موقعیت آرایه‌شناختی یک گونه‌ی بومی از جنس Feltiella Rübsaamen (Dip.: Cecidomyiidae) در ایران

نوع مقاله : مقالات پژوهشی


1 گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه فردوسی مشهد

2 دانشگاه فردوسی مشهد


موفقیت برنامه‌های مهار زیستی آفات به آرایه‌شناسی و شناسایی دقیق آفت و دشمنان طبیعی آن بسیار وابسته است. پشه‌های گال‌زای جنسRübsaamen, 1910  Feltiella از عوامل موثر مهار کنه‌های تارتن هستند، که علی‌رغم تنوع، گستردگی و توانایی مهار بالا تنها یک گونه از آن‌ها به‌صورت تجاری ثبت شده است. گونه‌های این جنس به دلیل داشتن هم‌نام‌های زیاد و پیچیده و نبود بررسی‌های جامع آرایه‌شناختی به سختی از یکدیگر قابل تفکیک می‌باشند. هدف از انجام این پژوهش، تعیین جایگاه آرایه‌بندی پشه  Feltiellaدر ایران بر اساس ویژگی‌های ریخت‌شناسی و مولکولی آن است. جایگاه آرایه‌بندی این پشه با استفاده از کلید‌های شناسایی موجود و عمدتا بر پایه ویژگی‌های دستگاه تناسلی حشرات نر و شناسایی مولکولی از طریق تعیین توالی زیرواحد یک ژن سازنده آنزیم سیتوکروم‌اکسیداز‌C  و مقایسه آن با نمونه‌های موجود در بانک ژن انجام شد. در بررسی ریخت‌شناختی شکارگر جمع‌آوری‌شده از کلنی کنه‌های تارتن به Feltiella acarisuga Vallot تعیین هویت شد. این در حالی بود که در بررسی‌های مولکولی نمونه جمع‏آوری شده از مشهد شباهت بسیار کمتری به گونه‌ی معروف F. acarisuga نسبت به گونه‌ی منتسب به F. tetranychi داشت. به‌طوری‌که مقایسه اختلافات نوکلئوتیدی بین گونه‌ای نمونه‌های گونه‌ی بومی حاضر با نمونه‌های F. acarisuga و F. tetranychi موجود در بانک ژن و همچنین نتایج به‌دست‌آمده از تحلیل اتصال- همسایگی نشان داد که نمونه‌های مشهد علی‌رغم شناسایی و تایید مورفولوژیکی به عنوان گونه F. acarisuga، احتمالا گونه‌ی F. tetranychi هستند، که به دلیل شباهت بسیار زیاد ریخت‌شناسی از آن به‌عنوان مترادف احتمالی F. acarisuga یاد می‌شود. با توجه به دقت بیشتر شناسایی مولکولی نسبت به ریخت‌شناختی، پژوهش حاضر تفاوت گونه‌ی مورد مطالعه‌ با گونه‌ی معروف تجاری F. acarisuga را نشان می‌دهد. اثبات یا رد فرضیه مترادف بودن این دو گونه نیازمند مطالعه‌ی نمونه‌های تیپ گونه‌های F. acarisuga و F. tetranychi و بررسی مولکولی و ریخت‌شناختی جمعیت‌های مختلف دنیا می‌باشد.



عنوان مقاله [English]

Taxonomic Status of a Native Species of the Genus Feltiella Rübsaamen (Dip.: Cecidomyiidae) in Iran

نویسندگان [English]

  • M. Mollaei 1
  • H Sadeghi Namaghi 2
1 Department of Plant Protection, Faculty of Africulture, Ferdowsi University of Mashhad
2 Ferdowsi University of Mashhad
چکیده [English]

