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Effects of Biotin and Ascorbic Acid Applications on Haploid Embryo Induction in Semisolid and Double Layer Nutrient Media in Pepper (Capsicum annuum L.) Anther Culture

Year 2021, Volume 5, Issue 2, 191 - 196, 28.06.2021
https://doi.org/10.31015/jaefs.2021.2.8

Abstract

Generation of homozygous double haploid (DH) lines by androgenesis is a promising alternative to selfpollination programs across generations. Despite the routine use of anther culture in peppers, there are still many bottlenecks and improvements in methodology are required. The aim of this study was to determine the effects of the structure of the nutrient medium (semi-solid and double layer) and the addition of biotin and ascorbic acid to the nutrient media on obtaining haploid embryos by anther culture method. MS (Murashige and Skoog 1962) medium containing 4 mg l-1 NAA, 0.1 mg l-1 BAP, 0.25% activated charcoal, 30 g l-1 sucrose, and 10 mg l-1 AgNO3 (silver nitrate) were used as the basal nutrient medium. A total of 8 nutrient media compounds were studied using 0.05 mg l-1 biotin and 0.5 mg l-1 ascorbic acid separately or together in semi-solid and bi-layer (double-phase) nutrient media. Solidification of nutrient media was achieved with 7 g 1-1 agar. The cultured anthers were subjected to high-temperature pre-treatments at 35 °C in continuous dark conditions for 2 days. Then they were taken to a climate chamber at of 25 °C temperature adjusted to 16/8 hour photoperiod. It has been observed that the success of obtaining embryos of semi-solid medium was higher than double-layer medium. The addition of biotin and ascorbic acid to the nutrient medium provided 8.8 fold increases in embryo regeneration compared to the control medium. In the presence of only one of biotin or ascorbic acid in the nutrient medium, the number of embryos increased compared to the control. 

