Tagging Pathogenicity Genes in Ustilago Maydis by Restriction Enzyme-mediated Integration (REMI)

Overview

Journal Mol Gen Genet

Specialties Genetics
Molecular Biology

Date 1995 Sep 20

PMID 7476854

Citations 48

Authors

M Bolker

H U Bohnert

K H Braun

J Gorl

R Kahmann

Affiliations

Soon will be listed here.

Abstract

In the maize pathogenic fungus Ustilago maydis integration of transforming DNA at homologous or heterologous sites is often accompanied by duplications of the DNA. We show that it is possible to generate single-copy integration events with high efficiency by restriction enzyme-mediated integration (REMI). In about 50% of cases, a plasmid that contains a single BamHI site is integrated at chromosomal BamH1 sites, if BamHI is added to the transformation mixtures. In the other cases it appears that integration events have also occurred preferentially at BamHI sites, but without restoration of the recognition sites. Using REMI we have generated approximately 1000 insertion mutants. Pathogenicity tests demonstrated that about 1-2% of these mutants were unable to induce symptoms when tested in planta. For two of the mutants we have shown that the phenotype is linked to the insertion event.

Citing Articles

Tetracycline-controlled (TetON) gene expression system for the smut fungus .

Ingole K, Nagarajan N, Uhse S, Giannini C, Djamei A Front Fungal Biol. 2023; 3:1029114.

PMID: 37746190 PMC: 10512375. DOI: 10.3389/ffunb.2022.1029114.

A non-JA producing oxophytodienoate reductase functions in salicylic acid-mediated antagonism with jasmonic acid during pathogen attack.

Huang P, Tate M, Berg-Falloure K, Christensen S, Zhang J, Schirawski J Mol Plant Pathol. 2023; 24(7):725-741.

PMID: 36715587 PMC: 10257049. DOI: 10.1111/mpp.13299.

Secreted Endo-Xylanases Are Involved in Fungal Filamentation and Proliferation on and Inside Plants.

Moreno-Sanchez I, Pejenaute-Ochoa M, Navarrete B, Barrales R, Ibeas J J Fungi (Basel). 2021; 7(12).

PMID: 34947062 PMC: 8706147. DOI: 10.3390/jof7121081.

Ustilago maydis: how its biology relates to pathogenic development.

Kahmann R, Kamper J New Phytol. 2021; 164(1):31-42.

PMID: 33873482 DOI: 10.1111/j.1469-8137.2004.01156.x.

N-glycosylation of the protein disulfide isomerase Pdi1 ensures full Ustilago maydis virulence.

Marin-Menguiano M, Moreno-Sanchez I, Barrales R, Fernandez-Alvarez A, Ibeas J PLoS Pathog. 2019; 15(11):e1007687.

PMID: 31730668 PMC: 6881057. DOI: 10.1371/journal.ppat.1007687.

References

Banuett F, Herskowitz I . Different a alleles of Ustilago maydis are necessary for maintenance of filamentous growth but not for meiosis. Proc Natl Acad Sci U S A. 1989; 86(15):5878-82. PMC: 297734. DOI: 10.1073/pnas.86.15.5878. View

Gillissen B, Bergemann J, Sandmann C, Schroeer B, Bolker M, Kahmann R . A two-component regulatory system for self/non-self recognition in Ustilago maydis. Cell. 1992; 68(4):647-57. DOI: 10.1016/0092-8674(92)90141-x. View

Hoffman C, Winston F . A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene. 1987; 57(2-3):267-72. DOI: 10.1016/0378-1119(87)90131-4. View

Kronstad J, Leong S . Isolation of two alleles of the b locus of Ustilago maydis. Proc Natl Acad Sci U S A. 1989; 86(3):978-82. PMC: 286602. DOI: 10.1073/pnas.86.3.978. View

Kamper J, Reichmann M, Romeis T, Bolker M, Kahmann R . Multiallelic recognition: nonself-dependent dimerization of the bE and bW homeodomain proteins in Ustilago maydis. Cell. 1995; 81(1):73-83. DOI: 10.1016/0092-8674(95)90372-0. View

