. The lethal effect（%）= （1-A experimentat well-A effector cell /A target cell well）×100%.⑤The expenmental data were presented as Mean ±SD, and paired t test was used. MAIN OUTCOME MEASURES： ① Morphological characters of dendritic cells in the process of induction and differentiation. ②CD1a^＋ cellular expression rate. ③Lethal effect of dendntic cells on neuroblastoma cells. RESULTS： ①Morphological characters of dendritic cells in the process of induction and differentiation： On the 15^th day after human umbilical cord blood mononuclear cells were induced by rhG-MCSF and rhlL-4, typical morphology of dendritic cells could be seen under an inverted microscope. ②Expression rate of CD1a^＋ cells was （43.12±5.83）%. ③Lethal effect of dendritic cells on neuroblastoma cells： Lethal effect of dendritic cells stimulated T cells in each experimental group （ dendritic cells： neuroblastoma cells=100：1,50：1, 20：1 respectively） on neuroblastoma cells was significantly higher than that in control group[（31.00 ±4.41 ）%, （30.92±5.27）%,（33.57±5.35）%,（26.23±5.20）%, t=3.51,2.98,4.24, P〈 0.01 ）; But the lethal effect of dendntic cells on neuroblastoma was significantly lower when their ratio was 100：1 and 50：1 in comparison with 20：1 （t=2.01,2.36, P 〈 0.05）, and no significant difference in lethal effect existed between the ratio at 100：1 and 50：1 （t=0.06,P 〉 0.05）. CONCLUSION： Dendritic cells differentiated from human umbilical cord blood mononuclear cells after in vitro induction of cytokines can promote the lethal effect of T cells on neuroblastoma cells. The lethal effect is associated with the concentration of dendritic cells within some range.展开更多
Desiccant dusts have been shown to be effective materials in the management of bed bugs(Cimex spp.).Past research primarily focused on exploring the direct lethal effect of dusts against bed bugs,whereas information o...Desiccant dusts have been shown to be effective materials in the management of bed bugs(Cimex spp.).Past research primarily focused on exploring the direct lethal effect of dusts against bed bugs,whereas information on their sublethal effects on bed bugs are limited.In this study,we evaluated the lethal effect of 3 dust products(Johnson's®Baby powder,Vazor DE powder^(TM),and CimeXaTM;abbreviated as Baby powder,DE,and TM CimeXa)against the tropical bed bug,Cimex hemipterus(F.),under laboratory conditions.Results showed that the 3 dust products caused 97%100%mortality to tropical bed bugs within 14 d,both in no-choice and choice experiments.However,in a brief exposure experiment(crossing a 2.5-cm-wide band),Baby powder exposure caused a significantly lower mortality(23%)than DE(88%)and CimeXa(100%).The more effective dusts(DE and CimeXa)were selected for further evaluation of their sublethal effects on C.hemipterus.DE and CimeXa caused significantly higher mortality(48%58%)through horizontal transfer than that of control(6%).Brief exposure to DE and CimeXa dusts did not affect tropical bed bug activity and their response to CO_(2).However,CimeXa-exposed tropical bed bugs exhibited reduced feeding and lowered survival rate after feeding.Moreover,both DE and CimeXa dust bands showed strong barrier effects on the crawling of tropical bed bugs.Our results indicate that both DE and CimeXa have great potential for preventing C.hemipterus from reaching protected areas and for controlling C.hemipterus.展开更多
This study evaluated the effects of purified paper wasp Ropalidia marginata venoms on various biomolecules in the blood serum of albino mice. Changes in the concentration of some important macromolecules, i.e., protei...This study evaluated the effects of purified paper wasp Ropalidia marginata venoms on various biomolecules in the blood serum of albino mice. Changes in the concentration of some important macromolecules, i.e., proteins, free amino acids, uric acid, cholesterol, pyruvic acid, total lipids and glucose were noted down. These alterations were measured after intraperitoneal injection of 40% and 80% 24-hour LD50 purified Ropalidia marginata venom