References

  • Açıkgöz, N., İlker E., Gökçöl, A. (2004). Biyolojik Araştırmaların Bilgisayarda Değerlendirilmeleri, Ege Üniversitesi Tohum Teknolojisi Uygulama. ve Araştırma Merkezi Yayınları No:2, Ege Üniversitesi Ziraat Fakültesi Ofset Atölyesi, Bornova-İzmir, 202 s (in Turkish).
  • Ahmadi, B., Ebrahimzadeh, H. (2020). In vitro androgenesis: spontaneous vs. artifcial genome doubling and characterization of regenerants. Plant Cell Reports. 39:299–316. Doi: https://doi.org/10.1007/s00299-020-02509-z
  • Al-Khayri, J.M. (2001). Optimization of biotin and thiamine requirements for somatic embryogenesis of date palm (Phoenix dactylifera L.). In Vitro Cell. & Dev. Biol.-Plant. 37, 453–456. Doi: https://doi.org/10.1007/s11627-001-0079-x
  • Becker, M. G., Chan, A., Mao X., Girard, I. J., Lee S., Elhiti M., Stasolla, C., Belmonte, M.F. (2014). Vitamin C deficiency improves somatic embryo development through distinct gene regulatory networks in Arabidopsis. Journal of Experimental Botany. 65(20) 5903–5918. 10.1093/jxb/eru330. Doi: https://doi.org/10.1093/jxb/eru330
  • Ceyhan, A.P. and Aktaş, H. (2020). Development of Three Dihaploid Nose and Bell Pepper Lines with Anther Culture Technique. Eurasian J Bio Chem Sci, 3:199-205, Doi: https://doi.org/10.46239/ejbcs.822593
  • Cheng, Y., Jiao, Y., Miao, R., Tian, R., Liang Y., Qiao, N. (2020). Exploring differentially expressed genes of microspore embryogenesis under heat stress in sweet pepper. African J. of Biotechnology. 19(9):661-674. Doi: https://doi.org/10.5897/AJB2020.17194
  • Çömlekçioğlu N., Ellialtıoğlu, Ş. Ş. (2018). Review on the Research carried out on in vitro Androgenesis of Peppers (Capsicum annuum L.) in Turkey. Review Paper: Research Journal of Biotechnology. 13(6): 75-84.
  • El sharabasy S.F., Bosila, H.A., Abdel-Aal, W.B., Mansour, B.M., Bana A.A. (2019). Effect of Vitamins (pyridoxine and nicotinic acid), Thiamine-Hcl and Myo-Inositol at Different Concentrations on Free Amino Acids and Indoles Content of Embryogenic Callus of in vitro Date Palm (Sakkoty and Bartamuda Cultivar) Materials Research Proceedings 11:244-252. Doi: https://doi.org/10.21741/9781644900178-20
  • Gill, S.S. Tuteja, N. (2010). Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemistry, 48(12): 909–930. Doi: https://doi.org/10.1016/j.plaphy.2010.08.016
  • Habibi, N., Suthar, P. K. Purohit, S. D. (2009). Role of PGRs and inhibitors in induction and control of somatic embryogenesis in Themeda quadrivalvis. Indian Journal of Experimental Biology, 47(3): 198–203.
  • Heidari-Zefreh, A.A., Shariatpanahi, M.E., Mousavi, A., Kalatejari, S. (2018). Enhancement of microspore embryogenesis induction and plantlet regeneration of sweet pepper (Capsicum annuum L.) using putrescine and ascorbic acid. Protoplasma. 256(1):13–24. Doi: https://doi.org/10.1007/s00709-018-1268-3
  • Hosp J., Tashpulatov, A., Roessner, U., Barsova, E., Katholnigg, H., Steinborn, R., Melikant, B., Lukyanov, S., Heberle-Bors, E., Touraev, E. (2007). Transcriptional and metabolic profiles of stress-induced, embryogenic tobacco microspores. Plant Molecular Biology. 63:137–149. Doi: https://doi.org/10.1007/s11103-006-9078-y
  • Irıkova, T., Grozeva, S., Popov, P., Rodeva, V., Todorovska, E. (2011). In vitro response of pepper anther culture (Capsicum annuum L.) depending on genotype, nutrient medium and duration of cultivation. Biotechnology and Biotechnological Equipment, 25(4): 2604-2609. Doi: https://doi.org/10.5504/BBEQ.2011.0090
  • Munoz-Amatriain, M., Svensson, J.T. Castillo, A.M. Cistue, L. Close, T.J. Valles, M.P. (2009). Expression profiles in barley microspore embryogenesis, in: A. Touraev, et al. (Eds.), Advances in Haploid Production in Higher Plants, Springer. pp. 127–134 Doi:https://doi.org/10.1007/978-1-4020-8854-4_9
