Theoretical and Applied Genetics

, Volume 122, Issue 3, pp 459–470 | Cite as

Characterization and genetic analysis of a low-temperature-sensitive mutant, sy-2, in Capsicum chinense

  • Song-Ji An
  • Devendra Pandeya
  • Soung-Woo Park
  • Jinjie Li
  • Jin-Kyung Kwon
  • Sota Koeda
  • Munetaka Hosokawa
  • Nam-Chon Paek
  • Doil Choi
  • Byoung-Cheorl Kang
Original Paper


A temperature-sensitive mutant of Capsicum chinense, sy-2, shows a normal developmental phenotype when grown above 24°C. However, when grown at 20°C, sy-2 exhibits developmental defects, such as chlorophyll deficiency and shrunken leaves. To understand the underlying mechanism of this temperature-dependent response, phenotypic characterization and genetic analysis were performed. The results revealed abnormal chloroplast structures and cell collapse in leaves of the sy-2 plants grown at 20°C. Moreover, an excessive accumulation of reactive oxygen species (ROS) resulting in cell death was detected in the chlorophyll-deficient sectors of the leaves. However, the expression profile of the ROS scavenging genes did not alter in sy-2 plants grown at 20°C. A further analysis of fatty acid content in the leaves showed the impaired pathway of linoleic acid (18:2) to linolenic acid (18:3). Additionally, the Cafad7 gene was downregulated in sy-2 plants. This change may lead to dramatic physiological disorder and alteration of leaf morphology in sy-2 plants by losing low-temperature tolerance. Genetic analysis of an F2 population from a cross between C. chinensesy-2’ and wild-type C. chinense ‘No. 3341’ showed that the sy-2 phenotype is controlled by a single recessive gene. Molecular mapping revealed that the sy-2 gene is located at a genomic region of the pepper linkage group 1, corresponding to the 300 kb region of the Ch1_scaffold 00106 in tomato chromosome 1. Candidate genes in this region will reveal the identity of sy-2 and the underlying mechanism of the temperature-dependent plant response.


Linolenic Acid Chloroplast Development Chloroplast Structure Photooxidative Damage COSII Marker 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This work was carried out with the support of “the BioGreen21 program (Code20070301034022)”, RDA, and by Technology Development Program for Agriculture and Forestry, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.

Supplementary material

122_2010_1460_MOESM1_ESM.doc (464 kb)
Supplementary material 1 (DOC 463 kb)


