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Screening and Field Evaluation of Synthetic Volatile Blends Attractive To Chemoecology DOI 10.1007/s00049-012-0110-1 CHEMOECOLOGY RESEARCH PAPER Screening and field evaluation of synthetic volatile blends attractive to adults of the tea weevil, Myllocerinus aurolineatus Xiao-Ling Sun • Guo-Chang Wang • Yu Gao • Zong-Mao Chen Received: 1 June 2012 / Accepted: 29 June 2012 Ó Springer Basel AG 2012 Abstract Plant volatiles are known to play a role in host host, and is a strong candidate for an attractant that could location in many herbivorous insects. Although a few be used to control the weevil. studies have determined the role of specific chemicals from herbivore-induced plant volatiles (HIPVs) in mediating Keywords Attractant Á Synthetic volatile blends Á interactions between conspecifics in insects belonging to Myllocerinus aurolineatus Á Tea plant Curculionidae, little is known about how this process works when different components are used. By measuring the behavioral responses of the tea weevil, Myllocerinus Introduction aurolineatus (Voss) (Coleoptera: Curculionidae), to a ser- ies of chemicals in a Y-tube olfactometer, we found that a Herbivore-induced plant volatiles (HIPVs) have both blend containing (E/Z)-b-ocimene and (Z)-3-hexenyl ace- repellent and attractive roles in host location by conspecific tate was attractive to male M. aurolineatus adults, and a and nonspecific herbivorous insects. For example, the two- blend containing 2-phenylethanol and (Z)-3-hexenyl ace- spotted spider mite Tetranychus urticae (Dicke 1986), the tate was attractive to female M. aurolineatus adults; both tobacco budworm Heliothis virescens (De Moraes et al. blends were as attractive to the insects as the volatiles 2001) and the western flower thrips Frankliniella occi- emitted by the tea plants infested with adult weevils. A net dentalis (Delphia et al. 2007) are reportedly repelled by cage experiment in the laboratory showed that traps baited plant volatiles induced by conspecifics. Ka¨nnaste et al. with (E/Z)-b-ocimene plus (Z)-3-hexenyl acetate attracted (2009) found that methyl salicylate and (-)-linalool, the male herbivores, whereas the traps baited with released from mite-infested spruce trees, significantly 2-phenylethanol and (Z)-3-hexenyl acetate did not. Field decreased the attractiveness of the tree to pine weevils experiments verified that tea plants exposed to a blend of Hylobius abietis (L.). The attractiveness of HIPVs to her- (E/Z)-b-ocimene plus (Z)-3-hexenyl acetate attracted sig- bivorous insects has been observed mainly for beetles nificantly more weevils than did the control plants or sticky (Coleoptera) with aggregative behavior, which has been traps baited with the above lure. These results suggest that discussed in detail by Sun et al. (2010). Surprisingly, most the blend of (E/Z)-b-ocimene and (Z)-3-hexenyl acetate is of these studies have measured only behavioral responses an important signal which the tea weevil uses to locate its in Curculionidae beetles. To our knowledge, only a few have determined the role of specific chemicals from HIPVs in mediating interactions between conspecifics. For instance, Diaprepes abbreviatus (Linnaeus) and Myllocer- X.-L. Sun (&) Á Y. Gao Á Z.-M. Chen Tea Research Institute, Chinese Academy of Agricultural inus aurolineatus Voss have been verified to be attracted Sciences, Hangzhou 310008, China by the specific volatiles emanating from host plants infes- e-mail: [email protected] ted by conspecific weevils in the laboratory (Ota´lora-Luna et al. 2009; Sun et al. 2010), while Sitophilus granarius G.-C. Wang Henan Institute of Science and Technology, (L.) adults were verified to have the ability to respond Xinxiang 453000, China behaviorally to a wide range of cereal volatiles. Moreover, 123 X.-L. Sun et al. that response may change as a function of concentration Experimental Plantation (Tea Research Institute, Chinese (Germinara et al. 2008). But whether these chemicals are Academy of Agricultural Sciences, Hangzhou Xihu still attractive to herbivores in the field remains unclear, Longjing First-degree Reserve, China). The insects were except that (E)-2-hexen-1-ol and (Z)-3-hexen-1-ol were separated according to their morphological characteristics found to be attractive to the gray corn weevil, Tanymecus into groups of males and females and maintained in sepa- (Episomecus) dilaticollis Gyllenhal, when presented indi- rate net cages (75 9 75 9 75 cm) with fresh tea shoots. vidually in the field (Toshova et al. 2010). Net cages were kept in a controlled climate room pro- The tea weevil, M. aurolineatus (Voss) (Coleoptera: grammed at 25 ± 2 °C, 70 ± 5 % r.h., and a photoperiod Curculionidae), is native to China. This species has one of 13:11 h (L:D). Two weeks later, weevils were chosen generation per year, and overwinters as larvae in