Kriopreservasi Tunas in Vitro Pepaya ‘Sukma’ Dengan Perlakuan Prakultur, Loading, Dan Dehidrasi Dengan Pvs2 Dan Modifikasinya

Fitri Fatma Wardani, Joko Ridho Witono, Darda Efendi, Diny Dinarti
10.14203/beritabiologi.v20i2.3999
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Abstract

Papaya has high genetic variability because it is an open-pollinated plant and has genotype and phenotypeare that are easily changed due to environment changes. Cryopreservation is a storing method of germplasm in liquid nitrogen (-196 oC) which can maintain the genotype and phenotype of germplasm. The experiment aimed to obtain the best preculture, loading, and dehydration for cryopreservation of papaya ‘Sukma’ in vitro shoots. For preculture, we planted shoots on MS media with 0.3 M and 0.4 M sucrose for 1, 2, and 3 days. In the loading treatment, we immersed shoots in loading solution (liquid MS+1.2M glycerol+0.4M sucrose) for 0, 10, 20, and 30 minutes. For dehydration, we immersed shoots in cryoprotectant (PVS2 and its modification) for 5, 10, and 15 minutes. Then, shoots were immersed in liquid nitrogen. The results showed thatshoots had the best survival rate while they had been precultured on MS medium with 0.3 M sucrose for 3 days. The best loading treatment time was 20–30 minutes. The best dehydration treatment was obtained by modification of PVS2 for 10 minutes. The shoots have not been able to recovery after cryopreservation, so it can be concluded that cryopreservation of in vitro papaya ‘Sukma’ shoots has not been successful.

Keywords

liquid nitrogen; long-term storage; vitrification

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References

Al-Shara, B., Taha, R.M., and Rashid, K., 2018. Biotechnological methods and limitations of micropropagation in papaya (Carica papaya L.) production. The Journal of Animal and Plant Science, 28(5), pp.1208-1226.

Azimi, M., Brien, C., Ashmore, S., and Drew, R., 2005. Cryopreservation of Papaya Germplasm. Proceeding Ilnd IS on Biotechnology of Tropical & Subtropical Species Acta Horticultura, 692. pp.43-50.

Burritt, D.J., 2012. Proline and the cryopreservation of plant tissues: functions and practical applications, current frontiers in cryopreservation. In Katkov, I. ed. Current Frontiers in Cryobiology. pp.415-430. InTech. Rijeka.

Cho, E.G., Hor, Y.L., Kim, H.H., Rao, V.R., and Engelmann, F., 2002. Cryopreservation of Citrus madurensis embryonic axes by vitrification: importance of loading and treatment with vitrification solution. Cryo Letters, 23, pp.317-324.

Elliot, G.D., Wang, S., and Fuller, B.J., 2017. Cryoprotectants: A review of the actions and applications of cryoprotective solutes that modulate cell recovery from ultra-low temperatures. Cryobiology, 76, pp.74-91.

Feng, C.H., Cui, Z.H., Li, B.Q., Chen, L., Ma, Y.L., Zhao, Y.H., and Wang, Q.C., 2012. Duration of sucrose preculture is critical for shoot regrowth of in vitro-grown apple shoot-tips cryopreserved by encapsulation-dehydration. Plant Cell Tissue Organ Culture. doi 10.1007/s11240-012-0245-3.

Hervani, D., Efendi, D., Suhartanto, M.R., and Purwoko, B.S., 2018. The preservation of somatic embryos of papaya derived from papaya lateral shoots after being stored in cryopreservation to maintain plant genetic information in the future. Biodiversitas, 19(3), pp.774-779.

Kaczmarczyk, A., Bryn, F., Menon, A., Phang, P.Y., Al-Hanbali, A., Bunn, E., and Mancera, L.R., 2012. Current issues in plant cryopreservation. In I. Katkov. Ed. Current Frontiers in Cryobiology. pp.417-438. InTech. Rijeka

Kim, H.H., Lee, Y.G., Park, S.U., Lee, S.C., Baek, H.J., Cho, E.G., and Engelmann, F., 2009. Development of alternative loading solution in droplet-vitrification procedures. Cryo Letters, 30(3), pp.291-299.

Markovic, Z., Chatelet, P., Sylvestre, I., Kontic, J.K., and Engelmann, F., 2013. Cryopreservation of grapevine (Vitis vinifera L.) in vitro shoot tips. Cent Eur J Biol, 8(10), pp.993-1000. doi: 10.2478/a11535-013-0223-8.

Mikula, A., Tomiczak, K., and Rybczynski, J.J., 2011. Cryopreservation enhances embryogenic capacity of Gentiana cruciata (L.) suspension culture and maintains (epi)genetic uniformity of regenerants. Plant Cell Reproduction, 30, pp.565-574. doi: 10.1007/s00299-010-0970-1.

Murashige, T., F. Skoog. 1962. A revised medium for rapid growth and bio assays with tobacco tissue culture. Physiol Plant, 15, pp.473-497.

Popova, E.V., Lee, E.J., Wu, C.H., Hahn, E.J., and Park, K.Y, 2009. A simple method for cryopreservation of Ginkgo biloba callus. Plant Cell Tissue Organ Culture, 97, pp.337-343.

Roostika, I., Darwati, I., dan Megia, R., 2007. Kriopreservasi tanaman purwoceng (Pimpinella pruatjan Molk.) dengan teknik vitrifikasi. Berita BiologiI, 8(6), pp.423-431.

Roostika, I., 2003. Studi penyimpanan kultur in vitro ubi jalar (Ipomea batatas (L.) Lam) secara kriopreservasi. Tesis. Sekolah Pasca Sarjana. Institut Pertanian Bogor. Bogor. Indonesia.

Sakai, A., Kobayashi, S., Oiyama, I., 1991. Cryopreservation of nucellar cells of navel orange (Citrus sinensis Osb.) by a simple freezing method. Plant Science, 74, pp.243-248.

Silva, J.A.T., 2014. Callus induction in papaya (Carica papaya L.) and synseed production for low temperature storage and cryopreservation. Folia Horticulturae, 26(2), pp.155-162. DOI: 10.1515/fhort-2015-0007.

Tsai, S.F., Yeh, S.D., Chan, C.F., and Liaw, S.I., 2009. High-efficiency vitrification protocols for cryopreservation of in vitro grown shoot tips of transgenic papaya lines. Plant Cell Tiss Organ Cult, 98(2), pp.157-164.

Wang, Y.L., Fab, M.J., and Liaw, S.I., 2005. Cryopreservation of in vitro-grown shoot tips of papaya (Carica papaya L.) by vitrification. Botany Bulletin Academy Singapore, 46, pp.29-34.

Wang, Q.C., Panis, B., Engelman, F., Ambardi, M., and Valkonen, J.P.T., 2008. Cryotherapy of shoot tips: a technique for pathogen eradication to produce healthy olanting materilas and prepare healthy plant genetic resources for cryopreservation. Annals Applied Biology, 154, pp.351-363.

DOI: https://doi.org/10.14203/beritabiologi.v20i2.3999

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