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Histo-morphological analysis of rice callus cultures reveals differential regeneration response with varying media combinations

  • Plant Tissue Culture
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Abstract

Genetic transformation of most indica rice (Oryza sativa) cultivars is hampered by poor in vitro culture performance and low regeneration potential. Histological study of primary calli can provide substantial information on their regeneration potential and can be used for early grading of calli expected to develop plantlets on regeneration media. The study was aimed to undertake histological analysis of primary calli derived from mature seeds of five indica rice cultivars viz. KSK-133, KS-282, Shaheen Basmati, Super Basmati, and DilRosh in order to assess their regeneration potential on different media combinations supplemented with various hormone concentrations (N6 + 2 mg/L 2,4-Dichlorophenoxyacetic acid; N6 + 2 mg/L 2–4 D + 2 mg/L Benzylaminopurine and MS + 2 mg/L 2,4-D). Calli with regeneration capability were subjected to histological assays by examining toulidine blue stained 5–8 μm thin sections for the presence of meristematic zones exhibiting embryogenic callus features. Based on our observations, formation of embryoids or embryoid-like structures was pronounced in KSK-133 and KS-282 calli. However, DilRosh, Super Basmati and Shaheen Basmati did not show these characteristic features. Three-week-old calli of all rice cultivars were transferred into regeneration medium (MS + 2 mg/L BAP + 1 mg/L Naphthaleneacetic acid). KSK-133 and KS-282 showed the highest regeneration potential (81% and 76%, respectively). These data were supported by histological observations where characteristic embryogenic units (EU) were noticed in these genotypes. These meristematic regions displayed high mitotic activity and stained relatively dark. The embryogenic calli cells were found heavily cytoplasmic with prominent nuclei and were located on the callus surface or inside surrounded by parenchymal cells.

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References

  • Afrasiab H, Jafar R (2011) Effect of different media and solidifying agents on callogenesis and plant regeneration from different plants of explants of rice (Oryza sativa L) varieties super basmati and IRRI-6. Pak J Bot 43(1):487–501

    CAS  Google Scholar 

  • Aldemita RR, Hodges TK (1996) Agrobacterium tumefaciens mediated transformation of japonica and indica varieties. Planta 199:612–617

    Article  CAS  Google Scholar 

  • Alfonso-Rubi J, Carbonero P, Diaz I (1999) Parameters influencing the regeneration capacity of calluses derived from mature indica and japonica rice seeds after micro projectile bombardment. Euphytica 107:115–122

    Article  Google Scholar 

  • Bano S, Jabeen M, Rahim F & Ilaho I. (2005) Callus induction and regeneration in seed explants of rice (Oryza sativa L.) cv. Swat-II. Pak.G.Bot. 37:829–836.

  • Bevitori R, Popielarska-Konieczna M, dos Santos EM, Grossi-de-Sá MF, Petrofeza R (2014) Morpho-anatomical characterization of mature embryo-derived callus of rice (Oryza sativa L.) suitable for transformation. Protoplasma 251:545–554

    CAS  PubMed  Google Scholar 

  • Cho MJ, Yano H, Okamoto HK, Kim HR, Jung K, Newcomb VK, Le HS, Yoo R, Langha BB, Buchanan, Lemaux PG (2004) Stable transformation of rice (Oryza sativa L) via micro projectile bombardment of highly regenerative, green tissues derived from mature seed. Plant Cell Rep 22:483–489

    Article  CAS  PubMed  Google Scholar 

  • Chu CC, Wang CS, Sun CC, Hsu C, Yin KC, Chu CY (1975) Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. SciSinica 18:659–668

    Google Scholar 

  • Dorosieve L (1996) Plant cell and tissue cultures: present state and future prospect. Genetika I Seleksi 19:356–362

    Google Scholar 

  • Ge X, Chu Z, Lin Y, Wang SA (2006) Tissue culture system for different germplasms of indica rice. Plant Cell Rep 25:392–402

    Article  CAS  PubMed  Google Scholar 

  • Ikeuchi M, Sugimoto K, Iwase A (2013) Plant callus: mechanisms of induction and repression. Plant Cell 25:3159–3173

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Joyia FA, Khan MS (2012) Reproducible and expedient rice regeneration system using in vitro grown plants. Afr J Biotechnol 11:138–144

    CAS  Google Scholar 

  • Khan MS, Maliga P (1999) Fluorescent antibiotic resistance marker for tracking plastid transformation in higher plants. Nat Biotechnol 17:910–915

