遗传 ›› 2021, Vol. 43 ›› Issue (5): 397-424.doi: 10.16288/j.yczz.20-448
王海燕1, 龚志云2, 蒋甲福1, 周宝良1, 娄群峰1, 曹清河3, 席梦利4, 陈佩度1, 顾铭洪2, 张天真6, 陈发棣1, 陈劲枫1, 李宗芸5, 王秀娥1
收稿日期:
2020-12-29
修回日期:
2021-02-25
出版日期:
2021-05-20
发布日期:
2021-03-16
作者简介:
作者简介:王海燕,博士,教授,研究方向:植物分子细胞遗传学。E-mail: 基金资助:
Haiyan Wang1, Zhiyun Gong2, Jiafu Jiang1, Baoliang Zhou1, Qunfeng Lou1, Qinghe Cao3, Mengli Xi4, Peidu Chen1, Minghong Gu2, Tianzhen Zhang6, Fadi Chen1, Jinfeng Chen1, Zongyun Li5, Xiue Wang1
Received:
2020-12-29
Revised:
2021-02-25
Online:
2021-05-20
Published:
2021-03-16
Supported by:
摘要:
20世纪初“遗传的染色体学说”的提出和证明标志着细胞遗传学交叉学科建立,伴随相关学科的发展,20世纪60年代末期细胞遗传学又与分子遗传学相结合,建立发展了分子细胞遗传学交叉学科。分子细胞遗传学以DNA分子原位杂交技术为核心,不断拓展应用领域,为生命科学研究提供了直观、高效的技术手段。原位杂交技术与基因组、细胞生物学等技术结合,被广泛应用于人类、动物、植物的起源、进化、驯化等基础研究和远缘杂交、染色体工程等应用研究。通过形象地展示DNA、RNA、蛋白质在细胞中的实际位置,揭示DNA序列之间的实际位置和顺序、亲缘物种间的进化关系和结构重排、基因组拼接序列的质量、转录水平RNA和翻译水平蛋白质的位置和数量变化等。江苏省遗传学会会员单位南京农业大学、扬州大学、南京林业大学、江苏师范大学、徐淮地区农科院等自20世纪中期开展细胞遗传学理论技术研究,伴随学科发展不断创新,建立了较完善的分子细胞遗传技术体系,并成功应用于开展植物系统进化、远缘杂交、染色体工程、基因组学等研究,取得了一批研究成果。本文将主要综述江苏省在该领域取得的重要进展,并展望未来发展方向。
王海燕, 龚志云, 蒋甲福, 周宝良, 娄群峰, 曹清河, 席梦利, 陈佩度, 顾铭洪, 张天真, 陈发棣, 陈劲枫, 李宗芸, 王秀娥. 江苏省植物细胞遗传学研究回顾与展望[J]. 遗传, 2021, 43(5): 397-424.
Haiyan Wang, Zhiyun Gong, Jiafu Jiang, Baoliang Zhou, Qunfeng Lou, Qinghe Cao, Mengli Xi, Peidu Chen, Minghong Gu, Tianzhen Zhang, Fadi Chen, Jinfeng Chen, Zongyun Li, Xiue Wang. Research progress of plant cytogenetics in Jiangsu province[J]. Hereditas(Beijing), 2021, 43(5): 397-424.
图3
FISH技术在棉属、菊属和杨树中的应用 A:澳洲棉-亚洲棉人工合成六倍体(2n=AADDGG=78)的多色FISH鉴定)。红色:Digoxigenin-dUTP标记的澳洲棉(GG) (Gossypium australe);Biotin绿色亚洲棉(AA) (G. arboreum)为探针;蓝色:DAPI套染;B:北京植物园野菊(C. indicum) 5S rDNA和45S rDNA的oligo-FISH检测。蓝色:DAPI套染;红色:TAMRA标记的5S rDNA信号;绿色:FAM标记的45S rDNA信号;C:毛果杨中期染色体的oligo-FISH。红色:地高辛标记的1号染色体;绿色:生物素标记的4号染色体,黄色:生物素和地高辛标记的13号染色体。"
表1
FISH技术及其应用"
名称 | 探针分子类型 | 探针标记物 | 杂交对象 | 研究目的 | 参考文献 |
---|---|---|---|---|---|
GISH 基因组原位杂交 | 总基因组DNA | 荧光素 | 有丝分裂中期染色体 减数分裂粗线期染色体 减数分裂中期I染色体 | 外源染色质鉴定 染色体结构重排 | [ |
FISH 荧光原位杂交 | (1)重复序列DNA (2) BAC克隆DNA (BAC-FISH) (3)质粒DNA (4)单拷贝基因DNA 序列(single gene-FISH) (5)寡核苷酸DNA (oligo-FISH) (6)单基因寡核苷酸探针库(unigene oligo-painting) | 荧光素/ 半抗原 | 有丝分裂中期染色体 减数分裂粗线期染色体 减数分裂中期I染色体 间期核DNA纤维 | 外源染色质鉴定 染色体上序列实际位置和顺序确定 物种间进化关系 染色体结构重排 物种核型构建 染色体构型分析 转基因鉴定 基因组拼接质量评价 | [ |
seqFISH RNA-seq荧光原位杂交 smFISH 单分子荧光原位杂交 | RNA | 荧光素 | 特定组织中的单个细胞 | 转录水平RNA位置和数量变化 | [ |
Immuno -staining 蛋白免疫共定位 | 蛋白抗体 | 荧光素 | 有丝分裂和减数分裂的细胞或者特定组织中的细胞 | 翻译水平蛋白质的位置和数量变化,表观基因组修饰 | [ |
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