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Arabidopsis lox3 lox4 double mutants are male sterile and defective in global proliferative arrest

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Fertility and flower development are both controlled in part by jasmonates, fatty acid-derived mediators produced via the activity of 13-lipoxygenases (13-LOXs). The Arabidopsis thaliana Columbia-0 reference genome is predicted to encode four of these enzymes and it is already known that one of these, LOX2, is dispensable for fertility. In this study, the roles of the other three 13-LOXs (LOX3, LOX4 and LOX6) were investigated in single and double mutants. Four independent lox3 lox4 double mutants assembled with different mutated lox3 and lox4 alleles had fully penetrant floral phenotypes, displaying abnormal anther maturation and defective dehiscence. The plants were no longer self-fertile and pollen was not viable. Fertility in the double mutant was restored genetically by complementation with either the LOX3 or the LOX4 cDNAs and biochemically with exogenous jasmonic acid. Furthermore, deficiency in LOX3 and LOX4 causes developmental dysfunctions, compared to wild type; lox3 lox4 double mutants are taller and develop more inflorescence shoots and flowers. Further analysis revealed that developmental arrest in the lox3 lox4 inflorescence occurs with the production of an abnormal carpelloid flower. This distinguishes lox3 lox4 mutants from the wild type where developmentally typical flower buds are the terminal inflorescence structures observed in both the laboratory and in nature. Our studies of lox3 lox4 as well as other jasmonic acid biosynthesis and perception mutants show that this plant hormone is not only required for male fertility but also involved in global proliferative arrest.

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Allene oxide synthase


Defective in anther dehiscence1


Jasmonic acid


α-linolenic acid

CaMV 35S promoter:

Cauliflower mosaic virus promoter


Wild type


Fatty acid


Global proliferative arrest


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We are grateful to Nicolas Guex for helping with sequence analysis and helpful comments, Shunping Yan for some photographs, Karolina Pajerowska-Mukthar and Ivan Acosta for constructive discussion. Funding was provided by NSF2010 grant to XD, Swiss NSF grant 3100A0_122441 to EEF, and by a Bourse pour l’Egalité des Chances at the University of Lausanne to DC. Author contributions: DC performed most experiments, GW prepared transgenic 35S::lox4 plants. DC, XD and EEF conceived experiments and DC and EEF wrote the paper, EEF made field observations.

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Correspondence to Daniela Caldelari.

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Suppl. Fig. 1

Amino acid sequence alignment of LOX3 and LOX4. [] Indicates the amino acids in the fatty acid binding pocket, * indicates identical amino acids, and: indicates amino acids that are similar. The frame shows the single amino acid change (leucine (L) to valine (V)) in the fatty acid binding pocket, between respectively, LOX3 and LOX4. (1_DOC 32 kb)

Suppl. Fig. 2

Phenotype of the entire plant. 2.S13 is the empty vector control of the silenced LOX2 line, 2.S12 is the silenced LOX2 line, lox6, WT (Col-0), lox3, lox4, lox3 lox4, aos. (TIFF 8131 kb)

Suppl. Fig. 3

Anther phenotype of WT (Col-0), 2.S12 is the silenced LOX2 line, lox6 mutant. (TIFF 6083 kb)

Suppl. Fig. 4

Inflorescence of LOX double mutants. WT, lox3B lox6B, lox4A lox6B, lox3B lox4A, aos. (TIFF 4708 kb)

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Caldelari, D., Wang, G., Farmer, E.E. et al. Arabidopsis lox3 lox4 double mutants are male sterile and defective in global proliferative arrest. Plant Mol Biol 75, 25–33 (2011).

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