Division of Biological Sciences, National Research Council of Canada, Ottawa, ON, Canada K1A OR6.
Proc Natl Acad Sci U S A. 1984 Feb;81(4):1164-8. doi: 10.1073/pnas.81.4.1164.
Differences in the distribution of Ava I and BstEII restriction sites in the chloroplast DNA (cpDNA) of Chlamydomonas eugametos and C. moewusii have been used to detect extensive cpDNA recombination in the hybrid progeny of these interfertile algae. In the present study, the inheritance of these restriction-site differences was tested for recombination with nonmendelian genetic markers for resistance to streptomycin and erythromycin in interspecific crosses and in hybrid backcrosses to C. moewusii. Most of the restriction-pattern markers appear linked to the antibiotic-resistance markers, thus supporting the chloroplast localization of the resistance markers. The streptomycin marker, in particular, shows perfect coordinate inheritance with an Ava I band containing one cpDNA fragment and a BstEII band containing two comigrating cpDNA fragments. Molecular hybridization experiments using DNA from the Ava I band as a probe show sequence homology between this DNA, the two comigrating BstEII fragments, and cpDNA fragments from C. reinhardtii containing the genes for 16S rRNA. The results show the feasibility of using C. eugametos-C. moewusii hybrids to identify cpDNA sequences that either contain or are closely linked to nonmendelian genetic markers.
在可育杂交种的叶绿体 DNA(cpDNA)中,衣藻(Chlamydomonas eugametos)和小眼衣藻(C. moewusii)的 Ava I 和 BstEII 限制位点的分布差异已被用于检测 cpDNA 重组。在本研究中,通过非孟德尔遗传标记(抗链霉素和红霉素)对同种间杂交和杂种回交到小眼衣藻中的这些限制位点差异的重组进行了检测。大多数限制图谱标记与抗生素抗性标记紧密连锁,从而支持了抗生素抗性标记的叶绿体定位。特别是,链霉素标记与含有一个 cpDNA 片段的 Ava I 带和含有两个共迁移 cpDNA 片段的 BstEII 带的抗生素标记表现出完全协调的遗传。使用来自 Ava I 带的 DNA 作为探针的分子杂交实验表明,该 DNA 与两个共迁移的 BstEII 片段以及含有 16S rRNA 基因的 cpDNA 片段之间具有序列同源性。这些结果表明,利用衣藻-小眼衣藻杂交种来鉴定含有或紧密连锁于非孟德尔遗传标记的 cpDNA 序列是可行的。