Polyethylene glycol effects on somatic embryogenesis of papaya hybrid UENF/CALIMAN 01 seeds

Heringer, Angelo Schuabb; Vale, Ellen Moura; Barroso, Tatiana; Santa-Catarina, Claudete; Silveira, Vanildo

Abstract

Among commercial fruits, papaya (Carica papaya L.) is notably one of the most common ones around the world. The aim of the current study was to establish a somatic embryogenesis protocol for the hybrid UENF/CALIMAN 01, elucidating the effects of sucrose and 2,4-dichlorophenoxyacetic acid (2,4-D) on the induction of embryogenic cultures and the effect of polyethylene glycol (PEG) on somatic embryo maturation. Immature zygotic embryos of C. papaya hybrid UENF/CALIMAN 01 were cultured in MS culture medium supplemented with different concentrations of sucrose (30 and 60 g L-1) and 2,4-D (0, 20, 40 and 80 μM). The combination of 30 g L-1 sucrose and 20 μM 2,4-D resulted in the highest induction rates and the largest callus diameters. Furthermore, this combination was associated with the greatest potential to form somatic embryos. To promote maturation, cultures were inoculated in MS medium and exposed to different concentrations of PEG (0, 30 and 60 g L-1). The 60 g L-1 PEG treatment group had the highest average for the number, maturation and conversion of somatic embryos that resulted in plantlets at the end of the process. The addition of PEG to MS medium led to increased protein synthesis. These results are unprecedented for C. papaya hybrid UENF/CALIMAN 01 as the current study demonstrates the development of somatic embryogenesis in this high quality hybrid.

Carica papaya L.; hybrid; induction; maturation; polyethylene glycol; protein analysis; somatic embryos

