Aislamiento, cultivo y fusión de protoplastos en el género Vasconcellea
Resumen
Las especies del género Vasconcellea, frecuentemente denominadas papayas de las tierras altas, son parientes silvestres de la papaya común (Carica papaya L.) ampliamente distribuida en la América tropical. Sus usos son varios entre los que se incluyen: materia prima en la industria de las frutas tropicales como fuente de papaína o su empleo en programas de mejoramiento genético. El presente trabajo se realizó con el objetivo de realizar una revisión de literatura científica sobre el aislamiento, purificación y fusión de protoplastos en el género Vasconcellea y sus usos en el mejoramiento genético. Además, se describen las principales fuentes de protoplastos del género Vasconcellea, los factores que influyen en el proceso de aislamiento, las condiciones de cultivo y la fusión de los protoplastos. Finalmente, se resumen los adelantos alcanzados hasta la fecha en la hibridación intergenérica con Carica papaya para generar mayor resistencia y productividad.
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Ahmed M, Miao M, Pratsinakis E, Zhang H, Wang W, Yuan Y, Lyu M, Iftikhar J, Ahmed F, Modesis P, Wu B (2021) Protoplast Isolation, Fusion, Culture and Transformation in the Woody Plant Jasminum spp. Journal of Agriculture 11(8): 699; doi: 10.3390/agriculture11080699
Amroune A (2011) Isolement, Purification et culture de protoplastos de Datura stramonium. Doctoral Thesis, Ecole National Supérieure Agronomique, El Harrach, Alger
Anand G, Yadav S, Gupta R, Yadav D (2020) Pectinases: from microbes to industries. En: Chowdhary P, Raj A, Verma D, Akhter Y (Eds). Microorganisms for Sustainable Environment and Health, pp. 287-313. Elsevier, London; doi: 10.1016/B978-0-12-819001-2.00014-0
Badillo VM (2000) Carica L. vs. Vasconcella St. Hil. (Caricaceae): con la rehabilitación de este último. Ernstia 10: 74–79
Barceló M, Wallin A, Medina J, Gil D, López G, Juarez J, Sánchez J, López C, López J, Mercado J, Pliego F (2019) Isolation and culture of strawberry protoplasts and field evaluation of regenerated plants. Scientia Horticulturae 256: 108552; doi: 10.1016/j.scienta.2019.108552
Bhatia S (2015) Application of Plant Biotechnology. En: Bhatia S, Sharma K, Dahiya R, Bera T (Eds). Modern Applications of Plant Biotechnology in Pharmaceutical Sciences, pp. 157-207. Elsevier, London; doi: 10.1016/C2014-0- 02123-5
Burris K, Dlugosz E, Collins A, Stewart C, Lenaghan S (2016) Development of a rapid, low-cost protoplast transfection system for switchgrass (Panicum virgatum L.). Plant Cell Reports 35: 693-704; doi: 10.1007/s00299-015-1913-7
Carvajal P (2020) Hibridación somática de Carica papaya var. pococi y Vasconcellea sp. (Brassicales: Caricaceae) mediante electrofusión de protoplastos. Tesis de Maestría, Universidad de Costa Rica, San José, Costa Rica
Carvalho F, Renner S (2014) The Phylogeny of the Caricaceae. En: Ming R, Moore P (Eds). Plant Genetics and Genomics, pp. 81-92; Springer, New York; doi: 10.1007/978-1-4614-8087-7_5
Chamani E, Tahami K, Zare N, Zakaria R (2012) Effect of Different Cellulase and Pectinase Enzyme Treatments on Protoplast Isolation and Viability in Lilium ledebeourii Bioss. Notulae Botanicae Horti Agrobotanici Cluj-Napoca 40(2): 123-128; doi: 10.15835/nbha4028055
Chawla H (2009) Introduction to Plant Biotechnology. CRC Press, New York
Cocking E (2000) Turning point article plant protoplasts. In Vitro Cell Dev Biol 36 (2): 1-6; doi: 10.1007/s11627-000-0018-2
Davey MR, Anthony P, Power JB, Lowe KC (2005) Plant protoplasts: status and biotechnological perspectives. Biotechnol Adv 23(2):131-71; doi: 10.1016/j.biotechadv.2004.09.008
