Identification of an AtCRN1-like chloroplast protein BeCRN1 and its distinctive role in chlorophyll breakdown during leaf senescence in bamboo (Bambusa emeiensisViridiflavus’)

  • Qiang Wei
  • Huiming Cao
  • Zhongru Li
  • Benke Kuai
  • Yulong DingEmail author
Original Paper


CRN1/PPH/NYC3 is one of the key genes responsible for chlorophyll degradation during senescence in both Arabidopsis and rice. In this study, BeCRN1 and its promoter, BeCRN1p, were isolated from, Bambusa emeiensis ‘Viridiflavus’, a bamboo variety, for the first time. BeCRN1 consists of 1,646 bp, with an open reading frame of 1,473 bp, encoding a predicted polypeptide of 490 amino acids. Besides, BeCRN1 contained 5 exons and 4 introns, and harbored a distinctive microsatellite in the fourth intron. Differences in BeCRN1p are observed primarily in the AtCRN1 promoter, while sharing similar cis element composition with the rice CRN1 promoter. BeCRN1 is localized within the chloroplast, and it is strongly induced by senescence signals. Although constitutive overexpression of BeCRN1 in crn1 has reversed the stay-green phenotype of crn1 to the wild type phenotype, its promoter has failed to do so with AtCRN1 following dark treatment. The efficiency of BeCRN1p is only about one-third of that of the AtCRN1 promoter.


Bamboo Chlorophyll degradation CRN1 Stay green 



This work was supported by a grant from the Natural Science Foundation of China (Grant No. 31000294), a grant from the National Science and Technology Support Program of China during the 12th Five-Year Plan Period (No. 2012BAD23B05), and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.