  1. Ahmad P, Sarwat M, Sharma S (2008) Reactive oxygen species, antioxidants and signaling in plants. J Plant Biol 51:167–173CrossRefGoogle Scholar
  2. Aluru MR, Bae H, Wu D, Rodermel SR (2001) The Arabidopsis immutans mutation affects plastid differentiation and the morphogenesis of white and green sectors in variegated plants. Plant Physiol 127:67–77CrossRefPubMedGoogle Scholar
  3. Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399CrossRefPubMedGoogle Scholar
  4. Blee E (2002) Impact or phyto-oxylipins in plant defense. Trends Plant Sci 7:315–321CrossRefPubMedGoogle Scholar
  5. Brash AR (1999) Lipoxygenase: occurrence, functions, catalysis, and acquisition of substrate. J Biol Chem 274:23679–23682CrossRefPubMedGoogle Scholar
  6. Browse J, Mccourt P, Somerville C (1986) A mutant of Arabidopsis deficient in C18-3 and C16-3 leaf lipids. Plant Physiol 81:859–864CrossRefPubMedGoogle Scholar
  7. Bryant DA, Frigaard NU (2006) Prokaryotic photosynthesis and phototrophy illuminated. Trends Microbiol 14:488–496CrossRefPubMedGoogle Scholar
  8. Chinnusamy V, Zhu J, Zhu JK (2007) Cold stress regulation of gene expression in plants. Trends Plant Sci 12:444–451CrossRefPubMedGoogle Scholar
  9. Cook D, Fowler S, Fiehn O, Thomashow MF (2004) A prominent role for the CBF cold response pathway in configuring the low-temperature metabolome of Arabidopsis. PNAS 101:15243–15248CrossRefPubMedGoogle Scholar
  10. Desai UJ, Pfaffle PK (1995) Single-step purification of a thermostable DNA polymerase expressed in Escherichia coli. Biotechniques 19:780–784PubMedGoogle Scholar
  11. Erickson AN, Markhart AH (2001) Flower production, fruit set, and physiology of bell pepper during elevate temperature and vapor pressure deficit. J Am Soc Hortic Sci 126:697–702Google Scholar
  12. Feussner I, Wasternack C (2002) The lipoxygenase pathway. Annu Rev Plant Biol 53:275–297CrossRefPubMedGoogle Scholar
  13. Fowler S, Thomashow MF (2002) Arabidopsis transcriptome profiling indicates that multiple regulatory pathways are activated during cold acclimation in addition to the CBF cold response pathway. Plant cell 14:1675–1690CrossRefPubMedGoogle Scholar
  14. Gechev TS, Breusegem FV, Stone JM, Denev I, Laloi C (2006) Reactive oxygen species as signals that modulate plant stress responses and programmed cell death. BioEssay 28:1091–1101CrossRefGoogle Scholar
  15. Givry S, Martin B, Patrick C, Denis M, Thomas S (2005) CARHTA GENE: multipopulation integrated genetic and radiation hybrid mapping. Bioinformatics 21:1703–1704CrossRefPubMedGoogle Scholar
  16. Gorsuch PA, Pandey S, Atkin OK (2010) Temporal heterogeneity of cold acclimation phenotypes in Arabidopsis leaves. Plant Cell Environ 33:244–258CrossRefPubMedGoogle Scholar
  17. Goulas E, Schubert M, Kieselbach T, Kleczkowski LA, Gardestrom P, Schroder W, Hurry V (2006) The chloroplast lumen and stromal proteomes of Arabidopsis thaliana show differential sensitivity to short- and long-term exposure to low temperature. Plant J 47:720–734CrossRefPubMedGoogle Scholar
  18. Gray GR, Heath D (2005) A global reorganization of the metabolome in Arabidopsis during cold acclimation is revealed by metabolic fingerprinting. Physiol Plant 124:236–248CrossRefGoogle Scholar
  19. Harvaux M, Kloppstech K (2001) The protective functions of carotenoid and flavonoid pigments against excess visible radiation at chilling temperature investigated in Arabidopsis npq and tt mutants. Planta 213:953–966CrossRefPubMedGoogle Scholar
  20. Herman EM, Rotter K, Premakumar R, Elwinger G, Bae R, Ehler-King L, Chen SX, Livingston DP (2006) Additional freeze hardiness in wheat acquired by exposure to −3°C is associated with extensive physiological, morphological, and molecular changes. J Exp Bot 57:3601–3618CrossRefPubMedGoogle Scholar