Dhekney S, Kandel R, Bergeya D, Sitther V, Soorianathasundaram K, Litz R (2016) Advances in papaya biotechnology. Biocatalysis and Agricultural Biotechnology 5: 133-142
Dinesh M, Rekha A, Ravishankar K, Praveen K, Santosh L (2007) Breaking the intergeneric crossing barrier in papaya using sucrose treatment. Scientia Horticulturae 114: 33–36; doi: 10.1016/j.scienta.2007.05.010
Djoudi N, Hadji B (2020) Application de la biotechnologie de la culture in vitro pour les plantes médicinales. Maestría Académica (En biotecnología vegetal), Université Mohamed Boudiaf - M’Sila, Algerienne, Italia
Dodds JH (1983) The use of protoplast technology in tissue culture of trees. En: Dodds JH (Eds). Tissue Culture of Trees , pp. 103-112. Springer, Boston
Drew R (2016) Applications of biotechnology to tropical fruit crops in australia and worldwide. Acta Horticulture 1-8; doi: 10.17660/ActaHortic.2008.787.18
Drew R, Kyndt T, Scheldeman X (2014) Vasconcellea for Papaya Improvement. En: Ming R, Moore PH (Eds). Genetics and Genomics of Papaya, pp. 47-79. Springer, Dordrecht; doi: 10.1007/978-1-4614-8087-7_4
Drew R, Siar S, Dillon S, Ramage C, O'Brien C, Sajise A (2007) Intergeneric hybridisation between Carica papaya and wild Vasconcellea species and identification of a PRSV-P resistance gene. Acta Hortic 738: 165-169; doi: 10.17660/ActaHortic.2007.738.14
Droogenbroeck B, Haegeman A, Kyndt T, O’Brien C, Drew R, Gheysen G (2005) Maternal inheritance of cytoplasmic organelles in intergeneric hybrids of Carica papaya L. and Vasconcellea spp. (Caricaceae Dumort., Brassicales). Euphytica 143: 161-168; doi: 10.1007/s10681-005-3156-0
Droogenbroeck B, Kyndt T, Romeijn P, Thuyne W, Motochi J, Gheysen G (2006) Evidence of Natural Hybridization and Introgression between Vasconcellea Species (Caricaceae) from Southern Ecuador Revealed by Chloroplast, Mitochondrial and Nuclear DNA Markers. Annals of Botany 97 (5): 793-805; doi: 10.1093/aob/mcl038
Duquenne B, Eeckhaut T, Werbrouck S, Huylenbroeck J (2007) Effect of enzyme concentrations on protoplast isolation and protoplast culture of Spathiphyllum and Anthurium. Plant Cell Tiss Organ Cult 91:165-173; doi: 10.1007/s11240-007-9226-3
Echeverri D, Romo J, Giraldo N, Atehortúa L (2019) Microalgae protoplasts isolation and fusion for biotechnology research. Revista Colombiana de Biotecnología 21(1): 101-112; doi: 10.15446/rev.colomb.biote.v21n1.80248
Fitch M (2022) Biotechnology of fruit and nut Crops. En: Litz RE (Ed).Caricaceae- Carica papaya, pp. 174-201. CAB International, London; ISBN: 9781780648279
Fowke L, Gamborg O (1980) Applications of Protoplasts to the Study of Plant Cells. International Review of Cytology68: 9-51; doi: 10.1016/S0074-7696(08)62306-2
Fuertes C (2019) Diversidad, distribución y uso del género vasconcellea (caricaceae) en el sur de los andes colombianos. Tesis de Pregrado, Universidad del Cauca, Popayán, Colombia
Gares M, Saidi S (2016) Etude de l’amélioration de la production d’éthanol par la technique de fusion des protoplastes entre deux souches fongiques. Mémoire présenté en vue de l’obtention du Diplôme de Master Domaine: Sciences de la Nature et de la Vie Filière, Université des Frères Mentouri Constantine. Constantina, Argelia
Graham J (2002) Purification of intact plant protoplasts by flotation at 1g. The Scientific World Journal (2002) 2: 1397-1399; doi: 10.1100/tsw.2002.835
Gray A (1993) Broccoli: Brassica oleracea L. (Italica group). En: Kalloo G, Bergh BO (Eds). Genetic Improvement of Vegetable Crops, pp. 61-86. Pergamon, Roma; doi: 10.1016/B978-0-08-040826-2.50010-2
Gilliard G, Huby E, Cordelier S, Ongena M, Dhondt-Cordelier S, Deleu M (2021) Protoplast: A Valuable Toolbox to Investigate Plant Stress Perception and Response. Front Plant Sci (5)12:749581; doi: 10.3389/fpls.2021.749581
