采用肾细胞体内注射法分析了中华草龟(Chinemys reevesii)(♀)、中华花龟(Ocadia sinensis)(♂)及其杂种F1代的染色体核型。三种龟染色体倍数为2 n=52,核型公式为18 m+10 st+24 t,臂数NF=80。采用聚丙烯酰胺垂直板电泳法检测了上述三种龟的肌肉、心脏、肝脏、脾脏和肾脏等5种组织的乳酸脱氢酶(LDH)、酯酶(EST)、过氧化物酶(POD)和醇脱氢酶(ADH),分析了三种龟的亲缘关系和遗传多样性。共检测50个基因座位,其中中华草龟15个,中华花龟16个,杂种龟19个,多态位点3个(Ldh-3,Pod-2和Pod-3),多态座位百分比P分别为13.3%、12.5%和15.8%,平均观测杂合度(H0)分别为0.025 6、0.041 7和0.102 3,平均预期杂合度(He)分别为0.041 6、0.040 7和0.074 6,遗传偏离指数(D)分别为-0.386 4、0.023 7和0.372 5。结果表明:杂种F1代群体遗传多样性较高,而亲本群体相对较低;在POD上,杂种F1代与母本中华草龟相近。对比发现,肝脏的POD和LDH及心脏的POD同工酶电泳图谱可以作为鉴别杂种F1代的生化遗传标记。
The karyotypes of hybrid(Chinemys reevesii♀×Ocadia sinensis♂) and their parents were studied from kidney cells by the method of PHA and colchicine injection,and air-dry technique.The results showed that their chromosome formula were 2 n=52,18 m +10 st +24 t,and arm number(NF) was 80.LDH,EST,POD and ADH of five tissues(muscle,heart,liver,spleen and kidney) of C.reevesii,O.sinensis and their hybrid were detected by using vertical polyacrylamide gel electrophoresis.The relationship and the genetic diversity were studied.In 4 isozymes(LDH,EST,POD,ADH),a total of 50 loci were detected,15 in C.reevesii,16 in O.sinensis and 19 in the hybrid.Ldh-3,Pod-2 and Pod-3 were polymorphic,with the mean proportion of polymorphic loci being 13.3%,12.5% and 15.8%.The observed heterozygosity were 0.0256,0.0417 and 0.1023,expected heterozygosity were 0.0416,0.0407 and 0.0746,and genetic deviation index were-0.3864,0.0237 and 0.3725,suggesting that the hybrid had a higher genetic diversity than their parents.Comparison showed that the hybrid was closer to the C.reevesii than to the other in POD.POD and LDH bands detected in liver,POD bands detected in heart could serve as the biochemical markers for distinguishing C.reevesii,O.sinensis and their hybrid.