  • Murashige, T. and Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures, Physiol. Plant 15: 473-497. Doi: https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  • Ozsan, T., Onus, A. N. (2017). In vitro Pepper (Capsicum annuum L.) Anther Culture: Can be Affected Via Vitamins B?. Biotechnology Journal International. 20(1):1-13. Doi: https://doi.org/10.9734/BJI/2017/37102
  • Perez-Pere,z Y., El-Tantawy, A.A., Solis, M.T., Risueno, M.C. Testillano, P.S. (2019) Stress-induced microspore embryogenesis requires endogenous auxin synthesis and polar transport in barley. Front. Plant Sci. 10:1200. Doi: https://doi.org/10.3389/fpls.2019.01200
  • Pinar, H., Mutlu, N., Yildiz, S., Simsek, D., Shams, M. (2020). Transferring the cultured anther to a medium without activated charcoal overcomes the recalcitrance in pepper genotypes. Canadian Journal of Plant Science. 12 August 2020. Doi: https://doi.org/10.1139/cjps-2020-0050
  • Rodriguez-Serrano, M., Barany, I., Prem, D., Coronado, M. J., Risueno, M. C. Testilano, P.S. 2012. NO, ROS, and cell death associated with caspase-like activity increase in stress induced microspore embryogenesis of barley. Journal of Experimental Botany. 63(5), 2007–2024. Doi: https://doi.org/org/10.1093/jxb/err400
  • Roje, S. (2007). Vitamin B biosynthesis in plants. Phytochemistry. 68(14):1904–1921. Doi: https://doi.org/10.1016/j.phytochem.2007.03.038
  • Sanchez, M.A., Coronado, Y.M., Coronado, A.C.M. (2020). Androgenic studies in the production of haploids and doubled haploids in Capsicum spp. Revista Facultad Nacional de Agronomia Medellin. 73(1):9047-9056.
  • Segui-Simarro J.M., Nuez, F. (2008). How microspores transform into haploid embryos: changes associated with embryogenesis induction and microspore derived embryogenesis. Physiologia Plantarum 134: 1–12. Doi: https://doi.org/10.1111/j.1399-3054.2008.01113.x
  • Shahinul Islam, S.M. Tuteja, N. (2012). Enhancement of androgenesis by abiotic stress and other pretreatments in majör crop species. Plant Science 182:134–144. Doi: https://doi.org/10.1016/j.plantsci.2011.10.001
  • Shariatpanahi, M.E., Bal, U., Heberle-Bors, E., Touraev, A. (2006). Stresses applied for the re-programming of plant microspores towards in vitro embryogenesis. Physiol Plant 127(4):519–534. Doi: https://doi.org/10.1111/j.1399-3054.2006.00675.x
  • Touraev, A., Vicente, O., Heberle-Bors E. 1997. Initiation of microspore embryogenesis by stress. Trends Plant Sci. 2(8):297–302. Doi: https://doi.org/10.1016/S1360-1385(97)89951-7
  • Varnier A.L., Jacquard C., Clement C. (2009) Programmed cell death and microspore embryogenesis. In: Touraev A., Forster B.P., Jain S.M. (eds) Advances in Haploid Production in higher plants. Springer, Dordrecht. 147–154. Doi: https://doi.org/10.1007/978-1-4020-8854-4_11
  • Zeng, A., Yan, J., Song, L., Gao, B., Li, J. (2015). Effects of ascorbic acid and embryogenic microspore selection on embryogenesis in white cabbage (Brassica oleracea L. var. capitata). The Journal of Horticultural Science and Biotechnology, 90(6): 607-612, Doi: https://doi.org/10.1080/14620316.2015.11668722
  • Zur, I., Dubas, E., Golemiec, E., Szechynska-Hebda, M., Golebiowska, G., Wedzony, M. (2009). Stress-related variation in antioxidative enzymes activity and cell metabolism efficiency associated with embryogenesis induction in isolated microspore culture of triticale (× Triticosecale Wittm.). Plant Cell Rep 28:1279–1287. Doi: https://doi.org/10.1007/s00299-009-0730-2

