Planta

, Volume 241, Issue 6, pp 1363–1379 | Cite as

Proteome analysis of pear reveals key genes associated with fruit development and quality

  • Jia Ming Li
  • Xiao San Huang
  • Lie Ting Li
  • Dan Man Zheng
  • Cheng Xue
  • Shao Ling Zhang
  • Jun Wu
Original Article

Abstract

Main conclusion

Comparative and association analyses of the proteome and transcriptome for pear fruit development were conducted for the first time in this study.

Pear fruit development involves complex physiological and biochemical processes, but there is still little knowledge available at proteomic and transcriptomic levels, which would be helpful for understanding the molecular mechanisms of fruit development and quality in pear. In our study, three important stages, including early development (S4-22), middle development (S6-27), and near ripening (S8-30), were investigated in ‘Dangshansuli’ by isobaric tags for relative and absolute quantitation (iTRAQ) labeling technology, identifying a total of 1,810 proteins during pear fruit development. The association analysis of proteins and transcript expression revealed 1,724, 1,722, and 1,718 associated proteins identified in stages S4-22, S6-27, and S8-30, respectively. A total of 237, 318, and 425 unique proteins were identified as differentially expressed during S4-22 vs S6-27, S6-27 vs S8-30, S4-22 vs S8-30, respectively, and the corresponding correlation coefficients of the overall differentially expressed proteins and transcripts data were 0.6336, 0.4113, and 0.7049. The phenylpropanoid biosynthesis pathway, which is related to lignin formation of pear fruit, was identified as a significantly enriched pathway during early stages of fruit development. Finally, a total of 35 important differentially expressed proteins related to fruit quality were identified, including three proteins related to sugar formation, seven proteins related to aroma synthesis, and sixteen proteins related to the formation of lignin. In addition, qRT-PCR verification provided further evidence to support differentially expressed gene selection. This study is the first to reveal protein and associated mRNA variations in pear during fruit development and quality conformation, and identify key genes and proteins helpful for future functional genomics studies, and provides gene resources for improvement of pear quality.

Keywords

Pear Fruit development Proteomic Transcriptomic Differential expression 

Abbreviations

DAFB

Day after full blooming,

iTRAQ

Isobaric tags for relative and absolute quantitation

SCX

Strong cation exchange

HPLC

High pressure liquid chromatography

AGC

Automatic gain control

HCD

High-energy collision dissociation

KEGG

Kyoto encyclopedia of genes and genomes

GO

Gene ontology

COG

Cluster of orthologous groups of proteins

PAL

Phenylalanineammonia-lyase

F5H

Ferulate-5-hydroxylase

4CL

4-coumarate-CoA ligase

CCR

Cinnamoyl-CoA reductase

CAD

Cinnamyl-alcoholdehydrogenase

POD

Peroxidase

HCT

Hydroxycinnamoyl transferase

MFS

Major facilitator super family transporter

Supplementary material

425_2015_2263_MOESM1_ESM.tif (4 mb)
Supplementary material 1 (TIFF 4067 kb)
425_2015_2263_MOESM2_ESM.tif (5.8 mb)
Supplementary material 2 (TIFF 5940 kb)
425_2015_2263_MOESM3_ESM.doc (158 kb)
Supplementary material 3 (DOC 158 kb)
425_2015_2263_MOESM4_ESM.xls (1.7 mb)
Supplementary material 4 (XLS 1734 kb)
425_2015_2263_MOESM5_ESM.xls (142 kb)
Supplementary material 5 (XLS 141 kb)