the soil. It for experiments. Before testing, the weevils were starved is a leaf-feeding pest of the tea plant Camellia sinensis O. for 2 h. Ktze., and has aggregative behavior that can seriously reduce tea yield and quality (Zhu et al. 1988). In recent Olfactometer test years, outbreaks of this weevil have occurred in eastern and western regions of China. Though chemical controls are Responses of M. aurolineatus adults to different odors effective against this weevil, they pose health and envi- released from different sources or combinations of syn- ronmental risks. Alternative strategies for controlling this thetic compounds were measured in a Y-tube olfactometer, weevil are urgently needed. Our previous study showed which was described in detail by Sun et al. (2010). Briefly, that the infestation of M. aurolineatus adults increased the the olfactometer consisted of a Y-shaped glass tube of emission of more than 40 volatiles. Among these, c-ter- 2.5 cm i.d. The base tube and two arms of the olfactometer pinene, benzyl alcohol, (Z)-3-hexenyl acetate, myrcene, were each 12 cm long. Each arm was connected to an odor benzaldehyde and (Z)-3-hexenal were found to be attrac- source container. An air stream was generated by a vacuum tive to both males and females, (E/Z)-b-ocimene and pump and divided, and each secondary air stream was sent (E,E)-a-farnesene were attractive only to male adults, and through a flowmeter, a tube with an active charcoal filter, a (E)-4,8-dimethyl-1,3,7-nonatriene, 2-phenylethanol, linal- humidifier bottle, and one of the odor containers. Subse- ool, and (Z)-3-hexenol were attractive only to females (Sun quently, the two airstreams passed through the two arms of et al. 2010). However, nothing is known about the attrac- the Y-tube olfactometer at 800 ml min-1. The Y-tube tive activity of these chemicals in the field. In order to olfactometer was placed in a green box (1.5 m long 9 1m explore the tea weevil’s attractants, we first determined the wide 9 1.5 m high). Each edge of the box top was illu- attractiveness of mixtures of some of these chemicals to the minated by a single 25 W lamp. All bioassays were herbivore and compared this attractiveness with those of conducted between 15:00 and 20:00. During experiments, the herbivore-induced tea volatiles in the lab. Then, we the temperature in the room was maintained at 25–27 °C. tested the attractiveness of the candidate mixtures to this Weevils were allowed to walk upwind towards the odor weevil in the field. sources. Each weevil was individually introduced into the base tube. An odor source was considered to have been chosen when a weevil crossed a line 8 cm after the division Methods and materials of the base tube and remained there for at least 1 min. A weevil that did not make a choice within 5 min was con- Tea plants sidered a non-responding individual and recorded as ‘‘no response’’. After 2 weevils were tested, the olfactometer Longjing tea plants were individually planted in plastic tube was washed with 98 % acetone and then heated at pots (14 cm i.d. 9 15 cm high), irrigated once every other 100 °C for 5 min. The position of odor source containers in day, and fertilized with rapeseed cake once a month. Plants relation to each arm was reversed after each replication to were kept in a controlled climate room, which was main- eliminate directional bias. To avoid contamination, the tained at 26 ± 2 °C, 70-80 % r.h., and 12-h photophase. connections of the odor chamber to the flow meter were The light intensity for plants was about 450 lmol pho- rinsed with 98 % acetone and then heated at 100 °C for tons m-2 s-1) during the photophase. One-year-old plants 5 min after each replication. were used for experiments. Response of M. aurolineatus adults to synthetic blends Insects of volatile compounds Adult weevils of mixed age and sex and of unmated status Our previous study showed that (E/Z)-b-ocimene and were collected from damaged plants on the Tea 2-phenylethanol were abundant components in the 123 Synthetic volatile blends attractive to adults of the tea weevil herbivore-induced tea volatiles. In search of potent attrac- 2-phenylethanol (10 mg); (6) PMC-F versus 2-phenyletha- tants, we chose (E/Z)-b-ocimene or 2-phenylethanol as a nol (10 mg); (7) PMC-M versus (E/Z)-b-ocimene (10 mg): control chemical, added one or two chemicals out of the 6 (Z)-3-hexenyl acetate (0.6 mg); (8) PMC-F versus (E/Z)- active chemicals in the control and then measured the b-ocimene (10 mg):(Z)-3-hexenyl acetate (0.6 mg); (9) difference in attractiveness of the mixtures compared to the PMC-M versus 2-phenylethanol (10 mg):(Z)-3-hexenyl control chemical. The behavioral responses of the adult acetate (0.7 mg); (10) PMC-F versus 2-phenylethanol weevils to the above pairs of odors, belonging to two (10 mg):(Z)-3-hexenyl acetate (0.7 mg).
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