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First published online 17 August 2005
doi: 10.1242/dev.01955

Development 132, 4107-4118 (2005)
Published by The Company of Biologists 2005
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Auxin response factors ARF6 and ARF8 promote jasmonic acid production and flower maturation

Punita Nagpal1, Christine M. Ellis1, Hans Weber2,*, Sara E. Ploense1, Lana S. Barkawi3, Thomas J. Guilfoyle4, Gretchen Hagen4, José M. Alonso5,{dagger}, Jerry D. Cohen3, Edward E. Farmer2, Joseph R. Ecker5 and Jason W. Reed1,{ddagger}

1 Department of Biology, University of North Carolina at Chapel Hill, CB #3280, Coker Hall, Chapel Hill, NC 27599-3280, USA
2 Gene Expression Laboratory, Plant Molecular Biology, University of Lausanne, Biology Building, 1015-Lausanne, Switzerland
3 Department of Horticultural Science, Center for Microbial and Plant Genomics, University of Minnesota, Saint Paul, MN 55108, USA
4 Department of Biochemistry, University of Missouri-Columbia, Columbia, MO 65211, USA
5 Plant Biology Laboratory, The Salk Institute, La Jolla, CA 92186-5800, USA



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  		Fig. 1. T-DNA insertion mutations in ARF6 and ARF8. (A) Locations of T-DNA insertions. Horizontal bars represent ARF6 (At1g30330) and ARF8 (At5g37020) -coding regions, with exons (black) and introns (white) indicated. (B) RNA gel blot hybridization using ARF6 and rDNA probes. (Left panel) RNA from 8-day-old wild-type seedlings, arf6-2 mutant seedlings or wild-type flowers, stems, leaves or roots. (Right panel) RNA from wild-type or mutant flowers. (C) RNA gel blot hybridization using ARF8 and rDNA probes. Left panel, RNA from wild-type seedlings, arf8-3 mutant seedlings or wild-type flowers, stems, leaves or roots. (Right panel) RNA from wild-type or mutant flowers.

 


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  		Fig. 2. Phenotypes of arf6-2, arf8-3 and arf6-2 arf8-3 double mutant plants. (A) 47-day-old plants. (B) Flowers of wild-type and arf6-2 arf8-3 double mutant plants. Flower buds from a single inflorescence are arranged from youngest to oldest. (C) Inflorescence stem of an arf6-2 arf8-3 plant showing arrested flower buds. (D) Scanning electron micrographs of apices of gynoecia of mature wild-type (left) and arrested arf6-2 arf8-3 (right) flowers of the same age. Scale bar: 0.1 mm. (E) Mature wild-type flower at the stage of self-fertilization. (F) arf6-2 arf8-3 flower bud untreated (left), or 2-3 days after application of linolenic acid, OPDA or jasmonic acid. In E and F, buds were dissected to reveal internal organs.

 


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  		Fig. 3. Lengths of wild-type, arf6-2, arf8-3 and arf6-2 arf8-3 floral organs±s.d. Data were gathered from flower bunches of the main inflorescence stem from seven 33-day-old plants of each genotype. The left panel shows organ lengths of the youngest flower bud of 2 mm length. The right panel shows organ lengths of the youngest flowers that were releasing pollen (anthesis). For the arf6-2 arf8-3 double mutant (which does not undergo anthesis), organs of the fourth flower after the youngest 2 mm bud were measured. For the other genotypes, numbers of flowers of different size classes are indicated in Table 1.

 


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  		Fig. 4. Expression of PARF6::GUS and PARF8::GUS fusions in flowers. (A-D,H) PARF6::GUS flowers. (E-G,I) PARF8::GUS flowers. (A) Stage 11. (B,C) Stage 12. (D) Stage 13. (E) Stage 11. (F) Stage 12. (G) Stage 13. (H,I) Higher magnification of images in A and E. Arrows indicate tapetum staining.

 


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  		Fig. 5. Effects of arf6-2 and arf8-3 mutations on gene expression in flowers. (A) Numbers of genes with expression that differs by at least 2.5-fold between different stages in wild-type flowers, grouped by stage of highest expression, together with numbers of genes in each wild-type expression class that were also developmentally regulated in arf6-2/arf6-2 arf8-3/ARF8 sesquimutant and arf6-2 arf8-3 double mutant flowers. (B,C) Venn diagrams showing numbers of genes that were underexpressed (B) or overexpressed (C) at different stages in arf6-2 arf8-3 double mutant flowers, and in arf6-2/arf6-2 arf8-3/ARF8 sesquimutant flowers (in parentheses). (D) Gene expression in response to auxin and jasmonic acid. Wild-type or arf6-2 arf8-3 mutant mature flowering plants were mock-treated or treated with IAA or JA for the times indicated, and blots of RNA from inflorescence apices were hybridized with the indicated probes.

 

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© The Company of Biologists Ltd 2005