Implication of the Whitefly, Bemisia tabaci, Collagen Protein in Begomoviruses Acquisition and Transmission.

Begomoviruses are the largest group of plant viruses transmitted exclusively by the whitefly, Bemisia tabaci (Gennadius), in a persistent, circulative, and nonpropagative manner. Begomoviruses in association with B. tabaci cause enormous loss to world agricultural crops. Transmission, retention, and circulation of begomovirus in B. tabaci are facilitated by its interaction with several proteins of the insect and its endosymbionts. However, very few such proteins have been identified from B. tabaci that are involved in this specific interaction. Here, we have performed yeast two-hybrid assay between B. tabaci complementary DNA expression library and the coat protein (CP) of tomato leaf curl New Delhi virus (ToLCNDV) and cotton leaf curl Rajasthan virus (CLCuV). Collagen was the common protein found to be interacting with both of the viruses. The collagen protein was found to be localized in gut layers of B. tabaci. Additionally, pull-down and dot-blot assays confirmed the association of endogenous collagen with ToLCNDV CP. Immunolocalization analysis also showed colocalization of ToLCNDV particles and collagen within insect gut. Finally, B. tabaci fed on anticollagen antibody and exhibited ∼46% reduction in ToLCNDV transmission, suggesting a supportive role for collagen in virus transmission.

Eusynnada, a resurrected genus of Ochyromerina (Coleoptera: Curculionidae: Curculioninae: Tychiini), with description of a second species from India and Thailand.

Eusynnada is resurrected as a valid genus of Ochyromerina and includes the type species E. plaxoides Heller and a related second new species, E. canariumi sp. nov., from Andaman Island (India) and Thailand.

Descriptions of three new species of Strattis Pascoe, 1883 (Coleoptera: Curculionidae: Cryptorhynchinae) from the Indian subcontinent.

The cryptorhynchine genus Strattis Pascoe originally described from Sri Lanka is recorded from India with the descriptions of three new species: S. pascoei sp. nov., S. maculatus sp. nov., and S. srilankaiensis sp. nov. from the Indian subcontinent. Strattis biguttatus is designated as type species for the genus. A key to species is provided. The character of a presumable stridulatory structure between the meta-femur and the first abdominal ventrite is discussed.

Insilco Prediction and Characterization of microRNAs from Oncopeltus fasciatus (Hemiptera: Lygaeidae) Genome.

For studies on functional genomics, small RNAs, especially microRNAs (miRNAs), have emerged as a hot topic due to their importance in cellular and developmental processes. Identification of insect miRNAs largely depends on the availability of genomic sequences in the public domain. The large milkweed bug, Oncopeltus fasciatus (Dallas) is a hemimetabolous insect which has become a model hemipteran system for various molecular studies. In this study, we identified 96 candidate mature miRNAs from O. fasciatus genome using a blast search with the previously reported animal miRNAs. The secondary structure of predicted miRNA sequences was determined online using "mfold" web server and verified by calculating the minimal free energy index (MFEI). Six miRNAs let-7e, miR-133c, miR-219b, mir-466d, mir-669f, and mir-669l are reported for the first time in Insecta. Comparison of O. fasciatus mir-2 and mir-71 family clusters to those of diverse insect species showed that they are highly conserved. The phylogenetic analysis of miRNAs revealed the evolutionary relationship of conserved miRNAs of O. fasciatus with other insect species. Using a classical rule-based algorithm method, we predicted the possible targets of the new miRNAs. Our study not only identified the list of miRNAs in O. fasciatus but also provides a basic platform for developing novel pest management strategies based on artificial miRNAs.

A Bemisia tabaci midgut protein interacts with begomoviruses and plays a role in virus transmission.

