Effect of various chemical solutions on micropropagation efficiency, antioxidant activities and clonal fidelity analysis of in vitro developed plantlets in pomegranate (Punica granatum L.) using SSR markers

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Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
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Doi: 10.1007/s42535-024-00846-y
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Keywords: Antioxidant, Biochemical analysis genetic fidelity, Micropropagation, Pomegranate, SSR markers


Abstract


This study aimed to develop an effective in vitro protocol for micropropagation of pomegranate (Punica granatum L.) var. Bhagwa using nodal explants. To induce shoot growth and cultivating the explants, Murashige and Skoog’s media were supplemented with benzyl amino purine (BAP), sodium nitroprusside (SNP), an approach demonstrated and experimented for the first time on Pomegranate and kinetin (Kn) in various concentrations and combinations. In the MS medium fortified with 1.5 mg/l BAP, 1.5 mg/l SNP, and 1.5 mg/l Kn, maximum shoot length, shoot per explants, and regeneration % were noted. For root induction, uniform micro shoots were separated and then placed in the rooting media (Murashige and Skoog medium at half strength), supplemented with different ratios of NAA and SNP with 2 mg/L NAA and 0.3 mg/L SNP yielding the highest culture rooting, number of roots, and root length. The described methodology permits the development of multiple pomegranate plants that have been micro propagated. Superoxide dismutase, catalase, and guaiacol peroxidase activities revealed the antioxidant defence mechanism of micro propagated pomegranate micro shoots, which increased shoot development and the number of shoots per explant. Using simple sequence repeat (SSR) markers, the mother plants and proliferating plantlets were checked for genetic integrity; the results showed that all of the plants were the same. The results of this study could be used commercially in the future to produce high-quality pomegranate planting material.


Antioxidant, Biochemical analysis genetic fidelity, Micropropagation, Pomegranate, SSR markers


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References


Abadi ZZB, Abad KKA, Saravi AT (2020) Comparison of different culture media and hormonal concentrations for In-vitro propagation of pomegranate. Int J Fruit Sci 20:S1721–S1728


Al-Wasel AS (1998) In Vitro Clonal Propagation of AL-Belehi Pomegranate (Punica granatum L). vitro Cell Dev Biology Anim 34:1031


Anon (1982) The wealth of India: a Dictionary of Indian raw materials and Industrial products. Vol. VIII, CSIR, New Delhi, pp 317–324


Arun M, Naing AH, Jeon SM, Ai TN, Aye T, Kim CK (2017) Sodium nitroprusside stimulates growth and shoot regeneration in chrysanthemum. Horti Environ Biotech 58:78–84


Bachake SS, Jadhav VB, Deshpande PP, Tele AA, Banda MA, Adki VS (2019) Standardization of in vitro propagation protocol for pomegranate cv. Super Bhagwa J Pharmacognosy Phytochemistry 8(3):2548–2553


Beers F, Sizer IF (1952) A spectrophotometric method for measuring breakdown of hydrogen peroxide by catalase. J Biol Chem 195:133–140


Cramer GR, Urano K, Delrot S, Pezzotti M, Shinozaki K (2011) Effects of abiotic stress on plants: a systems biology perspective. BMC pl Biology 11(1):1–14


Cuce M, Muslu AS (2022) Sodium nitroprusside mediates attenuation of paraquat-mediated oxidative stress in Eruca sativa in vitro. Physiol Mol Biol Plants 28(1):289–299


Dar SA, Nawchoo IA, Tyub S, Kamili AN (2022) In vitro culture and biochemical and antioxidant potential of the critically endangered medicinal plant Atropa acuminata Royle ex Lindl of Kashmir Himalaya. In Vitro Cellular & Developmental Biology-Plant 58(4): 540-550s


De Laia ML, Gomes EA, Esbrisse EJ, De Araujo EF (2000) Random amplified polymorphic DNA (RAPD) analysis of genotypic identities in Eucalyptus clones. Silvae Genetica 49(6):239–243


Deepika R, Kanwar K (2010) In vitro regeneration of Punica granatum L. plants from different juvenile explants. J Fruit Ornam Plant Res 18(1):5–22


Devarumath R, Nandy S, Rani V, Marimuthu S, Muraleedharan N, Raina S (2002) RAPD, ISSR and RFLP fingerprints as useful markers to evaluate genetic integrity of micropropagated plants of three diploid and triploid elite tea clones representing Camellia sinensis (China type) and C. Assamica ssp. Assamica (Assam-India type). Plant Cell Rep 21:166–173


Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15


El-Agamy SZ, Mostafa RA, Shaabanc MM, El-Mahdy MT (2009) In vitro propagation of Manfalouty and Nab El-gamal pomegranate cultivars. Res J Agric Biol Sci 5:1169–1175


Giannopolitis CN, Ries SK (1977) Superoxide dismutases: I. occurrence in higher plants. Pl Physiol 59(2):309–314


Goto S, Thakur RC, Ishii K (1998) Determination of genetic stability in long-term micropropagated shoots of Pinus thunbergii parl using RAPD markers. Plant Cell Rep 18:193–197


