An improved plant regeneration protocol for a popular Indian Madhubindu variety of papaya (Carica papaya L.) via somatic embryogenesis

, , , , , ,


Research Articles | Published:

DOI: 10.1007/s42535-024-00971-8
First Page: 250
Last Page: 259
Views: 2100

Keywords: n Carica papayan , Plant hormones, Seed germination, In vitro plant regeneration, Somatic embryogenesis


Abstract


Papaya (Carica papaya L.) is an important fruit crop, and it is highly nutritive and has several medicinal properties. In the present study, we have developed an improved protocol for in vitro plant regeneration through somatic embryogenesis. A  high seed germination rate was achieved, when the seeds were treated with 600 ppm of gibberellic acid to break the seed dormancy. Different explants, such as leaves, roots, hypocotyls, and mature zygotic embryos were inoculated on M1I2 medium supplemented with half-strength MS salts with full-strength vitamins, 400 mg/l glutamine, 100 mg/l proline, 160 mg/l adenine sulphate, 6% sucrose, 0.8% agar, and 4 mg/l 2,4-D for the induction of the embryogenic callus and further different stages of somatic embryos. Late torpedo and early cotyledonary stage SEs were regularly harvested and cultured on hormone-free MS medium in the dark till the roots appeared, and then transferred to light for further growth. For acclimatization, regenerated plants were transferred into earthen pots covered with polythene bags to maintain high humidity, grown for 2 months, and then transferred to greenhouse and then to the field conditions for their establishment.

n                     Carica papayan                  , Plant hormones, Seed germination, In vitro plant regeneration, Somatic embryogenesis


References


Al-Drishi E, Ibrahim M, Jasim A (2022) Explant type influences callus induction and shoots organogenesis in papaya under in vitro conditions. DYSONA Appl Sci 4:15–20


Al-Shara B, Taha RM, Mohamad J, Elias H, Khan A (2020) Somatic embryogenesis and plantlet regeneration in the Carica papaya L. cv. Eksotika. Plants 9:360


Aluthge ADRP, Nagahawaththa SMS, Basnayake BMVS, Hettiarachchi C, Senanayake DMJB (2021) Regeneration of Carica papaya “Rathna” variety through somatic embryogenesis from immature zygotic embryos. Asian J Res Rev Agric 3(1):185–192


Anandan R, Deenathayalan T, Kumar NS, Deepak KV (2018) An alternative in vitro plant regeneration system in papaya (Carica papaya L.) through callus derived nodular cultures. Meta Gene 17:147–152


Asano Y, Katsumoto H, Inokuma C, Kaneko S, Ito Y, Fujiie A (1996) Cytokinin and thiamine requirements and stimulative effects of riboflavin and α-ketoglutaric acid on embryogenic callus induction from the seeds of Zoysia japonica Steud. J Plant Physiol 149:413–417


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


Bhusare BP, John CK, Bhatt VP, Nikam TD (2020) Induction of somatic embryogenesis in leaf and root explants of Digitalis lanata Ehrh.: direct and indirect method. South Afr J Bot 130:356–365


Brar D, Jain SM (1998) Somaclonal variation: mechanism and applications in crop improvement. In: Ahloowalia BS (ed) Somaclonal variation and induced mutations in crop improvement. Kluwer Academic Publishers, Dordrecht, pp 15–37


Cai W, Gonsalves C, Tennant P, Fermin G, Souza M, Sarindu N, Jan FJ, Zhu HY, Gonsalves D (1999) A protocol for efficient transformation and regeneration of Carica papaya L. In Vitro Cell Dev Biol Plant 35:61–69


Carvalho F, Renner S (2012) A dated phylogeny of the papaya family (Caricaceae) reveals the crop’s closest relatives and the family’s biogeographic history. Mol Phylogenet Evol 65:46–53


Castillo B, Smith MA, Yadava UL (1998) Plant regeneration from encapsulated somatic embryos of Carica papaya L. Plant Cell Rep 17:172–176


Chatterjee A, Nirwan S, Sharma P, Agnihotri A, Shrivastava N (2021) Investigating the in vitro regeneration vigour of Brassica juncea var. varuna and its white rust tolerant mutant genotypes: conjugating the conventional and biotechnological approaches. Plant Cell Biotechnol Mol Biol 22:122–135


Chávez-Pesqueira M, Núñez-Farfán J (2017) Domestication and genetics of papaya: a review. Front Ecol Evol 5:155


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


Choudhury A, Rajam MV (2021) Genetic transformation of legumes: an update. Plant Cell Rep 40:1813–1830


Dantu PK, Tomar UK (2010) Somatic embryogenesis. In: Tripathi G (ed) Cellular and biochemical science. I.K. International House Pvt Ltd, New Delhi, pp 892–908


Dhekney SA, Kandel R, Bergey DR, Sitther V, Soorianathasundaram K, Litz RE (2016) Advances in papaya biotechnology. Biocat Agric Biotechnol 5:133–142


Dotto JM, Abihudi SA (2021) Nutraceutical value of Carica papaya: a review. Sci Afr 13:00933


El Hadrami I, Carron MP, D’Auzac J (1991) Influence of exogenous hormones on somatic embryogenesis in Hevea brasiliensis. Ann Bot 67:511–515


