Isolation and characterization of a cell wall-defective mutant of Chlamydomonas monoica (Chlorophyta)

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Cell wall-defective strains of Chlamydomonas have played an important role in the development of transformation protocols for introducing exogenous DNA (foreign genes or cloned Chlamydomonas genes) into C. reinhardtii. To promote the development of similar protocols for transformation of the distantly related homothallic species, C. monoica, we used UV mutagenesis to obtain a mutant strain with a defective cell wall. The mutant, cw-1, was first identified on the basis of irregular colony shape and was subsequently shown to have reduced plating efficiency and increased sensitivity to lysis by a non-ionic detergent as compared with wild-type cells. Tetrad analysis of crosses involving the cw-1 mutant confirmed 2:2 segregation of the cw:cw + phenotypes, indicating that the wall defect resulted from mutation of a single nuclear gene. The phenotype showed incomplete penetrance and variable expressivity. Although some cells had apparently normal cell walls as viewed by TEM, many cells of the cw-1 strain had broken cell walls and others were protoplasts completely devoid of a cell wall. Several cw-1 isolates obtained from crosses involving the original mutant strain showed a marked enhancement of the mutant phenotype and may prove especially useful for future work involving somatic cell fusions or development of transformation protocols.

Original languageEnglish (US)
Pages (from-to)1261-1267
Number of pages7
JournalJournal of Phycology
Volume39
Issue number6
DOIs
StatePublished - Dec 2003

Fingerprint

Chlamydomonas
Chlorophyta
cell walls
mutants
phenotype
gene
penetrance
cell fusion
genes
Chlamydomonas reinhardtii
cells
somatic cells
mutagenesis
detergents
lysis
protoplasts
detergent
defect
mutant
transmission electron microscopy

Keywords

  • Cell wall
  • Chlamydomonas monoica
  • Mutant
  • Protoplast
  • Transformation

ASJC Scopus subject areas

  • Aquatic Science
  • Plant Science

Cite this

Isolation and characterization of a cell wall-defective mutant of Chlamydomonas monoica (Chlorophyta). / Fuentes, Cesar; VanWinkle-Swift, Karen P.

In: Journal of Phycology, Vol. 39, No. 6, 12.2003, p. 1261-1267.

Research output: Contribution to journalArticle

@article{0fb03113e9ba482796502d3bf40c04e7,
title = "Isolation and characterization of a cell wall-defective mutant of Chlamydomonas monoica (Chlorophyta)",
abstract = "Cell wall-defective strains of Chlamydomonas have played an important role in the development of transformation protocols for introducing exogenous DNA (foreign genes or cloned Chlamydomonas genes) into C. reinhardtii. To promote the development of similar protocols for transformation of the distantly related homothallic species, C. monoica, we used UV mutagenesis to obtain a mutant strain with a defective cell wall. The mutant, cw-1, was first identified on the basis of irregular colony shape and was subsequently shown to have reduced plating efficiency and increased sensitivity to lysis by a non-ionic detergent as compared with wild-type cells. Tetrad analysis of crosses involving the cw-1 mutant confirmed 2:2 segregation of the cw:cw + phenotypes, indicating that the wall defect resulted from mutation of a single nuclear gene. The phenotype showed incomplete penetrance and variable expressivity. Although some cells had apparently normal cell walls as viewed by TEM, many cells of the cw-1 strain had broken cell walls and others were protoplasts completely devoid of a cell wall. Several cw-1 isolates obtained from crosses involving the original mutant strain showed a marked enhancement of the mutant phenotype and may prove especially useful for future work involving somatic cell fusions or development of transformation protocols.",
keywords = "Cell wall, Chlamydomonas monoica, Mutant, Protoplast, Transformation",
author = "Cesar Fuentes and VanWinkle-Swift, {Karen P}",
year = "2003",
month = "12",
doi = "10.1111/j.0022-3646.2003.03-087.x",
language = "English (US)",
volume = "39",
pages = "1261--1267",
journal = "Journal of Phycology",
issn = "0022-3646",
publisher = "Wiley-Blackwell",
number = "6",

}

TY - JOUR

T1 - Isolation and characterization of a cell wall-defective mutant of Chlamydomonas monoica (Chlorophyta)

AU - Fuentes, Cesar

AU - VanWinkle-Swift, Karen P

PY - 2003/12

Y1 - 2003/12

N2 - Cell wall-defective strains of Chlamydomonas have played an important role in the development of transformation protocols for introducing exogenous DNA (foreign genes or cloned Chlamydomonas genes) into C. reinhardtii. To promote the development of similar protocols for transformation of the distantly related homothallic species, C. monoica, we used UV mutagenesis to obtain a mutant strain with a defective cell wall. The mutant, cw-1, was first identified on the basis of irregular colony shape and was subsequently shown to have reduced plating efficiency and increased sensitivity to lysis by a non-ionic detergent as compared with wild-type cells. Tetrad analysis of crosses involving the cw-1 mutant confirmed 2:2 segregation of the cw:cw + phenotypes, indicating that the wall defect resulted from mutation of a single nuclear gene. The phenotype showed incomplete penetrance and variable expressivity. Although some cells had apparently normal cell walls as viewed by TEM, many cells of the cw-1 strain had broken cell walls and others were protoplasts completely devoid of a cell wall. Several cw-1 isolates obtained from crosses involving the original mutant strain showed a marked enhancement of the mutant phenotype and may prove especially useful for future work involving somatic cell fusions or development of transformation protocols.

AB - Cell wall-defective strains of Chlamydomonas have played an important role in the development of transformation protocols for introducing exogenous DNA (foreign genes or cloned Chlamydomonas genes) into C. reinhardtii. To promote the development of similar protocols for transformation of the distantly related homothallic species, C. monoica, we used UV mutagenesis to obtain a mutant strain with a defective cell wall. The mutant, cw-1, was first identified on the basis of irregular colony shape and was subsequently shown to have reduced plating efficiency and increased sensitivity to lysis by a non-ionic detergent as compared with wild-type cells. Tetrad analysis of crosses involving the cw-1 mutant confirmed 2:2 segregation of the cw:cw + phenotypes, indicating that the wall defect resulted from mutation of a single nuclear gene. The phenotype showed incomplete penetrance and variable expressivity. Although some cells had apparently normal cell walls as viewed by TEM, many cells of the cw-1 strain had broken cell walls and others were protoplasts completely devoid of a cell wall. Several cw-1 isolates obtained from crosses involving the original mutant strain showed a marked enhancement of the mutant phenotype and may prove especially useful for future work involving somatic cell fusions or development of transformation protocols.

KW - Cell wall

KW - Chlamydomonas monoica

KW - Mutant

KW - Protoplast

KW - Transformation

UR - http://www.scopus.com/inward/record.url?scp=0346361489&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0346361489&partnerID=8YFLogxK

U2 - 10.1111/j.0022-3646.2003.03-087.x

DO - 10.1111/j.0022-3646.2003.03-087.x

M3 - Article

AN - SCOPUS:0346361489

VL - 39

SP - 1261

EP - 1267

JO - Journal of Phycology

JF - Journal of Phycology

SN - 0022-3646

IS - 6

ER -