    Article  CAS  PubMed  Google Scholar 

  • Khanna HK, Raina SK (1998) Genotype x culture media interaction effects on regeneration response of three indica rice cultivars. Plant Cell Tiss Org Cult 52(3):145–153

    Article  CAS  Google Scholar 

  • Kumar K, Kumar M, Kim SR, Ryu H, Cho YG (2013) Insight into genomics of salt response in rice. Rice 6:27

    Article  PubMed  PubMed Central  Google Scholar 

  • Leng CE, Keng CL (2007) Induction of somatic embryogenic callus from the leaves of Pereskia grandifolia. Biotechnol 6(1):45–48

    Article  CAS  Google Scholar 

  • Mendoza AB, Hattori K, Futsuhara Y (1992) Shoot regeneration from the callus of immature primary leaves in Mungbean Vigna radiata(L.)Wilczek. Ikushugaku zasshi. 42:145–149. https://doi.org/10.1270/jsbbs1951.42.145.

  • Mohamed F, Mohamed I, Dermot P, Coyne, Paul ER (1993) Shoot organogenesis in callus induced from pedicel explants of common bean (Phaseolus vulgaris L). J Am Soc Hortic Sci 118(1):158–162

    Article  Google Scholar 

  • Molina MD, Aponte EM, Cortina H, Moreno G (2002) The effect of genotype and explant age on somatic embryogenesis of coffee. Plant Cell Tissue Org Cult 71:117–123

    Article  CAS  Google Scholar 

  • Murashige T, Skoog FA (1962) Revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plant 15:472–493

    Article  Google Scholar 

  • Naqvi SMS, Yasmin T, Mahmood T, Akhtar MS (2006) Efficient embryogenic system from tissue culture of mature embryos for some coarse varieties of rice (Oryza sativa L). Pak J Bot 38(4):969–975

    Google Scholar 

  • Narciso JO, Hattori K (2010) Genotypic differences in morphology and ultra structures of callus derived from selected rice varieties. Philipp Sci Lett 3:59–65

    Google Scholar 

  • Noor A, Rashid H, Chaudhry Z, Mirza B (2005) High frequency regeneration from scutellum derived calli of basmati rice cv Basmati 385 and Super Basmati. Pak J Bot 37(3):673–684

    Google Scholar 

  • Noor A, Rashid H, Khan MH, Chaudhry Z (2011) Studies on the effect of genotype and explants type on callogenesis and organogenesis in indica rice. Pak J Bot 43(5):2445–2449

    Google Scholar 

  • Ozawa K, Kawahigashi H, Kayano T, Ohkawa Y (2003) Enhancement of regeneration of rice (Oryza sativa L.) calli by integration of the gene involved in regeneration ability of the callus. Plant Sci 165:395–402

    Article  CAS  Google Scholar 

  • Peng J, Hodges TK (1989) Genetic analysis of plant regeneration in rice (Oryza sativa L). In Vitro Cell Dev B 25:91–94

    Article  Google Scholar 

  • Peng J, Kononowicz H, Hodges TK (1992) Transgenic indica rice plants. Theor Appl Genet 83:855–863

    Article  CAS  PubMed  Google Scholar 

  • Pravin VJ, Dudhare MS, Salujat T, Sarawgi AK, Saxena R, Girish C (2011) Assessment of critical factors influencing callus induction, in vitro regeneration and selection of bombarded indica rice genotypes. J Agric Biotechnol Sustain Dev 3:59

    Google Scholar 

  • Priya AM, Pandia SK, Ramesh M (2011) Efficient in vitro plant regeneration through leaf base derived callus cultures of abiotic stress sensitive popular Asian indica rice cultivar IR 64 (Oryza sativa L.). Acta Biol Hung 62(4):441–452

    Article  Google Scholar 

  • Qian H, Zhang X, Xue Q (2004) Factors affecting the CI and gustransient expression in indica rice Pei;ai 64s. Pak J Biol Sci 7(4):615–619

    Article  Google Scholar 

  • Quiroz-Figueroa FR, Rojas-Herrera R, Galaz-Avalos RM, Loyola-Vargas VM (2006) Embryo production through somatic embriogénesis can be used to study cell differentiation in plants. Plant Cell Tissue Org Cult 86:285–301

    Article  Google Scholar 

  • Rachmawati D, Anzai H (2006) Studies on callus induction, plant regeneration and transformation of Javanica rice cultivars. Plant Biotechnol 23:521–524

    Article  CAS  Google Scholar 

  • Rafique MZ, Rashid H, Chaudhary MF, Chaudhry Z, Cheema NM (2011) Study on callogenesis and organogenesis in local cultivars of rice (Oryza sativa L). Pak J Bot 34(1):191–203