  1. Alos E, Roca M, Iglesias DJ, Minguez-Mosquera MI, Damasceno CMB, Thannhauser TW, Rose JKC, Talon M, Cercos M (2008) An evaluation of the basis and consequences of a stay-green mutation in the navel negra citrus mutant using transcriptomic and proteomic profiling and metabolite analysis. Plant Physiol 147:1300–1315PubMedCrossRefGoogle Scholar
  2. Armstead I, Donnison I, Aubry S, Harper J, Hortensteiner S, James C, Mani J, Moffet M, Ougham H, Roberts L, Thomas A, Weeden N, Thomas H, King I (2006) From crop to model to crop: identifying the genetic basis of the staygreen mutation in the Lolium/Festuca forage and amenity grasses. New Phytol 172:592–597PubMedCrossRefGoogle Scholar
  3. Armstead I, Donnison I, Aubry S, Harper J, Hortensteiner S, James C, Mani J, Moffet M, Ougham H, Roberts L, Thomas A, Weeden N, Thomas H, King I (2007) Cross-species identification of Mendel’s/locus. Science 315:73PubMedCrossRefGoogle Scholar
  4. Barry CS (2009) The stay-green revolution: recent progress in deciphering the mechanisms of chlorophyll degradation in higher plants. Plant Sci 176:325–333CrossRefGoogle Scholar
  5. Barry CS, McQuinn RP, Chung MY, Besuden A, Giovannoni JJ (2008) Amino acid substitutions in homologs of the STAY-GREEN protein are responsible for the green-flesh and chlorophyll retainer mutations of tomato and pepper. Plant Physiol 147:179–187PubMedCrossRefGoogle Scholar
  6. Benedetti CE, Arruda P (2002) Altering the expression of the chlorophyllase gene ATHCOR1 in transgenic Arabidopsis caused changes in the chlorophyll-to-chlorophyllide ratio. Plant Physiol 128:1255–1263PubMedCrossRefGoogle Scholar
  7. Borovsky Y, Paran I (2008) Chlorophyll breakdown during pepper fruit ripening in the chlorophyll retainer mutation is impaired at the homolog of the senescence-inducible stay-green gene. Theor Appl Genet 117:235–240PubMedCrossRefGoogle Scholar
  8. Chen YX, Qiu K, Kuai BK, Ding YL (2011) Identification of an NAP-like transcription factor BeNAC1 regulating leaf senescence in bamboo (Bambusa emeiensisViridiflavus’). Physiol Plantarum 142:361–371CrossRefGoogle Scholar
  9. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J Cell Mol Biol 16:735–743CrossRefGoogle Scholar
  10. Fromm M, Taylor LP, Walbot V (1985) Expression of genes transferred into monocot and dicot plant cells by electroporation. Proc Natl Acad Sci 82:5824–5828PubMedCrossRefGoogle Scholar
  11. Holsters M, de Waele D, Depicker A, Messens E, van Montagu M, Schell J (1978) Transfection and transformation of Agrobacterium tumefaciens. Mol Gen Genet 163:181–187PubMedCrossRefGoogle Scholar
  12. Hortensteiner S (2006) Chlorophyll degradation during senescence. Annu Rev Plant Biol 57:55–77PubMedCrossRefGoogle Scholar
  13. Jiang HW, Li MR, Liang NB, Yan HB, Wei YL, Xu X, Liu JF, Xu Z, Chen F, Wu GJ (2007) Molecular cloning and function analysis of the stay green gene in rice. Plant J 52:197–209PubMedCrossRefGoogle Scholar
  14. Liu YG, Chen Y (2007) High-efficiency thermal asymmetric interlaced PCR for amplification of unknown flanking sequences. Biotechniques 43:649PubMedCrossRefGoogle Scholar
  15. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(−delta delta C) method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  16. Morita R, Sato Y, Masuda Y, Nishimura M, Kusaba M (2009) Defect in non-yellow coloring 3, an alpha/beta hydrolase-fold family protein, causes a stay-green phenotype during leaf senescence in rice. Plant J 59:940–952PubMedCrossRefGoogle Scholar
  17. Park SY, Yu JW, Park JS, Li J, Yoo SC, Lee NY, Lee SK, Jeong SW, Seo HS, Koh HJ, Jeon JS, Park YI, Paek NC (2007) The senescence-induced staygreen protein regulates chlorophyll degradation. Plant Cell 19:1649–1664PubMedCrossRefGoogle Scholar
  18. Ren G, An K, Liao Y, Zhou X, Cao Y, Zhao H, Ge X, Kuai B (2007) Identification of a novel chloroplast protein AtNYE1 regulating chlorophyll degradation during leaf senescence in Arabidopsis. Plant Physiol 144:1429–1441PubMedCrossRefGoogle Scholar
  19. Ren G, Zhou Q, Wu S, Zhang Y, Zhang L, Huang J, Sun Z, Kuai B (2010) Reverse genetic identification of CRN1 and its distinctive role in chlorophyll degradation in Arabidopsis. J Integr Plant Biol 52:496–504PubMedGoogle Scholar
  20. Sato Y, Morita R, Nishimura M, Yamaguchi H, Kusaba M (2007) Mendel’s green cotyledon gene encodes a positive regulator of the chlorophyll-degrading pathway. Proc Natl Acad Sci USA 104:14169–14174PubMedCrossRefGoogle Scholar
  21. Schelbert S, Aubry S, Burla B, Agne B, Kessler F, Krupinska K, Hortensteiner S (2009) Pheophytin pheophorbide hydrolase (Pheophytinase) is involved in chlorophyll breakdown during leaf senescence in arabidopsis. Plant Cell 21:767–785PubMedCrossRefGoogle Scholar
  22. Wei Q, Guo YJ, Kuai BK (2011) Isolation and characterization of a chlorophyll degradation regulatory gene from tall fescue. Plant Cell Rep 30:1201–1207PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Qiang Wei
    • 1
  • Huiming Cao
    • 1
  • Zhongru Li
    • 1
  • Benke Kuai
    • 2
  • Yulong Ding
    • 1
    Email author
  1. 1.Bamboo Research Institute and College of Forest Resources and EnvironmentNanjing Forestry UniversityNanjingChina
  2. 2.State Key Laboratory of Genetic Engineering and Institute of Plant Biology, School of Life SciencesFudan UniversityShanghaiChina

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