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Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 103, Issue 2, pp 165–174 | Cite as

Proteomic analysis of leaves from a diploid cybrid produced by protoplast fusion between Satsuma mandarin and pummelo

  • Lei Wang
  • Zhi-Yong Pan
  • Wen-Wu Guo
Original Paper

Abstract

In order to predict the performance of growth, development and resistance of a citrus diploid cybrid plant between Citrus unshiu Marc. cv. Guoqing No. 1 (G1) and Hirado Buntan pummelo (Citrus grandis (L.) Osbeck) (HBP), the proteomes of leaves from this cybrid and its parents were investigated. First, the diploid cybrid plant (G1 + HBP) created with its nuclear and chloroplast genomes from HBP and mitochondria genome from G1 was further verified by ploidy and molecular analysis (SSR, CAPS and cp-SSR). Then, 2-DE combined with MALDI-TOF/TOF MS were employed to analyze the variation of proteomes in the cybrid and its parental plants. Significant analysis allowed 90 (P < 0.05) differentially expressed protein spots between G1 and the cybrid, 20 between HBP and cybrid, and 116 between G1 and HBP. The comparative proteome patterns well validated the genetic background of the cybrid and its parental plants, suggesting that there is a good correlation between genome constitution and proteome expression in this cybrid. Seventy differentially expressed spots were selected for MALDI-TOF/TOF analysis and 25 proteins were identified. The identified proteins were mainly involved in photosynthesis, stress response, anti-oxidative stress and metabolism. Five proteins involved in photosynthesis such as Rubisco and Rubisco activase were significantly up-regulated in the cybrid, indicating the photosynthesis is enhanced in the cybrid. Expression analysis of proteins involved in stress response and anti-oxidative stress suggested that the resistance might be improved while anti-oxidative system exhibited a complex effect in the cybrid. The nitrogen, sulfur, carbon and energy metabolism might also be affected in the cybrid plant.

Keywords

Citrus Cybrid Proteomics 

Notes

Acknowledgments

This research was financially supported by the National Natural Science Foundation of China (Nos. 30921002, 30771481), the Ministry of Science & Technology of China (No. 2006AA100108) and the Science and Technology Department of Hubei province (No. 2008CDA069).

