Advertisement

Euphytica

, 215:5 | Cite as

Genetic demonstration of the involvement of WUSCHEL-related homeobox (WOX) genes in narrow-petal and narrow-leaf mutations in traditional Japanese azalea cultivars

  • Keisuke Tasaki
  • Akira Nakatsuka
  • Kyeong-Seong Cheon
  • Nobuo KobayashiEmail author
Article
  • 63 Downloads

Abstract

The spider-type azalea cultivars, Rhododendron kaempferi ‘Kin-kujyaku’ and Rhododendron macrosepalum ‘Seigaiha’, are characterized by extremely narrow leaves and choripetalous flowers with narrow petals. Previous morphological analysis of these cultivars revealed that narrowed organs would be a common mutation in the growth of lateral organs in the transverse plane. In the present study, we focused on the analysis of WUSCHEL-related homeobox (WOX) genes, WOX1 and WOX3, which regulate the laminar width and development of lateral organs. Sequence analysis of the two genes confirmed a fatal mutation in Rk/RmWOX3/SPIDER of ‘Kin-kujyaku’ and ‘Seigaiha’. To demonstrate the role of SPIDER in the spider-type phenotype in azalea, we performed genotyping using DNA markers developed for multiplex PCR that can distinguish between the wild-type and the spider-type allele based on the partial mutated region of RkWOX3/SPIDER in ‘Kin-kujyaku’. The investigation of BC1 population derived from ‘Kin-kujyaku’ confirmed complete co-segregation between spider-type mutation and Rkwox3/spider band pattern of DNA markers. Therefore, we concluded that SPIDER is responsible for the spider-type mutation in azalea.

Keywords

Choripetalous flower Molecular marker Narrow mutation WOX3 WUSCHEL-related homeobox gene 

Notes

Acknowledgements

This study was supported by Grant-in-Aid for Scientific Research (KAKENHI Nos. 26292017, 18K05617) from Japan Society for the Promotion of Science (JSPS). The authors thank the faculty of Life and Environmental Science at Shimane University for their help and financial support of this study.

Supplementary material

10681_2018_2323_MOESM1_ESM.pdf (997 kb)
Supplementary material 1 (PDF 997 kb)
10681_2018_2323_MOESM2_ESM.xlsx (16 kb)
Supplementary material 2 (XLSX 16 kb)

