D:\ 81(4) 19142--Selvaraj

Indian Journal of Entomology, 81(4): 000-000 (2019) DoI No.:

FIRST RECORD OF THE INVASIVE WOOLLY FLOCCOSUS (MASKELL) FROM INDIA

SUNDARARAJ, R*, SELVARAJ, K**, KALLESHWARASWAMY, C M***, RANJITH, M***, SUMALATHA, B V**

*Forest Protection Division, ICFRE-Institute of Wood Science and Technology, Bengaluru 560003 **Germplasm Conservation and Utilization, ICAR-National Bureau of Agricultural Resources, Bengaluru 560024 ***Department of Agricultural Entomology, College of Agriculture, University of Agricultural and Horticultural Sciences, Shivamogga 577204 *Email: [email protected] (corresponding author)

ABSTRACT

Occurrence of the highly polyphagous woolly whitefly, (Maskell) (: Aleyrodidae) on guava (Psidium guajava L.) was recorded as invasive in India. The whitefly is believed to be neotropical origin but is now found throughout the warmer parts of the world. The identification of A. floccosus was done through taxonomic morphological characters of puparium and reconfirmed by DNA barcoding of adult whitefly using partial (658 bp) mitochondrial cytochrome oxidase 1 (CO1) gene. This is the first report of the breeding of A. floccosus on guava in India. Identification details, diagnosis, symptoms of damage and impact of its infestation on guava and citrus are discussed.

Key words: Aleurothrixus floccosus, first record, invasive, guava,citrus, , diagnsotics, symptoms of damage, life stages, mtCo1, natural enemies,

Whiteflies (Hemiptera: Aleyrodidae) are distributed oriental, Neotropical and Nearctic Regions, Montenegro, throughout the major zoogeographical regions of the Croatia (Martin et al., 2000; Radonjic and Hrncic, 2003; world, with their greatest diversity in tropical and Simala and Milek, 2008). It is broadly polyphagous, south temperate regions. The trade of exotic plants is feeding on more than 20 plant families and exhibits a responsible for many accidental introductions of insects strong preference for citrus species (Malumphy et al., from their native area (Simala et al., 2015). Most alien 2015). In India it is found breeding on guava, Psidium species of were accidentally introduced along guajava L. (Myrtaceae) for the first time at Kerala. with their host plant through plant trade, small in size, Martin et al. (2000) indicated its presence in India, but cryptic nature and immature stages being attached to the it may not be authentic as there was no information on host-plant and due to these characteristics, they are one its host plant in India. of the most commonly transported groups. They are also among the most successful groups in Recently there is a surge in the invasion of exotic terms of invading new geographical areas. Moreover, whiteflies to India particularly through south India. The changes in climate and global warming could influence record of A. floccosus in Kerala confirms the fact that it new introductions and distribution of exotic whitefly is a species which has been slowly extending its range species and increase the chances of their survival in for many years, and has been the cause of economic new environment (Simala et al., 2015). problems in several parts of the world (Martin, 1999). PreviewThe occurrence of this whitefly on guava, P. guajava Aleurothrixus floccosus (Maskell) was first described was confirmed for the first time in India. from Jamaica in 1896 (Martin and Mound, 2007) and was noticed in Florida in 1909. This whitefly is native MATERIALS AND METHODS to the Neotropical region but is now found throughout Whitefly specimens were collected from infested the warmer parts of the world, wherever citrus is grown guava plants by Ranjith, M from several places in (Malumphy et al., 2015). It was reported from Europe and Kozhikode (11.58oN; 75.78o E) and Malappuram Mediterranean countries, Palaearctic, Ethiopian, Austro- (11.11o N; 76.17o E) districts in Kerala. Whitefly adult 2 Indian Journal of Entomology, 81(4), 2019 individuals were used for DNA based identification following standard protocols. The amplified products and adult emerged puparium processed for permanent were sequenced by M/s Chromous Biotech Pvt Ltd, mounting as per Dubey and David (2012) and Bengaluru and the same were uploaded to GenBank identification is confirmed based on the puparial after annotation. taxonomic characteristics specific to the species. The confirmed mounted slides were deposited in the RESULTS AND DISCUSSION collection of ICAR- NBAIR, Bengaluru. Heavily Taxonomy infested leaves were kept in separate containers to observe and study the emergence of parasitoids. The slide mounts were obtained from the puparia Population and intensity of damage of whitefly on five from which adults had emerged and the identity randomly selected guava tree at three strata (basal, was confirmed as Aleurothrixus floccosus (Maskell) medium, lower leaves) was assessed. (Hemiptera: Aleyrodidae). Pale white to brown puparia covered with flocculent wax; puparial margin not Samples of whitefly from which individuals had deflexed, overall concentric, regularly toothed, each been identified by senior author were used in barcode tooth with a basal gland, without any modifications at development. Genomic DNA was extracted from thoracic and tracheal pore regions; submargin wide individual whiteflies using the GIAGEN DNeasy® separated from dorsal disc by submarginal/ subdorsal kit, Germany following the manufacturer’s protocols. fold, which arranged in distinct sections and complete; The DNA barcode region was amplified from whitefly cephalic and first abdominal setae absent, submedian species using 10 µl of universal DNA primers LCO1490 metathoracic setae present, eight abdominal and caudal (5’-GGTCAACAAATCATAAAGATATTGG-3’) setae long extending beyond puparial margin; inner and HCO 2198 (5’-TAAACTTCAGGGTGACC submargin with a row of five pairs of minute setae in AAAAAATCA-3’) for the amplification of mitochondrial cephalothorax and anterior abdomen; submedian ridge cytochrome c oxidase subunit I (Folmer et al., 1994). prominent; vasiform orifice elevated, transversely The extracts were subjected to PCR amplification of elliptical, operculum similarly shaped, almost filling the a 658bp region near the 5’ terminus of the COI gene orifice and obscuring the lingula (Martin, 1987) (Fig. 1).