Fotheringham S, Holloman W . Cloning and disruption of Ustilago maydis genes. Mol Cell Biol. 1989; 9(9):4052-5. PMC: 362470. DOI: 10.1128/mcb.9.9.4052-4055.1989. View

Kronstad J, Wang J, Covert S, Holden D, McKnight G, Leong S . Isolation of metabolic genes and demonstration of gene disruption in the phytopathogenic fungus Ustilago maydis. Gene. 1989; 79(1):97-106. DOI: 10.1016/0378-1119(89)90095-4. View

Wang J, Holden D, Leong S . Gene transfer system for the phytopathogenic fungus Ustilago maydis. Proc Natl Acad Sci U S A. 1988; 85(3):865-9. PMC: 279656. DOI: 10.1073/pnas.85.3.865. View

Schiestl R, Petes T . Integration of DNA fragments by illegitimate recombination in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1991; 88(17):7585-9. PMC: 52346. DOI: 10.1073/pnas.88.17.7585. View

10.

Schulz B, Banuett F, Dahl M, Schlesinger R, Schafer W, Martin T . The b alleles of U. maydis, whose combinations program pathogenic development, code for polypeptides containing a homeodomain-related motif. Cell. 1990; 60(2):295-306. DOI: 10.1016/0092-8674(90)90744-y. View

11.

Kronstad J, Leong S . The b mating-type locus of Ustilago maydis contains variable and constant regions. Genes Dev. 1990; 4(8):1384-95. DOI: 10.1101/gad.4.8.1384. View

12.

Banuett F . Ustilago maydis, the delightful blight. Trends Genet. 1992; 8(5):174-80. DOI: 10.1016/0168-9525(92)90220-x. View

13.

Wahl G, Lewis K, Ruiz J, Rothenberg B, Zhao J, Evans G . Cosmid vectors for rapid genomic walking, restriction mapping, and gene transfer. Proc Natl Acad Sci U S A. 1987; 84(8):2160-4. PMC: 304608. DOI: 10.1073/pnas.84.8.2160. View

14.

Keon J, White G, Hargreaves J . Isolation, characterization and sequence of a gene conferring resistance to the systemic fungicide carboxin from the maize smut pathogen, Ustilago maydis. Curr Genet. 1991; 19(6):475-81. DOI: 10.1007/BF00312739. View

15.

Krieg P, Melton D . In vitro RNA synthesis with SP6 RNA polymerase. Methods Enzymol. 1987; 155:397-415. DOI: 10.1016/0076-6879(87)55027-3. View

16.

Spellig T, Bolker M, Lottspeich F, FRANK R, Kahmann R . Pheromones trigger filamentous growth in Ustilago maydis. EMBO J. 1994; 13(7):1620-7. PMC: 394992. DOI: 10.1002/j.1460-2075.1994.tb06425.x. View

17.

Fotheringham S, Holloman W . Pathways of transformation in Ustilago maydis determined by DNA conformation. Genetics. 1990; 124(4):833-43. PMC: 1203975. DOI: 10.1093/genetics/124.4.833. View

18.

Bolker M, Urban M, Kahmann R . The a mating type locus of U. maydis specifies cell signaling components. Cell. 1992; 68(3):441-50. DOI: 10.1016/0092-8674(92)90182-c. View

19.

Kuspa A, Loomis W . Tagging developmental genes in Dictyostelium by restriction enzyme-mediated integration of plasmid DNA. Proc Natl Acad Sci U S A. 1992; 89(18):8803-7. PMC: 50009. DOI: 10.1073/pnas.89.18.8803. View

20.

Tsukuda T, Carleton S, Fotheringham S, Holloman W . Isolation and characterization of an autonomously replicating sequence from Ustilago maydis. Mol Cell Biol. 1988; 8(9):3703-9. PMC: 365426. DOI: 10.1128/mcb.8.9.3703-3709.1988. View