RRC ID 50005
著者 Kanda N, Ichikawa M, Ono A, Toyoda A, Fujiyama A, Abe J, Tsuchikane Y, Nishiyama T, Sekimoto H.
タイトル CRISPR/Cas9-based knockouts reveal that CpRLP1 is a negative regulator of the sex pheromone PR-IP in the Closterium peracerosum-strigosum-littorale complex.
ジャーナル Sci Rep
Abstract Heterothallic strains of the Closterium peracerosum-strigosum-littorale (C. psl.) complex have two sexes, mating-type plus (mt+) and mating-type minus (mt-). Conjugation between these two sexes is regulated by two sex pheromones, protoplast-release-inducing protein (PR-IP) and PR-IP Inducer, which are produced by mt+ and mt- cells, respectively. PR-IP mediates the release of protoplasts from mt- cells during mating. In this study, we examined the mechanism of action of CpRLP1 (receptor-like protein 1), which was previously identified in a cDNA microarray analysis as one of the PR-IP-inducible genes. Using CRISPR/Cas9 technology, we generated CpRLP1 knockout mutants in mt- cells of the C. psl. complex. When the knockout mt- cells were mixed with wild-type mt+ cells, conjugation was severely reduced. Many cells released protoplasts without pairing, suggesting a loss of synchronization between the two mating partners. Furthermore, the knockout mutants were hypersensitive to PR-IP. We conclude that CpRLP1 is a negative regulator of PR-IP that regulates the timing of protoplast release in conjugating C. psl. cells. As the first report of successful gene knockout in the class Charophyceae, this study provides a basis for research aimed at understanding the ancestral roles of genes that are indispensable for the development of land plants.
巻・号 7(1)
ページ 17873
公開日 2017-12-19
DOI 10.1038/s41598-017-18251-8
PII 10.1038/s41598-017-18251-8
PMID 29259295
PMC PMC5736609
MeSH Amino Acid Sequence Arabidopsis Proteins / genetics* Biological Phenomena / genetics CRISPR-Cas Systems / genetics CRISPR-Cas Systems / physiology* Carrier Proteins / genetics* Closterium / genetics Closterium / physiology* Clustered Regularly Interspaced Short Palindromic Repeats / genetics* Clustered Regularly Interspaced Short Palindromic Repeats / physiology Gene Knockout Techniques / methods Protoplasts / physiology Reproduction / genetics Reproduction / physiology* Sex Attractants / genetics Sex Attractants / physiology*
IF 3.998
引用数 5
リソース情報
藻類 NIES-67 NIES-68