    Google Scholar 

  • Raina SK (1989) Tissue culture in rice improvement: status and potential. Adv Agron 42:339–398

    Article  Google Scholar 

  • Rashid H, Bokhari SNR, Chaudhry Z, Naqvi SM (2001) Studies on the genotype response to CI from three basmati cultivars of rice (Oryza sativa L.). Pak J Biol Sci 6(5):445–447

    Google Scholar 

  • Sahoo KK, Tripath AK, Pareek A, Sopory SK, Singla-Pareek SL (2011) An improved protocol for efficient transformation and regeneration of diverse indica rice cultivars. Plant Methods 7(1):49

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sangduen N, Klamsomboon P (2001) Histological and scanning electron observations on embryogenic and non-embryogenic calli of aromatic Thai rice (Oryza sativa L cv KhaoDaw Mali 105). Kasetsart J (Nat Sci) 35:427–432

    Google Scholar 

  • Sikder MBH, Sen PK, Al-Mamun MA, Ali RM, Rahman SM (2006) In vitro regeneration of aromatic rice (Oryza sativa L). Int J Agric Biol 6:759–762

    Google Scholar 

  • Tan Y, Hu W, Xu X, Zhou J, Wang C, Liu X, Cheng AG (2011) Ployamine plays a role in subculture growth of in vitro callus of indica rice. Acta Biologia Cracovinesia 59(1)

  • Toki S, Hara N, Ono K, Onodera H, Tagiri A, Oka S, Tanaka H (2006) Early infection of scutellum tissue with Agrobacterium allows high-speed transformation of rice. Plant J 47:969–976

    Article  CAS  PubMed  Google Scholar 

  • Vega R, Vásquez N, Espinoza AM, Gatica AM, Valdez-Melara M (2009) Histology of somatic embryogenesis in rice (Oryza sativa cv. 5272). Rev Biol Trop 57:141–150

    Google Scholar 

  • Vennapusa AR, Vemanna RS, Reddy BHR, Babitha KC, Kiranmai K, Nereshkumar A, Sudhakar C (2015) An efficient callus induction and regeneration protocol for drought tolerant rice indica genotype AC39020. J Plant Sci 3(5):248–254

    Google Scholar 

  • Visarada KBRS, Sailaja M, Sarma NP (2002) Effect of callus induction media on morphology of embryogenic calli in rice genotypes. Biol Plant 45:495–502

    Article  CAS  Google Scholar 

  • Wani SH, Sanger GS, Gosal SS (2011) An efficient and reproducible method for regeneration of whole plants from mature seeds of a high yielding Indica rice (Oryza sativa L.) variety PAU 201. New Biotechnol 28(4):418–422

    Article  CAS  Google Scholar 

  • Wanichananan P, Teerakathiti T, Roytrakal S, Kirdmanee C, Peyachoknagul SA (2010) Highly efficient method for agrobacterium mediated transformation in elite rice varieties (Oryza sativa L). Afr J Biotechnol 9(34):5488–5495

    CAS  Google Scholar 

Download references

Acknowledgments

We are thankful to Prof. Barbara Baldan, Department of Biology, University of Padua, Italy, for providing the microtome facility for histological analysis. We are also grateful to Dr. Tahir Abbas Shah, Department of Biosciences, CUI, Islamabad, Pakistan for assistance in the statistical analysis of data.

Funding

This work was supported by Erasmus Mundus Mobility for Asia (2014) grant no. ID06884 awarded to Ms Bushra Ijaz.

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Authors and Affiliations

  1. Department of Biosciences, COMSATS University Islamabad, Islamabad, Pakistan

    Bushra Ijaz, Muhammad Zeeshan Hyder, Sumaira Farrakh & Tayyaba Yasmin

  2. Department of Biology, University of Padua, Padua, Italy

    Cristina Sudiro & Fiorella Lo Schiavo

  3. Department of Plant Breeding and Genetics, PMAS Arid Agriculture University, Rawalpindi, Pakistan

    Saad Imran Malik

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  1. Bushra Ijaz
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  2. Cristina Sudiro
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  3. Muhammad Zeeshan Hyder
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Correspondence to Tayyaba Yasmin.

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Ijaz, B., Sudiro, C., Hyder, M.Z. et al. Histo-morphological analysis of rice callus cultures reveals differential regeneration response with varying media combinations. In Vitro Cell.Dev.Biol.-Plant 55, 569–580 (2019). https://doi.org/10.1007/s11627-019-09974-6

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  • DOI: https://doi.org/10.1007/s11627-019-09974-6

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