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Homoptera, Aphididae, Eriosomatinae) on Pistacia Trees from Kurdistan Region North of Iraq H

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Homoptera, Aphididae, Eriosomatinae) on Pistacia Trees from Kurdistan Region North of Iraq H Iraqi Journal of Agricultural Sciences –2020:51(Special Issue):184-190 Ali & et al. MOLECULAR PHYLOGENY OF GALL-FORMING APHIDS TRIBE FORDINI (HOMOPTERA, APHIDIDAE, ERIOSOMATINAE) ON PISTACIA TREES FROM KURDISTAN REGION NORTH OF IRAQ H. B. Ali R. S. Kamal H. Y. Fadhil Assist. Prof. Assist. Lecturer. Assist. Prof. Dept. Biol. Coll. Sci. University of Baghdad. Corresponds author e-mail: [email protected] ABSTRACT Four aphid species belonging to the Tribe Fordini (Homoptera, Aphididae, Eriosomatinae) induce galls on Pistacia trees in North Iraq (Kurdistan) were recorded in this study. Molecular phylogeny of Iraq Fordini species was constructed for These species (Forda riccobonii, Paracletus cimiciformis, Baizongia pistaciae and Forda marginata), based on sequences of mitochondrial Cytochrome Oxidase I (COI) gene of these aphid species to detect the relationship and taxonomy of Iraqi species for tribe Fordini. Our results confirmed that each of Forda and Paracletus are a sister genus, or monophyletic group by designing in same clade, while B. pistaciae is separate from other clades, supporting the taxonomic division of the tribe Fordini into two subtribes (Fordina and Baizongina). Key words: fordini, molecular phylogeny, (COI) gene, gall aphids, Pistacia مجلة العلوم الزراعية العراقية -2020 :51(عدد خاص(:184-190 علي وآخرون عﻻقات النشوء الجزيئية لحشرات المن المكونة للعقد قبيلة Homoptera, Aphididae, Eriosomatinae) Fordini) على أشجار الفستق من منطقة كردستان شمال العراق حيدر بدري علي رؤيا صفوان كمال ُحﻻ يونس فاضل استاذ مساعد مدرس مساعد استاذ مساعد قسم علوم الحياة/ كلية العلوم - جامعة بغداد المستخلص اربعةة انةوام مةن حشةرات المةن المكةون للعقةد علةى اشةجار الفسةتق تعةود لقبيلةة Homoptera, Aphididae, ( Fordini) (Eriosomatinae قةةد سةةجلت فةةي اةةذا الدراسةةةا عﻻقةةات النشةةوء الجزيئيةةة ينةةوام اةةذا القبيلةةة فةةي العةةراق تمةةت دراسةةت ا فةةي اينةوام (Forda riccobonii, Paracletus cimiciformis, Baizongia pistaciae and Forda marginata) اعتمادا على تتابعات الجين (mitochondrial Cytochrome Oxidase I (COI للتحةري عةن عﻻقةات اينةوام العراقيةة لقبيلة Fordini وتصنيف اا اكدت نتائج اذا الدراسة على ان كل من الجنس Forda و Paracletus اي اجناس شقيقة عةن طريق انضمام ا في مجموعةة وحيةدة ايصةل )clade( فةي حةين انفةراد النةوم B. pistaciae فةي مجموعةة اخةرم ممةا يةدعم تصنيف اذا القبيلة الى تحت قبيلتين اما (Fordina and Baizongina . الكلمات المفتاحية: قبيلة Fordini ، العﻻقات الجزيئية الجينية، جين COI ، المن المكون للعقد، الفستقا *Received:16/5/2019, Accepted:24/9/2019 184 Iraqi Journal of Agricultural Sciences –2020:51(Special Issue):184-190 Ali & et al. INTRODUCTION Pistacia sp. trees (9, 14, 36). In India The genus Pistacia belongs to a cosmopolitan Chakrabarti, (10, 11) listed four species of family “Anacardiaceae, this family include aphids on Pistacia trees included Forda about 70 genera and over 600 species (24). riccobonii; F. marginata; Baizongia pistaciae The species of the genus Pistacia are and Geoica urticularia. In Turkey 7 aphid dioecious trees or shrubs and characterized as species were Identified forming galls on xerophytic trees which and growing up to 8– Pistacia species. Forda formicaria, 10m height. In Iraq AL-Saghir and Porter (4) Smynthurodes betae, Aploneura lentisci, listed three Pistacia species, Pistacia atlantica Geoica utricularia, Baizongia pistaciae, F. Desf., Pistacia eurycarpa Yalt. and Pistacia Magenta and Rectinasus buxtoni. (12) Ortiz- khinjuk Stocks. The subfamily Eriosomatinae Rivas et al. (26) conducted a survey to explain has recently been placed with family the the phylogenetic relationships among 10 Aphididae (16, 17). It was previously placed Fordina species present in the Iberian in the family Pemphigidae (= Eriosomatidae). Peninsula and the Canary Islands when they There are about 19 aphid genera with 107 studied the implications for the taxonomy of valid species belong to the tribe Fordini genera Forda and Paracletus Mróz and Depa, (Subfamily Eriosomatinae) (5, 6, 28), (25) proved the usefulness of molecular according to Inbar, (19) about 30 of these analysis for solving taxonomic problems in species are gall-inducing aphids belong to this aphids when they studied the DNA sequences tribe. The division of subtribes in Fordini is of Mediterranean populations of Paracletus confusing. Some believe that Fordini includes cimiciformis in aphid specimens two subtribes, Fordina and Baizongina (14, morphologically identical with Forda rotunda 37), others suggest it is divided into Fordina Theobald, 1914. Li et al. (22) analyzed the and Melaphidina (8, 18, 34). Almost all the molecular phylogeny of the Subfamily species in Fordini are gall-forming and Eriosomatinae (also called