Selection and Optimization of Chromosome Preparation for Wheat Roots
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摘要: 为了获得小麦根尖细胞染色体的优良制片，清晰观测到小麦染色体，以Taichung 29、中国春、中4的根尖为材料，通过时间、染色、解离和制片4个方面对常规方法和酶解法进行比较，并且在这4个方面的一些细节方面进行了优化，使得在显微镜下能清晰地数出小麦染色体的数目。常规方法整体操作时间较长达到7 d，解离程度控制较精细，需要严格把控解离时间，并且对染色的时间长短要求比较严格，更需要在压片过程中严格控制力度单张玻片进行压制；而酶解法无须染色而且时间较短大约4~5 d，酶解过程较易控制并且只需要对根尖捣碎不需要进行压片，控制相对容易很多，一次可进行大量制片。对比2种制片方法可以发现酶解法比较适合进行大量染色体制片。Abstract: To obtain desirable chromosome samples for studying wheat roots, two preparation methods were compared for optimization. The root-tip cells of three wheat varieties (i.e., Taichung 29, Chinese Spring and Zhong 4) were collected for the study on the conventional and the enzymatic digestion methods. Aiming for microscopically clearly defined chromosome samples, the evaluation and optimization criteria encompassed the operation time, staining procedure, chromosome dissociation and sample preparation of the methods. The conventional method took up to a week to operate, and required precise controls in dissociating and staining the chromosomes as well as preparing microscope slides. On the other hand, the enzymatic method was conducted by pulverization without staining and slice-making, and the entire procedure could be completed in 4-5 days. Not only that the procedures were simpler and less prone to errors, the enzymatic method could generate many specimens in a single run. Consequently, it was considered superior to the conventional method for the chromosome sample preparation.
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Key words:
- wheat /
- root-tip /
- chromosome preparation /
- enzymatic digestion method
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图 1 不同染色时间的小麦根尖染色体
注：A为染色时间过短形态; B为染色时间正常形态; C为染色时间过久形态。
Figure 1. Chromosome morphology of wheat root-tip cells prepared by varied staining time
图 2 常规方法下小麦根尖染色形态
注：A为Taichung 29，2n=42; B为中4，2n=56。
Figure 2. Chromosome morphology of wheat root-tip cells prepared by conventional method
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