Ascencio-Cabral A, Rodríguez-Garay B, Gutiérrez-Pulido H, Gutiérrez-Mora A (2008) Plant regeneration of Carica papaya through somatic embryogenesis in response to light quality, gelling agent and phloridzin. Scientia Horticulturae 118:155-160.
Anandan R, Sudhakar D, Balasubramanian P, Gutieirrez-Mora A (2012) In vitro somatic embryogenesis from suspension cultures of Carica papaya L. Scientia Horticulturae 136:43-49.
Aroucha EMM, Silva RF, Oliveira JG, Viana AP, Gonzaga MP (2005) Época de colheita e período de repouso dos frutos de mamão (Carica papaya L.) cv Golden na qualidade fisiológica das sementes. Ciência Rural 35:537-543.
Bhattacharya J, Khuspe SS (2001) In vitro and in vivo germination of papaya (Carica papaya L.) seeds. Scientia Horticulturae 91:39-49.
Bhattacharya J, Renukdas NN, Khuspe SS, Rawal SK (2003) Multiple shoot regeneration from immature embryo explants of papaya. Biologia Plantarum 47:327-331.
Borisjuk L, Walenta S, Rolletschek H, Mueller-Klieser W, Wobus U, Weber H (2002) Spatial analysis of plant metabolism: Sucrose imaging with Vicia faba cotyledons reveals specific development patterns. The Plant Journal 29:521-530.
Borisjuk L, Rolletschek H, Wobus U, Weber H (2003) Differentiation of legume cotyledon as related to metabolism gradients and assimilate transport into seeds. Journal of Experimental Botany 54:503-512.
Bruijne E de, Langhe E de, Van Rijck R (1974) Actions of hormones and embryoid formation in callus cultures of Carica papaya In International Symposium on Crop Protection, Fytofarmacie en Fytiatrie Rijkslandsbouwhoogeschool Medelelingen 26:637-645.
Businge E, Bygdell J, Wingsle G, Moritz T, Egertsdotter U (2013) The effect of carbohydrates and osmoticum on storage reserve accumulation and germination of Norway spruce somatic embryos. Physiologia Plantarum doi:10.1111/ppl.12039.
Calic-Dragosavac D, Zdravkovi-Kora S, Bohanec B, Radojevi L, Vinterhalter B, Stevovi S, Cingel A, Savi J (2010) Effect of activated charcoal, abscisic acid and polyethylene glycol on maturation, germination and conversion of Aesculus hippocastanum androgenic embryos. African Journal of Biotechnology 9:3786-3793.
Chen MH, Wang PJ, Maeda E (1987) Somatic embryogenesis and plant regeneration in Carica papaya L. tissue culture derived from root explants. Plant Cell Reports 6:348-351.
Chen MH, Chen CC (1992). Plant regeneration from Carica protoplasts. Plant Cell Reports 11:404-407.
Coelho CMM, Benedito VA (2008) Seed development and reserve compound accumulation in common bean (Phaseolus vulgaris L.). Seed Science and Biotechnology 2:42-52.
Almeida PA, Oliveira RP, Dantas JLL (2001) Papaya callus and somatic embryo induction and development. Scientia Agricola 58: 51-54.
El-Tayeb MA, Hassanein AM (2000) Germination, seedling growth, some organic solutes and peroxidase expression of different Vicia faba lines as influenced by water stress. Acta Agronomica Hungarica 48:11-20.
FAO (Food and Agriculture Organization of the United Nations)/ FAOSTAT, 2010. http://faostat.fao.org/site/339/default.aspx
» link
Farzana ARF, Palkadapala PGVN, Meddegoda KMMN, Samarajeewa PK, Eeswara JP (2008) Somatic embryogenesis in papaya (Carica papaya L. cv. Rathna). Journal of the National Science Foundation of Sri Lanka 36:41-50.
Fehér A, Pasternak TP, Dudits D (2003) Transition of somatic plant cells to an embryogenic state. Plant Cell, Tissue and Organ Culture 74:201-228.
Ferreguetti GA (2003) Caliman 01 - O primeiro híbrido mamão Formosa brasileiro. In: Annals of the Papaya Brasil, Vitoria. pp. 211-218.
Find JI (1997) Changes in endogenous ABA levels in developing somatic embryos of Norway spruce (Picea abies (L.) Karst.) in relation to maturation medium, desiccation and germination. Plant Science 128:75-83.
Fitch M, Manshardt R (1990) Somatic embryogenesis and plant regeneration from immature zygotic embryos of papaya (Carica papaya L.). Plant Cell Reports 9:320-324.
Goyal K, Walton LJ, Tunnacliffe A (2005) LEA proteins prevent protein aggregation due to water stress. Biochemical Journal 388:151-157.
Jiménez VM, Bangerth F (2001) Endogenous hormone levels in explants and in embryogenic and non-embryogenic cultures of carrot. Physiologia Plantarum 111:389-395.
Keinonen-Metta La K, Jalonen P, Eurola P, Von Arnold S, Von Weissenberg K (1996) Somatic embryogenesis of Pinus sylvestris. Scandinavian Journal of Forest Research 11:242-250.
Koch KE (2004) Sucrose metabolism: regulatory mechanism and pivotal roles in sugar sensing and plant development. Current Opinion in Plant Biology 7:235-246.
Koehler AD, Carvalho CR, Abreu IS, Clarindo WR (2013) Somatic embryogenesis from leaf explants of hermaphrodite Carica papaya: A new approach for clonal propagation. African Journal of Biotechnology 12:2386-2391.