References

  1. Alba R, Payton P, Fei Z, McQuinn R, Debbie P, Martin GB, Tanksley SD, Giovannoni JJ (2005) Transcriptome and selected metabolite analyses reveal multiple points of ethylene control during tomato fruit development. Plant Cell 17(11):2954–2965CrossRefPubMedCentralPubMedGoogle Scholar
  2. Anterola AM, Lewis NG (2002) Trends in lignin modification: a comprehensive analysis of the effects of genetic manipulations/mutations on lignification and vascular integrity. Phytochemistry 61(3):221–294CrossRefPubMedGoogle Scholar
  3. Argenta LC, Fan X, Mattheis JP (2003) Influence of 1-methylcyclopropene on ripening, storage life, and volatile production by d’Anjou cv. pear fruit. J Agric Food Chem 51(13):3858–3864CrossRefPubMedGoogle Scholar
  4. Bain JM (1961) Some morphological, anatomical, and physiological changes in the pear fruit (Pyrus communis var. Williams Bon Chretien) during development and following harvest. Aust J Bot 9(2):99–123CrossRefGoogle Scholar
  5. Baucher M, Halpin C, Petit-Conil M, Boerjan W (2003) Lignin: genetic engineering and impact on pulping. Crit Rev Biochem Mol Biol 38(4):305–350CrossRefPubMedGoogle Scholar
  6. Bianco L, Lopez L, Scalone AG, Di Carli M, Desiderio A, Benvenuto E, Perrotta G (2009) Strawberry proteome characterization and its regulation during fruit ripening and in different genotypes. J Proteomics 72(4):586–607CrossRefPubMedGoogle Scholar
  7. Boerjan W, Ralph J, Baucher M (2003) Lignin biosynthesis. Annu Rev Plant Biol 54(1):519–546CrossRefPubMedGoogle Scholar
  8. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72(1):248–254CrossRefPubMedGoogle Scholar
  9. Cai Y, Li G, Nie J, Lin Y, Nie F, Zhang J, Xu Y (2010) Study of the structure and biosynthetic pathway of lignin in stone cells of pear. Sci Hortic 125(3):374–379CrossRefGoogle Scholar
  10. Cao H, Guo S, Xu Y, Jiang K, Jones AM, Chong K (2011) Reduced expression of a gene encoding a Golgi localized monosaccharide transporter (OsGMST1) confers hypersensitivity to salt in rice (Oryza sativa). J Exp Bot 62(13):4595–4604CrossRefPubMedCentralPubMedGoogle Scholar
  11. Chen JL, Wu JH, Wang Q, Deng H, Hu XS (2006) Changes in the volatile compounds and chemical and physical properties of Kuerle fragrant pear (Pyrus serotina Reld) during storage. J Agric Food Chem 54(23):8842–8847CrossRefPubMedGoogle Scholar
  12. Chen J, Wang Z, Wu J, Wang Q, Hu X (2007) Chemical compositional characterization of eight pear cultivars grown in China. Food Chem 104(1):268–275CrossRefGoogle Scholar
  13. Chervin C, Truett JK, Speirs J (1999) Alcohol dehydrogenase expression and alcohol production during pear ripening. J Am Soc Hortic Sci 124(1):71–75Google Scholar
  14. Choi J-H, Choi J-J, Hong K-H, Kim W-S, Lee S-H (2007) Cultivar differences of stone cells in pear flesh and their effects on fruit quality. Hortic Environ Biotechnol 48(1):27–31Google Scholar
  15. Christensen JH, Bauw G, Welinder KG, Van Montagu M, Boerjan W (1998) Purification and characterization of peroxidases correlated with lignification in poplar xylem. Plant Physiol 118(1):125–135CrossRefPubMedCentralPubMedGoogle Scholar
  16. Conde C, Agasse A, Silva P, Lemoine R, Delrot S, Tavares R, Gerós H (2007) OeMST2 encodes a monosaccharide transporter expressed throughout olive fruit maturation. Plant Cell Physiol 48(9):1299–1308CrossRefPubMedGoogle Scholar
  17. Dussi MC, Sugar D, Wrolstad RE (1995) Characterizing and quantifying anthocyanins in red pears and the effect of light quality on fruit color. J Am Soc Hortic Sci 120(5):785–789Google Scholar
