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Electrophysiology and Behaviour in Banana Pseudostem Weevil Odoiporus Longicollis Olivier to Conspecific Chemical Cues E:\ 82(1)

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Electrophysiology and Behaviour in Banana Pseudostem Weevil Odoiporus Longicollis Olivier to Conspecific Chemical Cues E:\ 82(1) E:\ 82(1) 19136-A T Rani 2020 Indian Journal of Entomology, 82(1): 000-000 (2020) DoI No.: ELECTROPHYSIOLOGY AND BEHAVIOUR IN BANANA PSEUDOSTEM WEEVIL ODOIPORUS LONGICOLLIS OLIVIER TO CONSPECIFIC CHEMICAL CUES A T RANI*, K R M BHANU**, VASUDEV KAMMAR*** AND A K CHakravartHY**** *Division of Vegetable Protection, ICAR-IIVR, Varanasi 221305, Uttar Pradesh **Biocontrol Research Laboratory, PCI, Bengaluru 562163 ***Department of Food and Public Distribution, Ministry of Consumer affairs and Food and Public Distribution, Govt. of India, Krishi Bhawan, New Delhi 110001 ****Division of Entomology and Nematology, Indian Institute of Horticulture Research, Hessaraghatta 560089 *Email: [email protected] (corresponding author) ABSTRACT An understanding of the role of semiochemicals in insect biology can be used for developing IPM tools. The olfactory and behavioural responses of banana pseudostem weevils (BPW) Odoiporus longicollis Olivier to semiochemicals isolated from conspecific insects were investigated by electroantennography (EAG) and Y-tube olfactometer bioassays. Volatiles were collected from male and female weevils, separately and together from both sexes by air entrainment. Dichloromethane extracts of male and female weevils were used as stimuli in electrophysiological study. The observations revealed that the antennae of both male and female produced significant EAG responses (1.378±0.350 mV and 1.704±0.246, respectively) to male volatile compared to other stimuli tested. In two-choice experiments, more males (p=0.001, χ2=11.84) and females (p= 0.033, χ2=4.57) were attracted to male volatiles. Significant responses of both sexes to headspace volatiles from male extracts during EAG and olfactometer experiments revealed that specific volatile compounds emitted by male played a significant role in aggregation. These results might enable identification of male specific pheromone components, which can enhance effectiveness of pheromone trapping for O. longicollis. Key words: Air entrainment, dichloromethane, semiochemicals, Odoiporus longicollis, electroantennography, Y-tube olfactometer, male and female volatiles, differences Banana Musa spp. is cultivated about 0.858 million that BPW incurs 10-90% yield oss (Padmanaban and ha with the production of 29.16 million MT and the Sathiamoorthy, 2001). productivity of 34 MT/ ha in India (Horticultural Statistics, 2017). The incidence of insect pests is a major The biology, ecology, seasonal incidence and constraint for banana cultivation. About 19 pests are chemical control of BPW had been previously found associated with banana in India (Padmanaban studied (Azam et al., 2010; Priyadarshini et al., 2014; et al., 2001). Of these, the banana pseudostem weevil Khairmode et al., 2015). Because of the long life (BPW) or banana pseudostem borer Odoiporus span of adults and endophytic behavior of the larvae, longicollis Olivier (Coleoptera: Curculionidae) is conventional methods of control, especially insecticides gaining importance as a serious pest and its occurrence were found less effective (Prasuna et al., 2008). Also, had been reported in nontraditional areas too (Justin et insecticides can be harmful to non-target species and al., 2008). The weevil is becoming a serious threat to leave residues. Hence, alternative control methods that banana crop in South India. BPW is a monophagous are safe and efficient are warranted. Semiochemicals pest and the adult femalesPreview lay eggs in the outermost like pheromones provide effective control (Welter leaf sheath of banana (Visalakshi et al., 1989). Larvae et al., 2005). Successful management of weevils in hatched from the eggs bore into the living tissue, cotton, coconut and sweet potato using aggregation producing frass- filled tunnels and permit the invasion pheromones had been reported (Dickerson et al., 1987; of fungal and bacterial pathogens. Mature larvae pupate Chinchilla et al., 1996; Downham et al., 2001). Sordidin, in cocoons made from plant fibres close to the exit holes. the aggregation pheromone of Cosmopolites sordidus The severity of the loss is greater when infestation Germar, another important pest of banana, had been occurs at the early vegetative stage. It is estimated successfully used (Jayaraman et al., 1997; Tinzaara et al., 2000). 