Krajnakova J, Haggman H, Gomory D (2009) Effect of sucrose concentration, polyethylene glycol and activated charcoal on maturation and regeneration of Abies cephalonica somatic embryos. Plant Cell, Tissue and Organ Culture 96:251-262.
Langhansov L, Konrdov H, Vanek T (2004) Polyethylene glycol and abscisic acid improve maturation and regeneration of Panax ginseng somatic embryos. Plant Cell Reports 22:725-730.
Lazzeri PA, Hildebrand DF, Sunega J, Williams EG, Collins GB (1988) Soybean somatic embryogenesis: interactions between sucrose and auxin. Plant Cell Reports 7:517-520.
Li XY, Huang FH, Murphy JB, Gbur JR. EE (1998) Polyethylene Glycol and Maltose Enhance Somatic Embryo Maturation in Loblolly Pine (Pinus taeda L.). In Vitro Cellular and Developmental Biology - Plant 34:22-26.
Litz RE, Conover RA (1982) In vitro somatic embryogenesis and plant regeneration from Carica papaya L. ovular callus. Plant Science Letters 26:153-158.
Litz RE (1986) Effect of osmotic stress on somatic embryogenesis in Carica suspension cultures. Journal of the American Society for Horticultural Science 111:969-972.
Lo Schiavo F, Pitto L, Giuliano G, Torti G, Nutironchi V, Marazziti D, Vergara R, Orselli S, Terzi M (1989) DNA methylation of embryogenic carrot cell cultures and its variations as caused by mutation, differentiation, hormones and hypomethylating drugs. Theoretical and Applied Genetics 77:325-331.
Malabadi RB, Kumar SV, Mulgund GS, Nataraja K (2011) Induction of somatic embryogenesis in Papaya (Carica papaya). Research in Biotechnology 2:40-55.
Martin AB, Cuadrado Y, Guerra H, Gallego P, Hita O, Martin L, Dorado A, Villalobos N (2000) Differences in the contents of total sugars, reducing sugars, starch and sucrose in embryogenic and non-embryogenic calli from Medicago arborea L. Plant Science 154:143-151.
Maruyama E, Hosoi Y (2007) Polyethylene glycol enhance somatic embryo production in Japanese cedar (Cryptomeria japonica D. Don). Propagation of Ornamental Plants 7:57-61.
Mishra M, Shukla N, Chandra R (2010) Role of polyethylene glycol in maturation and germination of trasformed somatic embryos of papaya (Carica papaya L.). Acta Horticulturae 851:227-230.
Morais PLD, Silva GG, Menezes JB, Maia FEN, Dantas DJ, Sales Junior R (2007) Postharvest shelf life of papaya UENF/CALIMAN 01 produced in Rio Grande do Norte. Revista Brasileira de Fruticultura 29:666-670.
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15:473-497.
Pinto DLP, Almeida AMR, Rêgo MM, Silva ML, Oliveira EJ, Otoni WC (2011) Somatic embryogenesis from mature zygotic embryos of commercial passion fruit (Passiflora edulis Sims) genotypes. Plant Cell, Tissue and Organ Culture 3:521-530.
Santa-Catarina C, Silveira V, Balbuena TS, Viana AM, Estelita MEM, Handro W, Floh EIS (2006) IAA, ABA, polyamines and free amino acids associated with zygotic embryo development of Ocotea catharinensis Plant Growth Regulation 49:237-247.
Schmildt ER, Amaral JAT, Schmildt O (2007) Sucrose on in vitro rooting phase of papaya tree 'Tainung 01'. Scientia Agraria 8:25-31.
Silva ML, Pinto DLP, Guerra MP, Floh EIS, Bruckner CH, Otoni WC (2009) A novel regeneration system for a wild passion fruit species (Passiflora cincinnata Mast.) based on somatic embryogenesis from mature zygotic embryos. Plant Cell, Tissue and Organ Culture 99:47-54.
Smulders MJM, de Klerk GJ (2011) Epigenetics in plant tissue culture. Plant Growth Regulation 63:137-146.
Sokal RR, Rohlf FJ (1995) Biometry: the principles of statistics in biological research. Freeman, New York.
Stasolla C, Yeung EC (2003) Recent advances in conifer somatic embryogenesis: improving somatic embryo quality. Plant Cell, Tissue and Organ Culture 74:15-35.
Walker DR, Parrott WA (2001) Effect of polyethylene glycol and sugar alcohols on soybean somatic embryo germination and conversion. Plant Cell, Tissue Organ and Culture 64:55-62.
Welin BV, Olson A, Nylander M, Palva ET (1994) Characterisation and differential expression of DHN/LEA/RAB -like genes during cold-acclimation and drought stress in Arabidopsis thaliana Plant Molecular Biology 26:131-144.
Wind J, Smeekens S, Hanson J (2010) Sucrose: metabolite and signaling molecule. Phytochemistry 71:1610-1614.
Vahdati K, Bayat S, Ebrahimzadeh H, Jariteh M, Mirmasoumi M (2008) Effect of exogenous ABA on somatic embryo maturation and germination in Persian walnut (Juglans regia L.). Plant Cell, Tissue and Organ Culture 93:163-171.
Vooková B, Kormuták A (2002) Some features of somatic embryo maturation of Algerian fir. In Vitro Cellular and Developmental Biology-Plant 38:549-551.

