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银钟花根尖染色体制片及核型分析

花匠小妙招
2025-05-02 12:05

摘要:

以银钟花种子的初生根尖为材料，探讨不同预处理试剂、预处理时间和解离方法对银钟花根尖染色体压片的影响，获得了一套银钟花染色体制片技术，并以此为基础进行核型分析，揭示其核型特征。结果表明：用0.003 mol/L的8-羟基喹啉避光处理6 h，卡诺氏固定液固定10 h，1 mol/L HCl在60℃的恒温水浴锅中解离12 min，20%的卡宝品红染色，可获得分散较好、形态清晰的高质量银钟花染色体压片。银钟花的染色体数目为2n=24，核型公式为2n=24=20m+4Sm，核型不对称系数为55.77%，核型分类属于1B型。

关键词: 银钟花 / 染色体 / 制片技术 / 核型分析 / 根尖

Abstract:

The root tips of P. macgregorii seeds was used as the research material to investigate the effects of different pretreatment reagents, pretreatment time and dissociation methods on the chromosome preparation of P. macgregorii root tips. A set of chromosome preparation technique of P. macgregorii was obtained and used as the basis for karyotype analysis to reveal its karyotype characteristics, providing a foundation for the conservation and genetic research of P. macgregorii. The results showed that pretreatment with 0.003 mol/L 8-hydroxyquinoline in the dark for 6 h, fixation with Carnoy's fixative for 10 h, dissociation with 1 mol/L HCl in a constant temperature water bath at 60 ℃ for 12 min, and staining with 20% carbol fuchsin could obtain high-quality chromosome squash of P. macgregorii with good dispersion and clear morphology. The chromosome number of P. macgregorii was 2n=24, and the karyotype formula was 2n=24=20m+4Sm, with an asymmetry coefficient of 55.77%. The karyotype was classified as type 1B.

图 1 不同预处理方法获得的银钟花根尖染色体形态

注: A—D: 0.003 mol/L 8-羟基喹啉处理2、4、6、8 h; E—H: 0.2%秋水仙素处理3、4、5、6 h; I—L: 饱和对二氯苯处理2、3、4、5 h。

Figure 1. Chromosome morphology of the root tips of P. macgregorii obtained by different pre-treatment methods

图 2 不同解离时间的银钟花染色体形态

注：A为解离10 min；B为解离12 min；B为解离14 min。

Figure 2. Chromosome morphology of P. macgregorii with different dissociation time

图 3 银钟花染色体核型分析及模式图

注：m为中着丝粒染色体；Sm为近中着丝粒染色体; L为长染色体; M1为中染色体; M2为中短染色体; S为短染色体。注: A为银钟花染色体中期分裂相; B为染色体核型分析; C为染色体核型模式。

Figure 3. Karyotype analysis and pattern diagram of P. macgregorii

表 1 根尖预处理方法

Table 1 The pretreatment methods of root tip

序号预处理试剂预处理时间/h 1 0.003 mol/L 8-羟基喹啉 2 4 6 8 2 0.2%秋水仙素 3 4 5 6 3 饱和对二氯苯 2 3 4 5

表 2 银钟花染色体核型参数

Table 2 Karyotypic parameters of P. macgregorii chromosomes

染色体编号相对长度(长臂+短臂)/% 臂比(长臂/短臂) 着丝点位置着丝点指数/% 相对长度系数相对长度类型 1 3.02+3.15=6.17 1.04 m 48.95 1.47 L 2 2.48+3.35=5.83 1.35 m 42.54 1.40 L 3 2.31+2.74=5.05 1.19 m 45.74 1.21 M2 4 1.53+2.81=4.85 1.84 Sm 31.55 1.16 M2 5 1.89+2.45=4.34 1.30 m 43.55 1.04 M2 6 1.53+2.56=4.09 1.67 m 37.41 0.98 M1 7 1.30+2.42=3.72 1.86 Sm 34.95 0.89 M1 8 1.71+1.96=3.67 1.15 m 46.59 0.88 M1 9 1.68+1.89=3.57 1.12 m 47.06 0.86 M1 10 1.74+1.80=3.54 1.03 m 49.15 0.85 M1 11 1.51+1.70=3.21 1.13 m 47.04 0.77 M1 12 0.94+1.10=2.04 1.17 m 46.08 0.49 S [1]

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