toxin. Serum total protein levels were found to decrease to 78% after 6 hrs, while serum free amino acid levels were significantly increased to 117% 6 hrs after venom injection compared to control. It was also found that serum uric acid levels increased to 138% after 8 hrs of venom injection compared to control. The increase in serum cholesterol i.e. (101% and 106%) and pyruvic acid increased significantly to a maximum value of 106% after 6 hrs of treatment at 40% LD<sub>50</sub>. Glycogen levels in the gastrocnemius muscle were found to decrease significantly (p-0.05) to 43% and 92% at LD<sub>50</sub> after injection of purified Ropalidia marginata venom after 8 h and 80% at LD<sub>50</sub> compared to control. Moreover, up to 71% and 81% were obtained at 10 hrs of treatment with the same dose. In the present study, the purified toxins significantly changed the levels of biomolecules in blood serum, indicating their wider effects on cellular physiology due to toxic effects and stress on the animal. These toxins can be good antigens and stimulate immune responses in experimental mice.展开更多
Termites are not only social insects but also significant global insect pests.Investigating the molecular mechanisms regulating immune defense response in termites is beneficial for developing novel approaches to term...Termites are not only social insects but also significant global insect pests.Investigating the molecular mechanisms regulating immune defense response in termites is beneficial for developing novel approaches to termite management.Currently,research mainly focuses on coding RNAs in termite immunity,with limited exploration of non-coding RNAs.Here,we identified miR-701,a markedly downregulated microRNA(miRNA)in the globally significant termite pest Coptotermes formosanus after Metarhizium anisopliae infection,which targets the immune gene Toll4.Transcriptome analysis of termites injected with miR-701 agomir revealed that miR-701 affects the immune-related response,growth,and development of termites.Treatment with miR-701 agomir,either through injection or ingestion,resulted in a notably reduced survival rate of termites infected with M.anisopliae compared to the control group infected with M.anisopliae alone.Additionally,termites injected with miR-701 agomir exhibited a significant decrease in the expression of antimicrobial peptide genes termicin and lysozyme,alongside a notable increase in the colony-forming units of M.anisopliae in the infected termites.Subsequent investigations demonstrated that miR-701 suppressed the expression of the target gene Toll4,consequently inhibiting the Toll signaling pathway and diminishing the expression of antimicrobial peptides.These findings suggest that termites can combat M.anisopliae by downregulating miR-701 expression to activate the Toll signaling pathway and enhance antimicrobial peptides synthesis.This discovery improves our comprehension of the role of miRNAs in termites'immune responses and the mechanism of termites managing miRNAs to boost their pathogen resistance.Additionally,it reveals a new molecular target for termite biological control.展开更多
基金funded by the National 973 Program of China (2007CB109204)the Major Projects of National Genetically Modified Organisms Breeding of China (2009ZX08011-007B, 2008ZX08011-001A)
文摘Lethal and sub-lethal effects of transgenic rice containing cry1Ac and CpTI genes on the pink stem borer, Sesamia inferens, were studied to collect information for ecological risk assessment on insect-resistance of transgenic rice. In vitro insect-feeding bioassays were conducted to evaluate the lethal and sub-lethal effects of transgenic rice lines （II YouKF6 and KF6） containing cry1Ac＋CpTI genes on S. inferens at four different growth stages, viz., seedling, tillering and elongation, booting, and milk and maturing. Transgenic rice at seedling stage showed significantly high lethal effect on S. inferens with the shortest lethal duration for 50 and 100% individuals and the highest corrected mortalities after feeding on