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Biochemical, ecological and molecular characterization of Xenorhabdus anantnagensis associated with Steinernema anantnagense from India: evaluating nematode efficacy against Helicoverpa armigera | ||
| Journal of Entomological Society of Iran | ||
| مقاله 9، دوره 45، شماره 1، فروردین 2025، صفحه 121-138 اصل مقاله (1.13 M) | ||
| نوع مقاله: Paper, English | ||
| شناسه دیجیتال (DOI): 10.61186/jesi.45.1.9 | ||
| نویسندگان | ||
| Krishnapriya Okram1؛ Aashaq Hussain Bhat* 2؛ Ladoi Drema3 | ||
| 1Department of Zoology, Manipur University, Canchipur 795003, Manipur, India | ||
| 2University Center for Research and Development, and Department of Biosciences, Chandigarh University Gharuan, Mohali, Punjab, 140413, India | ||
| 3Department of Life Science, Siliguri Terai B. ed College, Baba Saheb Ambedkar Educational University, University of Kolkata 734427, West Bengal, India | ||
| چکیده | ||
| This study provides a molecular and phenotypic characterization of an entomopathogenic nematode-bacterium complex isolated from agricultural soil and nematode efficacy against Helicoverpa armigera. The nematode, identified as Steinernema anantnagense KP_CU, was characterized using the internal transcribed spacer (ITS) region of rRNA, revealing 100% similarity with the type population of S. anantnagense. Phylogenetic analysis confirmed its conspecific status within a clade including S. kushidai, S. akhursti, and S. populi. Concurrently, the associated bacterium, identified as Xenorhabdus sp. KP_CU, exhibited 100% similarity in its 16S rRNA sequence with Xenorhabdus anantnagensis XENO-2T, suggesting conspecificity. Phenotypic characterization aligned the bacterium closely with X. anantnagensis, highlighting typical traits such as rod-shaped, gram-negative cells and absence of bioluminescence. Biochemical tests further supported this identification, distinguishing KP_CU from other Xenorhabdus species based on citrate utilization, gelatinase, lysine decarboxylase, urease, arginine dihydrolase, ornithine decarboxylase, glucose oxidation, cytochrome oxidase and indole production. Phylogenetic analysis based on 16S rRNA sequences placed Xenorhabdus sp. KP_CU within a monophyletic clade with X. anantnagensis, along with sister relationships to X. japonica and X. vietnamensis. The bacterial strains also exhibited larvicidal activity against Galleria mellonella and even at the lowest optical density (OD590 = 0.125) induced over 80% larval mortality within merely 24 h post-injection, emphasizing its elevated virulence. The strain KP_CU could kill the wax moth larvae with 38, 16 and 9 IJs at 24, 36 and 48 h, respectively. The nematode isolate KP_CU demonstrated high virulence against H. armigera larvae, with complete mortality achieved within 60 h across all tested inoculum levels. Mortality began at 36 h post-inoculation at 100 IJs/larva and was reached within 24 h at 200 IJs/larva. LD50 values decreased significantly from 38 IJs at 24 h to just 9 IJs at 48 h, indicating potent lethality. Additionally, progeny production showed a dose-dependent increase, though slightly reduced at higher doses, suggesting a trade-off between virulence and reproductive success. These results suggest that S. anantnagense KP_CU could hold potential as a biocontrol agent for H. armigera in agricultural settings in India. | ||
