بررسی اثر دما و سه زیر‌گونه از باکتری Bacillus thuringiensis بر تشکیل گره در سیستم ایمنی و برخی پارامتر‌های زیستی شب‌پره مدیترانه‌ای آرد Ephestia kuehniella Zeller (Lepidoptera: Pyralidae)

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه تنوع زیستی، پژوهشکده علوم محیطی، پژوهشگاه علوم و تکنولوژی پیشرفته و علوم محیطی، دانشگاه تحصیلات تکمیلی صنعتی و فناوری پیشرفته، کرمان، ایران

2 گروه بیوتکنولوژی، پژوهشکده علوم محیطی، پژوهشگاه علوم و تکنولوژی پیشرفته و علوم محیطی، دانشگاه تحصیلات تکمیلی صنعتی و فناوری پیشرفته، کرمان، ایران

چکیده

باکتری بیمارگر حشرات،Bacillus thuringiensis ، گرم مثبت و اسپورزا است که تعداد زیادی از زیر‌گونه‌های آن شناسایی و معرفی‌شده‌است. پاسخ‌های ایمنی حشرات، پس از بلعیدن Bt یا پروتئین‌های سمی آن فعال‌می‌شود. هدف از این تحقیق، بررسی اثر دما و سه زیر‌گونة B. thuringiensis subspecies thuringiensis (Btt) ، B. thuringiensis subspecies aizawai (Bta) و B. thuringiensis subspecies galleriae (Btg) بر تشکیل گره طی پاسخ ایمنی و برخی پارامتر‌های زیستی شب‌پرة Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) بود. برای آلوده‌سازی بستر غذایی لارو، از غلظت 109 اسپور در میلی­لیتر استفاده شد. این آزمایش در سه دمای مختلف (20، 25 و 30 درجة سلسیوس)، رطوبت نسبی 5±55 درصد و دورة نوری 16:8 (روشنایی: تاریکی) انجام شد. به‌طور معنی‌دار بیشترین تعداد گره‌ تحت تأثیر زیر‌گونه‌های Bta و Btg در دمای 30 درجة‌سلسیوس و کمترین میزان گره در شاهد در هر سه دما مشاهده‌شد. مدت زمان تا شفیره‌شدن، در هر دما بین سه زیرگونه‌ اختلاف معنی­دار نداشت. اما، بین دماهای مختلف در هر زیرگونه‌ اختلاف معنی­دار مشاهده‌شد. همچنین، این مدت زمان در شاهد به‌طور معنی‌دار تحت تأثیر دما قرار‌گرفت. بیشترین میزان تولید شفیره مربوط به دماهای 20 و 25 درجة‌سلسیوس در زیرگونة Bta و شاهد بود که با هم اختلاف معنی‌دار نداشتند. تعداد حشرات کامل بدست‌آمده در تیمار Bta مشابه شاهد تحت تأثیر دما قرار ‌نگرفت. از آنجا که دما بر تعداد گره‌ها در شاهد تأثیری نداشت، چنین استنباط می‌شود که افزایش دما با تأثیر بر باکتری Bt، تعداد گره‌های تشکیل‌شده را در لارو‌های زنده‌مانده افزایش ‌داده‌ و همچنین، نوع زیر‌گونة باکتری به‌طور معنی‌دار بر آن تأثیر داشته ‌است.

