Micro-morpho-anatomical changes in leaf structure of plantlets during in vitro propagation (micropropagation) of Gardenia jasminoides J. Ellis


Research Articles | Published:

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Website:www.vegetosindia.org
Pub Email: contact@vegetosindia.org
Doi: 10.1007/s42535-023-00577-6
First Page: 107
Last Page: 116
Views: 1419


Keywords: Acclimatization, Cape jasmine, Cytokinin, In vitro, Light microscopy, Mesophyll


Abstract


The present study aimed to determine the foliar micro-morpho-anatomical features of in vitro cultured Gardenia jasminoides J. Ellis (Rubiaceae) to compare the effect of exogenous supplementation of growth regulators (cytokinins; 6-benzylaminopurine and Kinetin), in order to attenuate heterotrophic nutrition (in vitro) induced structural disorders in the proliferating shoots. Murashige and Skoog’s (MS) medium supplemented with 2.0 mg L−1 6-benzylaminopurine (BAP) in combination with 0.15 mg L−1 indole-3-acetic acid (IAA) was detected optimal for axillary bud induction. Nutrient medium containing 0.5 mg L−1 BAP and 0.25 mg L−1 IAA was found appropriate combination for proliferation of multiple shoots, and yielded 37.2 shoots (per explant) with 7.45 cm average length after 2nd subculture (8 weeks). Supplementation of NAA with cytokinins resulted in callus formation. The proliferation of shoots on kinetin (Kn) and IAA combination resulted in the formation of fragile shoots and the leaves with increased structural impairments like underdeveloped photosynthetic, vascular, and ground tissue systems, non-functional stomata, and reduced vein density. Comparatively, BAP and IAA treatment favoured healthy shoot proliferation and development of stable tissue systems with reduced structural abnormalities. Half strength MS medium augmented with 3.0 mg L−1 indole-3-butytric acid (IBA) was the optimal medium for root induction (32.0 roots per shoot with 3.5 cm in length). The regenerated plantlets showed 97% survival success during acclimatization and exhibited normal growth characteristics and morphology. The light microscopic evaluation of foliages provided technical support to in vitro regeneration techniques to understand the structural adaptational mechanism of in vitro raised plantlets, thereby substantially contributed in the reduction of the rate of mortality of regenerated plants of G. jasminoides.


Acclimatization, Cape jasmine, Cytokinin, In vitro, Light microscopy, Mesophyll


*Get Access

(*Only SPR Members can get full access. Click Here to Apply and get access)

Advertisement

References


Ahmed ZS, Salim AM, Allawi AK (2021) Effect of growth regulators (BA, IBA) on micropropagation of Gardenia jasminoides Ellis. in vitro. Int J Agric Stat Sci 17:181–186


Al-Juboory KH, Skirvin RM, Williams DJ (1998) Callus induction and adventitious shoot regeneration of gardenia (Gardenia jasminoides Ellis) leaf explants. Sci Hortic 72:171–178


Amer EM, Fetouh MI, Rasha E-S, El-Laban HM (2019) Micropropagation and acclimatization of Gardenia jasminoides Ellis. J Biol Chem Envion Sci 14:107–120


Aremu AO, Plačková L, Bairu W, Novák O, Plíhalová L, Doležal K, Finnie JF, Van Staden J (2014) How does exogenously applied cytokinin type affect growth and endogenous cytokinins in micropropagated Merwilla plumbea? Plant Cell Tissue Organ Cult 118:245–256


Auer CA, Laloue M, Cohen JD et al (1992) Uptake and metabolism of benzyladenine during shoot organogenesis in Petunia leaf explants. Plant Growth Regul 11:105–114


Barupal M, Kataria V, Shekhawat NS (2018) In vitro growth profile and comparative leaf anatomy of the C3–C4 intermediate plant Mollugo nudicaulis Lam. In Vitro Cell Dev Biol Plant 54:689–700


Bidabadi SS, Jain SM (2020) Cellular, molecular and physiological aspects of in vitro plant regeneration. Plants 9:702