| کلیدواژه‌ها | ||
| Insect pathology؛ Entomopathogenic nematode؛ Biocontrol؛ Phylogenetic analysis؛ 16S rRNA | ||
| عنوان مقاله [English] | ||
| خصوصیات بیوشیمیایی، اکولوژیکی و مولکولی Xenorhabdus anantnagensis همزیست Steinernema anantnagense از هند: ارزیابی اثر نماتد روی Helicoverpa armigera | ||
| نویسندگان [English] | ||
| کریشناپریا اکرام1؛ عاشق حسین بهات2؛ لاوی درما3 | ||
| 1گروه جانورشناسی، دانشگاه مانیپور، کانچیپور 795003، مانیپور، هند | ||
| 2مرکز تحقیقات و توسعه دانشگاه، دانشگاه چندیگر، موهالی 140413، پنجاب، هند | ||
| 3گروه علوم زیستی، دانشکده سیلیگوری تیرایی B. ed، دانشگاه آموزشی بابا صاحب امبدکار، دانشگاه کلکته 734427، بنگال غربی، هند | ||
| چکیده [English] | ||
| این مطالعه، مشخصات مولکولی و فنوتیپی یک گونه نماتود بیمارگر حشرات و باکتری همزیستش که از خاک در هند جدا شده است را توصیف نموده و اطلاعات حشره کشی نماتود روی لارو Helicoverpa armigera نیز بررسی شده است. گونه نماتود به عنوان Steinernema anantnagense KP_CU معرفی می‌شود. توالی ناحیه ITS این نمونه، همسانی 100٪ با جمعیت تیپ گونه S. anantnagense داشت. تجزیه و تحلیل فیلوژنتیک نیز این نمونه را با گونه‌های S. kushidai، S. akhursti، و S. populi در یک شاخه قرار داد. همزمان،توالی ناحیه 16S باکتری همزیست این نماتود با نام Xenorhabdus sp. KP_CU،6S، شباهت 100٪ با Xenorhabdus anantnagensis XENO-2T نشان داد، که گویای همسانی است. خصوصیات فنوتیپی، نشان داد که باکتری قرابت بالایی با X. anantnagensis دارد و ویژگی‌های معمولی مانند سلول‌های میله‌ای شکل، گرم منفی و عدم وجود نور فلورسنت را برجسته کرد. آزمایش‌های بیوشیمیایی بیشتر از این شناسایی پشتیبانی کردند و KP_CU را از سایر گونه‌های Xenorhabdus متمایز کردند. تجزیه و تحلیل فیلوژنتیک بر اساس توالی 16S rRNA جدایه Xenorhabdus sp. KP_CU را با X. anantnagensis، در یک شاخه تک نیا قرار داد که گونه های X. japonica و X. vietnamensis.، آرایه های خواهری این جدایه بودند. جدایه باکتری همچنین فعالیت لاروکشی روی Galleria mellonella نشان داد و حتی در کمترین چگالی نوری (OD590 = 0.125) تنها 24 ساعت پس از تزریق بیش از 80 درصد لاروها دچار مرگ و میر شدند که توان باکتری را نشان می‌داد. جدایه KP_CU قادر بود روی لارو گالریا با غلظت های 9،16 و 38 لارو عفونت زا به ترتیب بعد از 24، 36 و 48 ساعت مرگ و میر ایجاد نماید. جدایه نماتد KP_CU توان حشره‌کشی بالایی روی لارو H. armigera نشان داد، مرگ و میر در 36 ساعت پس از تلقیح با تراکم لارو 0 IJs / لارو شروع شد و در 24 ساعت در 200 IJs / لارو به دست آمد. مقادیر LD50 به طور قابل توجهی از 38 IJ در 24 ساعت به فقط 9 IJ در 48 ساعت کاهش یافت که نشان دهنده کشندگی قوی است. علاوه بر این، تولید نتاج افزایش وابسته به دوز را نشان داد، اگرچه در دوزهای بالاتر کمی کاهش یافت، که نشان دهنده یک ارتباط بین زهراگینی و تولید مثل است. این نتایج نشان می دهد که S. anantnagense KP_CU پتانسیل آن را دارد تا به عنوان یک عامل کنترل زیست روی H. armigera در مزارع در هندوستان بکار رود. | ||
| کلیدواژه‌ها [English] | ||
| بیماری ناسی حشرات, نماتودشناسی حشرات, کنترل زیستی, آنالیز فیلوژنتیک, 16S rRNA | ||
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