کلیدواژه‌ها

موضوعات

Extended Abstract

Bacillus thuringiensis is a Gram-positive and spore-forming bacterium that has special toxicity for different orders of insects and can point out three subspecies B. thuringiensis subspecies thuringiensis (Btt), B. thuringiensis subspecies aizawai (Bta) and B. thuringiensis subspecies galleriae (Btg). Insects can activate their immune responses after consuming Bt or toxin proteins for defence. The purpose of this research is to investigate the effect of temperature and three subspecies of insect pathogenic bacteria including Btt, Bta and Btg were found to affect the production of nodules concerning the immune system of E. kuehniella larvae and some biological parameters of this moth such as the number of produced pupae, the period till pupation and the number of adults. This experiment was performed at three different temperatures (20, 25 and 30ºC), the relative humidity 55±5% and a photoperiod of 16:8 (L: D). The results showed that the interaction of bacterium and temperature on the number of nodules formed in larval hemolymph, the duration of 24-day-old larvae turning into pupae, and the number of pupae and adults produced were significant. In the case of all three subspecies of bacterium, the highest number of nodules was significantly formed at 30 ºC. However, the difference was not significant between the other two temperatures (20 and 25ºC). The lowest amount of nodules was significantly observed in the control and showed that temperature does not affect the number of nodules formed in larval hemolymph. There was no significant difference between the three subspecies in the time until the pupation of the larvae at any temperature. However, a significant difference was observed among different temperatures in each subspecies. Likewise, this duration in the control was significantly affected by temperature; therefore, this process took more time at 20 than 30ºC. The number of pupae produced in each of Btt and Btg treatments was the same at all three temperatures and no significant difference was observed between different temperatures. However, in Bta treatment at 30ºC, pupal production decreased significantly that was similar to the control. The highest amount of pupa production was related to the temperatures of 20 and 25 ºC in Bta and the control treatment did not have significant differences. At the temperature of 30ºC, there was no significant difference in the number of pupae produced between the three studied subspecies. The number of adults obtained in the Bta treatment was not affected by the temperature, similar to the control. While, in Btt and Btg subspecies, the lowest number was significantly obtained at 30 and 25ºC, respectively. In other words, a significant difference was revealed in the number of adults obtained among the three subspecies of the bacterium at two temperatures of 25 and 30ºC that were less than the control. In general, the average duration of transformation of bacteria-infected larvae to pupation was less than that of the control. The effects of environmental changes on insect immune responses are not strong. Since the temperature did not affect the number of nodules in the control, it is concluded that increasing the temperature by affecting the Bt bacterium increased the number of the nodule formation in the surviving larvae and also, the subspecies of the bacterium had a significant effect on it. It seems that for this reason, the treatment of 5th instar larvae with Bta and Btg subspecies at 30ºC caused the highest number of nodules in the hemolymph of larvae without significant difference between these two subspecies. Perhaps this has caused these two treatments to have no difference in the number of pupae and adults created compared to the control. Another result recorded in the present study showed that the increase in temperature caused the 24-day-old larvae to enter the pupal stage faster. This trend was observed in the bacterial treatment (in each of the subspecies) and the control. Of course, infected larvae entered the pupal stage even earlier than healthy larvae. This strategy is to escape from the adverse effects of bacterium and increase in temperature. Because it has been proven that the sensitivity of Lepidopteran larvae to Bt bacterial toxins is higher than that of adults (Lu et al. 2012). The results showed that no more pupae were formed in bacterial treatments from the 9th day after infection at high temperature and the 13th day after infection at low temperature. It seems that this issue happened due to the death of some larvae due to bacterial infection and it is more evident in the case of Btt subspecies in all three temperatures.