Pemphiginae) stimulate species-specific galls on their which traditionally consists of 3 tribes, primary hosts, whereas the primary hosts for Eriosomatini, Pemphigini, and Fordini based all species are full within two genera, Pistacia on morphological and biological evidence. In and Rhus, which belong to Anacardiaceae. Iraq, Aphid fauna induce species-specific gall Galls are the characteristic of aphids (29). almost poorly known, they are known mainly Their position, morphology and structure of by insufficient surveillance lists, three aphids these galls are important aphid appearing. forming galls on Pistacia khinjuk where listed Among the whole Aphididae, only by Daoud and El-Haidari, (13); Al-Ali, (1) in Eriosomatinae, Hormaphidinae and some North of Iraq the species are Baizongia species of Aphidinae produce galls (7), Galls pistaciae, Geoica articular and Forda of Fordini are significantly variable in formicary. The present study aimed to morphology and position, nevertheless determine forming galls aphid in the North of fascinatingly, are species-specific (32), since Iraq, and clarifying the evolutionary the gall founder, fundatrix forms a unique- relationships among the Tribe Fordini, and shaped gall in a species-specific manner, the further investigating the relationships within galling aphid species can usually be identified and between Forda, Paracletus and Baizongia. exclusively based on the gall morphology (21). This study represented the first phylogenetic Martinez, (23) and Wool, (33) reported study to this group in Iraq, preceded by few about 15 species of aphids on Pistacia trees studies on other groups such as: Ali et al., (2) induce species-specific gall. The more on the widow spider in Iraq; Al-Saad and common galls are those of Slavum wertheimae Aletby, (3) on the red palm weevil; Faraj et al., [H.R.L.], Smynthurodes betae West, Forda (15) on the protozoan parsites causing Equine riccobonii [Stefani] and Geoica sp,.The babesiosis in Baghdad city and few others. geographic distribution of these aphid species MATERIALS AND METHODS everywhere in the Mediterranean Basin, from Sampling Morocco and Spain to east Afghanistan and Specimens for DNA extraction and Iran, agrees with the distribution of host amplification of mitochondrial Cytochrome 185 Iraqi Journal of Agricultural Sciences –2020:51(Special Issue):184-190 Ali & et al. Oxidase I (COI) gene were collected from specimens were deposited in the Iraq National recent years, mostly from different types of History Research center and Museum, galls on the infected primary hosts Pistacia Department of Entomology, University of trees (figure 1). COI gene was applied widely Baghdad. The specimens were identified in the molecular phylogenetic studies of aphids according to their main morphological and are easy to use in PCR. (22). All diagnostic features (7, 31). The extraction of specimens were collected from different total genomic DNA using the Geneaid DNA locations in Duhok, Erbil and Sulaimaniya Mini extraction kit for (Tissue), according to provinces/ Kurdistan region North of Iraq the standard protocol recommended by the during Summer seasons 2012-2013. Collection manufacturer, with some modification. information of these specimens, including Extracted DNA was being stored at -20°C locations, sample numbers, and collection until use. the whole aphid individual was dates, (shown in table 1). Specimens were selected for molecular experiments in 10 stored at (95–100)% ethanol at 4°C. (25). All replicates for each per specimen. Figure 1. Shape of Pistacia trees-feeding aphids galls. (A,B and C), Pea/Margin galls on the blade of leaves by: A, Forda riccobonii from Erbil and Duhok provinces; B, Paracletus cimiciformis from Erbil, Sulaimaniya and Duhok provinces; and C, Forda marginata from Duhok province); D) Alate form of Paracletus cimiciformis from opening matured gall. Table 1. Summary of collecting data relative to species (Ford riccobonii, Paracletus cimiciformis, Baizongia pistaciae and F. marginata) included in the present study. Species Host Locality, Province Number of collecting galls date 6 14/05/2012 Forda riccobonii Pistacia khinjuk Erbil and Duhok Erbil (1) and Duhok (5) 10/6/2013 Forda marginata Pistacia Sp. Duhok 3 12/6/2013 15/5/2012 12 Paracletus cimiciformis Pistacia khinjuk Erbil and Duhok 13/6/2013 Erbil (4) and Duhok (8) 14/6/2013 10/5/2012 Paracletus cimiciformis Pistacia sp. Sulaimaniya 2 11/5/2012 Baizongia pistaciae Pistacia khinjuk Duhok 3 16/5/2012 Sequencing 3ʹ) (forward primer) and LepR (5 ʹ - The classical method of CTAB TAAACTTCTGGATGTCCAAAAAATCA- (hexadecyltrimethylammonium bromide) 5ʹ) (reverse primer) (22). Complete sequences extraction was reliable to obtain whole- were deposited in GenBank under accession genome DNA from each sample. The gene numbers given in table 2, together with the COI was amplified, and sequenced directly accession
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