  18. Eisen MB, Spellman PT, Brown PO, Botstein D (1998) Cluster analysis and display of genome-wide expression patterns. Proc Natl Acad Sci 95(25):14863–14868CrossRefPubMedCentralPubMedGoogle Scholar
  19. Fan G, Xu Y, Zhang X, Lei S, Yang S, Pan S (2011) Characteristics of immobilised β-glucosidase and its effect on bound volatile compounds in orange juice. Int J Food Sci Technol 46(11):2312–2320CrossRefGoogle Scholar
  20. Faurobert M, Mihr C, Bertin N, Pawlowski T, Negroni L, Sommerer N, Causse M (2007) Major proteome variations associated with cherry tomato pericarp development and ripening. Plant Physiol 143(3):1327–1346CrossRefPubMedCentralPubMedGoogle Scholar
  21. Feng S, Chen X, Zhang Y, Wang Y, Song Y, Chen X-L, Li X, Li M, Liu J, Wang Q (2011) Differential expression of proteins in red pear following fruit bagging treatment. Protein J 30(3):194–200CrossRefPubMedGoogle Scholar
  22. Feng C, Chen M, Xu C-J, Bai L, Yin X-R, Li X, Allan AC, Ferguson IB, Chen K-S (2012) Transcriptomic analysis of Chinese bayberry (Myrica rubra) fruit development and ripening using RNA-Seq. BMC Genom 13(1):19CrossRefGoogle Scholar
  23. Fernie AR, Stitt M (2012) On the discordance of metabolomics with proteomics and transcriptomics: coping with increasing complexity in logic, chemistry, and network interactions scientific correspondence. Plant Physiol 158(3):1139–1145CrossRefPubMedCentralPubMedGoogle Scholar
  24. Gasic K, Hernandez A, Korban SS (2004) RNA extraction from different apple tissues rich in polyphenols and polysaccharides for cDNA library construction. Plant Mol Biol Rep 22(4):437–438CrossRefGoogle Scholar
  25. Giribaldi M, Perugini I, Sauvage FX, Schubert A (2007) Analysis of protein changes during grape berry ripening by 2-DE and MALDI-TOF. Proteomics 7(17):3154–3170CrossRefPubMedGoogle Scholar
  26. Hudina M, Śtampar F (2000) Sugars and organic acids contents of European (Pyrus communis L.) and Asian (Pyrus serotina Rehd.) pear cultivars. Acta Alimentaria 29(3):217–230CrossRefGoogle Scholar
  27. Itai A, Kawata T, Tanabe K, Tamura F, Uchiyama M, Tomomitsu M, Shiraiwa N (1999) Identification of 1-aminocyclopropane-1-carboxylic acid synthase genes controlling the ethylene level of ripening fruit in Japanese pear (Pyrus pyrifolia Nakai). Mol Gen Genet 261(1):42–49CrossRefPubMedGoogle Scholar
  28. Jaillon O, Aury J-M, Noel B, Policriti A, Clepet C, Casagrande A, Choisne N, Aubourg S, Vitulo N, Jubin C (2007) The grapevine genome sequence suggests ancestral hexaploidization in major angiosperm phyla. Nature 449(7161):463–467CrossRefPubMedGoogle Scholar
  29. Kanehisa M, Goto S (2000) KEGG: kyoto encyclopedia of genes and genomes. Nucleic Acids Res 28(1):27–30CrossRefPubMedCentralPubMedGoogle Scholar
  30. Lara I, Miró R, Fuentes T, Sayez G, Graell J, López M (2003) Biosynthesis of volatile aroma compounds in pear fruit stored under long-term controlled-atmosphere conditions. Postharvest Biol Technol 29(1):29–39CrossRefGoogle Scholar
  31. Lee S-H, Choi J-H, Kim W-S, Han T-H, Park Y-S, Gemma H (2006) Effect of soil water stress on the development of stone cells in pear (Pyrus pyrifolia cv. ‘Niitaka’) flesh. Sci Hortic 110(3):247–253CrossRefGoogle Scholar
  32. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative pcr and the 2−ΔΔCT method. Methods 25(4):402–408CrossRefPubMedGoogle Scholar
  33. Lombard PB, Westwood MN (1987) Pear rootstocks. In: Rom RC, Carlson RF (eds) Rootstocks for fruit crops. Wiley, New York, pp 145–183Google Scholar
