Advertisement

Plant Molecular Biology Reporter

, Volume 34, Issue 1, pp 257–264 | Cite as

Knockdown of Carotenoid Cleavage Dioxygenase 4 (CCD4) via Virus-Induced Gene Silencing Confers Yellow Coloration in Peach Fruit: Evaluation of Gene Function Related to Fruit Traits

  • Songling Bai
  • Pham Anh Tuan
  • Miho Tatsuki
  • Hideaki Yaegaki
  • Akemi Ohmiya
  • Chihiro Yamamizo
  • Takaya Moriguchi
Original Paper

Abstract

Transgenic approach is an excellent way for the clarification of gene function, but it is generally difficult to create transgenic plants for most of the fruit trees including peach. Even if transgenic lines are successfully obtained, it will be extremely difficult to evaluate fruit traits due to the long juvenile phase of the plants. To overcome this problem, usage of virus vector is one of the excellent approaches. In this study, we evaluated gene function related to fruit traits in peaches via virus-induced gene silencing (VIGS). Carotenoid cleavage dioxygenase 4 (CCD4) is proposed to be the key factor responsible for carotenoid degradation in white flesh peaches. Then, we knocked down the CCD4 gene in the white flesh peaches (“Akatsuki” and “Manami”) via VIGS system. Resultantly, yellow pigmentation and increased contents of carotenoids including lutein, ß-carotene, ß-cryptoxanthin, zeaxanthin, and violaxanthin were observed in the agroinfiltration portions of the white flesh peaches, which is a direct evidence that CCD4 is a determinant for yellow flesh trait of peach. Our results suggested the possible application of VIGS system for functional studies of the genes related to fruit traits.

Keywords

Carotenoid cleavage dioxygenase 4 (CCD4) Peach fruits Transgenic fruit tress Tobacco rattle virus (TRV) Virus-induced gene silencing (VIGS) 

Abbreviations

CCD4

Carotenoid cleavage dioxygenase 4

qRT-PCR

Quantitative reverse transcription PCR

TRV

Tobacco rattle virus

VIGS

Virus-induced gene silencing

Notes

Acknowledgments

We would like to thank Dr. T. Peng (National Navel Orange Engineering Research Center, China) for her critical reading of the manuscript. We also express our thanks to Dr. S. P. Dinesh-Kumar for providing TRV1 and TRV2 vectors. This work was supported by a grant from the Ministry of Agriculture, Forestry, and Fisheries of Japan (Genomics-based Technology for Agricultural Improvement, DHR3).

Supplementary material

11105_2015_920_Fig8_ESM.jpg (45 kb)
Supplementary Fig. S1

HPLC chromatograms of the carotenoid extracts obtained from the white flesh of peach “Akatsuki” harvested in 7 July with (A) or without (B) saponification. HPLC analysis was carried out according to Yamamizo et al. (2010). The identified peaks include: (1) allE-violaxanthin, (2) cis-lutein, (3) allE-lutein, (4) zeaxanthin, (5) β-cryptoxanthin, (6) β-carotene. (JPEG 44 kb)

11105_2015_920_MOESM1_ESM.xlsx (10 kb)
Supplementary Table S1 (XLSX 10 kb)