  21. Iba K (2002) Acclimative response to temperature stress in higher plants: approaches of gene engineering for temperature tolerance. Annu Rev Plant Biol 53:225–245CrossRefPubMedGoogle Scholar
  22. Iba K, Takamiya KI, Toh Y, Satoh H, Nishimura M (1991) Formation of functionally active chloroplasts is determined at a limited stage of leaf development in virescent mutants of rice. Dev Genet 12:342–348CrossRefGoogle Scholar
  23. Kachroo P, Shanklin J, Shah J, Whittle EJ, Klessig DF (2001) A fatty acid desaturase modulates the activation of defense signaling pathways in plants. PNAS 98:9448–9453CrossRefPubMedGoogle Scholar
  24. Kang BC, Nahm SH, Huh JH, Yoo HS, Yu JW, Lee MH, Kim BD (2001) An interspecific (Capsicum annuum × C. chinense) F2 linkage map in pepper using RFLP and AFLP markers. Theor Appl Genet 102:531–539CrossRefGoogle Scholar
  25. Kargiotidou A, Deli D, Galanopoulou D, Tsaftaris A, Farmaki T (2008) Low temperature and light regulate delta 12 fatty acid desaturases (FAD2) at a transcriptional level in cotton (Gossypium hirsutum). J Exp Bot 59:2043–2056CrossRefPubMedGoogle Scholar
  26. Kariola T, Brader G, Li J, Palva ET (2005) Chlorophyllase 1, a damage control enzyme, affects the balance between defense pathways in plants. Plant Cell 17:282–294CrossRefPubMedGoogle Scholar
  27. Knoetzel J, Simpson D (1991) Expression and organization of antenna proteins in the light-and temperature-sensitive barley mutant chlorina. Planta 185:111–123CrossRefGoogle Scholar
  28. Koeda S, Hosokawa M, Kang BC, Yazawa S (2009) Dramatic changes in leaf development of the native Capsicum chinense from the Seychelles at temperature below 24°C. J Plant Res 122:623–631CrossRefPubMedGoogle Scholar
  29. Komagata K, Suzuki K (1987) Lipid and cell-wall analysis in bacterial systematic. Methods Microbiol 19:1–67Google Scholar
  30. Li J, Pandeya D, Nath K, Zulfugarov S, Yoo SC, Zhang H, Yoo JH, Cho SW, Koh HJ, Kim DS, Seo HS, Kang BC, Lee CH, Paek NC (2010) Zebra-Necrosis, a thylakoid-bound protein, is critical for photoprotection of developing chloroplasts during early leaf development. Plant J 62:713–725CrossRefPubMedGoogle Scholar
  31. Lichtenthaler HK (1987) Chlorophyll fluorescence signatures of leaves during the autumnal chlorophyll breakdown. J Plant Physiol 131:101–110Google Scholar
  32. Lynch DV, Steponkus PL (1986) Plasma membrane lipid alterations associated with cold acclimation of winter rye seedlings (Secale cereale L. cv Puma). Plant Physiol 83:761–767CrossRefGoogle Scholar
  33. Miquel M, James D, Dooner H, Browse J (1993) Arabidopsis requires polyunsaturated lipids for low-temperature survival. PNAS 90:6208–6212CrossRefPubMedGoogle Scholar
  34. Mou Z, He Y, Dai Y, Liu X, Jiayang Li (2000) Deficiency in fatty acid synthase leads to premature cell death and dramatic alterations in plant morphology. Plant Cell 12:405–418CrossRefPubMedGoogle Scholar
  35. Park SW, An SJ, Yang HB, Kwon JK, Kang BC (2009) Optimization of high resolution melting analysis and discovery of single nucleotide polymorphism in Capsicum. Hortic Environ Biotechnol 50:31–39Google Scholar
  36. Pasini L, Bruschini S, Bertoli A, Mazza R, Fracheboud Y, Marocco A (2005) Photosynthetic performance of cold sensitive mutants of maize at low temperature. Physiol Plant 124:362–370CrossRefGoogle Scholar
  37. Provart NJ, Gil P, Chen W, Han B, Chang HS, Wang X, Zhu T (2003) Gene expression phenotypes of Arabidopsis associated with sensitivity to low temperatures. Plant Physiol 132:893–906CrossRefPubMedGoogle Scholar