FAO (2018) FAOSTAT. Food and Agriculture Organization of the United Nations, Rome, Italy


Fitch MM (1993) High frequency somatic embryogenesis and plant regeneration from papaya hypocotyl callus. Plant Cell Tiss Org Cult 32:205–212


Fitch MM, Manshardt RM (1990) Somatic embryogenesis and plant regeneration from immature zygotic embryos of papaya (Carica papaya L.). Plant Cell Rep 9:320–324


George EF, Hall MA, De Klerk GJ (2008) Plant growth regulators I: introduction, auxins, their analogues and inhibitors. In: George EF, Hall MA, De Klerk G-J (eds) Plant propagation by tissue culture, 3rd edn. Springer, Netherlands, pp 45–65


Hossain M, Rahman SM, Islam R, Joarder OI (1993) High efficiency plant regeneration from petiole explants of Carica papaya L. through organogenesis. Plant Cell Rep 13:99–102


Kintzios S, Drossopoulos JB, Lymperopoulos C (2001) Effect of vitamins and inorganic micronutrients on callus growth and somatic embryogenesis from leaves of chilli pepper. Plant Cell Tiss Org Cult 67:55–62


Krishnanayak L, Kotiyal A, Koubouris G (2024) Optimized seed germination and adaptation of plantlets to a new environment of papaya cv. ‘Red Baby’ using organic media and plant growth regulators. Vegetos 37:355–362


Kumari R, Kundu M, Mir H, Kumar P (2023) In vitro regeneration technique of papaya (Carica papaya L.) cv. Pusa Dwarf through shoot tip culture. Natl Acad Sci Lett 46:1–5


Litz RE (1984) Papaya. In: Sharp WR, Evans DA, Amm Jrato PV, Yamada Y (eds) Handbook of plant cell culture, vol 2. MacMillan Publishing Co, New York, pp 349–368


Litz RE, Conover RA (1979) Development of systems for obtaining parasexual Carica hybrids. Proc Fl State Hortic Soc 92:180–182


Litz RE, O’Hair SK, Conover RA (1983) In vitro growth of Carica papaya L. cotyledons. Sci Hortic 19:287–293


Malabadi RB, Kumar SV, Mulgund GS, Nataraja K (2011) Induction of somatic embryogenesis in papaya (Carica papaya). Res Biotechnol 2:40–55


Manchanda P, Sidhu GS, Mankoo RK (2024) Bioactive compounds and antioxidant potential determination in callus tissue as compared to leaf, stem and root tissue of Carica papaya cv. Red Lady 786. J Food Meas Charact 37:1–14


Mishra R, Gaur RK, Patil BL (2016) Current knowledge of viruses infecting papaya and their transgenic management. In: Gaur R, Petrov N, Patil B, Stoyanova M (eds) Plant viruses: evolution and management. Springer, Singapore, pp 189–203


Möller B, Weijers D (2009) Auxin control of embryo patterning. Cold Spring Harb Perspect Biol 1:a001545


Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497


Ogita S, Sasamoto H, Yeung EC, Thorpe TA (2001) The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica. In Vitro Cell Dev Biol Plant 37:268–273


Palei S, Dash DK, Rout GR (2019) Standardization of in vitro protocol for plant regeneration of Carica papaya cv. Co 8 through indirect organogenesis. J Pharm Phytochem 8:1954–1956


Pang SZ, Sanford JC (1988) Agrobacterium-mediated gene transfer in papaya. J Amer Soc Hortic Sci 113:287–291


Priyanka, Rajam MV (2024) Improved, efficient and reliable plant regeneration protocol for a recalcitrant black rice (Oryza sativa cv. Chakhao amubi). Curr Trends Biotechnol Pharm 18(2):1669–1679


Rajesh CK, Kumar KK, Kavitha C, Karthikeyan G, Soorianathasundaram K (2020) Differential influence of growth regulators during somatic embryogenesis of gynodioecious papaya varieties ‘CO. 7’and ‘Red Lady.’ Adv Res 21:10–18


Randle M, Tennant P (2021) Transgenic papaya. In: Kavi Kishor PB, Rajam MV, Pullaiah T (eds) Genetically modified crops. Springer, Singapore, pp 129–160


Rizwan HM, Irshad M, He B, Liu S, Lu X, Sun Y, Qiu D (2020) Role of reduced nitrogen for induction of embryogenic callus induction and regeneration of plantlets in Abelmoschus esculentus L. South Afr J Bot 130:300–307


Singh SP, Kumar S, Mathan SV, Tomar MS, Singh RK, Verma PK, Kumar A, Kumar S, Singh RP, Acharya A (2020) Therapeutic application of Carica papaya leaf extract in the management of human diseases. DARU J Pharm Sci 28:735–744


Verma S, Attuluri VP, Robert HS (2021) An essential function for auxin in embryo development. Cold Spring Harb Perspect Biol 13:a039966


Vishal SGS, Gaikwad PN et al (2024) Optimized protocol for high-frequency papaya propagation: morpho-stereomicroscopic analysis and genetic fidelity assessment. Plant Cell Tiss Organ Cult 156:81


Yie S, Liaw SI (1977) Plant regeneration from shoot tips and callus of papaya. In Vitro 13:564–568

 


Author Information


Department of Genetics, University of Delhi, New Delhi, India