Molecular Genetics and Genomics

, Volume 279, Issue 5, pp 499–507 | Cite as

NARROW LEAF 7 controls leaf shape mediated by auxin in rice

  • Kenji Fujino
  • Yasuyuki Matsuda
  • Kenjirou Ozawa
  • Takeshi Nishimura
  • Tomokazu Koshiba
  • Marco W. Fraaije
  • Hiroshi Sekiguchi
Original Paper


Elucidation of the genetic basis of the control of leaf shape could be of use in the manipulation of crop traits, leading to more stable and increased crop production. To improve our understanding of the process controlling leaf shape, we identified a mutant gene in rice that causes a significant decrease in the width of the leaf blade, termed narrow leaf 7 (nal7). This spontaneous mutation of nal7 occurred during the process of developing advanced backcrossed progeny derived from crosses of rice varieties with wild type leaf phenotype. While the mutation resulted in reduced leaf width, no significant morphological changes at the cellular level in leaves were observed, except in bulliform cells. The NAL7 locus encodes a flavin-containing monooxygenase, which displays sequence homology with YUCCA. Inspection of a structural model of NAL7 suggests that the mutation results in an inactive enzyme. The IAA content in the nal7 mutant was altered compared with that of wild type. The nal7 mutant overexpressing NAL7 cDNA exhibited overgrowth and abnormal morphology of the root, which was likely to be due to auxin overproduction. These results indicate that NAL7 is involved in auxin biosynthesis.


Rice Narrow leaf FMO Auxin YUCCA 

Supplementary material

438_2008_328_MOESM1_ESM.tif (45 kb)
Frequency distribution of the width of the leaf blade in the flag leaf in the advanced backcrossed progeny. Arrowhead indicates the mean of Hayamasari (HY). Three classified genotypes, homozygous for the Italica Livorno allele (black), heterozygous (hatched), and homozygous for the Hayamasari allele (white), assessed using the marker GBR3003, are indicated (TIF 46 kb)
438_2008_328_MOESM2_ESM.tif (71 kb)
Sequence alignment of NAL7. The mutation identified in the nal7 mutant allele is indicated by bold type and by an arrowhead above the aligned sequences. The two conserved motifs are boxed (TIF 72 kb)
438_2008_328_MOESM3_ESM.tif (62 kb)
Phylogenetic tree of the YUCCA gene family. The phylogenetic tree of 14 rice and 11 Arabidopsis YUCCA genes was constructed using CLUSTAL W. Bootstrap analysis values are shown at the nodal branches. The indicated scale represents 0.1 amino acid substitution per site. The accession numbers and plant species are shown in green for Arabidopsis, blue for rice from Yamamoto et al. (2007), and red for rice from this study. Gene ID is according to RAP-DB ( (TIF 62 kb)
438_2008_328_MOESM4_ESM.tif (158 kb)
Expression of nal7 in transgenic plants measured by RT-PCR analysis. Ten independent lines (T1), and Hayamasari and the nal7 mutant as non transgenic plants were used. Total RNA was extracted from the whole shoot of the 6-day-old seedling. The numbers on the right indicate the number of PCR cycles. Ubi2 was used as a loading control (TIF 158 kb)
438_2008_328_MOESM5_ESM.tif (39 kb)
Supplementary material Table S1 (TIF 69 kb)
438_2008_328_MOESM6_ESM.tif (39 kb)
Supplementary material Table S2 (TIF 70 kb)
Supplementary material Table S3 (TIF 59 kb)