  38. Robinson SJ, Parkin IAP (2008) Differential SAGE analysis in Arabidopsis uncovers increased transcriptome complexity in response to low temperature. BMC Genomics 9:434CrossRefPubMedGoogle Scholar
  39. Rodríguez M, Canales E, Borroto CJ, Carmona E, López J, Pujol M, Borrás-Hidalgo O (2006) Identification of genes induced upon water-deficit stress in a drought-tolerant rice cultivar. J Plant Physiol 163:577–584CrossRefPubMedGoogle Scholar
  40. Routaboul JM, Fischer SF, Browse J (2000) Trienoic fatty acids are required to maintain chloroplast function at low temperatures. Plant Physiol 124:1697–1705CrossRefPubMedGoogle Scholar
  41. Sharma P, Sharma N, Deswal R (2005) The molecular biology of the low-temperature response in plants. BioEssays 27:1048–1059CrossRefPubMedGoogle Scholar
  42. Suzuki N, Mittler R (2006) Reactive oxygen species and temperature stresses—a delicate balance between signaling and destruction. Physiol Plant 126:45–51CrossRefGoogle Scholar
  43. Thakur P, Kumar S, Malik JA, Berger JD, Nayyar H (2010) Cold stress effects on reproductive development in grain crops: an overview. Environ Exp Bot 67:429–443CrossRefGoogle Scholar
  44. Thomashow MF (1998) Role of cold-responsive genes in plant freezing tolerance. Plant Physiol 118:1–8CrossRefPubMedGoogle Scholar
  45. Thomashow MF (1999) Plant cold acclimation: freezing tolerance genes and regulatory mechanisms. Annu Rev Plant Physiol Plant Mol Biol 50:571–599CrossRefPubMedGoogle Scholar
  46. Wang WX, Vinocur B, Altman A (2003) Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta 218:1–14CrossRefPubMedGoogle Scholar
  47. Wang Y, Diehl A, Wu F, Vrebalov J, Giovannoni J, Siepel A, Tanksley SD (2008) Sequencing and comparative analysis of a conserved syntenic segment in the Solanaceae. Genetics 180:391–408CrossRefPubMedGoogle Scholar
  48. Washid A, Gelani S, Ashraf M, Foolad MR (2007) Heat tolerance in plants: an overview. Environ Exp Bot 61:199–223CrossRefGoogle Scholar
  49. Wilhelm S, Helmut S (2003) Antioxidant activity of carotenoids. Mol Aspects Med 24:345–351CrossRefGoogle Scholar
  50. Wu J, Lightner J, Warwick N, Browse J (1997) Low-temperature damage and subsequent recovery of fab1 mutant arabidopsis exposed to 2°C. Plant Physiol 113:347–356CrossRefPubMedGoogle Scholar
  51. Wu F, Mueller LA, Crouzillat D, Pétiard V, Tanksley SD (2006) Combining bioinformatics and phylogenetics to identify large sets of single-copy orthologous genes (COSII) for comparative, evolutionary and systematic studies: a test case in the euasterid plant clade. Genetics 174:1407–1420CrossRefPubMedGoogle Scholar
  52. Wu F, Eannetta NT, Xu YM, Durrett R, Mazourek M, Jahn MM, Tanksley SD (2009) A COSII genetic map of the pepper genome provides a detailed picture of synteny with tomato and new insights into recent chromosome evolution in the genus Capsicum. Theor Appl Genet 118:927–935CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Song-Ji An
    • 1
  • Devendra Pandeya
    • 1
  • Soung-Woo Park
    • 1
  • Jinjie Li
    • 1
    • 2
  • Jin-Kyung Kwon
    • 1
  • Sota Koeda
    • 3
  • Munetaka Hosokawa
    • 3
  • Nam-Chon Paek
    • 1
  • Doil Choi
    • 1
  • Byoung-Cheorl Kang
    • 1
  1. 1.Department of Plant Science, Plant Genomics and Breeding Institute, Research Institute for Agriculture and Life SciencesSeoul National UniversitySeoulKorea
  2. 2.Key Laboratory of Crop Genomics and Genetic Improvement of Ministry of Agriculture, Beijing Key Lab of Crop Genetic ImprovementChina Agriculture UniversityBeijingChina
  3. 3.Department of Agronomy and Horticultural Science, Graduate School of AgricultureKyoto UniversityKyotoJapan

Personalised recommendations