Black MK, Osborne DJ (1965) Polarity of transport of benzyladenine, adenine and indole-3-acetic acid in petiole segments of Phaseolus vulgaris. Plant Physiol 40:676–680


Chen Q, Youn UJ, Min B, Bae KH (2008) Pyronane monoterpenoids from the fruit of Gardenia jasminoides. J Nat Prod 71(6):995–999


Chen QC, Zhang WY, Youn U, Kim H, Lee I, Jung HJ, Na M, Min B, Bae K (2009) Iridoid glycosides from Gardeniae fructus for treatment of ankle sprain. Phytochemistry 70(6):779–784


Chen L, Li M, Yang Z, Tao W, Tian P, Li X, Wang W (2020) Gardenia jasminoides Ellis: ethnopharmacology, phytochemistry, and pharmacological and industrial applications of an important traditional Chinese medicine. J Ethnopharmacol. https://doi.org/10.1016/j.jep.2020.112829


Chen B, Zhang J, Wu J, Li J, Fan H (2021) Optimizing the propagation techniques for Gardenia jasminoides Ellis. Pak J Bot. https://doi.org/10.30848/PJB2023-2(29)


Chirinea CF, Pasqual M, de Araujo AG, Pereira AR, de Castro EM (2012) Acclimatization and leaf anatomy of micropropagated fig plantlets. Rev Bras Fruticult 34:1180–1188


Chuenboonngarm N, Charoonsote S, Bhamarapravi S (2001) Effect of BA and 2iP on shoot proliferation and somaclonal variation of Gardenia jasminoides Ellis in vitro culture. Sci Asta 27:137–214


Cui Y, Deng Y, Zheng K, Hu X, Zhu M, Deng X, Xi R (2019) An efficient micropropagation protocol for an endangered ornamental tree species (Magnolia sirindhorniae Noot. & Chalermglin) and assessment of genetic uniformity through DNA markers. Sci Rep 9:9634


Duhoky MMS, Rasheed KA (2009) Micropropagation of Gardenia Gardenia jasminoides by using single nodes. Mesopotamia J Agric 37:1–12


Dumanois C, Godin B, Bigot C (1984) Multiplication végétative in vitro de Gardenia jasminoïdes Ellis: in vitro vegetative multiplication of Gardenia jasminoïdes Ellis. J Plant Physiol 116(5):389–407. https://doi.org/10.1016/S0176-1617(84)80131-5


Eburneo L, Ribeiro-Júnior NG, Karsburg IV, Rossi AAB, Silva IV (2017) Anatomy and micromorphometric analysis of leaf Catasetum × apolloi Benelli & Grill with addition of potassium silicate under different light sources. Braz J Biol 77:140–149


Economou AS, Spanoudaki MJ (1985) In vitro propagation of gardenia. Gortscience 20(2):213


El-Ashry AAE, Gabr AMM, Girgis ND, El-Bahr MK (2018) Influence of silver nitrate on enhancing in vitro rooting of Gardenia jasminoides Ellis. J Environ Sci Technol 11(5):238–245


Frick EM, Strader LC (2018) Roles for IBA-derived auxin in plant development. J Exp Bot 69(2):169–177


Gaber MK, Barakat AA (2019) Micropropagation and somatic embryogenesis induction of Gardenia jasminoides plants. Alex Sci Exch J 40:190–201


Glocke P, Collins G, Sedgley M (2006) 6-Benzylamino purine stimulates in vitro shoot organogenesis in Eucalyptus erythronema, E. stricklandii and their interspecific hybrids. Sci Hortic 109:339–344


Hong IK, Jeon H, Lee SB (2015) Extraction of natural dye from Gardenia and chromaticity analysis according to chi parameter. J Ind Eng Chem 24:326–332


Isah T (2015) Adjustments to in vitro culture conditions 367 and associated anomalies in plants. Acta Biol Crac Ser Bot 57(2):9–28


Iyer-Pascuzzi AS, Benfey PN (2009) Transcriptional networks in root cell fate specification. Biochim Biophys Acta 1789:315–325


