| Clostera restitura of Lepidoptera Notodontidae Clostera genus was chosen as the objective inthis article. The morphological differences to the sibling species C.anastomosis of egg, larva,pupae and adult were compared and the evolutionary relationship between them was analyzed. Theantennal morphology and sensilla of C. restitura were observed by scanning electron microscopy(SEM) and the morphology, number and distribution of various sensilla between two sexes werecompared and their possible functions were discussed. Besides, we constructed the full-lengthcDNA library of the leaves of Populus deltoids induced by C. restitura feeding with SMARTtechnique. Main results are as follows:1. Both C. restitura and C.anastomosis are important are significant defoliators of poplars, asthey are very similar in adult morphology, living habits and harmful way, and with overlappingdistribution, they are usually hard to be distinguished from each other. In order to accurately clarifytheir distribution and forecast their population dynamics, the morphological differences of egg,larva, pupae and adult were compared, and the distribution and biological characters wereinvestigated. Apart from some distinctionof male genitalia between C. restitura and C.anastomosis,the adult external morphology are extremely similar. Difference exists on egg arrangement, withthe eggs of C. restitura arranged in platy single layer, while the eggs of C.anastomosis arranged inpiles. Obvious distinction occurred in larva, the body color, body length and strumae distributionare all different. In pupa, C.anastomosis has distinct texture on the back, while C. restitura havenone. The living habits of both moths are very similar, but life cycle of C. restitura are muchshorter than C.anastomosis, thus genernation numbers are larger, too. Since the distribution area ofC. restitura are southern to C.anastomosis, the optimum temperature are higher,25～28℃and18～22℃, respectively. By amplifying part of mitochondria CO Ⅰ gene, the evolutionaryrelationship between them was analyzed. With the total DNA extracted, partial sequences ofmtDNA CO gene were amplified, the identity of the two moth reached up to93%after alignment.Combined with the reported sequences of CO gene from other Clostera insects, NJ tree and MLtree were constructed. From the above phylogenetic trees, C. restitura and C.anastomosis werefound gathered in the same branch, showing that the genetic relationship of the two moths are veryclose.2. Antennae of both sexes of C. restitura are bipectinate, feather-like in shape, consisting oftwo basal segments, scape and pedicel, followed by a long distal flagellum with more than30segments, each having a pair of lateral branches. The antennal spindle is covered by dense scales,sensilla are mainly distributed on the lateral branches. Twelve major sensillar types were identified:two types of sensilla trichodea (ST), two types of sensilla chaetica (SCh), three types of sensilla squamiformia (SSq), two types of sensilla basiconica (SB), two types of B hm Bristles (BB),sensilla coeloclnica (SCo), sensilla auricillica (SA), sensilla styloconica (SSt), sensilla cavitata-peg(SCP), sensilla gemmiformium(SG), sensilla basiconic-mastoid (SBM) and sensilla cylindrica(CY). Sensilla cylindrica and sensilla gemmiformium only exist on female antennae, while sensillabasiconic-mastoid merely distributes on male antennae. The other nine sensilla exist on both sexes,but the distributions and numbers of these sensella differs with each other, showing obviousdimorphism phenomenon.3. After P. deltoids895seedlings (1year old) were fed by the larva of C. restitura, total RNAof the leaves were extracted and the full-length cDNA were synthesized with SMART technique.The ds cDNA which are longer than500bp were collected and ligated to λTripIEx2vector andpackaged to bacteriophages. The induced resistance cDNA library induced by C. restitura wereconstructed by making the packaged bacteriophages infest E. Coli strain XL1-Blue. The titer ofthe original library was more than1×106pfu/ml, the amplified titer was over1010pfu/ml in totaland the recombination rate was more than80%and meets all the requirements for gene screening,which lays a great base for further study of the interaction between insects and plants. |
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