References

  1. Adami M, Franceschi PD, Brandi F, Liverani A, Giovannini D, Rosati C, Dondini L, Tartarini S (2013) Identifying a carotenoid cleavage dioxygenase (ccd4) gene controlling yellow/white fruit flesh color of peach. Plant Mol Biol Report 31:1166–1175CrossRefGoogle Scholar
  2. Bailey JS, French HP (1949) The inheritance of certain fruit and foliage characters in peach. Mass Agr Expt Sta Bul 452:2–31Google Scholar
  3. Brandi F, Einat Bar E, Mourgues F, Györgyi Horváth G, Erika Turcsi E, Giuliano G, Liverani A, Tartarini S, Lewinsohn E, Carlo Rosati C (2011) Study of ‘Redhaven’ and its white-fleshed mutant suggests a key role of CCD4 carotenoid dioxygenase in carotenoid and norisoprenoid volatile metabolism. BMC Plant Biol 11:24PubMedCentralCrossRefPubMedGoogle Scholar
  4. Bruun-Rasmussen M, Madsen CT, Jessing S, Albrechtsen M (2007) Stability of barley stripe mosaic virus-induced gene silencing in barley. Mol Plant Microbe Interact 20:1323–1331CrossRefPubMedGoogle Scholar
  5. Campbell R, Ducreux LJ, Morris WL, Morris JA, Suttle JC, Ramsay G, Bryan GJ, Hedley PE, Taylor MA (2010) The metabolic and developmental roles of carotenoid cleavage dioxygenase4 from potato. Plant Physiol 154:656–664PubMedCentralCrossRefPubMedGoogle Scholar
  6. Connors CH (1919) Some notes on the inheritance of unit characters in the peach. Proc Am Soc Hortic Sci 16:24–36Google Scholar
  7. Endo T, Shimada T, Fujii H, Kobayashi Y, Araki T, Omura M (2005) Ectopic expression of an FT homolog from Citrus confers an early flowering phenotype on trifoliate orange (Poncirus trifoliata L. Raf.). Transgenic Res 14:703–712CrossRefPubMedGoogle Scholar
  8. Falchi R, Vendramin E, Laura Zanon L, Scalabrin S, Cipriani G, Verde I, Vizzotto G, Morgante M (2013) Three distinct mutational mechanisms acting on a single gene underpin the origin of yellow flesh in peach. Plant J 76:175–187PubMedCentralPubMedGoogle Scholar
  9. Fu DQ, Zhu BZ, Zhu HL, Jiang WB, Luo YB (2005) Virus-induced gene silencing in tomato fruit. Plant J 43:299–308CrossRefPubMedGoogle Scholar
  10. Fukamatsu Y, Tamura T, Hihara S, Oda K (2013) Mutations in the CCD4 carotenoid cleavage dioxygenase gene of yellow-flesh peaches. Biosci Biotechnol Biochem 77:2514–2516CrossRefPubMedGoogle Scholar
  11. Hileman LC, Drea S, de Martino G, Litt A, Irish VF (2005) Virus-induced gene silencing is an effective tool for assaying gene function in the basal eudicot species Papaver somniferum (opium poppy). Plant J 44:334–341CrossRefPubMedGoogle Scholar
  12. Hornero-Méndez D, Mínguez-Mosquera MI (2000) Xanthophyll esterification accompanying carotenoid overaccumulation in chromoplast of Capsicum annuum ripening fruits is a constitutive process and useful for ripeness index. J Biol Chem 48:1617–1622Google Scholar
  13. Jia H, Guo J, Qin L, Shen Y (2010) Virus-induced PpCHLH gene silencing in peach leaves (Prunus persica). J Hortic Sci Biotechnol 85:528–532Google Scholar
  14. Kawai T, Gonoi A, Nitta M, Kaido M, Yamagishi N, Yoshikawa N, Tao R (2014) Virus-induced gene silencing in apricot (Prunus armeniaca L.) and Japanese apricot (P. mume Siebold & Zucc.) with the apple latent spherical virus vector system. J Jpn Soc Hortic Sci 83:23–31Google Scholar
  15. Lacomme C, Hrubikova K, Hein I (2003) Enhancement of virus-induced gene silencing through viral-based production of inverted-repeats. Plant J 34:543–553CrossRefPubMedGoogle Scholar
  16. Li Y-Y, Mao K, Zhao C, Zhao X-Y, Zhang H-L, Shu H-R, Hao Y-J (2012) MdCOP1 ubiquitin E3 ligases interact with MdMYB1 to regulate light-induced anthocyanin biosynthesis and red fruit coloration in apple. Plant Physiol 160:1011–1022PubMedCentralCrossRefPubMedGoogle Scholar
  17. Lu R, Martin-Hernandez AM, Peart JR, Malcuit I, Baulcombe DC (2003) Virus-induced gene silencing in plants. Methods 30:296–303CrossRefPubMedGoogle Scholar
  18. Ma J, Li J, Zhao J, Zhou H, Ren F, Wang L, Gu C, Liao L, Han Y (2014) Inactivation of a gene encoding carotenoid cleavage dioxygenase (CCD4) leads to carotenoid-based yellow coloration of fruit flesh and leaf midvein in peach. Plant Mol Biol Report 32:246–257CrossRefGoogle Scholar