 The idea of sustainable agriculture has been considered recently due to increasing knowledge and concerns about the destructive effects of chemical pesticides. Biological control is an ecologically based pest management strategy with an important role in achieving sustainable agriculture. The success of this beneficial method closely depends on taxonomy, since accurate identification of pests and their natural enemies has a great importance in biocontrol project’s achievement. The gall midges of the genus Feltiella are cosmopolitan species known as highly effective predators of tetranychid mites. Despite the high potential of Feltiella species as a biological control agent, F. acarisuga is the only species commercially available among eleven species of the genus. These predators are difficult to distinguish from each other because of the high similarity and low information about them. Comprehensive taxonomic studies are needed to identify promising species for the control of tetranychid mites. The aim of this study is to determine the status of the native Feltiella species in Iran emphasizing their molecular characteristics.
Materials and Methods
 The native predatory gall midges larvae and pupae were collected periodically from the spider mites colony on various host plants (Urtica dioica, Lactuca scariola and Rubus sp.) in countrysides around Mashhad during 2018-2019 and maintained in a growth chamber (LD 16:8, 21±1°C, RH 75±5%) until emerging adults. Adults were preserved in ethanol for further analysis and identified morphologically based on male genitalia and other structures used in taxonomic treatments of the genus. The molecular genetic analysis was included DNA extraction using the Chelex 100 method, PCR amplification of the mitochondrial COI gene using the LCO/HCO universal primer pair, sequencing the gene, and matching the sequence with those of the related species using BLAST. Nucleotide divergence between sequences was estimated by Maximum Composite Likelihood model and by the Pairwise deletion method in MEGA-X software. Intra- and interspecific distances were calculated using ExcaliBAR software and their frequency distribution histogram was plotted using Excel software. The sequence data were analyzed through the neighbor-joining method using MEGA-X software. Evolutionary distances for the NJ method were computed by Kimura’s two-parameter distances. The resulting tree was subjected to bootstrap analysis with 1000 pseudoreplications. The cecidomyiid genus Endaphis was employed as an outgroup taxon to construct the phylogenetic tree.
Results and Discussion
 Based on morphological studies, specimens of the native acarivorous gall midges from various localities in Mashhad were identified as Feltiella acarisuga Vallot. In spite of the morphological result, the DNA sequence of the native species was relatively different from the corresponding sequence of F. acarisuga available in GenBank. The sequence match between the two species was 92.74% in maximum. The match with F. acarivora sequences was also low (maximum 91.84%). Whereas the BLAST results of the indigenous species sequence matched the corresponding sequence of F. tetranychi with more than 99% homology. Comparing the nucleotide differences between the specimens of the present species with F. acarisuga and F. tetranychi also showed that our Feltiella is a distinct species from F. acarisuga, despite of morphological identification. In the histogram of nucleotide distances, intra and inter specific distances in the COI gene overlapped with each other which were related to the nucleotide distances between individuals of F. tetranychi species in the gene bank and individuals of the species collected in the present study. Based on the neighbor-joining tree inferred from partial sequences of the COI gene related to Feltiella species, Iranian indigenous species and F. tetranychi species were in the same ancestor, while individuals of F. acarisuga species were in separate ancestors from the native gall midges. Therefore, according to our molecular studies, the specimens of the native gall midges of Mashhad were F. tetranychi. The possible interpretation for the difference between morphological and molecular identification results in this study is the difficulty of distinguishing the two species from each other, due to their great morphological similarity. F. tetranychi has been mentioned as a possible synonym for F. acarisuga so far, because of the high morphological resemblance. Personal correspondence with international experts revealed that there are two taxa named F. tetranychi, one named by Rubsaamen and introduced as one of the synonymous names of F. acarisuga, and the other named by Kieffer which is an unknown species and mentioned as a possible synonym of F. acarisuga. To prove or disprove the hypothesis whether F. tetranychi is synonymous with F. acarisuga or completely separate from it, it is necessary to study voucher specimens of Feltiella species. Studying further populations of the gall midges on various hosts around the world through sequencing more than one molecular marker is also needed.
 In this study specimens of the native gall midges were identified as Feltiella acarisuga Vallot based on morphological identification, while molecular studies identified them as F. tetranychi. Since molecular identification is more accurate than morphological one, the present study can show how different the indigenous species is from the well-known commercial species F. acarisuga. The present native species probably has little ability to settle in artificial and manipulated environments despite of its activity in the nature of Mashhad. Its usage as a biological control agent for tetranychid mites requires further bio-ecological studies in the laboratory and its genetic comparison with known species in the world.

