An improved method for inducing prometaphase chromosomes in plants.
An improved method for inducing prometaphase chromosomes in plants.
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DOI：
10.1186/s13039-018-0380-6
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发表时间：
2018
影响因子：
1.3
通讯作者：
Koba T
中科院分区：
文献类型：
--
作者：
Setiawan AB;Teo CH;Kikuchi S;Sassa H;Koba T
甜瓜（Cucumis梅洛L.）仍然是一个挑战，因为他们的染色体尺寸小，染色体差。前中期染色体，这是两倍长，松散凝聚，提供了一个显着更好的分辨率比中期染色体荧光原位杂交（FISH）。然而，甜瓜前中期染色体的获取方法还不完善。在这项研究中，改良的Carnoy溶液II（MC II）[6：3：1（v/v）乙醇：乙酸：氯仿]被用作预处理溶液，以获得前中期染色体。我们证明，使用MC II方法获得的前中期染色体是非常好的核型分析和FISH分析。我们还观察到，MC II和改进的空气干燥（ADI）方法的组合提供了一个令人满意的减数分裂粗线期染色体制备减少细胞质背景和清晰的染色质扩散。此外，我们证明，粗线期和前中期染色体的甜瓜和六道木×大花产生显着更好的FISH图像时，使用所描述的方法制备。首次证实了大花六道木中期染色体短臂上存在强弱两对45 S核糖体DNA信号。MC Ⅱ法和ADI法是获取植物细胞质背景减少的前中期和粗线期染色体的简便有效的方法。我们的方法提供了高分辨率的FISH图像，可以帮助加速植物的分子细胞遗传学研究。
Detailed karyotyping using metaphase chromosomes in melon (Cucumis melo L.) remains a challenge because of their small chromosome sizes and poor stainability. Prometaphase chromosomes, which are two times longer and loosely condensed, provide a significantly better resolution for fluorescence in situ hybridization (FISH) than metaphase chromosomes. However, suitable method for acquiring prometaphase chromosomes in melon have been poorly investigated. In this study, a modified Carnoy’s solution II (MC II) [6:3:1 (v/v) ethanol: acetic acid: chloroform] was used as a pretreatment solution to obtain prometaphase chromosomes. We demonstrated that the prometaphase chromosomes obtained using the MC II method are excellent for karyotyping and FISH analysis. We also observed that a combination of MC II and the modified air dry (ADI) method provides a satisfactory meiotic pachytene chromosome preparation with reduced cytoplasmic background and clear chromatin spreads. Moreover, we demonstrated that pachytene and prometaphase chromosomes of melon and Abelia × grandiflora generate significantly better FISH images when prepared using the method described. We confirmed, for the first time, that Abelia × grandiflora has pairs of both strong and weak 45S ribosomal DNA signals on the short arms of their metaphase chromosomes. The MC II and ADI method are simple and effective for acquiring prometaphase and pachytene chromosomes with reduced cytoplasm background in plants. Our methods provide high-resolution FISH images that can help accelerate molecular cytogenetic research in plants.
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影响因子：
5.1
作者：
Ohmido, N;Akiyama, Y;Fukui, K
通讯作者：
Fukui, K
影响因子：
7.2
作者：
Koo, Dal-Hoe;Jiang, Jiming
通讯作者：
Jiang, Jiming
DOI：
10.1007/978-3-642-20450-0_6
发表时间：
2011-01-01
期刊：
WILD CROP RELATIVES: GENOMIC AND BREEDING RESOURCES - VEGETABLES
影响因子：
--
作者：
Chen, Jin-Feng;Zhou, Xiao-Hui
通讯作者：
Zhou, Xiao-Hui
影响因子：
5.2
作者：
EZURA, H;AMAGAI, H;OOSAWA, K
通讯作者：
OOSAWA, K
四倍体甜瓜改变形态特征并提高果实品质
影响因子：
4.3
作者：
Zhang, Wenqian;Hao, Hui;Yu, Xiyan
通讯作者：
Yu, Xiyan