| 摘要: |
| 为探索取材容易、结果稳定可靠的刺参染色体制备方法, 分别以刺参(Apostichopus japonicus)胚胎和成体为材料对刺参染色体制片方法进行了研究。结果表明, 以原肠后期胚胎为材料采用常规的热滴片法, 细胞有丝分裂指数最高, 获得分裂相效果最好; 以成体呼吸树、成体肠组织为材料时, 也能获得中期分裂相染色体, 但存在着细胞有丝分裂指数低、染色体易丢失等问题; 向刺参体内注射植物血球凝集素(PHA), 能有效促进刺参的有丝分裂; 缩短低渗处理的时间, 同时用甲醇进行第一次固定后再用卡诺氏固定液固定可有效避免刺参染色体的丢失; 实验得出刺参的染色体数目为2n = 46 条。为刺参进一步的遗传学研究如基因定位、染色体原位杂交等提供了基础。 |
| 关键词: 刺参(Apostichopus japonicus) 染色体制备 原肠后期 呼吸树 肠 |
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| 基金项目:农业结构调整重大技术研究专项(06-05-04B); 中华人民共和国水产行业标准(农业部农市发200507330); 中央级公益性科研院所基本科研业务费专项资金(2010-cb-03) |
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| A preliminary study on chromosome preparation of sea cucumber Apostichopus japonicus |
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| Abstract: |
| The metaphase chromosomes of sea cucumber Apostichopus japonicus were prepared from embryos and adults. The results indicate that the late gastrula is the best material for A. japonicus’ chromosomes preparation because it shows the highest mitotic division index and the best chromosomal figures. The chromosomes number of A. japonicus are 2n=46. The method for preparing A. japonicus chromosomes from respiratory tree and intestine of the adult was also established. The animals were injected in vivo with 10 μg of Phytohemagglutinin(PHA) per gram of body weight. After being cultured for 18 to 24 hours, the respiratory tree and intestine were taken out, treated with colchicines 0.08% for 1 hour, swollen with 7% sodium citrate for 40 minutes, fixed with methanol once and then with methanol: acetic acid (3 : 1) twice. Metaphase chromosomes were prepared using the conventional air-dry method. |
| Key words: Apostichopus japonicus chromosome preparation late gastrula respiratory tree intestine |
