Studies On The Contents Of Male Accessory Glands Of Apriona Germari(Hope),Batocera Lineolata Chevrolat,Anoplophora Chinensis(Forster)

Apriona germari(Hope),Batocera lineolata Chevrolat and Anoplophora chinensis(Forster) are included in the group of Lamiinae (Coleoptera, Cerambycida- e).These long-horned beetles are important stem borer pests in China. The whole larval stage was spended in their host trees. Not only the larvae damaged the xylem of the trees, but also the adult must feed on the bark for sexual mature. Male access- ory glands are important organ in male reproductive system. The contents take part in the process of mating and egg production by transferred together with sperm. For example, the contents can decrease female receptivity , increase egg production and sperm storage. It is valuable to study the ingredient and the physiological function of male accessory glands'contents. Furthermore, the results from the researching the contents of male accessory gland may provide a new method to control the population of beetles.The male accessory glands of three longe horned beetles were used in this paper. This research works included three parts: internal reproductive system study, the component analysis and physiological function analysis of contents of male access- ory glands. Main results are listed as following:1. The male internal reproductive of three long-horned beetles are composed of testes, vasa deferentia, seminal vesciel, ductus ejaculatorius duplex, and accessory glands. But the number of testes and the location of accessory glands are different. There are two testes in each side of male internal reproductive system of Apriona germari(Hope),Batocera lineolata Chevrol. The male accessory glands of Batocera horsfieldi(Hope)and Anoplophora chinensis(Forster) are opened in the posterior seminal vesciel. But the accessory glands of Apriona germari(Hope)are opened in the middle of seminal vesciel.2. The content of soluble protein, total sugar, trehalose and free amino acids(FAA) in the male accessory glands of three long-horned beetles was investigated, and showed different. For Apriona germari(Hope), this four secretions were listed as following: protein>sugar>treholase>FAA, according to the amount. After mating, the content of protein, sugar, FAA, trehalose were felled sharply, then, the amount increased gradually. But the recovered time of each secretion was different. For example, it took 48h for mated male to recover the content of protein.3. Male accessory gland proteins of three long-horned beetles were analyzed by PAGE and SDS-PAGE. The study revealed remarkable difference in the patterns. For Apriona germari(Hope), the protein fraction of big male accessory glands migrated in four groups. But the protein fraction of small accessory glands had only one group, consisting of high molecular weight fractions. This proteins are PAS positive. Analyzed by SDS-PAGE, the protein also could be categorized four groups, accordi- ng to the molecular weight. The protein fraction of 48h after mating showed the sam- e patterns of virgin male.4. The crude proteins, sugar and free acid amino were extracted from the contents of male accessory glands. Injecting the three crude extractions and total secretions into the female of Apriona germari(Hope) showed that total secretion deduced the longevity of female. The crude of sugar could stimulate the female'feeding activity, at the same time, the female beetles whose treated with it lived longest.5. There are a lot of spermatozeugma in the testes of Apriona germari(Hope). But this is individual sperm in seminal vesicle. The size of sperm is as follows: 79.2～94.1um long, 3umwide, and the head is 11.6～17.2um long. There were no significant differences between the sperm cultured in Grace's Insect Medium and in M199 culture medium. In fact, the sperm could survival until 6h in either of it. It was suggested that the secretions of male accessory glands may have the materials which can provide the energy to sperm or protect the sperm.6. The proteins were isolated and purified from the male accessory gland of Apriona germari(Hope) by a procedure involving Sephadex G-50 gel filtration chromatography and DEAE- A50 ion exchange chromatography. Fourth protein fractions were purified from gel filtration chromatography. After cultured with the fractionⅠ, the rate of mobile sperm was 37.1%. The acrosomal integnity of sperm was observed with fluorescence staining by CTC. The result showed that sperm was"F"type. Two protein fractions were purified from DEAE-A50 ion exchange chromatography. The fractionⅡalso promoted the activation of sperm, and the average rate was 42.1%. The sperm was"F"type too.