中国农业科学 ›› 2004, Vol. 37 ›› Issue (11): 1682-1698 .

• 畜牧.兽医.资源昆虫 • 上一篇    下一篇

家蚕微粒子虫(Nosema bombycis)与其形态变异株的侵染性及孢子表面蛋白的比较研究

黄少康,鲁兴萌   

  1. 浙江大学动物科学学院
  • 收稿日期:2004-03-01 修回日期:1900-01-01 出版日期:2004-11-20 发布日期:2004-11-20
  • 通讯作者: 鲁兴萌

Comparative Study on the Infectivity and Spore Surface Protein of Nosema bombycis and Its Morphological Variant Strain

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  1. 浙江大学动物科学学院
  • Received:2004-03-01 Revised:1900-01-01 Online:2004-11-20 Published:2004-11-20

摘要: 用家蚕微粒子虫(Nosema bombycis,Nb)连续感染桑尺蠖(Hemerophila atrilineata)24次后,获得孢子显著变粗短(P <0.01)的形态变异株(24Nbh)。Nb与24Nbh的侵染性及表面蛋白的比较研究表明,24Nbh感染2龄起蚕(Bombyx mori)的感染中浓度(IC50)为1.98×104spores/ml,而Nb为1.72×103 spores/ml,即Nb连续感染桑尺蠖24次后对家蚕的侵染性下降了11.5倍。24Nbh回复感染家蚕一次后的孢子,对2龄起蚕的IC50与Nb对照相差6.9倍,即对家蚕的侵染性又快速恢复;与24Nbh相比,孢子长径及长短径比虽然显著回复(P <0.01),但仍小于Nb(P <0.05)。SDS-PAGE显示,Nb和24Nbh孢子表面蛋白的带型基本一致,均有4条主带(12kD、17kD、30 kD、33 kD),但在量上差异明显。2D-PAGE显示,当上样量均为120μg时,二者孢子表面蛋白都以中偏酸性蛋白(pI5~7)为主,但表面蛋白在量和种类上均存在明显差异,发现5个可疑的差异蛋白点,推测这种差异与微孢子虫的侵染性变异有关。

关键词: 家蚕微粒子虫, 形态变异, 侵染性, 表面蛋白, 双向电泳

Abstract: A new morphological variant strain was produced by infecting the mulberry looper, Hemerophila atrilineata [Phthonandria atrilineata], with Nosema bombycis successively for 24 times, and named 24Nbh. Comparative studies on morphology, infectivity and spore surface protein were conducted. 24Nbh was short and wide, and had a significant difference (P<0.01) over the Nb spores. The infectivity tests conducted on second instar silkworm larvae showed that IC50 of 24Nbh was 1.98×104 spores/ml and of Nb was 1.72×103 spores/ml, thus indicating that the infectivity of Nb decreased 11.5 times after multiplying in mulberry looper for 24 times. The IC50 of spores from silkworm infected with 24Nbh was 6.9 time less than Nb, showing that the infectivity of 24Nbh spores rejuvenated very fast when reinfected to silkworms, furthermore, the length and width of such spore was larger (P<0.01) than 24Nbh and smaller (P<0.05) than Nb. The SDS-PAGE profiles of Nb and 24Nbh were generally the same, distinct 4 proteins of 12 kD, 17 kD, 30 kD, 33 kD were obtained with differences in quantity. When 120 μg of protein was used for 2D-PAGE, five suspected different proteins with difference in quantity was observed. These results demonstrate that these differential proteins maybe associated with variation in infectivity.

Key words: Nosema bombycis, Morphological variation, Infectivity, Surface protein, Two dimensional electrophoresis