Hayat S, Christias C (2010) Isolation and fusion of protoplasts from the phytopathogenic fungus. Sclerotium rolfsil. Brazilian Journal of Microbiology 41(1): 1-11; doi: 10.1590/S1517-838220100001000035
Ishii S (1989) Enzymes for the Isolation of Protoplasts. En: Bajaj YPS (eds). Plant Protoplasts and Genetic Engineering I, pp. 23-33. Springer, Berlín; doi: 10.1007/978-3-642-73614-8_2
Jadán M, Dorca C (2019) Propagation methods in Babaco plants (Vasconcella x helbornii). Tropical Plant Research 6(1): 37-45; doi: 10.22271/tpr.2019.v6.i1.007
Jayavalli R, Balamohan T, Manivannan N, Govindaraj M (2011) Breaking the intergeneric hybridization barrier in Carica papaya and Vasconcellea cauliflora. Scientia Horticulturae 130(7): 87-794; doi: 10.1016/j. scienta.2011.09.004
Jayavalli R, Balamohan T, Manivannan N, Shahidul IR (2012) Analysis of papaya intergeneric hybrids for morphological traits. Madras Agriculture Journal 99(4-6): 166-170
Jayavalli R, Balamohan T, Manivannan N, Rabindran R, Paramaguru P, Robin R (2015) Transmission of resistance to papaya ringspot virus (PRSV) in intergeneric populations s of Carica papaya and Vasconcellea cauliflora. Scientia Horticulturae 187: 10-14; doi: 10.1016/j.scienta.2015.01.020
Jordan M, Ciudad G, Roja M, Valverde F (1986) Isolation, culture and fusion of Carica candamarcencis and Carica papaya protoplasts. Gartenbauwissenschafl 51(4): 175-178
Kang H, Naing A, Kim C (2020) Protoplast Isolation and Shoot Regeneration from Protoplast-Derived Callus of Petunia hybrida Cv. Mirage rose. Biology: 1-8; doi: 10.3390/biology9080228
Kozlowski T, Pallardy S (1997) Growth Control in Woody Plants. Academic Press, California; ISBN: 0-12-424210-3
Kyndt T, Gheysen G (2007) Evolutionary relationships between and within the highland papayas (Genus Vasconcellea) and the common papaya (Carica papaya). ISHS Acta Horticulturae 740: 61-72; doi: 10.17660 / ActaHortic.2007.740.6
Lin HY, Chen JC, Fang SC (2018) A protoplast transient expression system to enable molecular, cellular, and functional studies in phalaenopsis orchids. Front Plant Sci 9:843; doi: 10.3389/fpls.2018.00843
Magdalita P, Adkins S, Godwin I, Drew R (1996) An Improved Embryo-Rescue Protocol for a Carica Interspecific Hybrid. Australian Journal of Botany 44: 343-353; doi: 10.1071/BT9960343
Mariño M (2019) Determinación del cariotipo dejigacho (Vasconcellea stipulata Badillo) mediante técnicas citogenéticas. Tesis de Pregrado, Universidad de las Fuerzas Armadas-ESPE, Sangolquí, Ecuador
Marques J, Paiva R, Atala L, Rodrigues M, Alves M, Carvalho F, Campos W (2012) Protoplast production and isolation from Etlingera elatior. Acta Scientiarum- Agronomy 1-17; doi: 10.1590/S1807-86212012000100007
Márquez F (2019) El cultivo de protoplastos como herramienta biotecnológica aplicable a las plantas medicinales. Tesis de Pregrado, Universidad Complutense, Facultad de Farmacia, Madrid, España
Mastuti R, Rosyidah M (2019) In vitro enzymatic isolation of protoplasts from tissues of the medicinal plant Physalis angulata L. AIP Conference Proceedings 2019 (1): 1-9; doi: 10.1063/1.5061838
Morales A, Medina D, Yaguache B (2004) Genetic diversity, phylogeny and geographic distribution of the genus Vasconcellea in Southern Ecuador. Lyonia 7: 4-15
Nagata T, Takebe I (1971) Planting of isolated tobacco mesophyll protoplasts on agar medium. Springer Planta (Berl) 99: 12-20
Naing A, Suleimon O, Kil C (2021) Protoplast technology in ornamental plants: Current progress and potential applications on genetic improvement. Scientia Horticulturae 283: 1-9; doi: 10.1016/j.scienta.2021.110043