Begomoviruses are a major group of plant viruses, transmitted exclusively by Bemisia tabaci (Gennadius) in a persistent circulative non-propagative manner. The information regarding molecular and cellular basis underlying Begomovirus - whitefly interaction is very scarce. Evidences have suggested that the insect gut possesses some crucial protein receptors that allow specific entry of virus into the insect haemolymph. We have performed yeast two hybrid gut cDNA expression library screening against coat protein of Tomato leaf curl New Delhi virus (ToLCV) and Cotton leaf curl Rajasthan virus (CLCuV) as bait. Midgut protein (MGP) was the common protein found interacting with both ToLCV and CLCuV. MGP was localized in whole mount B. tabaci as well as in dissected guts through confocal microscopy. Pull down and dot blot assays confirmed in vitro interaction between ToLCV/CLCuV coat protein and MGP. Immunolocalization analysis also showed colocalization of ToLCV/CLCuV particles and MGP within insect's gut. Finally, anti-MGP antibody fed B. tabaci, exhibited 70% reduction in ToLCV transmission, suggesting a supportive role for MGP in virus transmission.

A new species of the genus Acria Stephens, 1834 (Lepidoptera: Depressariidae: Acriinae) from India.

A new species, Acria meyricki sp. nov. (Lepidoptera: Depressariidae: Acriinae) occurring on oil palm, is described from India. The status and nomenclature of the genus is reviewed and an annotated checklist of species is given. A key to the seven species known so far from the Indian subcontinent is provided.

Taxonomy of Fulgoraecia melanoleuca (Fletcher, 1939), (Lepidoptera: Epipyropidae) in India, a biological control agent of Pyrilla perpusilla (Walker) (Hemiptera: Lophopidae).

The parasitic lepidopteran insect, Fulgoraecia melanoleuca (Fletcher) has been reported as an ectoparasitoid of Pyrilla perpusilla (Walker) from the Indian subcontinent. For the first time, the complete morphology, field biology, egg laying behavior, larval pupal, and adult morphology, including male and female gentialic features, are described and illustrated.

Distribution of Bemisia tabaci Genetic Groups in India.

The whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a phloem-feeding, economically important pest of crops worldwide. In addition to direct damage, it also vectors a number of plant viruses belonging to the family Geminiviridae. Its populations differ biologically with respect to insecticide resistance, virus transmission and host range. Therefore, understanding genetic variation among populations is important for management. We sequenced 850 bp of the mitochondrial COI (mtCOI) gene from B. tabaci populations surveyed across India. BLAST analysis of the mtCOI sequences generated in this study with sequences from the mtCOI dataset showed the presence of one invasive group, MEAM1, and eight other groups of B. tabaci in India. mtCOI sequence analyses showed the presence of Asia I, Asia I-India, Asia II-1, Asia II-5, Asia II-7, Asia II-8, and Asia II-11 genetic groups. We also found China-3 in a field in Birbhum district, West Bengal, India, suggesting a role of anthropogenic activities in the distribution of B. tabaci. Interestingly, more than one genetic group was found coexisting in the same field.

A new species of Magnimyiolia Shiraki (Diptera: Tephritidae: Trypetinae) and new records of Acanthonevrini from India.

A new species of the subfamily Trypetinae, Magnimyiolia perennifascia Singh & David, sp. nov. (Trypetini) is described from India. Two species of subfamily Phytalmiinae, Ectopomyia baculigera Hardy and Ptilona conformis Zia (Acanthonevrini) are recorded for the first time from India. An updated key to Oriental species of Magnimyiolia Shiraki is provided.

Flea weevils of the genus Megorchestes Kojima (Coleoptera, Curculionidae, Curculioninae, Rhamphini), with description of a second species from India.

A second species of Megorchestes Kojima, M. deccanensis sp. nov., is described from India and a key to the two species is provided.

Detection and localization of Rickettsia sp in mealybug.

