迪庆野生型百脉根的染色体制片优化与核型分析

Optimization of Chromosome Sectioning and Karyotype Analysis of Wild Type Lotus corniculatus in Diqing

采用常规压片法制片,以里奥百脉根为对照组,研究不同取材时间、预处理方法、解离方法以及染色时间对迪庆野生型百脉根根尖细胞染色体制片效果的影响,优化制片技术并进行核型分析。结果表明,于900-1100时段取材,0.02%放线菌酮预处理24h,在V_(无水乙醇)︰V_(浓盐酸)=1︰1解离液中解离10min,以卡宝品红染色8min染色体制片效果较好。核型分析表明,迪庆野生型百脉根核型公式2n=2x=12=8m+4sm,核型不对称系数为63%,为1A核型;栽培型百脉根核型公式2n=2x=24=16m+8sm,核型不对称系数为62%,为2B核型。迪庆野生型百脉根的核型为首次报道。

With Lotus corniculatus L.‘Leo'as control,the effects of different chromosome preparations to root-tip of wild type Lotus corniculatus in Diqing in terms of sampling time,the pretreatments,dissociations and staining time was studied,the chromosome sectioning technique was optimized and karyotype analysis was conducted. The results showed that optimal chromosome number could be obtained when Lotus corniculatus was sampled at 900-1100 am,pretreatments for 24 h with 0. 02% cycloheximide,which were dissociated for 10 min with dissociation liquid( the volume ratio of anhydrous alcohol and hydrochloric acid was 1 to 1),and stained for 8 min with carbol fuchsin solution. Karyotype analysis indicated that karyotype were 1A( 2n = 2x = 12 = 8m+4sm) and 2B( 2n = 2x= 24 = 16m+8sm),the index of karyotype asymmetry were 63% and 62% in wild type Lotus corniculatus in Diqing and cultivated Lotus corniculatus,respectively. It is for the first time that karyotype of wild type Lotus corniculatus in Diqing was reported.