transgenic lines at this stage for 3 and 6 d. Followed by tillering and elongation stage, 50 and 100% S. inferens were dead after feeding on transgenic lines at this stage for 4 and 10 d, respectively. Moreover, corrected mortalities for 6 d feeding on transgenic lines at this stage were significantly higher than that at booting, and milk and maturing stages. Lethal effect of KF6 on S. inferens decreased significantly at booting stage. Lethal duration for 50% S. inferens significantly extended and its corrected mortalities for 6 d feeding also declined remarkably. However, lethal effect of II YouKF6 on S. inferens did not decrease significantly at this stage. Transgenic rice at booting, and milk and maturing stages did not show significant lethal effect to S. inferens and it showed the longest lethal duration for 50% individuals and the lowest corrected mortalities for 3 and 6 d feeding. A few larvae of S. inferens could survive, pupate and emerge on these two transgenic lines at booting, and milk and maturing stages. Sub-lethal effect of two transgenic lines on S. inferens also differed significantly between different developmental stages. Continuously feeding on transgenic rice lines at seedling, and tillering and elongation stages delayed the development of larvae and pupae and decreased pupation rate, but no effect was observed on eclosion rate. Larval development was significantly inhibited after feeding on transgenic rice at booting stage, but no obvious effect was observed in pupal stage, pupation and eclosion rate. There were no significant differences for larval and pupal development, pupation, and eclosion rates between feeding on transgenic and control rice lines at milk and maturing stage. Larval and pupal weights significantly declined, but no influence was observed on fecundity when S. inferens infested on transgenic rice at any stage. These showed that there were significant differences in lethal and sub-lethal effects of transgenic rice on S. inferens among developmental stages, and the effects gradually decreased with the increase of growth stages of rice plant.
基金supported by 863 Project (2005AA601010-05)Student Innovative Experiment Fund of Shenzhen University
文摘[ Objective] The aim of this study was to provide the reference for the artificial breeding of aquatic animals. [ Method] Amoeba discoides and Trichodina were treated by neutral red solution with different concentrations to observe the physiological activities of organelles in their death processes. Effects of neutral red solution on the growth of common aquatic animals such as Paramecium caudatum, Euglena viridis and Brachionus plicatilis were analyzed, and the specific lethal mechanism of trace neutral red to A. discoides and Trichodina was also preliminarily studied. [ Re- suit] The neutral red solution at a concentration of 0.5 mg/L damaged the physiological function of contractile vacuole in Trichodina and also had the specific lethal effect on Tdchodina, but it had no effect on the growth and reproduction of non-parasitic protozoa and B. p/icatilis. Neutral red so- lution with certain concentration led to disorder of the physiological functions of A. discoides, such as assimilation and rejection, which was a main factor that caused the death of A. discoides. [ Conclusion] With the advantages such as targeting, safety and easiness to be oxidized and decom- posed, neutral red is an ideal drug for treating the diseases caused by A. discoides and Trichodina, and its suitable concentration is 0. 5 mg/L.