Guranna P, Hosamani I, Sathyanarayana R, Hegde R, Hipparagi K (2018) Micropropagation in pomegranate (Punica granatum L.) cv.‘Bhagwa’through indirect organogenesis and assessment of genetic fidelity by RAPD marker. Biotechnol J Int 20:1–8


Hasnaoui N, Buonamici A, Sebastiani F, Mars M, Trifi M, Vendramin GG (2010) Development and characterization of SSR markers for pomegranate (Punica granatum L.) using an enriched library. Conserv Genet Res 2:283–285


Hazarika BN (2006) Morpho-physiological disorders in in vitro culture of plants. Sci Hort 108(2):105–120


Hesami M, Daneshvar MH, Yoosefzadeh-Najafabadi M (2019) An efficient in vitro shoot regeneration through direct organogenesis from seedling-derived petiole and leaf segments and acclimatization of Ficus religiosa. J Forestry Res 30:807–815


Hristozkova M, Geneva M, Stancheva I, Iliev I, Azcon-Aguilar C (2017) Symbiotic association between golden berry (Physalis peruviana) and arbuscular mycorrhizal fungi in heavy metal-contaminated soil. J Plant Prot Res 57(2):173–184


Jin W, Wang Y, Wang H (2014) Adventitious shoot regeneration from leaves of apple rootstock ‘Pingyitiancha’(Malus hupehensis var. Pinyiensis) and genetic fidelity of regenerated plantlets using SSR markers. Can J Plant Sci 94(8):1345–1354


Jin W, Zhang Q, Liu S, Wei Q, Cheng Z, Xue X, Yang T (2012) Genetic diversity of 41 apple rootstocks based on simple sequence repeat markers. J Am Soc Horti Sci 137(1):51–56


Joyce SM, Cassells AC, Mohan, Jain S (2003) Stress and aberrant phenotypes in vitro culture. Planr Cell Tissue Organ Cult 74:103–121


Jurenka J (2008) Therapeutic applications of pomegranate (Punica granatum L.): a review. Alternat Med Rev 13:128–144


Kaji BV, Ershadi A, Tohidfar M (2013) In vitro propagation of pomegranate (Punica granatum L.) Cv.‘Males Yazdi. Albanian J Agri Sci 12(1):43


Kwak JM, Nguyen V, Schroeder JI (2006) The role of reactive oxygen species in hormonal responses. Pl Physiol 141(2):323–329


Lau SE, Hamdan MF, Pua TL, Saidi NB, Tan BC (2021) Plant nitric oxide signaling under drought stress. Plants 10(2):360


Martin KP, Pachathundikandi SK, Zhang CL, Slater A, Madassery J (2006) Rapd analysis of a variant of banana (musa sp.) cv. Grande naine and its propagation via shoot tip culture. Vitro Cell Dev Biology-Plant 42(2):188–192


Martins M, Sarmento D, Oliveira MM (2004) Genetic stability of micropropagated almond plantlets, as assessed by RAPD and ISSR markers. Plant Cell Rep 23(7):492–496


Maurya RL, Kumar M, Sirohi U, Chaudhary V, Sharma VR, Datta SK, Yadav MK (2021) An effective micropropagation protocol and determination of the clonal fidelity of in vitro developed microshoots of carnation (Dianthus caryophyllus L.) using SSR markers. Nucleus 65(1):49–55


Molassiotis AN, Dimassi K, Diamantidis G, Therios I (2004) Changes in peroxidases and catalase activity during in vitro rooting. Biol Plant 48:1–5


Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497


Murkute AA, Patil S, Singh SK (2004) In vitro regeneration in pomegranate cv. Ganesh from mature trees. Indian J Hortic 61(3):206–208


Naik SK, Pattnaik S, Chand PK (1999) In vitro propagation of pomegranate (Punica granatum L. Cv. Ganesh) through axillary shoot proliferation from nodal segments of mature tree. Sci Hort 79(3–4):175–183


Naik SK, Chand PK (2011) Tissue culture-mediated biotechnological intervention in pomegranate: a review. Plant Cell Rep 30(5):707–721


Naik SK, Pattnaik S, Chand PK (2000) High frequency axillary shoot proliferation and plant regeneration from cotyledonary nodes of pomegranate (Punica granatum L). Sci Hort 85(4):261–270


Niki E (2011) Antioxidant capacity: which capacity and how to assess it. J Berry Res, (4): 169–176


Patil VM, Dhande GA, Thigale DM, Rajput JC (2011) Micropropagation of pomegranate (Punica granatum L.) Bhagava’cultivar from nodal explant. Afr J Biotechnol 10(79):18130–18136


Rani V, Parida A, Raina SN (1995) Random amplified polymorphic DNA (RAPD) markers for genetic analysis in micropropagated plants of Populus deltoides Marsh. Plant Cell Rep 14:459–462


Sichanova M, Geneva M, Petrova M, Miladinova-Georgieva K, Kirova E, Nedev T, Tsekova D, Iwanov I, Dochev K, Ivanova V, Trendafilova A (2022) Improvement of Stevia rebaudiana Bertoni in vitro propagation and steviol glycoside content using aminoacid silver nanofibers. Plants 11(19):2468