References

  1. Cheon KS, Nakatsuka A, Kobayashi N (2016) Mutant PI/GLO homolog confers the hose-in-hose flower phenotype in Kurume azaleas. Hortic J 85:380–387CrossRefGoogle Scholar
  2. Cheon KS, Nakatsuka A, Gobara Y, Kobayashi N (2017a) Mutant RoPI-1 allele-based marker development for selection of the hose-in-hose flower phenotype in Rhododendron obtusum cultivars. Euphytica 213:3CrossRefGoogle Scholar
  3. Cheon KS, Nakatsuka A, Tasaki K, Kobayashi N (2017b) Floral morphology and MADS gene expression in double-flowered Japanese evergreen azalea. Hortic J 86:269–276CrossRefGoogle Scholar
  4. Cheon KS, Nakatsuka A, Tasaki K, Kobayashi N (2018) Long-lasting Corolla cultivars in Japanese azaleas: a mutant AP3/DEF homolog identified in traditional azalea cultivars from more than 300 years ago. Front Plant Sci.  https://doi.org/10.3389/fpls.2017.02239 CrossRefPubMedPubMedCentralGoogle Scholar
  5. Gobara Y, Nakatska A, Cheon KS, Kobayashi N (2017) Floral morphology and inheritance of the long-lasting flower trait (misome-sho) in Japanese evergreen azalea cultivars. Hortic Res (Jpn) 16:383–390 (In Japanese with English abstract) CrossRefGoogle Scholar
  6. Haecker A, Gross-Hardt R, Geiges B, Sarkar A, Breuninger H, Herrmann M, Laux T (2004) Expression dynamics of WOX genes mark cell fate decisions during early embryonic patterning in Arabidopsis thaliana. Development 131:657–668CrossRefGoogle Scholar
  7. Ikeda M, Mitsuda N, Ohme-Takagi M (2009) Arabidopsis WUSCHEL is a bifunctional transcription factor that acts as a repressor in stem cell regulation and as an activator in floral patterning. Plant Cell 21:3493–3505CrossRefGoogle Scholar
  8. Ito I, Creech JL (1984) A brocade pillow: azaleas of old Japan. John Weatherhill Inc, New York and TokyoGoogle Scholar
  9. Kobayashi N, Horikoshi T, Katsuyama H, Handa T, Takayanagi K (1998) A simple and efficient DNA extraction method from the plants, especially from woody plants. Plant Tissue Cult Biotechnol 4:76–80Google Scholar
  10. Kobayashi N, Handa T, Yoshimura Y, Tsumura Y, Arisumi K, Takayanagi K (2000) Evidence for introgressive hybridization based on chloroplast DNA polymorphisms and morphological variation in wild evergreen azalea populations of the Kirishima mountains, Japan. Edinb J Bot 57:209–219CrossRefGoogle Scholar
  11. Kurashige Y, Kobayashi N (2008) Evergreen azalea cultivars and breeding trends in the Taisho era inferred by discovered ‘‘azalea research notes’’ of Kanagawa agricultural research station. Hortic Res (Jpn) 7:323–328 (In Japanese with English abstract) CrossRefGoogle Scholar
  12. Lin H, Niu L, McHale NA, Ohme-Takagi M, Mysore KS, Tadege M (2013) Evolutionarily conserved repressive activity of WOX proteins mediates leaf blade outgrowth and floral organ development in plants. Proc Natl Acad Sci 110:366–371CrossRefGoogle Scholar
  13. Matsumoto N, Okada K (2001) A homeobox gene, PRESSED FLOWER, regulates lateral axis-dependent development of Arabidopsis flowers. Gene Dev 15:3355–3364CrossRefGoogle Scholar
  14. McHale NA, Marcotrigiano M (1998) LAM1 is required for dorsoventrality and lateral growth of the leaf blade in Nicotiana. Development 125:4235–4243PubMedGoogle Scholar
  15. Nakata M, Matsumoto N, Tsugeki R, Rikirsch E, Laux T, Okada K (2012) Roles of the middle domain-specific WUSCHEL-RELATED HOMEOBOX genes in early development of leaves in Arabidopsis. Plant Cell 24:519–535CrossRefGoogle Scholar
  16. Nardmann J, Ji J, Werr W, Scanlon MJ (2004) The maize duplicate genes narrow sheath1 and narrow sheath2 encode a conserved homeobox gene function in a lateral domain of shoot apical meristems. Development 131:2827–2839CrossRefGoogle Scholar
  17. Ochman H, Gerber AS, Hart DL (1988) Genetic applications of an inverse polymerase chain reaction. Genetics 120:621–623PubMedPubMedCentralGoogle Scholar
  18. Shimizu R, Ji J, Kelsey E, Ohtsu K, Schnable PS, Scanlon MJ (2009) Tissue specificity and evolution of meristematic WOX3 function. Plant Physiol 149:841–850CrossRefGoogle Scholar
  19. Tadege M, Lin H, Bedair M, Berbel A, Wen J, Rojas CM, Niu L, Tang Y, Sumner L, Ratet P, McHale NA, Madueño F, Mysore KS (2011) STENOFOLIA regulates blade outgrowth and leaf vascular patterning in Medicago truncatula and Nicotiana sylvestris. Plant Cell 23:2125–2142CrossRefGoogle Scholar
  20. Tasaki K, Nakatsuka A, Kobayashi N (2012a) Morphological analysis of narrow-petaled cultivars of Rhododendron macrosepalum Maxim. J Jpn Soc Hortic Sci 81:72–79CrossRefGoogle Scholar
  21. Tasaki K, Nakatsuka A, Cheon KS, Koga M, Kobayashi N (2012b) Morphological and expression analyses of MADS genes in Japanese traditional narrow- and or staminoid- petaled cultivars of Rhododendron kaempferi Planch. Sci Hortic 134:191–199CrossRefGoogle Scholar
  22. Tasaki K, Nakatsuka A, Cheon KS, Kobayashi N (2015) Inheritance of the narrow leaf mutation in traditional Japanese evergreen azaleas. Euphytica 206:649–656CrossRefGoogle Scholar
  23. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680CrossRefGoogle Scholar
  24. Vandenbussche M, Horstman A, Zetho JF, Koes R, Rijpkema AS, Gerats T (2009) Differential recruitment of WOX transcription factors for lateral development and organ fusion in Petunia and Arabidopsis. Plant Cell 21:2269–2283CrossRefGoogle Scholar
  25. Wan CY, Wilkins TA (1994) A modified hot borate method significantly enhances the yield of high-quality RNA from cotton (Gossypium hirsutum L.). Anal Biochem 223:7–12CrossRefGoogle Scholar
  26. Yoshikawa T, Tanaka SY, Masumoto Y, Nobori N, Ishii H, Hibara K, Itoh J, Tanisaka T, Taketa S (2016) Barley NARROW LEAFED DWARF1 encoding a WUSCHEL-RELATED HOMEOBOX 3 (WOX3) regulates the marginal development of lateral organs. Breed Sci 66:416–424CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Faculty of Life and Environmental SciencesShimane UniversityMatsueJapan
  2. 2.Department of Molecular and Functional Genomics, Center for Integrated Research in ScienceShimane UniversityMatsueJapan
  3. 3.Faculty of AgricultureTokyo University of AgricultureAtsugiJapan
  4. 4.Gene Engineering Division, RDANational Institute of Agricultural SciencesJeonjuRepublic of Korea

Personalised recommendations