Year 2021, Volume 5, Issue 2, 191 - 196, 28.06.2021
https://doi.org/10.31015/jaefs.2021.2.8

Abstract

References

  • Açıkgöz, N., İlker E., Gökçöl, A. (2004). Biyolojik Araştırmaların Bilgisayarda Değerlendirilmeleri, Ege Üniversitesi Tohum Teknolojisi Uygulama. ve Araştırma Merkezi Yayınları No:2, Ege Üniversitesi Ziraat Fakültesi Ofset Atölyesi, Bornova-İzmir, 202 s (in Turkish).
  • Ahmadi, B., Ebrahimzadeh, H. (2020). In vitro androgenesis: spontaneous vs. artifcial genome doubling and characterization of regenerants. Plant Cell Reports. 39:299–316. Doi: https://doi.org/10.1007/s00299-020-02509-z
  • Al-Khayri, J.M. (2001). Optimization of biotin and thiamine requirements for somatic embryogenesis of date palm (Phoenix dactylifera L.). In Vitro Cell. & Dev. Biol.-Plant. 37, 453–456. Doi: https://doi.org/10.1007/s11627-001-0079-x
  • Becker, M. G., Chan, A., Mao X., Girard, I. J., Lee S., Elhiti M., Stasolla, C., Belmonte, M.F. (2014). Vitamin C deficiency improves somatic embryo development through distinct gene regulatory networks in Arabidopsis. Journal of Experimental Botany. 65(20) 5903–5918. 10.1093/jxb/eru330. Doi: https://doi.org/10.1093/jxb/eru330
  • Ceyhan, A.P. and Aktaş, H. (2020). Development of Three Dihaploid Nose and Bell Pepper Lines with Anther Culture Technique. Eurasian J Bio Chem Sci, 3:199-205, Doi: https://doi.org/10.46239/ejbcs.822593
  • Cheng, Y., Jiao, Y., Miao, R., Tian, R., Liang Y., Qiao, N. (2020). Exploring differentially expressed genes of microspore embryogenesis under heat stress in sweet pepper. African J. of Biotechnology. 19(9):661-674. Doi: https://doi.org/10.5897/AJB2020.17194
  • Çömlekçioğlu N., Ellialtıoğlu, Ş. Ş. (2018). Review on the Research carried out on in vitro Androgenesis of Peppers (Capsicum annuum L.) in Turkey. Review Paper: Research Journal of Biotechnology. 13(6): 75-84.
  • El sharabasy S.F., Bosila, H.A., Abdel-Aal, W.B., Mansour, B.M., Bana A.A. (2019). Effect of Vitamins (pyridoxine and nicotinic acid), Thiamine-Hcl and Myo-Inositol at Different Concentrations on Free Amino Acids and Indoles Content of Embryogenic Callus of in vitro Date Palm (Sakkoty and Bartamuda Cultivar) Materials Research Proceedings 11:244-252. Doi: https://doi.org/10.21741/9781644900178-20
  • Gill, S.S. Tuteja, N. (2010). Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemistry, 48(12): 909–930. Doi: https://doi.org/10.1016/j.plaphy.2010.08.016
  • Habibi, N., Suthar, P. K. Purohit, S. D. (2009). Role of PGRs and inhibitors in induction and control of somatic embryogenesis in Themeda quadrivalvis. Indian Journal of Experimental Biology, 47(3): 198–203.
  • Heidari-Zefreh, A.A., Shariatpanahi, M.E., Mousavi, A., Kalatejari, S. (2018). Enhancement of microspore embryogenesis induction and plantlet regeneration of sweet pepper (Capsicum annuum L.) using putrescine and ascorbic acid. Protoplasma. 256(1):13–24. Doi: https://doi.org/10.1007/s00709-018-1268-3
  • Hosp J., Tashpulatov, A., Roessner, U., Barsova, E., Katholnigg, H., Steinborn, R., Melikant, B., Lukyanov, S., Heberle-Bors, E., Touraev, E. (2007). Transcriptional and metabolic profiles of stress-induced, embryogenic tobacco microspores. Plant Molecular Biology. 63:137–149. Doi: https://doi.org/10.1007/s11103-006-9078-y
  • Irıkova, T., Grozeva, S., Popov, P., Rodeva, V., Todorovska, E. (2011). In vitro response of pepper anther culture (Capsicum annuum L.) depending on genotype, nutrient medium and duration of cultivation. Biotechnology and Biotechnological Equipment, 25(4): 2604-2609. Doi: https://doi.org/10.5504/BBEQ.2011.0090
  • Munoz-Amatriain, M., Svensson, J.T. Castillo, A.M. Cistue, L. Close, T.J. Valles, M.P. (2009). Expression profiles in barley microspore embryogenesis, in: A. Touraev, et al. (Eds.), Advances in Haploid Production in Higher Plants, Springer. pp. 127–134 Doi:https://doi.org/10.1007/978-1-4020-8854-4_9
  • Murashige, T. and Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures, Physiol. Plant 15: 473-497. Doi: https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  • Ozsan, T., Onus, A. N. (2017). In vitro Pepper (Capsicum annuum L.) Anther Culture: Can be Affected Via Vitamins B?. Biotechnology Journal International. 20(1):1-13. Doi: https://doi.org/10.9734/BJI/2017/37102