文摘BACKGROUND： Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human granulocyte-macrophage colony stimulating factor （rhG-MCSF） and recombinant human interleukin-4 （rhlL-4） can generate a great many dendritic cells and promote the lethal effect of T cells on human neuroblastoma, but it is unclear that whether the lethal effect is associated with the most proper concentration of dendritic cells. OBJEETIVE： To investigate the lethal effect of human umbilical cord blood mononuclear cells induced in vitro by cytokines differentiating into dendritic cells on human neuroblastoma, and its best concentration range. DESIGN ： Open experiment SEI-FING： Department of Pediatrics, the Medical School Hospital of Qingdao University MATERIALS ： The study was carried out in the Shandong Provincial Key Laboratory （Laboratory for the Department of Pediatrics of the Medical School Hospital of Qingdao University） during September 2005 to May 2006. Human umbilical cord blood samples were taken from the healthy newborn infants of full-term normal delivery during October to November 2005 in the Medical School Hospital of Qingdao University, and were voluntarily donated by the puerperas. Main instruments： type 3111 CO2 incubator （Forma Scientific, USA）, type 550 ELISA Reader （Bio-Rad, USA）. Main reagents： neuroblastoma cell line SK-N-SH （Shanghai Institute of Life Science, Chinese Academy of Sciences）, RPMI-1640 culture fluid and fetal bovine serum （Hyclone）, rhlL-4 （Promega, USA）, rhG-MCSF （Harbin Pharmaceutic Group Bioengineering Co.Ltd）, rat anti-human CDla monoclonal antibody and FITC-labeled rabbit anti-rat IgG （Xiehe Stem cell Gene Engineering Co.Ltd）. METHODS： ① Human umbilical cord blood mononuclear cells obtained with attachment methods differentiated into human umbilical cord blood dendritic cells, presenting typical morphology of dendritic cells after in vitro induction by rhG-MCSF and rhlL-4. ② Different concentrations of dendritic cells[ dendritic cells： neuroblastoma cells=20：1,50：1,100：1 （2×10^8 L^-1,5×10^8 L^-1,1×10^9 L^-1）], 1×10^9 L^-1 T cells and 1×10^7 L^-1 neuroblastoma cells were added in the experimental group. 1 ×10^9 L^-1 T cells and 1 ×10^7 L^-1 neuroblastoma cells were added in the control group. ③ Main surface marker CDla molecules of dendritic cells were detected with indirect immunofluorescence, and the percent rate of dendritic cells was counted with ultraviolet light and expressed as the expression rate of CD1a^＋ cells. ④Single effector cells and target cells were respectively set in the experimental group and control group to obtain the lethal effect. The lethal effect of dendritic cells on neuroblastoma cells was indirectly evaluated by detecting cellular survival with MTT assay. The lethal effect（%）= （1-A experimentat well-A effector cell /A target cell well）×100%.⑤The expenmental data were presented as Mean ±SD, and paired t test was used. MAIN OUTCOME MEASURES： ① Morphological characters of dendritic cells in the process of induction and differentiation. ②CD1a^＋ cellular expression rate. ③Lethal effect of dendntic cells on neuroblastoma cells. RESULTS： ①Morphological characters of dendritic cells in the process of induction and differentiation： On the 15^th day after human umbilical cord blood mononuclear cells were induced by rhG-MCSF and rhlL-4, typical morphology of dendritic cells could be seen under an inverted microscope. ②Expression rate of CD1a^＋ cells was （43.12±5.83）%. ③Lethal effect of dendritic cells on neuroblastoma cells： Lethal effect of dendritic cells stimulated T cells in each experimental group （ dendritic cells： neuroblastoma cells=100：1,50：1, 20：1 respectively） on neuroblastoma cells was significantly higher than that in control group[（31.00 ±4.41 ）%, （30.92±5.27）%,（33.57±5.35）%,（26.23±5.20）%, t=3.51,2.98,4.24, P〈 0.01 ）; But the lethal effect of dendntic cells on neuroblastoma was significantly lower when their ratio was 100：1 and 50：1 in comparison with 20：1 （t=2.01,2.36, P 〈 0.05）, and no significant difference in lethal effect existed between the ratio at 100：1 and 50：1 （t=0.06,P 〉 0.05）. CONCLUSION： Dendritic cells differentiated from human umbilical cord blood mononuclear cells after in vitro induction of cytokines can promote the lethal effect of T cells on neuroblastoma cells. The lethal effect is associated with the concentration of dendritic cells within some range.
基金supported by the Ningbo Key Research and Development Plan and"Jie Bang Gua Shuai"Program(2022Z124)the Foreign Expert Project of the Ministry of Science and Technology of China(G2023030035).