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Fig. 1. Aleurothriocus floccosus: a. Dorsal view of mounted puparium, b. Margin, c. Posterior region showing elongated caudal and 8th abdominal setae, d. Submargin with submarginal seta First record of the invasive woolly whiteflyAleurothrixus floccosus (Maskell) from india 3 Sundararaj, R et. al.

This species is quite distinct from A. trichoides, the approximately 1.5 mm in length, with a yellow-white other already invaded known species of Aleurothrixus in body and wings that are covered with a white waxy India by the submargin without band of microtubercles; powder (Fig.3 b-d) and they prefer to feed and oviposit submarginal/ subdorsal fold almost concentric with on the undersides of young leaves that sprout when the margin, arranged in distinct sections and complete host trees are actively growing. between vasiform orifice and puparial margin; comparatively longer 8th abdominal and caudal setae, Natural enemies 8th abdominal setae reaching beyond puparial margin; No natural parasitism either of the indigenous inner submargin with a row of five pairs of tiny setae parasitoid or its potential parasitoid such as Cales noacki in cephalothorax and anterior abdomen and lingula tip (Hymenoptera: Aphelinidae) was observed, though to not bilobed. The determined slides are deposited in the the extent of 82% and 88% parasitism in Greece and collection of ICAR-NBAIR (Acc. No NBAII/HEM- Turkey, respectively were reported (Katsoyannos et ALEU/8-12/2019). al., 1998; Telli and Yiğit, 2012). Few predators such as Acletoxenus sp. (Diptera: Drosophilidae and Scymnus The nucleotide sequence obtained with 637 bp (Pullus) nr. utilis Hoang (Coleoptera: Coccinellidae) size from M/s Chromous Biotech Pvt Ltd, Bengaluru were found associated with A. floccosus colony under were edited and then subjected for BLAST with NCBI field conditions. Yu et al. (2011) recorded predaceous nucleotide sequence data library. The sequence was drosophilid Acletoxenus indicus Malloch preying on submitted to GenBank (acc. no MN027508) and larvae of A. dispersus and Aleurocanthus sp. in South further BLAST analysis revealed that our sequence China. was matched 98% nucleotide identity with available sequence (KF059956) of A. floccosus at NCBI Impact on guava and citrus submitted by Ovalle et al (2013) from Colombia and A. floccosus may become invasive pest and cause whiteflies collected inCitrus sinensis and thus, confirm economic loss for horticulture farmers not only on the identity of the species. P. guajava but also on Citrus spp. as it is a serious Symptoms and severity of damage pest on Citrus in many countries. In India, guava and citrus are two economically important fruit crops with Nymphs and adults suck phloem sap from host rich source of vitamin C and widely cultivated. In plants and indirectly damage by copious execration of guava, about 80 species of insects have been recorded honey dew which attract and support for development to affecting yield and quality of fruits which include of the sooty mould on infested plants and blackening three invasive whiteflies viz., A. dispersus, P. bondari of entire leaves (Fig. 2a). Under severe infestation, the and A. rugioperculatus especially in South India. sooty mold might reduce the photosynthetic ability Invasion and infestation of A. floccosus would further of infested plants significantly, leading to leaf fall, increase pest load on guava, inter-specific competition senescence and eventual death. This also reduces among whitefly species and yield loss. Similarly, other photosynthesis, resulting in decreased fruit size and important likely host is citrus which commercially also contaminates the fruits as well. Severe infestation grown important crop in India. (60-100% plants) of A. floccosus on P. guajava with the nymphal population of 59-363 and adult population The study conclude that more than 250 species 8-27 /leaf was observed alone or in association with of insects were found to feed on citrus including spiralling whiteflyAleurodicus dispersus Russell colonies a dozen whiteflies viz., A. dispersus Russell, A. (Fig.2b). Heaviest infestation was noticed to extent of rugioperculatus Martin, Acaudaleyrodes rachipora 54.2 nymphs/ sq.cm and mostly colonizes on younger (Singh), Aleurocanthus citriperdus Quaintance & leaves than mature leaves (Fig. 2c). Baker, A. husaini Corbett, A. shillongensis Jesudasan & PreviewDavid, A. spiniferus (Quaintance), A. woglumi Ashby, Life stages Aleuroclava citri Jesudasan & David, Alwurolobus The sausage shaped, pale white eggs were deposited marlatti (Quaintance), Dialeurodes citri (Ashmead) in a circular pattern and attached to the leaf with stalks and Dialeurolonga elongata (Dozier). The polyphagous which changes to brownish before hatching (Fig.3a). nature of the non-native A. floccosus warrants stringent The first nymphal instar is light green in colour and quarantine protocols to prevent its further spread to other subsequent stages turn brownish in colour with slight guava and citrus growing areas in India immediately. to heavy wool like wax filaments cover. The adult is Furthermore, continuous monitoring through regular 4 Indian Journal of Entomology, 81(4), 2019

Fig. 2. Aleurothriocus floccosus: a. Typical symptoms of damage, b. eggs, first and second instar nymphs, c. adults

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Fig. 3. Life stages of Aleurothriocus floccosus: a. eggs, b. first and second instar nymphs, c. third instar nymphs, d. puparium First record of the invasive woolly whiteflyAleurothrixus floccosus (Maskell) from india 5 Sundararaj, R et. al.

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(Manuscript Received: Augsut, 2019; Revised: October, 2019; Accepted: November, 2019; Online Published: November, 2019) Preview