麦类作物体细胞基因组原位杂交(GISH)效果影响因素的分析  

ANALYSIS OF FACTORS INFLUENCING GISH USED IN TRITICEAE CROPS'BODY CELLS

作　　者：林小虎[1] 李兴锋[1] 王黎明[1] 陆文辉[1] 王洪刚[1] 

机构地区：[1]山东农业大学农学院国家小麦改良中心泰安分中心

出　　处：《实验生物学报》2005年第2期126-132,共7页Acta Biologiae Experimentalis Sinica

基　　金：国家自然科学基金(39970458)

摘　　要：以八倍体小滨麦、八倍体小黑麦、八倍体小偃麦、小麦-中间偃麦草双体异附加系为实验材料对影响麦类作物体细胞GISH技术实验效果的因素进行分析,研究结果表明:细胞分裂相多、染色体分散良好、无杂质影响的高质量的染色体制片是取得理想实验效果的基础;探针DNA浓度与封阻DNA浓度的比例及杂交后洗脱条件的控制是取得理想实验效果的关键。此外,还对麦类作物体细胞基因组原位杂交实验中出现的染色体丢失、外源染色体无杂交信号、杂交信号的强弱、杂交信号过多(杂交背景重)或过少、噪音信号及杂交污点产生的原因进行了分析,并提出了相应的解决方法或注意事项。Factors influencing genomic in situ hybridization (GISH) used in the body cells of Triticeae crops including Octoploid Tritileymus, Octoploid Trititrigia, Octoploid Tritelytrigia and wheat—Elytrigia intermedium alien disomic addition line were analyzed. The results indicated that ideal GISH effects were based on the high quality chromosome preparations containing more cell mitotic metaphase, better dispersed chromosomes and no foreign matter. The critical factors were suitable ratio of probe DNA to blocker DNA, and the control of elution condition. In addition, some abnormal phenomena, such as chromosome loss, no hybridization signal in alien chromosome, strong or weak hybridization signal, poor or overabundant hybridization signal (high hybridization background), noisy hybridization signal and hybridization spots, were preliminarily analyzed. Pro- posals and solutions for these problems were further brought forth.

关 键 词：麦类作物 基因组原位杂交 体细胞 DNA浓度 八倍体小滨麦 八倍体小黑麦 八倍体小偃麦 双体异附加系 杂交信号 理想实验 中间偃麦草 染色体制片 染色体丢失 外源染色体 GISH 实验材料 细胞分裂 研究结果 杂质影响 洗脱条件 

分 类 号：S512[农业科学—作物学]