Singh P, Patel RM, Kadam S (2013a) In vitro mass multiplication of pomegranate from cotyledonary nodal explants cv. Ganesh Afr J Biotechnol 12(20):2863–2868


Singh SR, Dalal S, Singh R, Dhawan AK, Kalia RK (2013b) Evaluation of genetic fidelity of in vitro raised plants of Dendrocalamus asper (Schult. & Schult. F.) Backer ex K. Heyne using DNA-based markers. Acta Physiol Plant 35(2):419–430


da Silva JAT, Rana TS, Narzary D, Verma N, Meshram DT, Ranade SA (2013b) Pomegranate biology and biotechnology: a review. Sci Hort 160:85–107


Tian M, Gu Q, Zhu M (2003) The involvement of hydrogen peroxide and antioxidant enzymes in the process of shoot organogenesis of strawberry callus. Plant Sci 165:701–707


Tubić L, Savić J, Mitić N, Milojević J, Janošević D, Budimir S, Zdravković-Korać S (2016) Cytokinins differentially affect regeneration, plant growth and antioxidative enzymes activity in chive (Allium schoenoprasum L). Planr Cell Tissue Organ Cult 124:1–14


Urbanek H, Kuzniak-Gebarowska E, Herka K (1991) Elicitation of defense responses in bean leaves by Botrytis Cinerea polygalacturonase. Acta Physiol Plant 13:43–50


Wojtania A, Skrzypek E (2014) Effects of cytokinins on antioxidant enzymes in in vitro grown shoots of Pelargonium hortorum LH Bayley. Acta Agrobotanica 67(4)


Xu J, Yin H, Wang W, Mi Q, Liu X (2009) Effects of sodium nitroprusside on callus induction and shoot regeneration in micro-propagated Dioscorea opposita. Plant Growth Regul 59(3):279–285


Yassin MT, Mostafa AA-F, Al Askar AA (2021) In Vitro Evaluation of Biological Activities and Phytochemical Analysis of different solvent extracts of Punica granatum L. (pomegranate) peels. Plants 10(12):2742. https://doi.org/10.3390/plants10122742


Zayova E, Geneva M, Stancheva I, Dimitrova L, Petrova M, Hristozkova M, Salamon I (2018) Evaluation of the antioxidant potential of in vitro propagated hyssop (Hyssopus officinalis L.) with different plant growth regulators. Med Plants-International J Phytomedicines Relat Industries 10(4):295–304


Zayova EG, Geneva MP, Miladinova-Georgieva KD, Hristozkova MG, Stancheva IV (2019) Impact of plant growth regulators on Greek Oregano micropropagation and antioxidant activity. Biosciences, Biotechnology Research Asia 16: 297–305


Singh B, Singh JP, Kaur A, Singh N (2018). Phenolic compounds as beneficial phytochemicals in pomegranate (Punica granatum L.) peel: a review. Food Chem 261:75–86. https://doi.org/10.1016/j.foodchem.2018.04.039


Shelar SD, Kad VP, Kanawade VL, Patil AP, Unde PA (2017) Ready-to-serve beverage from spray dried pomegranate juice powder. J Krishi Vigyan 6(1):166–171. https://doi.org/10.5958/2349-4433.2017.00073.3


Kaur A, Kumar A (2020) The effect of gelling agent, medium pH and silver nitrate on adventitious shoot regeneration in Solanum tuberosum. bioRxiv. https://doi.org/10.1101/2020.01.03.894063


Kumar R, Meena R, Sharma D, Saroj P (2018) Production technology of pomegranate in arid region. CIAH/Tech./bull. No. 65, ICAR-central Institute for arid horticulture, Bikaner, Rajasthan, India


Debnath SC (2017). Molecular markers, clonal fidelity and antioxidant properties of micropropagated horticultural crops. Acta Horticulturae 1155:261–268. https://doi.org/10.17660/ActaHortic.2017.1155.38

 


Acknowledgements


Authors are thankful to Director Research, SVPUAT for providing all the necessary support for conducting the present study.


Author Information


Pal Devendra
College of Horticulture, Sardar Vallabhbhai Patel University of Agriculture & Technology, Meerut, India

Kumar Mukesh
College of Horticulture, Sardar Vallabhbhai Patel University of Agriculture & Technology, Meerut, India
dr.mukesh123@gmail.com

Yadav Manoj Kumar
College of Agriculture, Sardar Vallabhbhai Patel University of Agriculture & Technology, Meerut, India


Chauhan Chetan
College of Horticulture, Sardar Vallabhbhai Patel University of Agriculture & Technology, Meerut, India


Kumar Arvind
College of Horticulture, Sardar Vallabhbhai Patel University of Agriculture & Technology, Meerut, India

Sirohi Ujjwal
College of Agriculture, Sardar Vallabhbhai Patel University of Agriculture & Technology, Meerut, India

Sharma V. Rakesh
College of Horticulture, Sardar Vallabhbhai Patel University of Agriculture & Technology, Meerut, India

,
Chaudhary Veena
Department of Chemistry, Meerut College, Meerut, India