Publication Dates

Publication in this collection
15 Oct 2013
Date of issue
2013

History

Received
31 Jan 2013
Accepted
15 July 2013

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] Ascencio-Cabral A, Rodríguez-Garay B, Gutiérrez-Pulido H, Gutiérrez-Mora A (2008) Plant regeneration of Carica papaya through somatic embryogenesis in response to light quality, gelling agent and phloridzin. Scientia Horticulturae 118:155-160.

[2] Anandan R, Sudhakar D, Balasubramanian P, Gutieirrez-Mora A (2012) In vitro somatic embryogenesis from suspension cultures of Carica papaya L. Scientia Horticulturae 136:43-49.

[3] Aroucha EMM, Silva RF, Oliveira JG, Viana AP, Gonzaga MP (2005) Época de colheita e período de repouso dos frutos de mamão (Carica papaya L.) cv Golden na qualidade fisiológica das sementes. Ciência Rural 35:537-543.

[4] Bhattacharya J, Khuspe SS (2001) In vitro and in vivo germination of papaya (Carica papaya L.) seeds. Scientia Horticulturae 91:39-49.

[5] Bhattacharya J, Renukdas NN, Khuspe SS, Rawal SK (2003) Multiple shoot regeneration from immature embryo explants of papaya. Biologia Plantarum 47:327-331.

[6] Borisjuk L, Walenta S, Rolletschek H, Mueller-Klieser W, Wobus U, Weber H (2002) Spatial analysis of plant metabolism: Sucrose imaging with Vicia faba cotyledons reveals specific development patterns. The Plant Journal 29:521-530.

[7] Borisjuk L, Rolletschek H, Wobus U, Weber H (2003) Differentiation of legume cotyledon as related to metabolism gradients and assimilate transport into seeds. Journal of Experimental Botany 54:503-512.

[8] Bruijne E de, Langhe E de, Van Rijck R (1974) Actions of hormones and embryoid formation in callus cultures of Carica papaya In International Symposium on Crop Protection, Fytofarmacie en Fytiatrie Rijkslandsbouwhoogeschool Medelelingen 26:637-645.

[9] Businge E, Bygdell J, Wingsle G, Moritz T, Egertsdotter U (2013) The effect of carbohydrates and osmoticum on storage reserve accumulation and germination of Norway spruce somatic embryos. Physiologia Plantarum doi:10.1111/ppl.12039.

[10] Calic-Dragosavac D, Zdravkovi-Kora S, Bohanec B, Radojevi L, Vinterhalter B, Stevovi S, Cingel A, Savi J (2010) Effect of activated charcoal, abscisic acid and polyethylene glycol on maturation, germination and conversion of Aesculus hippocastanum androgenic embryos. African Journal of Biotechnology 9:3786-3793.

[11] Chen MH, Wang PJ, Maeda E (1987) Somatic embryogenesis and plant regeneration in Carica papaya L. tissue culture derived from root explants. Plant Cell Reports 6:348-351.

[12] Chen MH, Chen CC (1992). Plant regeneration from Carica protoplasts. Plant Cell Reports 11:404-407.

[13] Coelho CMM, Benedito VA (2008) Seed development and reserve compound accumulation in common bean (Phaseolus vulgaris L.). Seed Science and Biotechnology 2:42-52.

[14] Almeida PA, Oliveira RP, Dantas JLL (2001) Papaya callus and somatic embryo induction and development. Scientia Agricola 58: 51-54.