2 Indian Journal of Entomology, 82(1), 2020 It had been reported that male BPW produced a pheromone, 2-methyl-4-heptanol which attracted both male and female weevils (Gunawardena and Dissanayake, 2000a). Ravi and Palaniswami (2002) reported the presence of its female sex pheromone. The aggregation pheromones of several weevil species consist of multiple behaviorally active components. The advantage of multicomponent pheromones is that they convey more information than a component (Silverstein, and Young, 1976). The combination of one or more pheromone components increases the attractiveness of insects to pheromone baited traps. Gunawardena and Dissanayake, (2000) reported that 2-methyl-4-heptanol works only with food bait under field conditions. The single component aggregation pheromone ineffective in trapping more weevils. Therefore, the present study to determine the presence of other pheromone components to improve the existing pheromone trapping method by increasing the attractiveness to the sexes and also to develop multicomponent pheromone- based control method for BPW. MATERIALS AND METHODS Odoiporus longicollis adults of mixed age and sex and of unknown mating status were collected from damaged banana plants around Tubgere hobli, Fig. 1. Banana pseudostem weevil (A) male (B) female Doddaballapur Taluk (1306’, 77036’, 930 masl) and Hesaraghatta hobli, Bengaluru (1305’,77035’, 930 m every 5 min for a 12 hr period both during scotophase AMSL). Males and females were separated based (6 pm to 6 am) and photophase (6 am to 6 pm). During on the rostrum characteristics (Fig. 1). Weevils were the scotophase, observations were facilitated using a maintained in separate plastic containers (29×17×33 red bulb (5 W). Observations on number of copulation cm) in laboratory under a photoperiod of 12L: 12D events/ pair and mean number of mating pairs were at 25± 20 C, 70± 10% RH at the Biocontrol Research recorded and the observations repeated twice. laboratories (BCRL), Pest Control (India) Pvt. Ltd., Prior to insect volatile collection, preliminary trials Sriramanahalli, Bengaluru. Weevils were provided with were conducted to determine the peak time of mating freshly cut pseudostem pieces as food. Unfed pieces activity of male and female BPWs during scotophase contaminated with fecal matter were replaced weekly. and photophase under caged conditions. The volatile A laboratory study was conducted to determine the collection was carried out between 1100 to 1400 hr of peak time of mating activity and timing of pheromone photophase by air entrainment method. Groups of 50 release. Experiments were conducted during scotophase males and 50 females were maintained separately in and photophase of weevils in the Biocontrol Research different glass aeration chambers (6 x 19 cm). Thirty Laboratory, Bengaluru to understand the time of activity. males and 30 females were held separately in wire mesh In this study, number of mating pairs and frequency of and wooden cages (0.3 m3) and both cages were kept Previewin a single plastic acrylic container (6.5×12×17 cm) mating in each hour were recorded. Five pairs of male and female in three replicates (n= 15 pairs) were placed for volatile collection from male and female BPWs. in separate transparent acrylic boxes (19×15×9 cm) A charcoal filtered humidified airstream was pushed covered by a muslin cloth tied with an elastic ribbon. through the aeration system at 1.8 L min-1 (Fig. 3). Weevils were provided with fresh pieces of banana Emitted volatiles were collected daily in glass column pseudostem (3×2 cm). The study was conducted at trap filled with a 10 mg of Porapak-Q adsorbent. The 25± 20 C, 70± 10% RH. Weevil pairs were analyzed weevil volatiles were collected from the Porapak-Q with Electrophysiology and behaviour in banana pseudostem weevil odoiporus longicollis olivier to conspecific chemical cues 3 A T Rani et al Fig. 2. Frequency of matings (n=15 pairs); estimated hourly in 1st to 12th hr Fig. 3. Insect volatile collection (air entrainment method) 1.5 ml of HPLC grade Dichloromethane (DCM) for × 0.25 μm film (J and W Scientific, Folsom, CA, USA). 15 min. The extracts were concentrated under nitrogen The GC was run at a flow rate of 5 ml min-1 with a stream and reduced to 1 ml so that the volatiles are in oven temperature from 60 to 220 °C with a hold of 2 diluted form for direct Previewuse for different studies. Extracts min at 60°C. The chromatograms were analysed for were stored at -200C in a refrigerator when not in use. the presence of common and sex-specific pheromone components. Gas chromatographic (GC) analyses of the contents of volatiles collected from both sexes of O. longicollis The electroantennographic system used for the study separately were performed on Agilent 7890A GC system was Syntech, Hilversum, The Netherlands. Antenna equipped with flame ionization detector. The GC was of the weevils was excised using a micro scissor in a fitted with a DB-5 capillary column (30 m × 0.25 mm way that all the antennal segments and the basal nerve 4 Indian Journal of Entomology, 82(1), 2020 remained intact. An excised antenna was mounted female volatile in DCM (100 μl) vs DCM (100 μl) and on to an antenna holder for the EAG probe in such a (4) only DCM (100 μl) (Control). The bioassays were way that the tip of the antenna was in contact with -ve carried out between 1100 to 1400 h during photophase. electrode and base i.e. scape touched +ve electrode. This was accomplished using electroconductivity gel Electroantennogram responses were compared (Sigma-gel, Parker Laboratory Fories. Inc. USA). using a one-way ANOVA followed by Honestly The different stimuli tested were (1) male volatile, (2) Significant Different (HSD) Tukey test (SPSS 16.0 female volatile, (3) male + female volatile and (4) DCM version software). Results of all olfactometer bioassays 2 solvent (control).
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