  34. Lu X-P, Liu Y-Z, An J-C, Hu H-J, Peng S-A (2011) Isolation of a cinnamoyl CoA reductase gene involved in formation of stone cells in pear (Pyrus pyrifolia). Acta Physiologiae Plantarum 33(2):585–591CrossRefGoogle Scholar
  35. Lücker J, Laszczak M, Smith D, Lund ST (2009) Generation of a predicted protein database from EST data and application to iTRAQ analyses in grape (Vitis vinifera cv. Cabernet Sauvignon) berries at ripening initiation. BMC Genom 10(1):50CrossRefGoogle Scholar
  36. Maier T, Schmidt A, Güell M, Kühner S, Gavin AC, Aebersold R, Serrano L (2011) Quantification of mRNA and protein and integration with protein turnover in a bacterium. Mol Syst Biol 7(1):511CrossRefPubMedCentralPubMedGoogle Scholar
  37. Marsh E, Alvarez S, Hicks LM, Barbazuk WB, Qiu W, Kovacs L, Schachtman D (2010) Changes in protein abundance during powdery mildew infection of leaf tissues of Cabernet Sauvignon grapevine (Vitis vinifera L.). Proteomics 10(10):2057–2064CrossRefPubMedGoogle Scholar
  38. Martínez-Esteso MJ, Vilella-Antón MT, Pedreño MÁ, Valero ML, Bru-Martínez R (2013) iTRAQ-based protein profiling provides insights into the central metabolism changes driving grape berry development and ripening. BMC Plant Biol 13(1):167CrossRefPubMedCentralPubMedGoogle Scholar
  39. Ming R, Hou S, Feng Y, Yu Q, Dionne-Laporte A, Saw JH, Senin P, Wang W, Ly BV, Lewis KL (2008) The draft genome of the transgenic tropical fruit tree papaya (Carica papaya Linnaeus). Nature 452(7190):991–996CrossRefPubMedCentralPubMedGoogle Scholar
  40. Morales AL, Duque C (2002) Free and glycosidically bound volatiles in the mammee apple (Mammea americana) fruit. Eur Food Res Technol 215(3):221–226CrossRefGoogle Scholar
  41. Mounet F, Moing A, Garcia V, Petit J, Maucourt M, Deborde C, Bernillon S, Le Gall G, Colquhoun I, Defernez M (2009) Gene and metabolite regulatory network analysis of early developing fruit tissues highlights new candidate genes for the control of tomato fruit composition and development. Plant Physiol 149(3):1505–1528CrossRefPubMedCentralPubMedGoogle Scholar
  42. Nogueira SB, Labate CA, Gozzo FC, Pilau EJ, Lajolo FM, Oliveira do Nascimento JR (2012) Proteomic analysis of papaya fruit ripening using 2DE-DIGE. Journal of proteomics 75(4):1428–1439CrossRefPubMedGoogle Scholar
  43. Osorio S, Alba R, Damasceno CM, Lopez-Casado G, Lohse M, Zanor MI, Tohge T, Usadel B, Rose JK, Fei Z (2011) Systems biology of tomato fruit development: combined transcript, protein, and metabolite analysis of tomato transcription factor (nor, rin) and ethylene receptor (Nr) mutants reveals novel regulatory interactions. Plant Physiol 157(1):405–425CrossRefPubMedCentralPubMedGoogle Scholar
  44. Palma JM, Corpas FJ, del Río LA (2011) Proteomics as an approach to the understanding of the molecular physiology of fruit development and ripening. J Proteomics 74(8):1230–1243CrossRefPubMedGoogle Scholar
  45. Pan Z, Zeng Y, An J, Ye J, Xu Q, Deng X (2012) An integrative analysis of transcriptome and proteome provides new insights into carotenoid biosynthesis and regulation in sweet orange fruits. J Proteomics 75(9):2670–2684CrossRefPubMedGoogle Scholar
  46. Pan X, Zhu B, Zhu H, Chen Y, Tian H, Luo Y, Fu D (2014) iTRAQ protein profile analysis of tomato green-ripe mutant reveals new aspects critical for fruit ripening. J Proteome Res 13(4):1979–1993CrossRefPubMedGoogle Scholar
  47. Pao SS, Paulsen IT, Saier MH (1998) Major facilitator superfamily. Microbiol Mol Biol Rev 62(1):1–34PubMedCentralPubMedGoogle Scholar
