橡胶树叶片染色体制片方法的优化

Optimization of Chromosome Preparations of Hevea Leaves

采用压片法、酶解去壁低渗法和悬液法等方法,对橡胶树热研7-33-97古铜期嫩叶染色体制片及其过程中预处理药剂、预处理时间、前低渗温度、酶解时间等因子进行了探讨。结果表明:悬液法更适合橡胶树叶片染色体制片。最佳预处理药剂为0.1%的秋水仙素和0.002mol/L8-羟基喹啉混合液,预处理时间为2h。酶解温度37℃,时间为5～7h。

Chromosome preparation is of great significance for chromosome and karyotype analysis in ploidy breeding, interspecific hybridization and germplasm analysis of Hevea brasiliensis Muell. Arg. Chromosome preparations of bronze-coloured tender leaves of Reyan 7-33-97 were made by using the squash method, the enzyme and low osmotic method and suspension method, and some important factors during the preparation, such as the chemicals for pretreatment, pretreatment time, early low osmosis temperature, enzymolysis time, etc. were discussed to optimize the chromosome preparation. The result showed that the suspension method was the best one for chromosome preparation of the tender rubber leaves. The optimum pretreatment chemical was the mixture of 0.1% colchicine and 0.002mol/L 8- Hydroxyquinoline（1：1） , and the pretreatment time 2 hrs. Enzymolysis lasted 5 -7 hrs at 37 ℃.