Protoplast Isolation from In Vitro Leaf of Pineapple (Ananas comosus) cv. Phulae

Main Article Content

วุฒิชัย จิตดิลกธรรม
เยาวพา จิระเกียรติกุล
ธราธร ทีรฆฐิติ

Abstract

Protoplast isolation is one of the important steps for protoplast fusion in pineapple breeding program. Condition for protoplast isolation is different in each plant species. The objective of this study was to establish appropriate condition for protoplast isolation from in vitro leaf of Pineapple cv. Plulae. Mannitol concentrations (0.1 – 0.5 M), cellulase concentrations (Cellulase Onuzuga R-10, 1 – 2.5% (w/v)), macerozyme concentrations (Macerozyme Onozuga R-10, 0.5 – 2% (w/v)), digestion time (4 – 6 h) and centrifugation speeds (700 – 1000 rpm) were investigated. The results showed that 0.4 M mannitol, 2% (w/v) cellulase, 1% (w/v) macerozyme, 6 h digestion time and centrifugation speed at 800 rpm were the appropriate condition for protoplast isolation from in vitro leaf of Pineapple cv. Plulae. The protoplast yield was 3.82x106±0.02/ g FW.

Article Details

How to Cite
จิตดิลกธรรม ว. ., จิระเกียรติกุล เ., & ทีรฆฐิติ ธ. (2021). Protoplast Isolation from In Vitro Leaf of Pineapple (Ananas comosus) cv. Phulae. Thai Journal of Science and Technology, 10(2), 185–195. https://doi.org/10.14456/tjst.2021.15
Section
วิทยาศาสตร์ชีวภาพ

References

Chaudhary, V., Kumar, V., Sunil, Vaishali, Kavindra, S., Kumar, R. and Kumar, V., 2019, Pineapple (Ananas cosmosus) product processing: A review, J. Pharmacogn. Phytochem. 8(3): 4642-4652.

Chamani, E. Tahami, S. K., Zare, N., Zakaria, R. A., Mohebodini, M. and Joyce, D., 2012, Effect of different cellulase and pectinase enzyme treatments on protoplast isolation and viability in Lilium ledebeourii Bioss, Not. Bot. Horti. Agrobot Cluj-Napoca. 40(2): 123-128.

Diab, S. M. A. M., 2009, In Vitro Rooting and Acclimatization of Pineapple (Ananas comosus L. Merr.) “Smooth Cayenne” cultivar, M.Sc. Thesis, University of Khartoum, Sudan, 138 p.

Farahani, F., 2014, Micropropagation and growth of in vitro pineapple (Ananas cosmosus L. Merr) in Iran, Plant Arch. 14 (1): 337-341.

Hagagy, N. A. A., Mohamed, G. E. A., Mougheith, M. G. E. A., Saleh, M. A. E, Gendiah, H. M., Mohamed, S. K. and Hassan, S. A. M., 2007, Isolation and culture of pineapple (Ananas comnosus) protoplast, 2 (1): 97 -103. In Proceedings of the Second International Conference on The Role of Genetics and Biotechnology in Conservation of Natural Resources, 9-11 July 2007, Ismailia, Egypt.

Jia, X., Zang, X., Qu, J. and Han, R., 2016, Optimization conditions of wheat mesophyll protoplast isolation, Agricultural Sciences. 7: 850-858.

Joy, P. P and Anjana, R., 2016, Evolution of pineapple, pp. 263-296, In Peter, K. V. (Ed.), The Genesis and Evolution of Horticultural Crops, Kruger Brentt Publishers, UK.

Kang, H. H., Naing, A. H. and Kim, C. K., 2020, Protoplast isolation and shoot regeneration from protoplast-derived callus of Petunia hybrida cv. Mirage Rose, Biology. 9: 228 doi: 10.3390/biology9080228.

Kanchanapoom, K., 2002, Plant Protoplast Technology, Chulalongkorn University Printing House, Bangkok, 116 p. (in Thai)

Li, J., Liao, X., Zhou, S., Liu, S., Jiang, L. and Wang, G., 2018, Efficient protoplast isolation and transient gene expression system for Phalaenopsis hybrid cultivar ‘Ruili Beauty’, In Vitro Cell. Dev. Biol. Plant. 54: 87–93.

Mastuti, R. and Rosyidah, M., 2018, In vitro enzymatic isolation of protoplasts from tissues of the medicinal plant Physalis angulata L, AIP Conference Proceedings 2019, 020002 (2018). https://doi.org/10.1063/1.5061838.

Phonyiam, O., Kongsuwan, A. and Setha, S., 2016, Effect of short-term anoxic treatment on internal browning and antioxidant ability in pineapple cv. Phulae, Int. Food Res. J. 23(2): 521-527.

Priyadarshani, S. V. G. N., Hu, B., Li, W., Ali, H., Jia, H., Zhao, L., Peter, S., Syed, O., Azam, M., Xiong, J., Yan, M., Rahman, Z., Wu, Q. and Qin, Y., 2018, Simple protoplast isolation system for gene expression and protein interaction studies in pineapple (Ananas comosus L.), Plant Methods. 14: 95.

Sanputawong, S., 2015, Influence of culture media and concentrations of BA on shoot multiplication of pineapple cv. Phulae. Wichcha Journal. 34(1): 77-83. (in Thai)

Suwanno, S., 2010, Isolation and Culture of Protoplasts from Cell Suspension Culture of Oil Palm cv. Tenera. Master Thesis, Prince of Songkla University.

Tomar, U. K. and Dantu, P.K., 2010, Protoplast culture and somatic hybridization, pp. 876-891, In Tripathi G. (Ed). Cellular and Biochemical Science. I.K. International PVT Ltd, New Delhi.

Wongwirattana, S., laksanalamai, V., Teerakathiti, T. and Leelapon, O., 2016, In vitro embryo culture in pineapple hybrids. The 6th STOU National Research Conference. 25 November 2016. Nonthaburi, Thailand.

Zulkarnain, Z., Neliyati, N., and Eliyanti, E., 2018, Plantlets regeneration from crown bud slicing of pineapple (Ananas comosus). Biosaintifika: Journal of Biology & Biology Education 10(3): 484-490.

Zhao, W., Yang, W., Wei, C. and Sun, G., 2011, A simple and efficient method for isolation of pineapple protoplasts. Biotechnol. & Biotechnol Eq. 25(3): 2464–2467.