[15] El-Tayeb MA, Hassanein AM (2000) Germination, seedling growth, some organic solutes and peroxidase expression of different Vicia faba lines as influenced by water stress. Acta Agronomica Hungarica 48:11-20.

[16] FAO (Food and Agriculture Organization of the United Nations)/ FAOSTAT, 2010. http://faostat.fao.org/site/339/default.aspx
» link

[17] Farzana ARF, Palkadapala PGVN, Meddegoda KMMN, Samarajeewa PK, Eeswara JP (2008) Somatic embryogenesis in papaya (Carica papaya L. cv. Rathna). Journal of the National Science Foundation of Sri Lanka 36:41-50.

[18] Fehér A, Pasternak TP, Dudits D (2003) Transition of somatic plant cells to an embryogenic state. Plant Cell, Tissue and Organ Culture 74:201-228.

[19] Ferreguetti GA (2003) Caliman 01 - O primeiro híbrido mamão Formosa brasileiro. In: Annals of the Papaya Brasil, Vitoria. pp. 211-218.

[20] Find JI (1997) Changes in endogenous ABA levels in developing somatic embryos of Norway spruce (Picea abies (L.) Karst.) in relation to maturation medium, desiccation and germination. Plant Science 128:75-83.

[21] Fitch M, Manshardt R (1990) Somatic embryogenesis and plant regeneration from immature zygotic embryos of papaya (Carica papaya L.). Plant Cell Reports 9:320-324.

[22] Goyal K, Walton LJ, Tunnacliffe A (2005) LEA proteins prevent protein aggregation due to water stress. Biochemical Journal 388:151-157.

[23] Jiménez VM, Bangerth F (2001) Endogenous hormone levels in explants and in embryogenic and non-embryogenic cultures of carrot. Physiologia Plantarum 111:389-395.

[24] Keinonen-Metta La K, Jalonen P, Eurola P, Von Arnold S, Von Weissenberg K (1996) Somatic embryogenesis of Pinus sylvestris. Scandinavian Journal of Forest Research 11:242-250.

[25] Koch KE (2004) Sucrose metabolism: regulatory mechanism and pivotal roles in sugar sensing and plant development. Current Opinion in Plant Biology 7:235-246.

[26] Koehler AD, Carvalho CR, Abreu IS, Clarindo WR (2013) Somatic embryogenesis from leaf explants of hermaphrodite Carica papaya: A new approach for clonal propagation. African Journal of Biotechnology 12:2386-2391.

[27] Krajnakova J, Haggman H, Gomory D (2009) Effect of sucrose concentration, polyethylene glycol and activated charcoal on maturation and regeneration of Abies cephalonica somatic embryos. Plant Cell, Tissue and Organ Culture 96:251-262.

[28] Langhansov L, Konrdov H, Vanek T (2004) Polyethylene glycol and abscisic acid improve maturation and regeneration of Panax ginseng somatic embryos. Plant Cell Reports 22:725-730.

[29] Lazzeri PA, Hildebrand DF, Sunega J, Williams EG, Collins GB (1988) Soybean somatic embryogenesis: interactions between sucrose and auxin. Plant Cell Reports 7:517-520.

[30] Li XY, Huang FH, Murphy JB, Gbur JR. EE (1998) Polyethylene Glycol and Maltose Enhance Somatic Embryo Maturation in Loblolly Pine (Pinus taeda L.). In Vitro Cellular and Developmental Biology - Plant 34:22-26.

[31] Litz RE, Conover RA (1982) In vitro somatic embryogenesis and plant regeneration from Carica papaya L. ovular callus. Plant Science Letters 26:153-158.

[32] Litz RE (1986) Effect of osmotic stress on somatic embryogenesis in Carica suspension cultures. Journal of the American Society for Horticultural Science 111:969-972.

[33] Lo Schiavo F, Pitto L, Giuliano G, Torti G, Nutironchi V, Marazziti D, Vergara R, Orselli S, Terzi M (1989) DNA methylation of embryogenic carrot cell cultures and its variations as caused by mutation, differentiation, hormones and hypomethylating drugs. Theoretical and Applied Genetics 77:325-331.