کلیدواژه‌ها [English]

  • Biological control
  • Gall midge
  • Molecular identification
  • Tetranychid mite
1- Abe, J., Ganaha-Kikumura, T., & Yukawa, J. (2011). Morphological features, distribution, prey mites, and life history traits of Feltiella acarisuga (Vallot)(Diptera: Cecidomyiidae) in Japan. Applied Entomology and Zoology 46(2): 271-279. https://doi.org/10.1007/s13355-011-0038-x.
2- Aliabadian, M., Nijman, V., Mahmoudi, A., Naderi, M., Vonk, R., & Vences, M. (2014). ExcaliBAR: a simple and fast software utility to calculate intra-and interspecific distances from DNA barcodes. Contributions to Zoology, 83(1), 79-84d. https://doi.org/10.1163/18759866-08301004.
3- Armstrong, K., & Ball, S. (2005). DNA barcodes for biosecurity: invasive species identification. Philosophical Transactions of the Royal Society B: Biological Sciences, 360(1462): 1813-1823. https://doi.org/10.1098/rstb.2005.1713.
4- Baker, B.P., Green, T.A., & Loker, A.J. (2020). Biological control and integrated pest management in organic and conventional systems. Biological Control 140: 104095. https://doi.org/10.1016/j.biocontrol.2019.104095.
5- Baker, K. & Cook, R.J. (1974). Biological control of plant pathogens. WH Freeman and Company.
6- Barratt, B.I., Cock, M.J., & Oberprieler, R.G. (2018). Weevils as targets for biological control, and the importance of taxonomy and phylogeny for efficacy and biosafety, Diversity 10(3): 73. https://doi.org/10.3390/d10030073.
7- Fedotova, Z. & Kozlova, E. (2019). Gall Midges of the Genus Feltiella Rübsaamen (Diptera, Cecidomyiidae) in the Northwest of Russia, with Description of a New Species, Entomological Review 99(9): 1359-1381.
8- Folmer, O., Black, M., Hoeh, W., Lutz, R., & Vrijenhoek, R. (1994). DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates, Molecular Marine Biology and Biotechnology 3(5): 294–299.
9- Gagné, R.J. (1995). Revision of tetranychid (Acarina) mite predators of the genus Feltiella (Diptera: Cecidomyiidae), Annals of the Entomological Society of America 88(1): 16-30. https://doi.org/10.1093/aesa/88.1.16.
10- Gagné, R.J. & Jaschhof, M. (2021). A Catalog of the Cecidomyiidae (Diptera) of the World. Fifth Edition. Digital. 816 pp.
11- Ganaha-Kikumura, T., Yukawa, J., Tokuda, M., Ohno, S., & Abe, J. (2012). Occurrence of two acarivorous species of the genus Feltiella (Diptera: Cecidomyiidae) in Okinawa, southern Japan, and redescription of F. acarivora (Zehntner), Applied Entomology and Zoology 47(4): 319-329. https://doi.org/10.1007/s13355-012-0122-x.
12- Gillespie, D.R., Roitberg, B., Basalyga, E., Johnstone, M., Opit, G., Rodgers, J., & Sawyer, N. (1998). Biology and Application of Feltiella acarisuga (Vellot)(Diptera: Cecidomyiidae) for Biological Control of Twospotted Spider Mites on Greenhouse Vegetable Crops. Technical Report 145 Agriculture and Agri-Food Canada, Pacific Agri-Food Research Centre: Agassiz, BC, Canada.
13- Harris, K. (1966). Gall midge genera of economic importance (Diptera: Cecidomyiidae) Part 1: Introduction and subfamily Cecidomyiinae; supertribe Cecidomyiidi, Transactions of the Royal Entomological Society of London 118(10): 313-358. https://doi.org/10.1111/j.1365-2311.1966.tb03156.x.
14- Hewitt, G.M., Johnston, A.W.B., & Young, J.P.W. (1991). Molecular Techniques in Taxonomy. Berlin, Germany: Springer.
15- Hoddle, M.S., & Van Driesche, R.G. (2009). Biological control of insect pests. p. 91–100. In: Resh, V.H., Cardé, R.T. (Eds.) Encyclopedia of Insects, second ed. Academic Press, San Diego.
