Mandipropamid targets the cellulose synthase-like PiCesA3 to inhibit cell wall biosynthesis in the oomycete plant pathogen, Phytophthora infestans.
Oomycete plant pathogens cause a wide variety of economically and environmentally important plant diseases. Mandipropamid (MPD) is a carboxylic acid amide (CAA) effective against downy mildews, such as Plasmopara viticola on grapes and potato late blight caused by Phytophthora infestans. Historically, the identification of the mode of action of oomycete-specific control agents has been problematic. Here, we describe how a combination of biochemical and genetic techniques has been utilized to identify the molecular target of MPD in P. infestans. Phytophthora infestans germinating cysts treated with MPD produced swelling symptoms typical of cell wall synthesis inhibitors, and these effects were reversible after washing with H(2)O. Uptake studies with (14)C-labelled MPD showed that this oomycete control agent acts on the cell wall and does not enter the cell. Furthermore, (14)C glucose incorporation into cellulose was perturbed in the presence of MPD which, taken together, suggests that the inhibition of cellulose synthesis is the primary effect of MPD. Laboratory mutants, insensitive to MPD, were raised by ethyl methane sulphonate (EMS) mutagenesis, and gene sequence analysis of cellulose synthase genes in these mutants revealed two point mutations in the PiCesA3 gene, known to be involved in cellulose synthesis. Both mutations in the PiCesA3 gene result in a change to the same amino acid (glycine-1105) in the protein. The transformation and expression of a mutated PiCesA3 allele was carried out in a sensitive wild-type isolate to demonstrate that the mutations in PiCesA3 were responsible for the MPD insensitivity phenotype.
Blum
Mathias
M
Syngenta Crop Protection AG, CH-4332 Stein, Switzerland.
Boehler
Martine
M
Randall
Eva
E
Young
Vanessa
V
Csukai
Michael
M
Kraus
Sabrina
S
Moulin
Florence
F
Scalliet
Gabriel
G
Avrova
Anna O
AO
Whisson
Stephen C
SC
Fonne-Pfister
Raymonde
R
D020418:Algal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000577:Amides / Q000494:pharmacology*; D000595:Amino Acid Sequence; D002264:Carboxylic Acids / Q000494:pharmacology*; D002473:Cell Wall / Q000187:drug effects / Q000378:metabolism*; D002482:Cellulose / Q000096:biosynthesis; D003433:Crosses, Genetic; D005020:Ethyl Methanesulfonate; D018628:Gene Dosage / Q000235:genetics; D005947:Glucose / Q000378:metabolism; D005964:Glucosyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000187:drug effects; D009154:Mutation / Q000235:genetics; D055750:Phytophthora infestans / Q000166:cytology / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D010944:Plants / Q000187:drug effects / Q000382:microbiology*; D014170:Transformation, Genetic / Q000187:drug effects
D020418:Algal Proteins; D000577:Amides; D002264:Carboxylic Acids; C550943:mandipropamid; D002482:Cellulose; D005020:Ethyl Methanesulfonate; D005964:Glucosyltransferases; C478648:cellulose synthase; D005947:Glucose
Cell Microbiol. 2007 Jan;9(1):31-9
17081190
Phytopathology. 2000 Nov;90(11):1201-8
18944421
Plant Dis. 2008 May;92(5):675-683
30769584
Plant Cell. 2008 Mar;20(3):720-38
18349153
Mol Plant Microbe Interact. 2006 Dec;19(12):1348-58
17153919
Genetics. 2003 Jun;164(2):731-40
12807792
Plant Dis. 2004 Sep;88(9):930-934
30812244
Nature. 2009 Sep 17;461(7262):393-8
19741609
Plant Physiol. 1998 Mar;116(3):1043-51
9501137
Genome Biol. 2009;10(4):R36
19368726
Protoplasma. 2003 Mar;220(3-4):201-7
12664284
Eur J Biochem. 2004 Sep;271(18):3635-45
15355340
Pest Manag Sci. 2001 Nov;57(11):1081-7
11721527
Mol Microbiol. 2006 Oct;62(2):552-65
16978258
Pest Manag Sci. 2008 Mar;64(3):255-61
18095384
Plant J. 1993 Jan;3(1):1-30
8401598
Science. 1988 Jan 29;239(4839):487-91
2448875
Mol Plant Pathol. 2005 Mar 1;6(2):153-63
20565646
Annu Rev Microbiol. 1968;22:87-108
4879523
Phytochemistry. 2002 Apr;59(7):689-96
11909624
Plant Physiol. 1990 Jun;93(2):695-700
16667525
Plant Physiol. 1977 Jun;59(6):1088-97
16660000
Anal Biochem. 1969 Dec;32(3):420-4
5361396
Genetics. 1996 Nov;144(3):1005-13
8913745
Proc Natl Acad Sci U S A. 2005 May 24;102(21):7766-71
15894622
Microbiology (Reading). 2008 Dec;154(Pt 12):3743-3751
19047742
Nature. 2007 Nov 1;450(7166):115-8
17914356
Cell Microbiol. 2008 Nov;10(11):2271-84
18637942
Plant Physiol. 2001 Jul;126(3):981-92
11457949
Mol Plant Microbe Interact. 1991 Nov-Dec;4(6):602-7
1804404
FEMS Microbiol Lett. 2007 Sep;274(1):1-8
17559387
Trends Parasitol. 2008 Nov;24(11):502-8
18805734
Proc Natl Acad Sci U S A. 2001 Aug 28;98(18):10079-84
11517344
Mol Microbiol. 2005 May;56(3):638-48
15819621
J Exp Bot. 2008;59(14):3963-74
18832186
New Phytol. 2009;184(1):114-126
19645738
Plant Physiol. 2002 Feb;128(2):482-90
11842152
Phytopathology. 2007 May;97(5):643-9
18943584
Mol Plant Microbe Interact. 2008 Apr;21(4):433-47
18321189
Fungal Genet Biol. 2003 Oct;40(1):4-14
12948509
Plant Physiol. 2001 Apr;125(4):2040-52
11299383
Plant Cell Physiol. 2002 Dec;43(12):1407-20
12514238