  1. Bartel B (1997) Auxin biosynthesis. Annu Rev Plant Physiol Plant Mol Biol 48:51–66PubMedCrossRefGoogle Scholar
  2. Byrne ME (2005) Networks in leaf development. Curr Opin Plant Biol 8:59–66PubMedCrossRefGoogle Scholar
  3. Castellano MM, Sablowski R (2005) Intercellular signaling in the transition from stem cells to organogenesis. Curr Opin Plant Biol 8:26–31PubMedCrossRefGoogle Scholar
  4. Cheng Y, Dai X, Zhao Y (2006) Auxin biosynthesis by the YUCCA flavin monooxygenases controls the formation of floral organs and vascular tissues in Arabidopsis. Gene Dev 20:1790–1799PubMedCrossRefGoogle Scholar
  5. Cheng Y, Dai X, Zhao Y (2007) Auxin synthesized by the YUCCA flavin monooxygenases is essential for embryogenesis and leaf formation in Arabidopsis. Plant Cell 19:2430–2439PubMedCrossRefGoogle Scholar
  6. Cohen JD, Slovin JP, Hendrickson AM (2003) Two gentically discrete pathways convert tryptophan to auxin: more redundancy in auxin biosysnthesis. Trends Plant Sci 8:197–199PubMedCrossRefGoogle Scholar
  7. Cui KH, Peng SB, Xing YZ, Yu SB, Xu CG et al (2003) Molecular dissection of the genetic relationships of source, sink and transport tissue with yield traits in rice. Theor Appl Genet 106:649–658PubMedGoogle Scholar
  8. Fleming AJ (2005) Formation of primordia and phyllotaxy. Curr Opin Plant Biol 8:53–58PubMedCrossRefGoogle Scholar
  9. Fujino K, Sekiguchi H, Sato T, Kiuchi H, Nonoue Y et al (2004) Mapping of quantitative trait loci controlling low-temperature germinability in rice (Oryza sativa L.). Theor Appl Genet 108:794–799PubMedCrossRefGoogle Scholar
  10. Fujino K, Sekiguchi H, Kiguchi T (2005) Identification of an active transposon in intact rice plants. Mol Genet Genomics 273:150–157PubMedCrossRefGoogle Scholar
  11. Hake S, Smith HMS, Holtan H, Magnani E, Mele G et al (2004) The role of knox genes in plant development. Annu Rev Cell Dev Biol 20:125–51PubMedCrossRefGoogle Scholar
  12. Jackson RG, Kowalczyk M, Li Y, Higgins G, Ross J et al (2002) Over-expression of an Arabidopsis gene encoding a glucosyltransferase of indole-3-acetic acid: phenotypic characterisation of transgenic lines. Plant J 32:573–583PubMedCrossRefGoogle Scholar
  13. Kepinski S (2006) Integrating hormone signaling and patterning mechanisms in plant development. Curr Opin Plant Biol 9:28–34PubMedCrossRefGoogle Scholar
  14. Kessler S, Sinha N (2004) Shaping up: the genetic control of leaf shape. Curr Opin Plant Biol 7:65–72PubMedCrossRefGoogle Scholar
  15. Klee HJ, Horsch RB, Hinchee MA, Hein MB, Hoffmann NL (1987) The effects of overproduction of two Agrobacterium tumefaciens T-DNA auxin biosynthetic gene products in transgenic petunia plants. Gene Dev 1:86–96CrossRefGoogle Scholar
  16. Li Z, Pinson SRM, Stansel JW, Paterson AH (1998) Genetic dissection of the source–sink relationship affecting fecundity and yield in rice (Oryza sativa L.). Mol Breed 4:419–426CrossRefGoogle Scholar
  17. Ljung K, Bhalerao RP, Sandberg G (2001) Sites and homeostatic control of auxin biosynthesis in Arabidopsis during vegetative growth. Plant J 28:465–474PubMedCrossRefGoogle Scholar
  18. Malito E, Alfieri A, Fraaije MW, Mattevi A (2004) Crystal structure of a Baeyer–Villiger monooxygenase. Proc Natl Acad Sci USA 101:13157–13162PubMedCrossRefGoogle Scholar
  19. Nishimura T, Mori Y, Furukawa T, Kadota A, Koshiba T (2006) Red light causes a reduction in IAA levels at the apical tip by inhibiting de novo biosynthesis from tryptophan in maize coleoptiles. Planta 224:1427–1435PubMedCrossRefGoogle Scholar