Navrátilova B (2004) Protoplast cultures and protoplast fusion focused on Brassicaceae – a review. Hort Sci (PRAGUE) 31(4): 140-157; doi: 10.17221/3809-HORTSCI
O’Brien C, Drew R (2009) Potential for using Vasconcellea parviflora as a bridging species in intergeneric hybridisation between V. pubescens and Carica papaya. Australian Journal of Botany 57(7): 592-601; doi: 10.1071/BT09111
O'Brien C, Drew R (2010) Marker-assisted hybridisation and backcrossing between Vasconcellea species and Carica papaya for PRSV-p resistance. ISHS Acta Horticulturae 859: 361-368; doi: 10.17660/ActaHortic.2010.859.43
ONDŘEJ M (1985) Cytogenetika a molekulární genetika rostlin. Academia, Praha
Ondráčková E, Ludvíková M, Griga M (2020) Developments in fibrous flax and linseed breeding and cultivation. n: Kozłowski RM, Mackiewicz-Talarczyk M (Eds). Handbook of Natural Fibres, pp. 605-692. Woodhead Publishing, Cambridge; doi: 10.1016/b978-0-12-818398-4.00019-0
Pavelek M, Prokopová V, Ondrackova M, Ludvikova M, Griga M (2020) Developments in fibrous flax and linseed breeding and cultivation. En: Kozłowski RM, Mackiewicz-Talarczyk M (Eds). Handbook of Natural Fibres, pp. 605-692. Woodhead Publishing, Cambridge; doi: 10.1533/9780857095503.2.391
Poddar S, Tanaka J, Cate J, Staskawicz B, Cho M (2020) Efficient isolation of protoplasts from rice calli with pause points and its application in transient gene expression and genome editing assays. Plant Methods 16: 151; doi: 10.1186/s13007-020-00692-4
Restrepo M, Eeckenbrugge G, Jiménez D, Veja J (2004) Morphological diversity of cultivated mountain papayas (Vasconcellea spp.) in Ecuador. Proceedings of the Interamerican Society for Tropical Horticulture 48: 119-123
Rivera U, Jadán M (2010) Aislamiento, purificación y fusión de protoplastos, obtenidos a partir de hojas in vitro y callos friables de babaco (Vasconcellea heibornii) y jigacho (Vasconcellea stipulata). EIDOS 3-7; doi: 10.29019/eidos.v
Roberts AV, Schum A (2003) Cell, tissue and organ culture. En: In
Roberts AV, Debener T, Gudin S (eds). Encyclopedia of
rose science, 57-66. Elsevier Academic Press, Pune; doi: 10.1016/B0-12-227620-5/00124-5
Salvatierra M, Jana C (2015) Floral expression and pollen germination ability in productive mountain papaya (Vasconcellea pubescens A.DC.) orchards. Revista Chilena de Investigación Científica 76(2): 136-142; doi: 10.4067/S0718-58392016000200001
Scheldeman X, Kyndt T, d’Eeckenbrugge G, Ming R, Drew R, Van Droogenbroeck B, Van Damme P, Moore P (2011) Vasconcellea. En: Kole C (ed). Wild Crop Relatives: Genomic and Breeding Resources, pp. 213-249. Springer-Berlin-Heidelberg, Dordrecht; doi: 10.1007/978-3-642-20447-0_11
Scheldeman X, Willemen L, Coppens G, Romeijn E, Restrepo M, Romero J, Jiménez D, Lobo M, Medina C, Reyes C, Rodriguez D, Ocampo Jhon, Van Damme P, Goetgebeur P (2007) Distribution, diversity and environmental adaptation of highland papaya (Vasconcellea spp.) in tropical and subtropical America. En: Hawksworth DL, Bull AT (eds). Biodiversity and Conservation, pp. 293-310. Springer, Dordrecht; doi: 10.1007/978-1-4020-6444-9_19
Sharma SK, Tripathi S (2014) Papaya ringspot virus-P In Plant Virus-Host Interaction: Molecular Approaches and Viral Evolution. Academic Press, Pune; doi: 10.1016/B978-0-12-411584-2.00009-3
Sharma SK, Tripathi S (2021) Overcoming limitations of resistance breeding in Carica papaya L. against papaya ringspot virus—Recent approaches En: Lashmangarh S (Ed).Plant Virus-Host Interaction Molecular Approaches and Viral Evolution, pp. 489-506. Academic Press, Pune; doi: 10.1016/B978-0-12-821629-3.00008-7