مراجع

Alves, S.B. (1998). Controle microbiano de insetos, Fundação de Estudos Agrários Luiz de Queiroz, Piracicaba.
Bajwa, W. I., & Kogan, M. (2000) An interactive knowledge-based system for integrated codling moth management. In M. Shenk & M. Kogan (Eds.), IPM in Oregon: Achievements and future directions (pp. 211-226). Corvallis, Oregon: Oregon State University.
Bauerfeind, S. S., & Fischer, K. (2014). Integrating temperature and nutrition-environmental impacts on an insect immune system. Journal of Insect Physiology, 64, 14–20.
BenFarhat-Touzri, D., Driss, F., & Tounsi, S. (2016). A promising HD133-like strain of Bacillus thuringiensis with dual efficiency to the two Lepidopteran pests: Spodoptera littoralis (Noctuidae) and Ephestia kuehniella (Pyralidae). Toxicon, 118, 112-120.
Borges, A., Santos, P.N., Furtado, A.F., &  Figueiredo, R.C.B.Q. (2008). Phagocytosis of latex beads and bacteria by hemocytes of the triatomine bug Rhodnius prolixus (Hemiptera: Reduvidae). Micron, 39(4), 486-494.
Carriere, Y., Showalter, A. M., Fabrick, J. A., Sollome, J., Ellers–Kirk, C., & Tabashnik B. E. (2009). Cadherin gene expression and effects of Bt resistance on sperm transfer in pink bollworm. Journal of Insect Physiology, 55, 1058-1064.
Contreras, E., Benito-Jardon, M., Lopez-Galiano, M. J., Real, M. D., & Rausell, C., 2015. Tribolium castaneum immune defense genes are differentially expressed in response to Bacillus thuringiensis toxins sharing common receptor molecules and exhibiting disparate toxicity. Developmental and Comparative Immunology, 50, 139–145.
Curwen, B., Palmer, S., & Ruddell, P. (2000). Brief cognitive behaviour therapy (Brief Therapies Series). London: Sage.
Dequech, S. T. B., Silva, R. F. P., & Fiuza, L. M. (2005). Interação entre Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), Campoletis flavicincta (Ashmead) (Hymenoptera: Ichneumonidae) e Bacillus thuringiensis aizawai, em laboratório. Neotropical Entomology, 34(6), 937–944.
Dubovskiy, I. M., Grizanova, E. V., Whitten, M. M. A., Mukherjee, K., Greig, C., Alikina, T., Kabilov, M., Vilcinskas, A., Glupov, V. V., & Butt, T. M. (2016). Immuno-physiological adaptations confer wax moth Galleria mellonella resistance to Bacillus thuringiensis. Virulence 7, 860–870.
El Khoury, M., Azzouz, H., Chavanieu, A., Abdelmalak, N., Chopineau, J., & Awad, M. K. (2014). Isolation and characterization of a new Bacillus thuringiensis strain Lip harboring a new cry1Aa gene highly toxic to Ephestia kuehniella (Lepidoptera: Pyralidae) larvae. Archives of Microbiology, 196(6), 435-444.
Franssens, V., Smagghe, G.,  Simonet, G.,  Claeys, I.,  Breugelmans, B.,  De Loof, A., &  Broeck, J. V. (2006). 20-Hydroxyecdysone and juvenile hormone regulate the laminarin-induced nodulation reaction in larvae of the flesh fly, Neobellieria bullata. Developmental and Comparative Immunology, 30(9), 735-740.
Hagstrum, D., Klejdysz, T., Subramanyam, B., & Nawrot, J. (2013). Atlas of stored-product insects and mites, advancing grain science worldwide, Minnesota, USA: AACC International Press.
Itoua-Apoyolo, C., Drif, L., Vassal, J. M., Debarjac, H., Bossy, J. P., Leclant, F., & Frutos, R. (1995) Isolation of Multiple Subspecies of Bacillus thuringiensis from a Population of the European Sunflower Moth, Homoeosoma nebulella. Applied and Environmental Microbiology, 61(12): 4343-4347.
Kalman, S. K., Kiehne, L., Libs, J. L., & Yamamoto, T. (1993). Cloning of a novel cryIC-type gene from a strain of Bacillus thuringiensis subsp. galleriae. Applied and Environmental Microbiology, 59 (4), 1131-1137.
Korkmaz, E., Altun, N., & Faiz, Ö. (2022). Effects of diet on phenoloxidase activity and development of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) Larvae. Biological Bulletin of Russian Academy of Science, 49 (Supplementary 1), S189–S197.
Kurtulus¸ A., Pehlivan, S., Achiri, T. D., & Atakan, E. (2020). Influence of different diets on some biological parameters of the Mediterranean flour moth, Ephestia kuehniella Zeller (Lepidoptera: Pyralidae). Journal of Stored Products Research 85, 101554.
Li, L., Chen, Z., & Yu, Z. (2017). Mass production, application and market development of Bacillus thuringiensis biopesticides in China. In L. M. Fiuza, R. A. Polanczyk & N. Crickmore (Eds.), Bacillus thuringiensis and Lysinibacillus sphaericus characterization and use in the field of biocontrol (pp. 185-212). Switzerland: Springer.
Lu, Y., Wu, K., Jiang, Y., Guo, Y., & Desneux, N. (2012). Widespread adoption of Bt cotton and insecticide decrease promotes biocontrol services. Nature, 487, 362-365.
Mostafa, A., Fields, P., & Holliday, N. (2005). Effect of temperature and relative humidity on the cellular defense response of Ephestia kuehniella larvae fed Bacillus thuringiensis. Journal of invertebrate pathology, 90 (2), p. 79-84.
Nation J. L. (2016). Insect physiology and biochemistry. Third Edition. Florida: CRC Press, Boca Raton.
Panwar, B.S., Kaur, J., Kumar, P., & Kaur, S. (2018). A novel cry52Ca1 gene from an Indian Bacillus thuringiensis isolate is toxic to Helicoverpa armigera (cotton boll worm). Journal of Invertebrate Pathology, 159, 137–140.