Mealybug, Phenacoccus solenopsis Tinsley, is a sap-sucking hemipteran insect. It is an agricultural pest that is now widely distributed in India. In this study we report the presence of Rickettsia from P. solenopsis. We constructed a 16S rRNA gene library to study the bacterial diversity associated with this insect and we found that all the clones from the library were only of Candidatus Tremblaya phenacola. This study also highlights that the normal protocol adopted to study the bacterial diversity from environmental sample, by preparation of a 16S rRNA gene library, does not work when the bacterial population is highly skewed in favor of one bacteria (primary endosymbiont in this case). Hence, we used bacterial genus specific polymerase chain reaction primers to test the presence of any of the widely known secondary endosymbionts associated with insects. We tested for the presence of Cardinium, Rickettsia, Wolbachia, and Arsenophonus in P. solenopsis collected from 10 different locations across India. Only Rickettsia was detected from four locations while we were not able to find any other bacteria. We confirmed the presence of these bacteria by localizing Rickettsia and the primary endosmbiont, Candidatus Tremblaya sp. to the bacteriocyte of P. solenopsis using fluorescent in situ hybridization.

DNA barcoding and elucidation of cryptic aphid species (Hemiptera: Aphididae) in India.

Rapid, precise and timely identification of invasive pest insects such as aphids is important and a challenge worldwide due to their complex life cycles, parthenogenetic reproduction, sex and colour morphs. In this respect, DNA barcoding employing a 658 bp fragment of 5' region of the mitochondrial cytochrome oxidase I (CO-I) gene is an effective tool in addressing the above. In the present study, we employed CO-I for discriminating 142 individuals representing 32 species of aphids from India. Sequence analyses revealed that the intraspecific and interspecific distances ranged from zero to 3.8% and 2.31 to 18.9%, respectively. In addition, the study also showed for the first time the prevalence of three cryptic species, namely Brevicoryne brassicae (Linnaeus), Hyperomyzus carduellinus (Theobald) and Brachycaudus helichrysi (Kaltenbach) from India. Our work has clearly demonstrated that DNA barcoding is an efficient and accurate method for identification of aphid species (including cryptic species), an approach that potentially could play an important role in formulating viable pest management strategies, more especially biocontrol.

Description of a new evacanthine leafhopper species Sophonia chandrai sp. nov., with new record of Chudania axona (Hemiptera: Cicadellidae) from India.

A new evacanthine leafhopper, Sophonia chandrai sp. nov., is described from Manipur, India. The Chinese species, Chudania axona Zhang and Yang is recorded from Meghalaya, India and redescribed. A detailed morphological description, illustrations and photographs are provided. Material is deposited in the National Pusa Collection, Division of Entomology, Indian Agricultural Research Institute, New Delhi, India. A key to the known species of Chudania from the Indian sub-continent is also included.

Dialeurolonga re-defined (Hemiptera: Aleyrodidae): with a new genus and species from India, two new genera from Australia, and discussion of host-correlated puparial variation.

Aleuropositus Dubey gen. n., Australeurodes Dubey gen. n. and Septemaleurodes Dubey gen. n. are proposed with their respective type species as A. sinus Dubey sp. n. from India, D. operculobata Martin & Carver from Australia, and D. swainei Martin from Australia. A. sinus sp. n. is described from Kerala, India, illustrated with line drawings, microphotographs and SEM images. The puparia are asymmetric in taxonomic characters and shape, and variation in puparia associated with a single host is discussed. A generic diagnosis of Dialeurolonga is provided based on SEM study of the type species, D. elongata. Australian species placed in this genus have puparial characteristics that distinguish them from Afrotropical assemblages, and are here referred to two new genera.

Three new species of Kerria (Hemiptera: Coccoidea: Tachardiidae) from India.

Three new species of Kerria Targioni-Tozzetti from India, namely Kerria pennyae Ahmad & Ramamurthy sp. nov. on Schleichera oleosa from Orissa, Kerria dubeyi Ahmad & Ramamurthy sp. nov. on Ficus bengalensis from Bangalore and Kerria varshneyi Ahmad & Ramamurthy sp. nov. on Ziziphus mauritiana from Punjab are described and illustrated, and a key is provided to species of Kerria known from India.

Three new species of Kerria (Hemiptera: Sternorrhyncha: Coccoidea: Tachardiidae), a redesciption of K. yunnanensis Ou & Hong, and a revised key to species of Kerria.

