木耳菜染色体制片优化及核型分析
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四川农业大学本科质量工程项目(04052109);四川农业大学本科教学改革研究项目(X2015020)


Optimization of chromosome preparation and karyotype analysis of Basella alba
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    【目的】优化木耳菜 Basella alba 染色体制片条件,并了解其核型特征,为今后研究不同类型木耳菜的亲缘关系和系统进化提供理论基础。【方法】以木耳菜品种‘大叶木耳菜’为材料,对影响染色体制片效果的取材部位、预处理时间和解离时间等条件进行优化,并进行核型分析。【结果】木耳菜主根根尖的分裂相细胞和中期分裂相细胞比例最多; 0.002 mol·L-1 的8-羟基喹啉预处理6 h,木耳菜染色体收缩性最好,形态最佳,分散性好;在60 ℃下1 mol·L-1 的HCl解离8 min,木耳菜染色体着色良好且细胞质透明,对比度高。木耳菜的核型公式为2n=2x=44=38m (2SAT)+6sm,染色体相对长度组成为20 M2+22 M1+2 S,染色体长度比为1.93,染色体相对长度变化范围为3.13%~6.06%,着丝粒指数变化范围为35.19%~47.86%,臂比值变化范围为1.09~1.84,第10、16、17对染色体为近中部着丝粒染色体,其余均为中部着丝粒染色体,第13对染色体具有随体,木耳菜核型不对称系数为57.89%,核型按分类标准属1A型。【结论】明确了木耳菜染色体制片的最佳条件参数,并从细胞遗传学角度揭示了木耳菜的核型特征。

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    【Objective】To optimize the chromosome preparation procedure and investigate the karyotype of malabar spinach (Basella alba) and to provide a theoretical basis for studying genetic and phylogenetic relationships of different types of malabar spinach.【Method】The cultivar ‘large leaf malabar spinach’ was used as plant material. Factors that affected chromosome preparation including organ selection, pretreatment and dissociation durations were investigated. With the optimized protocol, the karyotypic characteristics were analyzed.【Result】The proportions of mitotic cells and metaphase cells of the main root tip samples were the highest compared to other sampled organs. Six-hour pretreatment using 0.002 mol·L-1 8-hydroxyquinoline resulted in the highest degree of chromosome contraction, the best shape and evenly spreading of chromosomes. Eight-minute dissociation at 60 ℃ using 1 mol·L-1 HCl resulted in the highest contrast with clear chromosome staining and transparent cytoplasm. The karyotype formula of malabar spinach is 2n=2x=44=38m (2SAT)+6sm. The constitution of relative length is 20 M2+22 M1+2 S, and the ratio of chromosome length to width is 1.93. The relative length ranges from 3.13% to 6.06%, the centromeric index ranges from 3519% to 47.86%, and the arm ratio ranges from 1.09 to 1.84. The 10th, 16th and 17th pairs of chromosomes belong to submetacentric chromosomes and the rest are metacentric chromosomes. Two satellites were observed at the 13th pair of chromosomes. The karyotype asymmetry index is 57.89%, and malabar spinach has the 1A karyotype according to Stebbins’ classification criteria.【Conclusion】This study established an optimal chromosome preparation procedure for malabar spinach,and revealed its karyotypic characteristics from the cytogenetic aspects.

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孙勃,辜金花,童袁桃,张芬,汤浩茹.木耳菜染色体制片优化及核型分析[J].华南农业大学学报,2016,37(6):105-109

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  • 收稿日期:2016-02-05
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  • 在线发布日期: 2016-11-02