  48. Pedreschi R, Hertog M, Robben J, Noben J-P, Nicolaï B (2008) Physiological implications of controlled atmosphere storage of ‘Conference’pears (Pyrus communis L.): a proteomic approach. Postharvest Biol Technol 50(2):110–116CrossRefGoogle Scholar
  49. Pesis E (2005) The role of the anaerobic metabolites, acetaldehyde and ethanol, in fruit ripening, enhancement of fruit quality and fruit deterioration. Postharvest Biol Technol 37(1):1–19CrossRefGoogle Scholar
  50. Ross PL, Huang YN, Marchese JN, Williamson B, Parker K, Hattan S, Khainovski N, Pillai S, Dey S, Daniels S (2004) Multiplexed protein quantitation in Saccharomyces cerevisiae using amine-reactive isobaric tagging reagents. Mol Cell Proteomics 3(12):1154–1169CrossRefPubMedGoogle Scholar
  51. Sauer N, Tanner W (1989) The hexose carrier from Chlorella: cDNA cloning of a eucaryotic H+-cotransporter. FEBS Lett 259(1):43–46CrossRefPubMedGoogle Scholar
  52. Schwab W, Davidovich-Rikanati R, Lewinsohn E (2008) Biosynthesis of plant-derived flavor compounds. Plant J 54(4):712–732CrossRefPubMedGoogle Scholar
  53. Seymour GB, Taylor JE, Tucker GA (1993) Biochemistry of fruit ripening. Chapman & Hall, LondonCrossRefGoogle Scholar
  54. Shulaev V, Sargent DJ, Crowhurst RN, Mockler TC, Folkerts O, Delcher AL, Jaiswal P, Mockaitis K, Liston A, Mane SP (2011) The genome of woodland strawberry (Fragaria vesca). Nat Genet 43(2):109–116CrossRefPubMedCentralPubMedGoogle Scholar
  55. Soglio V, Costa F, Molthoff J, Weemen-Hendriks W, Schouten H, Gianfranceschi L (2009) Transcription analysis of apple fruit development using cDNA microarrays. Tree Gene Geno 5(4):685–698CrossRefGoogle Scholar
  56. Suwanagul A, Richardson DG (1997) Identification of headspace volatile compounds from different pear (Pyrus communis L.) varieties. In: VII International Symposium on Pear Growing 475 605–624Google Scholar
  57. Tao S, Khanizadeh S, Zhang H, Zhang S (2009) Anatomy, ultrastructure and lignin distribution of stone cells in two Pyrus species. Plant Sci 176(3):413–419CrossRefGoogle Scholar
  58. Tatusov RL, Natale DA, Garkavtsev IV, Tatusova TA, Shankavaram UT, Rao BS, Kiryutin B, Galperin MY, Fedorova ND, Koonin EV (2001) The COG database: new developments in phylogenetic classification of proteins from complete genomes. Nucleic Acids Res 29(1):22–28CrossRefPubMedCentralPubMedGoogle Scholar
  59. Velasco R, Zharkikh A, Affourtit J, Dhingra A, Cestaro A, Kalyanaraman A, Fontana P, Bhatnagar SK, Troggio M, Pruss D (2010) The genome of the domesticated apple (Malus domestica Borkh.). Nat Genet 42(10):833–839CrossRefPubMedGoogle Scholar
  60. Verde I, Abbott AG, Scalabrin S, Jung S, Shu S, Marroni F, Zhebentyayeva T, Dettori MT, Grimwood J, Cattonaro F (2013) The high-quality draft genome of peach (Prunus persica) identifies unique patterns of genetic diversity, domestication and genome evolution. Nat Genet 45(5):487–494CrossRefPubMedGoogle Scholar
  61. Washburn MP, Koller A, Oshiro G, Ulaszek RR, Plouffe D, Deciu C, Winzeler E, Yates JR (2003) Protein pathway and complex clustering of correlated mRNA and protein expression analyses in Saccharomyces cerevisiae. Proc Natl Acad Sci 100(6):3107–3112CrossRefPubMedCentralPubMedGoogle Scholar
  62. Whetten RW, MacKay JJ, Sederoff RR (1998) Recent advances in understanding lignin biosynthesis. Annu Rev Plant Biol 49(1):585–609CrossRefGoogle Scholar
