Vegetos
(An International Journal of Plant Research & Biotechnology)
Somatic embryogenesis in N22 variety of rice (Oryza sativa L.) with drought and heat tolerance
*Article not assigned to an issue yet
Research Articles | Published: 26 June, 2025
First Page: 0
Last Page: 0
Views: 274
Keywords: Somatic embryogenesis, Explant, Subculturing, Plantlets, Nagina 22
Abstract
Standardized in vitro regeneration protocols are essential for effective crop improvement through transformation, particularly for the challenging indica subspecies of rice. Somatic embryogenesis serves as a promising method for genetic transformation and for the production of a large number of plantlets. This research aimed to establish a protocol for somatic embryogenesis and regeneration in Nagina 22 (N22), an aus type rice variety known for its drought and heat tolerance, which originated in India. The study examined the effects of two media (MS and N6), two carbon sources (sucrose and maltose), and 2, 4-D (at concentrations of 0, 1, 2, and 3 µM) on callus induction using mature rice seeds as explants. Somatic embryo formation was achieved in the calli obtained by subculturing on MS medium enriched with 2, 4-D (at 0 and 0.4 µM) and kinetin (0, 0.4, 1, 1.5 and 2 µM). Regeneration experiments were conducted using MS medium, incorporating NAA and BAP to initiate shoot formation, along with IBA to enhance root development in the plantlets produced. The highest frequency of callus induction (100%) was achieved in MS medium with maltose as the carbon source, supplemented with 2, 4-D at 3 µM. Regeneration rates were maximized in MS medium enriched with NAA (both 0.2 and 1 µM) and BAP (0.8 and 3.5 µM) from the developed somatic embryos. The resulting plantlets were effectively hardened and successfully established in pot culture. This finding holds significant importance for future rice enhancement and gene editing initiatives globally, using N22 as a donor for introducing drought and heat tolerance.
References
Bin-Rahman AR, Zhang J (2022) Trends in rice research: 2030 and beyond. Food Energy Security e390.
Carneros E, Sánchez-Muñoz J, Pérez-Pérez Y, Pintos B, Gómez-Garay A, Testillano PS (2023) Dynamics of endogenous auxin and its role in somatic embryogenesis induction and progression in Cork Oak. Plants 12(7):1542
Chu CC (1975) Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Sci Sin 18:659–668
Da Silva GJ, Schreinert R, Medeiros T, Dienes N, Peters JA, Costa A (2015) Somatic embryogenesis and plant regeneration in Brazilian rice genotypes. Aus J Crop Sci 9:1126–1130
GRiSP (Global Rice Science Partnership). (2013) Rice Almanac. 4th Edition, International Rice Research Institute, Los Baños.
Hiei Y, Komari T (2008) Agrobacterium-mediated transformation of rice using immature embryos or calli induced from mature seed. Nat Protoc 3:824–834
Hiei Y, Ishida Y, Kasaoka K, Komari T (2006) Improved frequency of transformation in rice and maize by treatment of immature embryos with centrifugation and heat prior to infection with Agrobacterium tumefaciens. Plant Cell Tissue Organ Cult 87:233–243
Herve P, Kayano T (2006) Japonica rice varieties (Oryza sativa, Nipponbare, and others). Methods Mol Biol 343:213–222
Husna SF, Dewanti P, Sugiharto B, Fanata WID (2022) Expression of cytokinin responsive and ethylene biosynthesis genes in rice callus with different regeneration rates. Indonesian J Biotech 27(2):65–72
Indoliya Y, Tiwari P, Chauhan AS, Goel R, Shri M, Bag SK, Chakrabarty D (2016) Decoding regulatory landscape of somatic embryogenesis reveals differential regulatory networks between japonica and indica rice subspecies. Sci Rep 6(1):1–15
Khatun MM, Ali MH, Desamero NV (2003) Effect of genotype and culture media on callus formation and plant regeneration from mature seed scutella culture in rice. Plant Tissue Cult 13(2):99–107
Krishnan SR, Priya AM, Ramesh M (2013) Rapid regeneration and ploidy stability of ‘cv IR36’ indica rice (Oryza sativa. L.) confers efficient protocol for in vitro callus organogenesis and Agrobacterium tumefaciens mediated transformation. Bot Stud 54(1):1–12