16- Honarmand, A., Sadeghi Namaghi, H., & Fekrat, L. (2015). First report of Feltiella acarisuga Vallot (Diptera: Cecidomyiidae) in Iran, Plant Protection 28(3): 434-436. https://doi.org/10.22067/JPP.V28I3.27134.
17- Kimura, M. (1980). A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences, Journal of Molecular Evolution 16(2): 111-120. https://doi.org/10.1007/BF01731581.
18- Lee, H.-S., Chung, B.-K., & Kim, K.-J. (2004). First report of Feltiella acarisuga Vallot (Diptera: Cecidomyiidae) in Korea, Korean Journal of Applied Entomology 43(3): 185-188.
19- Muraji, M., Kawasaki, K., Shimizu, T., & Noda, T. (2004). Discrimination among Japanese species of the Orius flower bugs (Heteroptera: Anthocoridae) based on PCR-RFLP of the nuclear and mitochondrial DNAs, Japan Agricultural Research Quarterly: JARQ 38(2): 91-95.
20- Opit, G., Roitberg, B., & Gillespie, D. (1997). The functional response and prey preference of Feltiella acarisuga (Vallot)(Diptera: Cecidomyiidae) for two of its prey: male and female twospotted spider mites, Tetranychus urticae Koch (Acari: Tetranychiidae), The Canadian Entomologist 129(2): 221-227. https://doi.org/10.4039/Ent129221-2.
21- Rosen, D. (1986). The role of taxonomy in effective biological control programs, Agriculture, Ecosystems and Environment 15(2-3): 121-129. https://doi.org/10.1016/0167-8809(86)90085-X.
22- Saxena, A., Mishra, S., Raghuwanshi, R., & Singh, H.B. (2013). Biocontrol agents: Basics to biotechnological applications in sustainable agriculture. p. 141-164. In: Tiwari S.P., Sharma R., Gaur R. (Eds.) Recent Advances in Microbiology, Vol 2. Nova Science Publishers Inc., U.S.A.
23- Sayed, S.M., Montaser, M.M., Elsayed, G., Amer, S.A., & Iatrou, K. (2013). Preliminary molecular identification of a predatory bug, Orius albidipennis, collected from ornamental plants, Journal of Insect Science 13: 11. https://doi.org/10.1673/031.013.1101.
24- Sharaf, N. (1984). Studies on natural enemies of tetranychid mites infesting eggplant in the Jordan Valley, Journal of Applied Entomology 98(1‐5): 527-533. https://doi.org/10.1111/j.1439-0418.1984.tb02745.x.
25- Sharma, A., Diwevidi, V.D., Singh, S., Pawar, K.K., Jerman, M., Singh, L., Singh, S., & Srivastawa, D. (2013). Biological Control and its Important in Agriculture. International Journal of Biotechnology and Bioengineering Research 4(3): 175-180. https://www.ripublication.com/ijbbr_spl/ijbbrv4n3spl_03.pdf.
26- Singh, H. (2006). Achievements in biological control of diseases with antagonistic organisms at National Botanical Research Institute, Lucknow, Current status of biological control of plant diseases using antagonistic organisms in India. Proceedings of the group meeting on antagonistic organisms in plant disease management held at Project Directorate of Biological Control, Bangalore, India on 10-11th July 2003, pp. 329-336. https://eurekamag.com/research/012/763/012763166.php.
27- Skuhravá, M. & Skuhravý, V. (2020). The Gall Midges of Europe. Prague: Maxdorf Jesenius.
28- Wardlow, L. & Jobin, A. (1990). Potential new additions to the armoury of natural enemies for protected tomatoes. In Integrated control in glasshouses, Bulletin SROP (Vol. 13, Num. 5, pp. 225-227).
29- Watson, G. (1997). The Role of Taxonomy in Biological Control, With Special Reference to Insects. In Prociding Kongres Perhimpunan Entomologi Indonesia, Universitas Padjajaran, Bandung (Vol. V, pp. 24-26).