  63. Wilkins MR, Sanchez J-C, Gooley AA, Appel RD, Humphery-Smith I, Hochstrasser DF, Williams KL (1996) Progress with proteome projects: why all proteins expressed by a genome should be identified and how to do it. Biotechnol Genet Eng Rev 13(1):19–50CrossRefPubMedGoogle Scholar
  64. Wu J, Wang Z, Shi Z, Zhang S, Ming R, Zhu S, Khan MA, Tao S, Korban SS, Wang H, Chen NJ, Nishio T, Xu X, Cong L, Qi K, Huang X, Wang Y, Zhao X, Wu J, Deng C, Gou C, Zhou W, Yin H, Qin G, Sha Y, Tao Y, Chen H, Yang Y, Song Y, Zhan D, Wang J, Li L, Dai M, Gu C, Wang Y, Shi D, Wang X, Zhang H, Zeng L, Zheng D, Wang C, Chen M, Wang G, Xie L, Sovero V, Sha S, Huang W, Zhang S, Zhang M, Sun J, Xu L, Li Y, Liu X, Li Q, Shen J, Wang J, Paull RE, Bennetzen JL, Wang J, Zhang S (2013) The genome of the pear (Pyrus bretschneideri Rehd.). Genome Res 23(2):396–408CrossRefPubMedCentralPubMedGoogle Scholar
  65. Wu H-X, Jia H-M, Ma X-w, Wang S-B, Yao Q-S, Xu W-t, Zhou Y-G, Gao Z-S, Zhan R-L (2014a) Transcriptome and proteomic analysis of mango (Mangifera indica Linn) fruits. J Proteomics 105:19–30CrossRefPubMedGoogle Scholar
  66. Wu J, Xu Z, Zhang Y, Chai L, Yi H, Deng X (2014b) An integrative analysis of the transcriptome and proteome of the pulp of a spontaneous late-ripening sweet orange mutant and its wild type improves our understanding of fruit ripening in citrus. J Exp Bot 65(6):1651–1671CrossRefPubMedCentralPubMedGoogle Scholar
  67. Xie M, Huang Y, Zhang Y, Wang X, Yang H, Yu O, Dai W, Fang C (2013) Transcriptome profiling of fruit development and maturation in Chinese white pear (Pyrus bretschneideri Rehd). BMC Genom 14(1):823CrossRefGoogle Scholar
  68. Xu Q, Chen L-L, Ruan X, Chen D, Zhu A, Chen C, Bertrand D, Jiao W-B, Hao B-H, Lyon MP (2013) The draft genome of sweet orange (Citrus sinensis). Nat Genet 45(1):59–66CrossRefPubMedGoogle Scholar
  69. Yamada K, Kojima T, Bantog N, Shimoda T, Mori H, Shiratake K, Yamaki S (2007) Cloning of two isoforms of soluble acid invertase of Japanese pear and their expression during fruit development. J Plant Physiol 164(6):746–755CrossRefPubMedGoogle Scholar
  70. Yang Y-N, Zhao G, Yue W-Q, Zhang S-L, Gu C, Wu J (2013) Molecular cloning and gene expression differences of the anthocyanin biosynthesis-related genes in the red/green skin color mutant of pear (Pyrus communis L.). Tree Genet Genomes 9(5):1351–1360CrossRefGoogle Scholar
  71. Yu K, Xu Q, Da X, Guo F, Ding Y, Deng X (2012) Transcriptome changes during fruit development and ripening of sweet orange (Citrus sinensis). BMC Genom 13(1):10CrossRefGoogle Scholar
  72. Zhou YS, Lamrani M, Chan-Park MB, Leong SSJ, Wook MC, Chen WN (2010) iTRAQ-coupled two-dimensional liquid chromatography/tandem mass spectrometric analysis of protein profile in Escherichia coli incubated with human neutrophil peptide 1-potential in antimicrobial strategy. Rapid Commun Mass Spectrom 24(18):2787–2790CrossRefPubMedGoogle Scholar
  73. Zhu M, Simons B, Zhu N, Oppenheimer DG, Chen S (2010) Analysis of abscisic acid responsive proteins in Brassica napus guard cells by multiplexed isobaric tagging. J Proteomics 73(4):790–805CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Jia Ming Li
    • 1
  • Xiao San Huang
    • 1
  • Lie Ting Li
    • 1
  • Dan Man Zheng
    • 2
  • Cheng Xue
    • 1
  • Shao Ling Zhang
    • 1
  • Jun Wu
    • 1
  1. 1.State Key Laboratory of Crop Genetics and Germplasm Enhancement, Centre of Pear Engineering Technology ResearchNanjing Agricultural UniversityNanjingChina
  2. 2.Roy J. Carver Biotechnology CenterUniversity of Illinois Urbana-ChampaignUrbanaUSA

Personalised recommendations