  19. MacFarlane SA (1999) Molecular biology of the tobraviruses. J Gen Virol 80:2799–2807CrossRefPubMedGoogle Scholar
  20. Matsuda N, Ikeda K, Kurosaka M, Takashina T, Isuzugawa K, Endo T, Omura M (2009) Early flowering phenotype in transgenic pears (Pyrus communis L.) expressing the CiFT gene. J Jpn Soc Hortic Sci 78:410–416CrossRefGoogle Scholar
  21. Pérez-Clemente RM, Pérez-Sanjuán A, García-Férriz L, Beltrán J-P, Cañas LA (2004) Transgenic peach plants (Prunus persica L.) produced by genetic transformation of embryo sections using the green fluorescent protein (GFP) as an in vivo marker. Mol Breed 14:419–427CrossRefGoogle Scholar
  22. Pérez-Jiménez M, Carrillo-Navarro A, Cos-Terrer J (2012) Regeneration of peach (Prunus persica L. Batsch) cultivars and Prunus persica × Prunus dulcis rootstocks via organogenesis. Plant Cell Tissue Organ Cult 108:55–62CrossRefGoogle Scholar
  23. Pérez-Jiménez M, Cantero-Navarro E, Acosta M, Cos-Terrer J (2013) Relationships between endogenous hormonal content and direct somatic embryogenesis in Prunus persica L. Batsch cotyledons. Plant Growth Regul 71:219–224CrossRefGoogle Scholar
  24. Rubio-Moraga A, Rambla JL, Fernández-de-Carmen A, Trapero-Mozos A, Ahrazem O, Orzáez D, Granell A, Gómez-Gómez L (2014) New target carotenoids for CCD4 enzymes are revealed with the characterization of a novel stress-induced carotenoid cleavage dioxygenase gene from Crocus sativus. Plant Mol Biol 86:555–569CrossRefPubMedGoogle Scholar
  25. Sasaki S, Yamagishi N, Yoshikawa N (2011) Efficient virus-induced gene silencing in apple, pear and Japanese pear using Apple latent spherical virus vectors. Plant Methods 7:15PubMedCentralCrossRefPubMedGoogle Scholar
  26. Schery RW (1972) Plants for man. Prentice Hall, Englewood CliffsGoogle Scholar
  27. Tanaka N, Ayano Ureshino A, Shigeta N, Mimida N, Komori S, Takahashi S, Tanaka-Moriya Y, Wada M (2014) Overexpression of Arabidopsis FT gene in apple leads to perpetual flowering. Plant Biotechnol 31:11–20CrossRefGoogle Scholar
  28. Tsuda T, Yamaguchi M, Honda C, Moriguchi T (2004) Expression of anthocyanin biosynthetic genes in the skin of peach and nectarine fruit. J Am Soc Hortic Sci 129:857–862Google Scholar
  29. Vishnevetsky M, Ovadis M, Vainstein A (1999) Carotenoid sequestration in plants: the role of carotenoid-associated proteins. Trends Plant Sci 4:232–235CrossRefPubMedGoogle Scholar
  30. Wan C, Wilkins TA (1994) A modified hot-borate method significantly enhances the yield of high-quality RNA from cotton Gossypium hirsutum L. Anal Biochem 223:7–12Google Scholar
  31. Wang Z, Meng D, Wang A, Li T, Jiang S, Cong P, Li T (2013) The methylation of the PcMYB10 promoter is associated with green-skinned sport in Max Red Bartlett pear. Plant Physiol 162:885–896PubMedCentralCrossRefPubMedGoogle Scholar
  32. Williamson J, Peace C, Bliss F, Garner D, Crisosto C (2006) Evidence for a single locus controlling flesh color, senescent leaf color, and hypanthium color in peach. J Am Soc Hortic Sci 131:256–260Google Scholar
  33. Yamamizo C, Kishimoto S, Ohmiya A (2010) Carotenoid composition and carotenogenic gene expression during Ipomoea petal development. J Exp Bot 61:709–719PubMedCentralCrossRefPubMedGoogle Scholar
  34. Yamamizo C, Hirashima M, Kishimoto S, Ohmiya A (2011) Carotenoid composition in the yellow and pale green petals of Primula species. Bull Natl Inst Flor Sci 11:67–72Google Scholar
  35. Zhou H, Li M, Zhao X, Fan X, Guo A (2010) Plant regeneration from in vitro leaves of the peach rootstock ‘Nemaguard’ (Prunus persica × P. davidiana). Plant Cell Tissue Organ Cult 101:79–87CrossRefGoogle Scholar
  36. Zhou H, Lin-Wang K, Wang H, Gu C, Dare AP, Espley RV, He H, Allan AC, Han Y (2015) Molecular genetics of blood-fleshed peach reveals activation of anthocyanin biosynthesis by NAC transcription factors. Plant J 82:105–121CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Songling Bai
    • 1
  • Pham Anh Tuan
    • 1
  • Miho Tatsuki
    • 1
  • Hideaki Yaegaki
    • 1
  • Akemi Ohmiya
    • 2
  • Chihiro Yamamizo
    • 2
  • Takaya Moriguchi
    • 1
  1. 1.NARO Institute of Fruit Tree ScienceTsukubaJapan
  2. 2.NARO Institute of Floricultural ScienceTsukubaJapan

Personalised recommendations