四川农业大学学报 ›› 2007, Vol. 25 ›› Issue (03): 277-281.doi: 10.16036/j.issn.1000-2650.2007.03.009

• 研究论文 • 上一篇    下一篇

根尖压片法制备树莓染色体标本的研究

王小蓉, 汤浩茹, 李玲, 段娟, 邓群仙   

  1. 四川农业大学 林学园艺学院, 四川 雅安 625014
  • 收稿日期:2007-05-08 出版日期:2007-09-30 发布日期:2017-03-04
  • 基金资助:
    国家自然科学基金(30671454);四川省教育厅项目(2004A025)。

Studies on Root-tip Squashing Method for Chromosome Preparation of Rubus L.

WANG Xiao-rong, TANG Hao-ru, LI Ling, DUAN Juan, DENG Qun-xian   

  1. College of Forestry and Horticulture, Sichuan Agricultural University, Yaan 625014, Sichuan, China
  • Received:2007-05-08 Online:2007-09-30 Published:2017-03-04

摘要: 以红泡刺藤(R. niveus)、栽秧泡(R. ellipticusvar.obcordatus)和插田泡(R. coreanus)绿枝或硬枝扦插生根的根尖为材料,通过取材时季与扦插环境气温,8-羟基喹啉、秋水仙素和对二氯苯3种药剂在不同的温度下预处理不同时间,混合酶液和盐酸在不同温度下解离,醋酸洋红、卡宝品红、席夫试剂和铁矾-苏木精各染色剂染色压片等的对比研究,并以细胞中有丝分裂中期分裂相的数目、染色体的清晰性、分散程度和聚缩程度以及染色体的形态为衡量指标,探索出适于树莓属植物核型分析的根尖压片方法是:当根尖长至1~2 cm、扦插环境气温17~23℃时取材,卡诺氏固定液I低温固定24 h,用0.002 mol/L 8-羟基喹啉在0~4℃下预处理24 h,再用1 mol/L HCl(60±1)℃恒温解离4~7 min,最后用卡宝品红染色压片能获得效果好树莓染色体标本,该染色体标本完全适用于核型分析。

关键词: 树莓, 根尖压片法, 染色体标本

Abstract: Using the root-tips of Rubus niveus, R. ellipticus var. obcordatus and R. coreanus as materials, the key factors, such as the sampling times and environmental temperatures when sampling materials, pretreatment agents (8-hydroxyquinoline, 8-hydroxyquinoline plus colchicine and saturated p-dichlorobenzene), temperatures and times, dissociating agents (25 g/L cellulase plus 25 g/L pectic enzyme and hydrochloric acid), processes and times and staining agents (acetocarmine, carbol fuchsin, schiff' solution and ferriammonium sulfate-haematoxylin), and methods, which influences the preparation of chromosome, had been studied. Based on the contrasts of numbers of mitosis metaphase cells, the difficulty of techniques, the degree of treatment complications and the fact that chromosomes are distinct and countable from their background under microscope, the suitable root-tip squashing method for chromosome preparation of Rubusis as follows: sampling materials when the roots are 1~2 cm in length and the environrmental temperature is 17~23℃ and fixating with Carnoy's I, pretreating with 0.002 mol/L 8-hydroxyquinoline at 0~4℃ for 24 h, dissociating with 1 mol/L HCl at (60±1)℃ for 4~7 m, and staining with carbol fuchsin solution. The chromosomes obtained by above preparation method could be used for karyotype analysis.

Key words: Rubus, root-tip squashing method, chromosome preparation

中图分类号: 

  • S663.2