Katsiotis A, Schmidt T, Heslop-Harrison J S
Karyobiology Group, John Innes Centre, Norwich, U.K.
Genome. 1996 Apr;39(2):410-7. doi: 10.1139/g96-052.
A cloned repetitive sequence, pAvKB30, obtained from an Avena vaviloviana (AB genome) genomic library, along with two polymerase chain reaction products derived from the conserved region of the reverse transcriptase (RT) gene of retrotransposons, were characterized molecularly and cytologically. The cloned DNA fragment was a dispersed repeat present in all Avena species used in this study (A. strigosa, A. clauda, A. vaviloviana, A. magna, and A. sativa). The fragment was sequenced (210 bp) and found to be 69.5% homologous to part of WIS-2-1A, and 60.5% homologous to the leader sequence of BARE-1; both of these elements have been characterized as Ty1-copia-like retrotransposons in wheat and barley, respectively. In situ hybridization of pAvKB30 to diploid, tetraploid, and hexaploid oat species revealed that the probe is present on both arms of all chromosomes (A, B, C, and D genomes) but is excluded from their centromeric and nucleolar organizer regions. By using double in situ hybridization in hexaploid A. sativa (ACD genome), pAvKB30 was found to be present in lower copy numbers in C-genome chromosomes compared with A- and D-genome chromosomes. Furthermore, under low stringency conditions, pAvKB30 hybridized on Southern blots containing barley, wheat, rye, and Arrhenatherum DNA. However, under high stringency conditions, it hybridized only on Arrhenatherum DNA, which is considered to be the genus most closely related to Avena. All Avena species included in this study yielded a PCR product when the primers from the RT domain of retrotransposons were used. Two products, rtA, obtained by using A. strigosa (A(s) genome) as template, and rtC, obtained by using A. clauda (Cp genome) as template, gave Southern and in situ hybridization results similar to pAvKB30, but each was more abundant in its genome of origin.
从瓦维洛夫燕麦(AB基因组)基因组文库中获得的一个克隆重复序列pAvKB30,以及来自反转录转座子反转录酶(RT)基因保守区的两个聚合酶链反应产物,进行了分子和细胞学特征分析。克隆的DNA片段是本研究中所用所有燕麦物种(六倍体燕麦、克劳德燕麦、瓦维洛夫燕麦、大燕麦和普通燕麦)中都存在的分散重复序列。对该片段进行了测序(210 bp),发现其与WIS-2-1A的部分序列有69.5%的同源性,与BARE-1的前导序列有60.5%的同源性;这两个元件在小麦和大麦中分别被鉴定为Ty1-copia类反转录转座子。pAvKB30与二倍体、四倍体和六倍体燕麦物种的原位杂交显示,该探针存在于所有染色体(A、B、C和D基因组)的两条臂上,但被排除在着丝粒和核仁组织区之外。通过在六倍体普通燕麦(ACD基因组)中进行双重原位杂交发现,与A基因组和D基因组染色体相比,pAvKB30在C基因组染色体中的拷贝数较低。此外,在低严谨条件下,pAvKB30在含有大麦、小麦、黑麦和燕麦草DNA的Southern杂交印迹上杂交。然而,在高严谨条件下,它仅与燕麦草DNA杂交,燕麦草被认为是与燕麦关系最密切的属。当使用来自反转录转座子RT结构域的引物时,本研究中包括的所有燕麦物种都产生了一个PCR产物。以六倍体燕麦(A(s)基因组)为模板获得的两个产物rtA和以克劳德燕麦(Cp基因组)为模板获得的rtC,其Southern杂交和原位杂交结果与pAvKB30相似,但各自在其起源基因组中更为丰富。