References

  1. Audrius AZ, Andrew PB (2005) Extraction methods for analysis of Citrus leaf proteins by two-dimensional gel electrophoresis. J Chromatogr A 1078:201–205CrossRefGoogle Scholar
  2. Badger MR, Price GD (1994) The role of carbonic anhydrase in photosynthesis. Annu Rev Plant Physiol Plant Mol Biol 45:369–392CrossRefGoogle Scholar
  3. Baker RL, Brown RL, Chen ZY, Cleveland TE, Fakhoury AM (2009) A maize lectin-like protein with antifungal activity against Aspergillus flavus. J Food Protection 72:120–127Google Scholar
  4. Bassene JB, Berti L, Carcouet E, Dhuique-Mayer C, Fanciullino AL, Bouffin J, Ollitrault P, Froelicher Y (2008) Influence of mitochondria origin on fruit quality in a citrus cybrid. J Agric Food Chem 56:8635–8640CrossRefPubMedGoogle Scholar
  5. Buchanan BB, Gruissem W, Jones RL (2004) Biochemistry and molecular biology of plants. Science Press, BeijingGoogle Scholar
  6. Cabasson CM, Luro F, Ollitrault P, Grosser JW (2001) Non-random inheritance of mitochondrial genomes in Citrus hybrids produced by protoplast fusion. Plant Cell Rep 20:604–609Google Scholar
  7. Cai XD, Fu J, Deng XX, Guo WW (2007) Production and molecular characterization of potential seedless cybrid plants between pollen sterile Satsuma mandarin and two seedy Citrus cultivars. Plant Cell Tiss Org Cult 90:275–283CrossRefGoogle Scholar
  8. Cantú MD, Mariano AG, Palma MS, Carrilho E, Wulff NA (2008) Proteomic analysis reveals suppression of bark chitinases and proteinase inhibitors in citrus plants affected by the citrus sudden death disease. Phytopathology 98:1084–1092CrossRefPubMedGoogle Scholar
  9. Cheng YJ, Guo WW, Deng XX (2003) Molecular characterization of cytoplasmic and nuclear genomes in phenotypically abnormal Valencia orange (Citrus sinensis) + Meiwa kumquat (Fortunella crassifolia) intergeneric somatic hybrids. Plant Cell Rep 21:445–451PubMedGoogle Scholar
  10. Dai SJ, Chen TT, Chong K, Xue YB, Liu SQ, Wang T (2007) Proteomic identification of differentially expressed proteins associated with pollen germination and tube growth reveals characteristics of germinated Oryza sativa pollen. Mol Cell Proteomics 6:207–230PubMedGoogle Scholar
  11. Davletova S, Rizhsky L, Liang H, Shengqiang Z, Olive DJ, Coutu J, Shulaev V, Schlauch K, Mittle R (2005) Cytosolic ascorbate peroxidase 1 is a central component of the reactive oxygen gene network of Arabidopsis. Plant Cell 17:268–281CrossRefPubMedGoogle Scholar
  12. Depege N, Drevet J, Boyer N (1998) Molecular cloning and characterization of tomato cDNAs encoding glutathione peroxidase-like proteins. Eur J Biochem 253:445–451CrossRefPubMedGoogle Scholar
  13. Downton J, Slatyer RO (1972) Temperature dependence of photosynthesis in cotton. Plant Physiol 50:518–622CrossRefPubMedGoogle Scholar
  14. Fanciullino AL, Gancel AL, Froelicher Y, Luro F, Ollitrault P, Brillouet JM (2005) Effects of nucleo-cytoplasmic interactions on leaf volatile compounds from citrus somatic diploid hybrids. J Agric Food Chem 53:4517–4523CrossRefPubMedGoogle Scholar
  15. Gancle AL, Grimplet J, Sauvage FX, Ollitrault P, Brillouet JM (2006) Predominant expression of diploid mandarin leaf proteome in two citrus mandarin-derived somatic allotetraploid hybrids. J Agric Food Chem 54:6212–6218CrossRefGoogle Scholar
  16. Giovanni C, Maurizio B, Luca M, Laura S, Gian Marco G, Barbara C, Paola O, Luciano Z, Pier GR (2004) Blue silver: a very sensitive colloidal coomassie G-250 staining for proteome analysis. Electrophoresis 25:1327–1333Google Scholar
  17. Grosser JW, Gmitter FG Jr (2005) Application of somatic hybridization and cybridization in crop improvement, with citrus as a model. In Vitro Cell Dev Biol Plant 41:220–225CrossRefGoogle Scholar
  18. Grosser JW, Gmitter FG Jr, Tusa N, Reforgiato Recupero G, Cucinotta P (1996) Further evidence of a cybridization requirement for plant regeneration from citrus leaf protoplasts following somatic fusion. Plant Cell Rep 15:672–676CrossRefGoogle Scholar
  19. Guo WW, Prasad D, Cheng YJ, Serrano P, Deng XX, Grosser JW (2004) Targeted cybridization in citrus: transfer of Satsuma cytoplasm to seedy cultivars for potential seedlessness. Plant Cell Rep 22:752–758CrossRefPubMedGoogle Scholar
  20. Guo WW, Wu RC, Cheng YJ, Deng XX (2008) Regeneration and molecular characterisation of two interspecific somatic hybrids of Citrus for potential rootstock improvement. J Hortic Sci Biotechnol 83:407–410Google Scholar
  21. Holland D, Ben-Hayyim G, Faltin Z, Camoin L, Strosberg AD, Eshdat Y (1993) Molecular characterization of salt-stress-associated protein in citrus: protein and cDNA sequence homology to mammalian glutathione peroxidases. Plant Mol Biol 21:923–927CrossRefPubMedGoogle Scholar
  22. Isaacon T, Damasceno CM, Saravanan RS, He YH, Catala C, Saladie M, Rose JK (2006) Sample extraction techniques for enhanced proteomic analysis of plant tissues. Nat Protoc 1:769–774CrossRefGoogle Scholar
