title: The active digestion of uniparental chloroplast DNA in a single zygote of Chlamydomonas reinhardtii is revealed by using the optical tweezer.
abstract: The non-Mendelian inheritance of organelle genes is a phenomenon common to almost all eukaryotes, and in the isogamous alga Chlamydomonas reinhardtii, chloroplast (cp) genes are transmitted from the mating type positive (mt(+)) parent. In this study, the preferential disappearance of the fluorescent cp nucleoids of the mating type negative (mt(-)) parent was observed in living young zygotes. To study the change in cpDNA molecules during the preferential disappearance, the cpDNA of mt(+) or mt(-) origin was labeled separately with bacterial aadA gene sequences. Then, a single zygote with or without cp nucleoids was isolated under direct observation by using optical tweezers and investigated by nested PCR analysis of the aadA sequences. This demonstrated that cpDNA molecules are digested completely during the preferential disappearance of mt(-) cp nucleoids within 10 min, whereas mt(+) cpDNA and mitochondrial DNA are protected from the digestion. These results indicate that the non-Mendelian transmission pattern of organelle genes is determined immediately after zygote formation.
authors.affiliation: Department of Biological Sciences, Graduate School of Science, University of Tokyo, Hongo, Tokyo 113-0033, Japan. yoshiki@biol.s.u-tokyo.ac.jp
authors.forename: Y
authors.initials: Y
authors.lastname: Nishimura
authors.forename: O
authors.initials: O
authors.lastname: Misumi
authors.forename: S
authors.initials: S
authors.lastname: Matsunaga
authors.forename: T
authors.initials: T
authors.lastname: Higashiyama
authors.forename: A
authors.initials: A
authors.lastname: Yokota
authors.forename: T
authors.initials: T
authors.lastname: Kuroiwa
mesh_terms: D000818:Animals; D001483:Base Sequence; D019470:Biolistics; D016825:Chlamydomonas reinhardtii / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers; D018742:DNA, Chloroplast / Q000378:metabolism*; D009713:Nucleotidyltransferases / Q000235:genetics; D055095:Optics and Photonics; D010940:Plant Proteins / Q000235:genetics; D012273:Ribulose-Bisphosphate Carboxylase; D015053:Zygote / Q000378:metabolism
chemical_list: D017931:DNA Primers; D018742:DNA, Chloroplast; D010940:Plant Proteins; D009713:Nucleotidyltransferases; C022678:streptomycin 3''-adenylyltransferase; C101625:RbcL protein, plastid; D012273:Ribulose-Bisphosphate Carboxylase