  20. Ozawa K, Kawahigashi H (2006) Positional cloning of the nitrite reductase gene associated with good growth and regeneration ability of calli and establishment of a new selection system for Agrobacterium-mediated transformation in rice (Oryza sativa L.). Plant Sci 170:384–393CrossRefGoogle Scholar
  21. Perez-Perez JM, Serrano-Cartagena J, Micol JL (2002) Genetic analysis of natural variations in the architecture of Arabidopsis thaliana vegetative leaves. Genetics 162:893–915PubMedGoogle Scholar
  22. Reinhardt D, Pesce ER, Stieger P, Mandel T, Baltensperger K et al (2003) Regulation of phyllotaxis by polar auxin transport. Nature 426:255–260PubMedCrossRefGoogle Scholar
  23. Tax FE, Durbak A (2006) Meristems in the movies: live imaging as a tool for decoding intercellular signaling in shoot apical meristems. Plant Cell 18:1331–1337PubMedCrossRefGoogle Scholar
  24. Tobene-Santamaria R, Bliek M, Ljung K, Sandberg G, Mol JNM et al (2002) FLOOZY of petunia is a flavin mono-oxygenase-like protein required for the specification of leaf and flower architecture. Gene Dev 16:753–763CrossRefGoogle Scholar
  25. Tsukaya H (2006) Mechanism of leaf-shape determination. Annu Rev Plant Biol 57:477–496PubMedCrossRefGoogle Scholar
  26. van Berkel WJH, Kamerbeek NM, Fraaije MW (2006) Flavoprotein monooxygenases, a diverse class of oxidative biocatalysts. J Biotechnol 124:670–689PubMedCrossRefGoogle Scholar
  27. Woo YM, Park HJ, Suudi M, Yang JI, Park JJ et al (2007) Constitutively wilted 1, a member of the rice YUCCA gene family, is required for maintaining water homeostasis and an appropriate root to shoot ratio. Plant Mol Biol 65:125–136PubMedCrossRefGoogle Scholar
  28. Woodward C, Bemis SM, Hill EJ, Sawa S, Koshiba T et al (2005) Interaction of auxin and ERECTA in elaborating Arabidopsis inflorescence architecture revealed by the activation tagging of a new member of the YUCCA family putative flavin monooxygenases. Plant Physiol 139:192–203PubMedCrossRefGoogle Scholar
  29. Yamamoto Y, Kamiya N, Morinaka Y, Matsuoka M, Sazuka T (2007) Auxin biosynthesis by the YUCCA genes in rice. Plant Physiol 143:1362–1371PubMedCrossRefGoogle Scholar
  30. Zazimalova E, Napier RM (2003) Points of regulation for auxin action. Plant Cell Rep 21:625–634PubMedGoogle Scholar
  31. Zhao Y, Christensen SK, Fankhauser C, Cashmen JR, Cohen JD et al (2001) A role for flavin monooxygenase-like enzymes in auxin biosynthesis. Science 291:306–309PubMedCrossRefGoogle Scholar
  32. Zhao Y, Hull AK, Gupta NR, Goss KA, Alonso J et al (2002) Trp-dependent auxin biosynthesis in Arabidopsis: involvement of cytochrome P450s CYP79B2 and CYP79B3. Gene Dev 16:3100–3112PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Kenji Fujino
    • 1
  • Yasuyuki Matsuda
    • 1
  • Kenjirou Ozawa
    • 2
  • Takeshi Nishimura
    • 3
  • Tomokazu Koshiba
    • 3
  • Marco W. Fraaije
    • 4
  • Hiroshi Sekiguchi
    • 1
  1. 1.Agricultural Research InstituteHOKUREN Federation of Agricultural CooperativesHokkaidoJapan
  2. 2.Department of Low-Temperature SciencesNational Agricultural Research Center for Hokkaido RegionSapporo, HokkaidoJapan
  3. 3.Department of Biological SciencesTokyo Metropolitan UniversityHachioji, TokyoJapan
  4. 4.Biochemical Laboratory, Groningen Biomolecular Sciences and Biotechnology InstituteUniversity of GroningenGroningenThe Netherlands

Personalised recommendations