  • Perez-Pere,z Y., El-Tantawy, A.A., Solis, M.T., Risueno, M.C. Testillano, P.S. (2019) Stress-induced microspore embryogenesis requires endogenous auxin synthesis and polar transport in barley. Front. Plant Sci. 10:1200. Doi: https://doi.org/10.3389/fpls.2019.01200
  • Pinar, H., Mutlu, N., Yildiz, S., Simsek, D., Shams, M. (2020). Transferring the cultured anther to a medium without activated charcoal overcomes the recalcitrance in pepper genotypes. Canadian Journal of Plant Science. 12 August 2020. Doi: https://doi.org/10.1139/cjps-2020-0050
  • Rodriguez-Serrano, M., Barany, I., Prem, D., Coronado, M. J., Risueno, M. C. Testilano, P.S. 2012. NO, ROS, and cell death associated with caspase-like activity increase in stress induced microspore embryogenesis of barley. Journal of Experimental Botany. 63(5), 2007–2024. Doi: https://doi.org/org/10.1093/jxb/err400
  • Roje, S. (2007). Vitamin B biosynthesis in plants. Phytochemistry. 68(14):1904–1921. Doi: https://doi.org/10.1016/j.phytochem.2007.03.038
  • Sanchez, M.A., Coronado, Y.M., Coronado, A.C.M. (2020). Androgenic studies in the production of haploids and doubled haploids in Capsicum spp. Revista Facultad Nacional de Agronomia Medellin. 73(1):9047-9056.
  • Segui-Simarro J.M., Nuez, F. (2008). How microspores transform into haploid embryos: changes associated with embryogenesis induction and microspore derived embryogenesis. Physiologia Plantarum 134: 1–12. Doi: https://doi.org/10.1111/j.1399-3054.2008.01113.x
  • Shahinul Islam, S.M. Tuteja, N. (2012). Enhancement of androgenesis by abiotic stress and other pretreatments in majör crop species. Plant Science 182:134–144. Doi: https://doi.org/10.1016/j.plantsci.2011.10.001
  • Shariatpanahi, M.E., Bal, U., Heberle-Bors, E., Touraev, A. (2006). Stresses applied for the re-programming of plant microspores towards in vitro embryogenesis. Physiol Plant 127(4):519–534. Doi: https://doi.org/10.1111/j.1399-3054.2006.00675.x
  • Touraev, A., Vicente, O., Heberle-Bors E. 1997. Initiation of microspore embryogenesis by stress. Trends Plant Sci. 2(8):297–302. Doi: https://doi.org/10.1016/S1360-1385(97)89951-7
  • Varnier A.L., Jacquard C., Clement C. (2009) Programmed cell death and microspore embryogenesis. In: Touraev A., Forster B.P., Jain S.M. (eds) Advances in Haploid Production in higher plants. Springer, Dordrecht. 147–154. Doi: https://doi.org/10.1007/978-1-4020-8854-4_11
  • Zeng, A., Yan, J., Song, L., Gao, B., Li, J. (2015). Effects of ascorbic acid and embryogenic microspore selection on embryogenesis in white cabbage (Brassica oleracea L. var. capitata). The Journal of Horticultural Science and Biotechnology, 90(6): 607-612, Doi: https://doi.org/10.1080/14620316.2015.11668722
  • Zur, I., Dubas, E., Golemiec, E., Szechynska-Hebda, M., Golebiowska, G., Wedzony, M. (2009). Stress-related variation in antioxidative enzymes activity and cell metabolism efficiency associated with embryogenesis induction in isolated microspore culture of triticale (× Triticosecale Wittm.). Plant Cell Rep 28:1279–1287. Doi: https://doi.org/10.1007/s00299-009-0730-2

Details

Primary Language English
Subjects Horticulture
Published Date June 2021
Journal Section Research Articles
Authors

Burcu DEMİRKAYA This is me
Eskişehir Osmangazi Üniversitesi Ziraat Fakültesi Bahçe Bitkileri Bölümü
0000-0002-3390-4669
Türkiye


Nuray ÇÖMLEKÇİOĞLU (Primary Author)
ESKİŞEHİR OSMANGAZİ ÜNİVERSİTESİ
0000-0001-7189-613X
Türkiye

Supporting Institution No financial support was received for this study.
Thanks The authors would like to thank the Faculty of Agriculture, Eskisehir Osmangazi University for providing their greenhouse to cultivation of donor plants
Publication Date June 28, 2021
Application Date March 3, 2021
Acceptance Date May 12, 2021
Published in Issue Year 2021, Volume 5, Issue 2

Cite

APA Demirkaya, B. & Çömlekçioğlu, N. (2021). Effects of Biotin and Ascorbic Acid Applications on Haploid Embryo Induction in Semisolid and Double Layer Nutrient Media in Pepper (Capsicum annuum L.) Anther Culture . International Journal of Agriculture Environment and Food Sciences , 5 (2) , 191-196 . DOI: 10.31015/jaefs.2021.2.8