Pinos, D., Andrés-Garrido, A., Ferré, J., & Hernández-Martíneza, P. (2021). Response mechanisms of invertebrates to Bacillus thuringiensis and its pesticidal proteins. Microbiology and Molecular Biology Reviews, 85(1), e00007-20.
Rabinovitch, L., Vivoni, A. M., Machado, V., Knaak, N., Berlitz, D. L., Polanczyk, R. A., & Fiuza, L. M. (2017). Bacillus thuringiensis characterization: morphology, physiology, biochemistry, pathotype, cellular, and molecular aspects. In L. M. Fiuza, R. A. Polanczyk & N. Crickmore (Eds.), Bacillus thuringiensis and Lysinibacillus sphaericus characterization and use in the field of biocontrol (pp. 1-18). Switzerland: Springer.
Rahman, M. M., Roberts, H.L.S., & Schmidt, O. (2007). Tolerance to Bacillus thuringiensis endotoxin in immune-suppressed larvae of the flour moth Ephestia kuehniella. Journal of Invertebrate Pathology, 96(2), 125-132.
Seifinejad, A., Salehi Jouzani, G. R., Hosseinzadeh, A., & Abdmishani, C. (2008). Characterization of Lepidoptera-active Cry and Vip genes in Iranian Bacillus thuringiensis strain collection. Biological Control, 44, 216–226.
Shapiro-Ilan, D. I., Fuxa, J. R., Lacey, L. A., Onstad, D. W., & Kayae, H. K. (2005). Definitions of pathogenicity and virulence in invertebrate pathology. The Journal of Invertebrate Pathology, 88:1–7.
Silva, M. C., Siqueira, H. A. A., Marques, E. J., Silva, L. M., Barros, R., Lima Filho, J. V. M.,  & Silva, S. M. F.A.  (2012). Bacillus thuringiensis isolates from northeastern Brazil and their activities against Plutella xylostella (Lepidoptera: Plutellidae) and Spodoptera frugiperda (Lepidoptera: Noctuidae). Biocontrol Science and Technology, 22(5), 583-599.
Silva, M.C., Siqueira, H. A. A., Silva, L. M., Marques, E. J., & Barros, R. (2015). Cry proteins from Bacillus thuringiensis active against diamondback moth and fall armyworm. Neotropical Entomology, 44(4), 392-401.
Stanley, D. W., & Miller, J. S. (2006). Eicosanoid actions in insect cellular immune functions. Entomologia Experimentalis et Applicata, 119, 1–13.
Tabashnik, B., Van Rensburg, J. B. J., & Carriere, Y. (2009). Field-evolved insect resistance to Bacillus thuringiensis crops: definition, theory and data. Journal of Economic Entomology, 92, 2011-2025.
Vilcinskas, A., & Matha, V. (1997). Effects of entomophatogenic fungus Metarhizium anisopliae and its secondary metabolites on morphology and cytosceleton of plasmatocytes isolated from the greater wax moth Galleria mellonella. Journal of Insect Physiology, 43, 1149-1159.
Washburn, J. O., Haas-Stapleton, E. J., Tan, F. F., Beckage, N. E., & Volkman, L. E. (2000). Co-infection of Manduca sexta larvae with polydnavirus from Cotesia congregata increases susceptibility to fatal infection by Autographa californica M Nucleopolyhedrovirus. Journal of Insect Physiology, 46, 179–190.
Wei, J., Yang, S., Chen, L., Liu, X., Du, M., An, S., & Liang, G. (2018). Transcriptomic responses to different Cry1Ac selection stresses in Helicoverpa armigera. Frontiers in Physiology, 9, 1653.
Wraight, S. P., Molloy, D., Jamnback, H., & McCoy, P.  (1981). Effects of temperature and instar on the efficacy of Bacillus thuringiensis var. israelensis and Bacillus sphaericus strain 1593 against Aedes stimulans larvae. Journal of Invertebrate Pathology, 38(1), 78-87.
Wu, G., & Yi, Y. (2018). Transcriptome analysis of differentially expressed genes involved in innate immunity following Bacillus thuringiensis challenge in Bombyx mori larvae. Molecular Immunology, 103, 220–228.
Yılmaz, S., Ayvaz, A., Akbulut, M., Azizoglu, U., & Karabörklü, S. (2012). A novel Bacillus thuringiensis strain and its pathogenicity against three important pest insects. Journal of Stored Products Research, 51, 33-40.
Yilmaz, S., Karabörklü, S., Azizoğlu, U., Ayvaz, A., Akbulut, M., & Yildiz, M. (2013). Toxicity of native Bacillus thuringiensis isolates on the larval stages of pine processionary moth Thaumetopoea wilkinsoni at different temperatures. Turkish Journal of Agriculture and Forestry, 37, 163-172.
Zhang, Y., Ma, Y., Wan, P.-J., Mu, L.-L., & Li, G.-Q. (2013). Bacillus thuringiensis insecticidal crystal proteins affect lifespan and reproductive performance of Helicoverpa armigera and Spodoptera exigua adults. Journal of Economic Entomology, 106(2), 614-621.
Zhao, M., Yuan, X., Wei, J., Zhang, W., Wang, B., Khaing, M. M., & Liang, G. (2017). Functional roles of cadherin, aminopeptidase-N and alkaline phosphatase from Helicoverpa armigera (Hübner) in the action mechanism of Bacillus thuringiensis Cry2Aa. Science Report, 7, 46555.
Zhu, L., Tian, L. J., Zheng, J., Gao, Q. L., Wang, Y. Y., Peng, D. H., & Sun, M. (2015). Complete genome sequence of Bacillus thuringiensis serovar galleriae strain HD-29, a typical strain of commercial biopesticide. Journal of Biotechnology, 19, 108-109.
کنترل بیولوژیک آفات و بیماری های گیاهی
دوره 11، شماره 2
دی 1401
صفحه 135-152

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