Three new species of lac insect, Kerria Targioni-Tozzetti viz., Kerria manipurensis Ahmad & Ramamurthy sp. nov., Kerria maduraiensis Ahmad & Ramamurthy sp. nov., and Kerria thrissurensis Ahmad & Ramamurthy sp. nov. are described and illustrated. Kerria yunnanensis Ou & Hong is illustrated and redescribed, and a key to the known species of Kerria is provided. The usefulness of star pores as a diagnostic character in the subgeneric divisions of Kerria is also discussed.

DNA barcoding and development of species-specific markers for the identification of tea mosquito bugs (Miridae: Heteroptera) in India.

Rapid, accurate, and timely identification of insects as a group is important and challenging worldwide, as they outnumber all other animals in number and diversity. DNA barcoding is a method for the identification of species in a wide range of animal taxa, which uses the 5' region of the mitochondrial cytochrome c oxidase-I (CO-I). Yet another easy, accurate, and economical method of species discrimination is by developing species-specific markers, which produce specific amplicon for the species in question. The method is handy because it is not limited by life stages, sex, polymorphism, and other factors. Herein, we measured the usefulness of CO-I for the species discrimination of mirids in India viz. Helopeltis antonii Signoret, H. thievora Waterhouse, H. bradyi Waterhouse, and Pachypeltis maesarum Kirkaldy in their various life stages. Furthermore, our study showed the utility of species-specific markers in differentiating H. antonii (295) and H. bradyi (514) regardless of their life stages. Analysis of CO-I gene revealed <1% intraspecific divergence for all four species examined, whereas the interspecific distances ranged from 7 to 13%. This study showed that the DNA barcode and species-specific markers will aid the identification of mirids in India and will stand as a decisive tool in formulating integrated pest management (IPM) strategy, quick identification of invasive and cryptic species, haplotypes, biotypes, and other factors, if any.

Mouthparts and stylet penetration of the lac insect Kerria lacca (Kerr) (Hemiptera:Tachardiidae).

Hitherto less known aspects on mouthpart morphology and penetration mechanism of the lac insect Kerria lacca have been explored. Unique details of the mouthparts, i.e. morphology of labium and stylets and salivary sheath have been brought out. The gross morphology of the mouthparts though resembled other plant sucking homopterans; a two-segmented labium with symmetrically distributed six pairs of contact-chemoreceptors on its surface was distinct; the mandibular stylets had serrations on its extreme apical region, while the maxillary stylets had their external surface smooth with parallel longitudinal grooves on their inner surface. Formation of flanges, salivary sheath and penetration pathway observed along with probing and penetration of the stylets intracellularly up to the phloem cells, as illustrated herein, are the addition to the existing knowledge on the structural details of the mouthparts and the feeding behavior thereupon.

Biology of the mealybug, Phenacoccus solenopsis on cotton in the laboratory.

Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) has been the current topic of research for insect taxonomists and applied entomologists in India due to its invasiveness, rapid spread, morphological and biological variations and the need for establishing an effective control strategy. The biology of the mealybug P. solenopsis was studied on cotton under laboratory conditions between August and October of 2009 with mean temperature and relative humidity of 23.3­30.2° C and 40.5­92.5% RH, respectively, in central India. Neonate crawlers that emerged from a field population were collected and constituted the study population. The developmental period from immature crawler to adult stage was greater for males (18.7 ± 0.9 days) compared to females (13.2 ± 1.8 days), probably due to the additional molt to the pupal stage in males. Survival of second instars was lower (45.5%) than first and third instars (71.4%). Females showed dynamic patterns of fecundity with the number of crawlers produced per female ranging between 128 and 812, with a mean of 344 ± 82. The reproductive period lasted 30.2 ± 8.2 days. Parthenogenesis with ovoviviparity (96.5%) was dominant over the oviparous (3.5%) mode of reproduction. Adult females lived 42.4 ± 5.7 days. Males accounted for less than 5% of the population, and lived 1.5 ± 0.1 days. The life history parameters of P. solenopsis adult females are discussed relative to the appearance of symptoms on the cotton crop, and the importance of making management interventions during the effective reproductive period of the insect.