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作者	单位
张蔚，熊自颖，段延玲，罗涛，罗杰，唐宗祥，符书兰	四川农业大学农学院/四川农业大学植物遗传育种省级重点实验室，四川成都 611130

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中文摘要:

黑麦(Secale cereale L.)基因资源在小麦(Triticum aestivum L.)品种改良方面具有很大的利用潜力。建立一种高效鉴定小麦背景中黑麦染色体的细胞学标记方法，有利于黑麦优良基因/遗传物质的追溯和高效利用。本研究利用核仁组织区（NOR）核糖体DNA（rDNA）串联重复序列，开发了一种新的寡核苷酸探针Oligo-1RNOR。非变性荧光原位杂交(ND-FISH)试验表明，该探针可专化性识别黑麦1R染色体的NOR区域，并产生强于Oligo-5BL.46的信号。同时，为了提高外源遗传物质细胞学鉴定的效率，建立了一种快速、简便制备小麦间期细胞的方法。利用基于Oligo-1RNOR、Oligo-pSc200和Oligo-pSc250探针的ND-FISH对制备的间期细胞滴片进行分析，结果与中期染色体细胞滴片的FISH结果相同，均能够清楚地判定材料中是否含有黑麦染色体。该方法无需制备中期染色体，叶片和根尖均可用于制备间期细胞，避免了常规细胞学根尖组织制片的预处理和固定程序，为检测小麦背景中的黑麦遗传物质或小麦-黑麦易位染色体提供了更方便的技术手段。本研究建立的方法不足之处在于，要求小麦背景中的黑麦片段必须含有高拷贝数的串联重复序列。

英文摘要:

Genes from rye(Secale cereale L.) have great potential in improvement of wheat (Triticum aestivum L.) cultivars. An efficient method based on cytological marker for identifying rye chromosomes in wheat background is beneficial to trace and utilize the elite genes of rye. Ribosomal DNA tandem repeats in nucleolar organizer region(NOR) were used to develop a new oligonucleotide(oligo) probe Oligo-1RNOR. Non-denaturing fluorescence in situ hybridization(ND-FISH) analysis indicated that this probe can specifically identify the NOR of rye 1R chromosome, and its signal was stronger than that of Oligo-5BL.46. Meanwhile, in order to improve the efficiency of identifying alien chromatins in wheat, a rapid and simple method for the preparation of wheat interphase cells was established. ND-FISH using Oligo-1RNOR, Oligo-pSc200 and Oligo-pSc250 as probes was used to analyze interphase and metaphase cells, and rye chromosomes in both of the two kinds of cells can be clearly distinguished. In this method, both leaves and roots can be used to prepare interphase cells; The preparation of metaphase cells is not necessary, and the procedures of preprocessing and fixation of root tip tissue are also omitted. It provides a more convenient technique for the detection of rye chromatins or wheat-rye translocation chromosomes in wheat background. The limitation of the method in this study is that only the rye segments containing tandem repeats with a high copy number can be detected in wheat background.

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