文摘Desiccant dusts have been shown to be effective materials in the management of bed bugs(Cimex spp.).Past research primarily focused on exploring the direct lethal effect of dusts against bed bugs,whereas information on their sublethal effects on bed bugs are limited.In this study,we evaluated the lethal effect of 3 dust products(Johnson's®Baby powder,Vazor DE powder^(TM),and CimeXaTM;abbreviated as Baby powder,DE,and TM CimeXa)against the tropical bed bug,Cimex hemipterus(F.),under laboratory conditions.Results showed that the 3 dust products caused 97%100%mortality to tropical bed bugs within 14 d,both in no-choice and choice experiments.However,in a brief exposure experiment(crossing a 2.5-cm-wide band),Baby powder exposure caused a significantly lower mortality(23%)than DE(88%)and CimeXa(100%).The more effective dusts(DE and CimeXa)were selected for further evaluation of their sublethal effects on C.hemipterus.DE and CimeXa caused significantly higher mortality(48%58%)through horizontal transfer than that of control(6%).Brief exposure to DE and CimeXa dusts did not affect tropical bed bug activity and their response to CO_(2).However,CimeXa-exposed tropical bed bugs exhibited reduced feeding and lowered survival rate after feeding.Moreover,both DE and CimeXa dust bands showed strong barrier effects on the crawling of tropical bed bugs.Our results indicate that both DE and CimeXa have great potential for preventing C.hemipterus from reaching protected areas and for controlling C.hemipterus.
文摘This study evaluated the effects of purified paper wasp Ropalidia marginata venoms on various biomolecules in the blood serum of albino mice. Changes in the concentration of some important macromolecules, i.e., proteins, free amino acids, uric acid, cholesterol, pyruvic acid, total lipids and glucose were noted down. These alterations were measured after intraperitoneal injection of 40% and 80% 24-hour LD50 purified Ropalidia marginata venom toxin. Serum total protein levels were found to decrease to 78% after 6 hrs, while serum free amino acid levels were significantly increased to 117% 6 hrs after venom injection compared to control. It was also found that serum uric acid levels increased to 138% after 8 hrs of venom injection compared to control. The increase in serum cholesterol i.e. (101% and 106%) and pyruvic acid increased significantly to a maximum value of 106% after 6 hrs of treatment at 40% LD<sub>50</sub>. Glycogen levels in the gastrocnemius muscle were found to decrease significantly (p-0.05) to 43% and 92% at LD<sub>50</sub> after injection of purified Ropalidia marginata venom after 8 h and 80% at LD<sub>50</sub> compared to control. Moreover, up to 71% and 81% were obtained at 10 hrs of treatment with the same dose. In the present study, the purified toxins significantly changed the levels of biomolecules in blood serum, indicating their wider effects on cellular physiology due to toxic effects and stress on the animal. These toxins can be good antigens and stimulate immune responses in experimental mice.
基金supported by GDAS Special Project of Science and Technology Development(2022GDASZH-2022010106)Science and Technology Projects in Guangzhou(2023A04J0147).
文摘Termites are not only social insects but also significant global insect pests.Investigating the molecular mechanisms regulating immune defense response in termites is beneficial for developing novel approaches to termite management.Currently,research mainly focuses on coding RNAs in termite immunity,with limited exploration of non-coding RNAs.Here,we identified miR-701,a markedly downregulated microRNA(miRNA)in the globally significant termite pest Coptotermes formosanus after Metarhizium anisopliae infection,which targets the immune gene Toll4.Transcriptome analysis of termites injected with miR-701 agomir revealed that miR-701 affects the immune-related response,growth,and development of termites.Treatment with miR-701 agomir,either through injection or ingestion,resulted in a notably reduced survival rate of termites infected with M.anisopliae compared to the control group infected with M.anisopliae alone.Additionally,termites injected with miR-701 agomir exhibited a significant decrease in the expression of antimicrobial peptide genes termicin and lysozyme,alongside a notable increase in the colony-forming units of M.anisopliae in the infected termites.Subsequent investigations demonstrated that miR-701 suppressed the expression of the target gene Toll4,consequently inhibiting the Toll signaling pathway and diminishing the expression of antimicrobial peptides.These findings suggest that termites can combat M.anisopliae by downregulating miR-701 expression to activate the Toll signaling pathway and enhance antimicrobial peptides synthesis.This discovery improves our comprehension of the role of miRNAs in termites'immune responses and the mechanism of termites managing miRNAs to boost their pathogen resistance.Additionally,it reveals a new molecular target for termite biological control.