[34] Malabadi RB, Kumar SV, Mulgund GS, Nataraja K (2011) Induction of somatic embryogenesis in Papaya (Carica papaya). Research in Biotechnology 2:40-55.

[35] Martin AB, Cuadrado Y, Guerra H, Gallego P, Hita O, Martin L, Dorado A, Villalobos N (2000) Differences in the contents of total sugars, reducing sugars, starch and sucrose in embryogenic and non-embryogenic calli from Medicago arborea L. Plant Science 154:143-151.

[36] Maruyama E, Hosoi Y (2007) Polyethylene glycol enhance somatic embryo production in Japanese cedar (Cryptomeria japonica D. Don). Propagation of Ornamental Plants 7:57-61.

[37] Mishra M, Shukla N, Chandra R (2010) Role of polyethylene glycol in maturation and germination of trasformed somatic embryos of papaya (Carica papaya L.). Acta Horticulturae 851:227-230.

[38] Morais PLD, Silva GG, Menezes JB, Maia FEN, Dantas DJ, Sales Junior R (2007) Postharvest shelf life of papaya UENF/CALIMAN 01 produced in Rio Grande do Norte. Revista Brasileira de Fruticultura 29:666-670.

[39] Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15:473-497.

[40] Pinto DLP, Almeida AMR, Rêgo MM, Silva ML, Oliveira EJ, Otoni WC (2011) Somatic embryogenesis from mature zygotic embryos of commercial passion fruit (Passiflora edulis Sims) genotypes. Plant Cell, Tissue and Organ Culture 3:521-530.

[41] Santa-Catarina C, Silveira V, Balbuena TS, Viana AM, Estelita MEM, Handro W, Floh EIS (2006) IAA, ABA, polyamines and free amino acids associated with zygotic embryo development of Ocotea catharinensis Plant Growth Regulation 49:237-247.

[42] Schmildt ER, Amaral JAT, Schmildt O (2007) Sucrose on in vitro rooting phase of papaya tree 'Tainung 01'. Scientia Agraria 8:25-31.

[43] Silva ML, Pinto DLP, Guerra MP, Floh EIS, Bruckner CH, Otoni WC (2009) A novel regeneration system for a wild passion fruit species (Passiflora cincinnata Mast.) based on somatic embryogenesis from mature zygotic embryos. Plant Cell, Tissue and Organ Culture 99:47-54.

[44] Smulders MJM, de Klerk GJ (2011) Epigenetics in plant tissue culture. Plant Growth Regulation 63:137-146.

[45] Sokal RR, Rohlf FJ (1995) Biometry: the principles of statistics in biological research. Freeman, New York.

[46] Stasolla C, Yeung EC (2003) Recent advances in conifer somatic embryogenesis: improving somatic embryo quality. Plant Cell, Tissue and Organ Culture 74:15-35.

[47] Walker DR, Parrott WA (2001) Effect of polyethylene glycol and sugar alcohols on soybean somatic embryo germination and conversion. Plant Cell, Tissue Organ and Culture 64:55-62.

[48] Welin BV, Olson A, Nylander M, Palva ET (1994) Characterisation and differential expression of DHN/LEA/RAB -like genes during cold-acclimation and drought stress in Arabidopsis thaliana Plant Molecular Biology 26:131-144.

[49] Wind J, Smeekens S, Hanson J (2010) Sucrose: metabolite and signaling molecule. Phytochemistry 71:1610-1614.

[50] Vahdati K, Bayat S, Ebrahimzadeh H, Jariteh M, Mirmasoumi M (2008) Effect of exogenous ABA on somatic embryo maturation and germination in Persian walnut (Juglans regia L.). Plant Cell, Tissue and Organ Culture 93:163-171.

[51] Vooková B, Kormuták A (2002) Some features of somatic embryo maturation of Algerian fir. In Vitro Cellular and Developmental Biology-Plant 38:549-551.

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Polyethylene glycol effects on somatic embryogenesis of papaya hybrid UENF/CALIMAN 01 seeds

Abstract

Publication Dates

History