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摘要：蚕豆（Vicia faba L.）是我国重要的食用豆科作物，籽粒营养丰富，属高蛋白低脂肪作物,具有较大的研究和应用潜力。然而，目前定位在蚕豆染色体上的细胞学标记仍然不足，无法对同一物种不同染色体臂或区段进行有效的区分与鉴别。本文利用南京农业大学开发出的寡核苷酸探针对蚕豆染色体进行了分析,获得的主要结果如下：利用多色-顺序ND-FISH，成功筛选出12个寡聚核苷酸探针包括（AAT）19、（ATT）10、（ATT）19、（ACC）19、(AGG)10、（GAA）10、（GA）29、（AT）29、18s-1、AFA-4、pSc119.2-1A-2和TEL-FF,在蚕豆染色体上产生清晰、稳定的信号。通过比较发现，(AAT)19、(ATT)19、(ATT)10、(AGG)(10)、AFA-4和pSc119.2-1A-2在染色体上具有相似的信号分布位置，位于染色体中部和近端部，ACC(19)和AT（29）信号相似，位于染色体中部、近端部和端部，而TEL-FF不但分布在所有染色体两端，而且在2对染色体短臂近端部具有很强的扩增信号，并在全部染色体着丝粒附近具有弱但清晰的杂交新号。以AFA-4和TEL-FF探针信号为基础，获得了蚕豆核型，可以准确识别蚕豆全部染色体，为蚕豆染色体工程提供了简单、经济的新方法。25542
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毕业论文关键词：蚕豆；寡核苷酸探针；多色-顺序ND-FISH；核型
Karyotype analysis of Faba bean chromosomes based on FISH
Abstract: Faba bean is an important edible legume crop in China, its seeds are nutritious and have high-protein and low-fat which has a large research and application potential. However, the cytological markers in this species are very few and not effective for distinction and identification of the same species of different chromosome or arms. In this paper, the oligonucleotide probes developed by Nanjing Agricultural University were analyzed and the major results indicated that twelve oligonucleotide probes produced stable and clear signals which included（AAT）19,（ATT）10,（ATT）19,（ACC）19, (AGG)10,（GAA）10,（GA）29,（AT）29, 18s-1, AFA-4, pSc119.2-1A-2 and TEL–FF. By comparison, we found that (AAT)19, (ATT)19, (ATT)10, (AGG)10, AFA-4 and pSc119.2-1A-2 produced similar signals in the middle and proximal centromeric region of most chromosomes, while (ACC)19 and (AT)29 produced signals in the middle, proximal region and the ends of chromosomes, TEL-FF not only distributed across all chromosome ends, but also has strong amplification signals at the proximal telomeres of two pairs of chromosome short arms and has weak but clear hybrid signals in the proximal centromeres of all chromosomes. Based on the distribution of AFA-4 and TEL-FF, the karyotype of the broad bean was developed which could accurately identify all the chromosomes of broad beans and provides a simple, economical and new method for broad bean chromosomes engineering.
Key words: faba bean; oligonucleotides; multicolor-sequential ND- FISH; karyotype
目 录
摘要3
关键词3
Abstract3
Key words4
引言4
1材料与方法5
1.1材料 6
1.1.1植物材料6
1.1.2探针6
1.2方法7
1. 2. 1材料处理与取材7
1 .2. 2根尖的固定7
1.2.3根尖细胞有丝分裂中期染色体制片7
1.2.4非变性FISH7
1.2.5染色体分析7
2结果与分析7
2. 1蚕豆根尖细胞染色体制片方法7
2. 2寡聚核苷酸探针的筛选8
2.3蚕豆核型的建立.11
3讨论.12
致谢13
参考文献13
基于FISH的蚕豆染色体核型分析
引言：
蚕豆（Vicia faba L.）是我国重要的食用豆科作物。蚕豆籽粒营养丰富，属高蛋白低脂肪作物，是一种重要的植物蛋白资源。近年来，国内外学者们对蚕豆的营养特点、种质资源、品种改良进行了大量研究。但是,研究发现，由于目前栽培利用的蚕豆品种单一、遗传基础较狭窄、育种方法单一、基础研究较为薄弱，所以蚕豆遗传改良工作尚具有很大的发展空间[1]。