Manickavasagam M, Pavan G, Vasudevan V (2019) A comprehensive study of the hormetic influence of biosynthesized AgNPs on regenerating rice calli of indica cv. IR64. Sci Rep 9(1):1–11
Méndez-Hernández HA, Ledezma-Rodríguez M, Avilez-Montalvo RN, Juárez-Gómez YL, Skeete A, Avilez-Montalvo J, De-La-Peña C, Loyola-Vargas VM (2019) Signaling overview of plant somatic embryogenesis. Front Plant Sci 10:77
Mesbah HA, Nassar AESM, El-Sheikh MA (2021) Histological studies on somatic embryogenesis in rice (Oryza sativa L.). Afr J Biol Sci 17(1):73–81
Ming NJ, Mostafiz SB, Johon NS, Zulkifli NSA, Wagiran A (2019) Combination of plant growth regulators, maltose, and partial desiccation treatment enhance somatic embryogenesis in selected Malaysian rice cultivar. Plants 8(6):144
Molina-Risco M, Ibarra O, Faion-Molina M, Kim B, Septiningsih EM, Thomson MJ (2021) Optimizing Agrobacterium-mediated transformation and CRISPR-Cas9 gene editing in the tropical japonica rice variety Presidio. Int J Mol Sci 22:10909
Mostafiz SB, Wagiran A (2018) Efficient callus induction and regeneration in selected indica rice. Agronomy 8(5):77
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15(3):473–497
Nic-Can GI, Loyola-Vargas VM (2016) The Role of the auxins during somatic embryogenesis. In: Loyola-Vargas V, Ochoa-Alejo N (eds) Somatic embryogenesis: fundamental aspects and applications. Springer, Cham, pp 171–182
Niroula RK, Sah BP, Bimb HP, Nayak S (2005) Effect of genotype and culture media on callus induction and plant regeneration from matured rice grain culture. J Inst Agric Animal Sci 26:21–26
Noor W, Lone R, Kamili AN, Husaini AM (2022) Callus induction and regeneration in high-altitude Himalayan rice genotype SR4 via seed explant. Biotech Rep 36:e00762
Repalli SK, Geda CK, NSN P, Rao GJN (2019) Influence of media type and carbon source on callus induction and regeneration response of different indica rice genotypes. Adv Crop Sci Tech 7(448):2
Sahoo KK, Tripathi AK, Pareek A, Sopory SK, Singla-Pareek SL (2011) An improved protocol for efficient transformation and regeneration of diverse indica rice cultivars. Plant Meth 7(1):1–11
Shetty PK, Hegde MR, Mahadevappa M (2013) Innovations in Rice Production (NIAS Books and Special Publications No. SP1–2013. NIAS, Bangalore. [24 September, 2015]. http://eprints.nias.res.in/id/eprint/369
Shri M, Rai A, Verma PK, Misra P, Dubey S, Kumar S, Verma S, Gautam N, Tripathi RD, Trivedi PK et al (2013) An improved Agrobacterium-mediated transformation of recalcitrant indica rice (Oryza sativa L.) cultivars. Protoplasma 250:631–636
Swapna TS, Padma N (2000) Generation of variability for salt tolerance in rice using tissue culture techniques. Doctoral Thesis, Department of Biotechnology, CUSAT, Kerala, India. p.178 http://dyuthi.cusat.ac.in/ purl/915
Verma D, Joshi R, Shukla A, Kumar P (2011) Protocol for in vitro somatic embryogenesis and regeneration of rice (Oryza sativa L.). Indian J Exp Biol 49(12):958–963
Barrera M, Olmedo B, Zúñiga C, Cepeda M, Olivares F, Vergara R, Cordero-Lara K, Prieto H (2024) Somatic embryogenesis and Agrobacterium-mediated gene transfer procedures in chilean temperate Japonica rice varieties for precision breeding. Plants 13:416. https://doi.org/10.3390/plants13030416
Huang X, Kurata N, Wei X, Wang ZX, Wang Z, Zhao Q, Zhao Y, Liu K, Lu H, Li W, Guo Y, Lu Y, Zhou C, Fan D, Weng Q, Zhu C, Huang T, Zhang L, Wang Y, Feng L, Furuumi H, Kubo T, Miyabayashi T, Yuan X, Xu Q, Dong G, Zhna Q, Li C, Fujiyama A, Lu T, Feng Q, Qian Q, Li J, Han B. A map of rice genome variation reveals the origin of cultivated rice. Nature. 2012;490:497–503. https://doi.org/10.1038/nature11532
Zuraida A, Naziah B, Zamri Z, Sreeramanan S (2011) Efficient plant regeneration of Malaysian indica rice MR 219 and 232 via somatic embryogenesis system. Acta Physiol Plant 33:1913–1921. https://doi.org/10.1007/s11738-011-0739-3
Author Information
Department of Plant Breeding and Genetics, College of Agriculture, Vellanikkara, KAU, Thrissur, India