Siar SV, Beligan GA, Sajise AJ, Villegas VN, Drew RA (2011) Papaya ringspot virus resistance in Carica papaya via introgression from Vasconcellea quercifolia. Euphytica 181: 159-168; doi: 10.1007/s10681-011-0388-z
Sudha R, Balamohan T, Soorianathasundaram K, Manivannan N, Rabindran R (2013) Evaluation of F2 intergeneric population of papaya (Carica papaya L.) for resistance to papaya ringspot virus (PRSV). Scientia Horticulturae 158: 68-74; doi: 10.1016/j.scienta.2013.04.031
Sudha R, Balamohan T, Soorianathasundaram K, Rabindran R, Manoranjitham S (2015) Studies on resistance to papaya ring spot virus (PRSV) in intergeneric populations of Carica papaya L. and Vasconcellea cauliflora. SABRAO Journal of Breeding and Genetics 47(2): 113-123
Szabados L (1991) Protoplasto: aislamiento, cultivo y regeneración de plantas. En: Roca WM, Mrogisnski LA (Eds). Cultivo de tejidos en la agricultura, Fundamentos y aplicaciones, 239-270. CIAT, Cali; ISBN: 958-9183-15-8
Szabados L, Narváez J, Roca W (1987) Técnicas para el Aislamiento y Cultivo de Protoplastos de Yuca (Manihot esculenta Crantz). CIAT, Cali
Tapingkae T, Zulkarnain Z, Ikeda T, Kawaguchi M (2012) Somatic (asexual) procedures (haploids, protoplasts, cell selection) and their applications. Plant Biotechnology and Agriculture 141-162; doi: 10.1016/B978-0-12-381466-1.00001-8
Tarora K, Shudo A, Kawano S, Yasuda K, Ueno H, Matsumura H, Urasaki N (2016) Development of plants resistant to Papaya leaf distortion mosaic virus by intergeneric hybridization between Carica papaya and Vasconcellea cundinamarcensis. Japanese Society of Breeding 66(5): 734-741; doi: 10.1270/jsbbs.16107
Torora K, Irei A, Tamaki M, Kawano S, Yasuda K, Shoda M, Urasaki N, Matsumura H (2018) Production of backcross plants between intergeneric hybrids (Carica papaya × Vasconcellea cundinamarcensis) and C. papaya, as a novel breeding material for Papaya leaf distortion mosaic virus resistance. Breeding Research 20(2): 115-123; doi: 10.1270/jsbbr.18j08
Teixeira Da Silva J, Rashid Z, Tan D, Sivakumar D, Gera A, Texeira M, Tennant P (2007) Papaya (Carica papaya L.) Biology and Biotechnology. Tree and Forestry Science and Biotechnology 1(1): 47-73
Tineo D, Bustamante D, Calderón M, Mendoza J, Huaman E, Oliva M (2020) An integrative approach reveals five new species of highland papayas (Caricaceae, Vasconcellea) from northern Peru. PLOS ONE 15(12): e0242469; doi: 10.1371/journal.pone.0242469
Tomar U, Dantu P (2010) Protoplast Culture and Somatic Hybridization. In: Tripathi GIK, (Ed). Cellular and Biochemical Sciences, pp. 876-891. International Publishing House, New Delhi
Wiszniewska A, Pindel A (2009) Protoplast culture utilization in studies on legume crops. Acta Agriculturae Scandinavica , Section B — Soil & Plant Science 60(5): 389–399; doi: 10.1080/09064710903179545
Wua J, Choub J (2019) Optimization of Protoplast Preparation and Regeneration of a Medicinal Fungus Antrodia cinnamomea. Journal of Mycobiology 47(4): 483–493; doi: 10.1080/12298093.2019.1687252
Zhang J, Shen W, Yan P, Li X, Zhou P (2011) Factors that influence the yield and viability of protoplasts from Carica papaya L. African Journal of Biotechnology 10(26): 5137-5142; doi: 10.5897/AJB10.1156
Zhao F, Li Y, Yang H, Gao R, Zang X, Qin D, Wang J, Qiang Y (2016) A highly efficient grapevine mesophyll protoplast system for transient gene expression and the study of disease resistance proteins. Plant Cell Tiss Organ Cult 125: 43-57; doi: 10.1007/s11240-015-0928-7
Zhao C, Randall D, Holford P, Haigh A M, Chen ZH (2019) Isolation of high purity guard cell protoplasts of Arabidopsis thaliana for omics research. Plant Growth Regul 89(1): 37-47; doi: 10.1007/s10725-019-00520-3
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