  23. Kwon M, Davin LB, Lewis NG (2001) In situ hybridization and immunolocalization of lignan reductases in woody tissues: implications for heartwood formation and other forms of vascular tissue preservation. Phytochemistry 57:899–914CrossRefPubMedGoogle Scholar
  24. Lang P, Zhang CK, Ebel RC, Dane F, Dozier WA (2005) Identification of cold acclimated genes in leaves of Citrus unshiu by mRNA differential display. Gene 359:111–118CrossRefPubMedGoogle Scholar
  25. Levine A, Tenhaken R, Dixon R, Lamb C (1994) H2O2 from oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell 79:583–593CrossRefPubMedGoogle Scholar
  26. Li CJ, Lin QH, Zhang CF (1999) Influence of NaCl on Gln synthetase and its isozymes in rice. J Wuhan University 45:497–500 In ChineseGoogle Scholar
  27. Li XM, Deng XX, Deng BX (2000) Study on the photosynthetic trait of somatic hybrid in citrus. J Agric Huazhong University 19:585–588 In ChineseGoogle Scholar
  28. Lliso I, Tadeo FR, Phinney BS, Wilkerson CG, Talón M (2007) Protein changes in the albedo of citrus fruits on postharvesting storage. J Agric Food Chem 55:9047–9053CrossRefPubMedGoogle Scholar
  29. Lyou SH, Park HJ, Jung C, Sohn HB, Lee G, Kim CH, Kim M, Choi YD, Cheong JJ (2009) The Arabidopsis AtLEC gene encoding a lectin-like protein is up-regulated by multiple stimuli including developmental signal, wounding, jasmonate, ethylene, and chitin elicitor. Mol Cell 27:75–81CrossRefGoogle Scholar
  30. Mittler R, Vanderauwera S, Gollery M, Van Breusegem F (2004) Reactive oxygen gene network of plants. Trends Plant Sci 9:490–498CrossRefPubMedGoogle Scholar
  31. Moreira CD, Chase CD, Gmitter FG Jr, Grosser JW (2000) Inheritance of organelle genomes in citrus somatic cybrids. Mol Breed 6:401–405CrossRefGoogle Scholar
  32. Moriguchi T, Hidaka T, Omura M, Motomura T, Akihama T (1996) Genotypes and parental combination influence efficiency of cybrid induction in Citrus by electrofusion. HortSci 31:275–278Google Scholar
  33. Nam MH, Heo EJ, Kim JY, Kim SI, Kwon KH, Seo JB, Kwon O, Yoo JS, Park YM (2003) Proteome analysis of the responses of Panax ginseng C.A. Meyer leaves to high light: use of electrospray ionization quadruple-time of flight mass spectrometry and expressed sequence tag data. Proteomics 3:2351–2367CrossRefPubMedGoogle Scholar
  34. Pan ZY, Guan R, Zhu SP, Deng XX (2009) Proteomic analysis of somatic embryogenesis in Valencia sweet orange (Citrus sinensis Osbeck). Plant Cell Rep 28:281–289CrossRefPubMedGoogle Scholar
  35. Parry MA, Andralojc PJ, Khan S, Lea PJ, Keys AJ (2002) Rubisco activity: effects of drought stress. Ann Bot (Lond) 89:833–839CrossRefGoogle Scholar
  36. Romero-Aranda R, Bondana BR, Syvertsen JP, Grosser JW (1997) Leaf characteristics and net gas exchange of diploid and autotetraploid citrus. Ann Bot 79:153–160CrossRefGoogle Scholar
  37. Salekdeh GH, Siopongco J, Wade LJ, Ghareyazie B, Bennett J (2002) Proteomic analysis of rice leaves during drought stress and recovery. Proteomics 2:1131–1145CrossRefPubMedGoogle Scholar
  38. Sasaki H, Samejima M, Ishii R (1996) Analysis by 13C measurement on mechanism of cultivar difference in leaf photosynthesis of rice (Oryza sativa L.). Plant Cell Physiol 37:1161–1166Google Scholar
  39. Schlumbaum A, Mauch F, Voegeli U, Boller T (1986) Plant chitinases are potent inhibitors of fungal growth. Nature 324:365–367CrossRefGoogle Scholar
  40. Tanou G, Job C, Rajjou L, Arc E, Belghazi M, Diamantidis G, Molassiotis A, Job D (2009) Proteomics reveals the overlapping roles of hydrogen peroxide and nitric oxide in the acclimation of citrus plants to salinity. Plant J 60:795–804CrossRefPubMedGoogle Scholar
  41. Tsukuda S, Gomi K, Yamamoto H, Akimitsu K (2006) Characterization of cDNAs encoding two distinct miraculin-like proteins and stress-related modulation of the corresponding mRNAs in Citrus jambhiri Lush. Plant Mol Biol 60:125–136CrossRefPubMedGoogle Scholar
  42. Vander Mijnsbrugge K, Beeckman H, De Rycke R, Van Montagu M, Engler G, Boerjan W (2000) Phenylcoumaran benzylic ether reductase, a prominent poplar xylem protein, is strongly associated with phenylpropanoid biosynthesis in lignifying cells. Planta 211:502–509CrossRefPubMedGoogle Scholar
  43. Wang W, Vignani R, Scali M, Cresti M (2006) A universal and rapid protocol for protein from recalcitrant plant tissues for proteomic analysis. Electrophoresis 27:2782–2786CrossRefPubMedGoogle Scholar
  44. Wu YY, Wu DY, Zhang HP, Liu J, Li GX (2007) Study on the relationship of carbonic anhydrase activity and photosynthetic trait in soybean. Agric Sci Henan 2:43–45 (In Chinese)Google Scholar
  45. Yamamoto M, Kobayashi S (1995) A cybrid plant produced by electrofusion between Citrus unshiu and C. sinensis. Plant Tiss Cult Lett 12:131–137Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.Key Laboratory of Horticultural Plant Biology, Ministry of Education; National Key Laboratory of Crop Genetic ImprovementHuazhong Agricultural UniversityWuhanChina

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