摘要
以生物素标记的簇毛麦基因组ＤＮＡ作探针与小麦－簇毛麦杂种双二倍体及异附加系染色体进行原位杂交，旨在探索一种鉴定小麦中簇毛麦染色质的分子细胞遗传学方法。首先试验了标记的簇毛麦ＤＮＡ与小麦ＤＮＡ之含量比对原位杂交效果的影响，发现探针混合液中不加未标记的小麦ＤＮＡ时，获得的原位杂交结果较理想。在八倍体小麦－簇毛麦双二倍体（２ｎ＝５６）中，１４对簇毛麦染色体出现明显的棕褐色原位杂交信号，而小麦染色体不显标记，使这２个物种的染色体很易相互区分开来。进一步用生物素标记的簇毛麦ＤＮＡ与小麦－簇毛麦６Ｖ染色体附加系体细胞杂交，也可检测出其中１对附加的６Ｖ染色体。由于该方法标记的簇毛麦ＤＮＡ覆盖各簇毛麦染色体的整个染色体臂，由此，用其鉴定小片段小麦－簇毛麦染色体易位是有效的。
Although DNA In situ hybridization has been widely used to identify alien chrornosomes inwheat background, there has been few reports on its application in Haynaldia Villosa chromosome detectiondue to the absence of suitable H. villosa-speclfic cloned DNA probes. In the present study, biotin-labeledgenomic DNA of H.villosa was used as probe to hybridize with soniatic chromosomes of hybrids betweenwheat and H. villosa with an attempt to provide a simple and effective molecular-cytogenetic method for H.villosa chromatin characterization in wheat. We checked 1:0,1:1,1: 4 and 1:10 ratios of labeled H.villosa DNA to unlabeled wheat DNA (used as a blocker)in experiments with 8X Triticum aestivum-H. vil-losa amphiploid,and an 1 : 0 ratio showed better contrast between the H. villosa and wheat chromosomes.The brown in situ slgnals appeared in all 14 H.villosa chromosomes of the amphlploid, where wheat chromo somes were not labed and appeared blue as a result of wright’s counterstain,when total genornic biotin=la-beled DNA of H. villosa was used to hybridlze to metaphase spreads of H. villosa chrornosome addition line6V, the chromosomes added to wheat could also be easily identified based on their labeling signals。 It shouldbe posslble to detect small H. villosa chromosome segments translocated into wheat with this mothod since al-most all parts of each H. villosa chromosome could be labeled.
出处
《江苏农学院学报》
CSCD
1994年第3期6-8,共3页
Jiangsu Agricultural Research
基金
国家科学基金
农业部资金资助
关键词
小麦
杂交
簇毛麦
杂种
染色体
wheat
genome+DNA+probes
biotin
in situ hybridzation/Haymaldia villosa
