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Molecular identification of Heliothine species by nuclear and mitochondrial PCR-RFLP profile | ||
| Journal of Entomological Society of Iran | ||
| دوره 44، شماره 4، 2024، صفحه 477-486 اصل مقاله (957.64 K) | ||
| نوع مقاله: Paper, English | ||
| شناسه دیجیتال (DOI): 10.61186/jesi.44.4.10 | ||
| نویسندگان | ||
| Paulo Queiroz* 1؛ Elias Ferreira Sabiá Júnior2؛ Erica Martins3؛ Rose Monnerat4 | ||
| 1University Center of Brasília - CEUB - SEPN, 707/907, Via W 5 Norte, 70790-075, Asa Norte/Brasília (Distrito Federal), Brazil | ||
| 2Department of Physiological Sciences, Institute of Biological Sciences, University of Brasília - UnB, 70910-900, Brasília (Distrito Federal), Brazil. | ||
| 3State Department of Education – SEE – DF. Undersecretary of Planning, Monitoring and Educational Assessment – SUPLAV. SBN Quadra 02 Bloco C, Edifício Phenícia, 11º andar, 70.040-020, Brasília (Distrito Federal), Brazil | ||
| 4Embrapa Genetic Resources and Biotechnology – Laboratory of Entomopathogenic Bacteria. Parque Estação Biológica, PqEB, Av. W5 Norte (final) CP 02372, 70770-917, Brasília (Distrito Federal), Brazil | ||
| چکیده | ||
| The taxonomy of the Heliothinae subfamily is complex, often becoming an obstacle in the identification of these species, especially during the larval stage. An alternative is the use of molecular methods such as the PCR-RFLP technique (or Restriction Fragment Length Polymorphism), which helps to aid and complement the identification of some heliothine species. The aim of this study was to establish the PCR-RFLP profile using molecular markers for the identification of four heliothine species. Specific primers were constructed for the amplification of the cytochrome oxidase I (COI) and elongation factor 1 alpha (EF-1) genes for identifying Helicoverpa armigera, H. zea, H. gelotopoeon, and Chloridea virescens species. In the digestion of the COI gene with the BfaI enzyme, fragments of 294 bp and 321 bp were obtained for H. armigera, while there was no digestion for H. zea, H. gelotopoeon, and C. virescens. The HpaI enzyme produced fragments of 58 bp and 557 bp for H. gelotopoeon, while there was no digestion for H. armigera, H. zea, and C. virescens. The digestion of the EF-1 gene with the EcoRV enzyme produced fragments of 142 bp and 964 bp for C. virescens, while there was no cleavage for the other heliothines. These molecular markers can help the entomologists, aiding in the discrimination between H. zea, H. armigera, H. gelotopoeon, and C. virescens, and can also be used as a tool for monitoring the spread of these pests in agricultural regions. | ||
| کلیدواژه‌ها | ||
| molecular entomology, Commodities؛ Cytochrome oxidase I؛ Helicoverpa؛ Nuclear elongation factor 1 alpha | ||
| عنوان مقاله [English] | ||
| شناسایی مولکولی گونه های آفت Heliothine (Lep. Noctuidae) با مشخصات PCR-RFLP هسته‌ای و میتوکندریایی | ||
| نویسندگان [English] | ||
| پائولو روبرتو مارتینز کیروش1؛ الیاس فریرا سابیا جونیور2؛ اریکا مارتینوس3؛ رز مونرات4 | ||
| 1مرکز دانشگاهی برزیل، برازیلیا، برزیل | ||
| 2گروه علوم فیزیولوژیکی، موسسه علوم زیستی، دانشگاه برازیلیا، برازیلیا، برزیل | ||
| 3اداره دولتی آموزش. معاون برنامه ریزی، نظارت و ارزیابی آموزشی، سوپلاو، برزیل | ||
| 4اتحادیه تحقیقات کشاورزی برزیل (Embrapa)، بخش منابع ژنتیکی و بیوتکنولوژی – آزمایشگاه باکتری های بیمارگر حشرات، برازیلیا ، برزیل | ||
| چکیده [English] | ||
| آرایه بندی زیرخانواده Heliothinae پیچیده است، به‌ویژه شناسایی این گونه‌ها در مرحله لاروی با مانع مواجه میشود. روش جایگزین، استفاده از روش‌های مولکولی مانند تکنیک PCR-RFLP (یا پلی‌مورفیسم طول قطعه محدود) است که به کمک و تکمیل شناسایی برخی از گونه‌های Heliothinae کمک می‌کند. هدف از این مطالعه ایجاد مشخصات PCR-RFLP با استفاده از نشانگرهای مولکولی برای شناسایی چهار گونه Heliothinae بود. آغازگرهای اختصاصی برای تکثیر ژن‌های سیتوکروم اکسیداز زیر واحد یک (COI) و ژن EF-1a برای شناسایی گونه‌های Helicoverpa armigera، H. zea، H. gelotopoeon و Chloridea virescens ساخته شدند. در هضم ژن COI با آنزیم BfaI، قطعات 294 جفت باز و 321 جفت باز برای H. armigera به دست آمد، در حالی که برای H. zea، H. gelotopoeon و C. virescens هیچ نقطه برشی وجود نداشت. آنزیم HpaI قطعات 58 جفت باز و 557 جفت باز برای H. gelotopoeon ایجاد نمود، در حالی که برای گونه های H. armigera، H. zea و C. virescens هیچ گونه برشی رخ نداد. برش ناحیه ژنی EF-1a با آنزیم EcoRV برای C. virescens دو قطعه به طول 142 و 964 باز ایجاد کرد، در حالی که برای سایر گونه ها هیچ نقطه برشی نداشت. مارکر مولکولی مبتنی بر برش آنزیمی نواحی ژنی مذکور می توانند به حشره شناسان برای تمایز بین گونه‌های H. zea، H. armigera، H. gelotopoeon و C. virescens کمک کنند و همچنین می توانند به عنوان ابزاری برای نظارت بر محدوده پراکنش این آفات در مناطق کشاورزی استفاده شوند. | ||
| کلیدواژه‌ها [English] | ||
| حشره شناسی مولکولی, سیتوکروم اکسیداز زیرواحد I, تنوع ژنتیکی, Helicoverpa, ژن EF-1α | ||
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