Jogam P, Sandhya D, Shekhawat MS, Alok A, Manokari M, Abbagani S, Allini VR (2020) Genetic stability analysis using DNA barcoding and molecular markers and foliar micro-morphological analysis of in vitro regenerated and in vivo grown plants of Artemisia vulgaris L. Ind Crops Prod 151:112476


Johansen DA (1940) Plant microtechnique, 1st edn. McGraw Hill Book Co., New York, pp 182–197


Kadhim ZK, Nayyef MN, Awadh HAA, Jaafar HM, Abdulhussein MAA (2020) Impact of plant growth regulators and adenine sulphate on Gardenia jasminoides micropropagation. Plant Arch 20:71–75


Kannan N, Manokari M, Shekhawat MS (2021) Induction of adventitious roots from leaf explants of Morinda coreia Buch. and ham.: an important dye yielding plant. Plant Cell Tissue Organ Cult. https://doi.org/10.1007/s11240-021-02016-3


Keswick M (2003) The Chinese garden, 2nd edn. Frances Lincoln, London, p 63


Khare C (2007) Gardenia jasminoides Ellis. In: Khare C (ed) Indian medicinal plants. Springer, New York. https://doi.org/10.1007/978-0-387-70638-2_671


Kobayashi KD, Kaufman AJ (2006) Common Gardenia. College of Tropical Agriculture and Human Resources (CTAHR). University of Hawaii at Manoa, Honolulu, pp 1–7


Koo HJ, Lim KH, Jung HJ, Park EH (2006) Anti-inflammatory evaluation of Gardenia extract geniposide and genipin. J Ethnopharmacol 103(3):496–500


Kwak SC, Lee C, Kim JY, Oh HM, So HS, Lee MS, Rho MC, Oh J (2013) Chlorogenic acid inhibits osteoclast differentiation and bone resorption by down-regulation of receptor activator of nuclear factor kappa-B ligand-induced nuclear factor of activated T cells c1 expression. Biol Pharm Bull 36(11):1779–1786


Li HB, Yu Y, Wang ZZ, Dai Y, Gao H, Xiao W et al (2013) Iridoid and bis-iridoid gluco sides from the fruit of Gardenia jasminoides. Fitoterapia 88:7–11


Luo YJ, Zuo YM, Zhang ZL, Cai MT, Luo GM (2014) Study on chemical constituents of Gardenia jasminoides (III). J Chin Med Mat 37(7):1196–1199


Manokari M, Cokul Raj M, Priyadharshini S, Phulwaria M, Shekhawat MS (2020) Foliar micro-morphology—a promising tool to improve survival percentage of tissue culture raised plantlets with special reference to in vitro propagation of Vitex negundo L. Vegetos 33:504–515


Manokari M, Priyadharshini S, Shekhawat MS (2021a) Micro-structural stability of micropropagated Vitex negundo L. Microsc Microanal. https://doi.org/10.1017/S1431927621000283


Manokari M, Priyadharshini S, Shekhawat MS (2021b) Influence of physio-chemical factors on high throughput plant regeneration and micro-morpho-anatomy of shoots of Ormocarpum sennoides (Willd.) DC. Acta Physiol Plant. https://doi.org/10.1007/s11738-020-03180-3


Martins JPR, Schimildt ER, Alexandre RS, Castro EM, Nani TF, Pires MF, Pasqual M (2014) Direct organogenesis and leaf anatomy modifications in vitro of Neoregelia concentric (Vellozo) L.B. Smith (Bromeliaceae). Pak J Bot 46:2179–2187


Martins JPR, Santos ER, Rodrigues LCA, Gontijo ABPL, Falqueto AR (2018) Effects of 6-benzylaminopurine on photosystem II functionality and leaf anatomy of in vitro cultivated Aechmea blanchetiana. Biol Plant 62:793–800


Mizukami H (1989) Gardenia jasminoides Ellis: in vitro propagation and the formation of iridoid glucosides. In: Bajaj YPS (ed) Medicinal and aromatic plants II. Biotechnology in agriculture and forestry, vol 7. Springer, Berlin. https://doi.org/10.1007/978-3-642-73617-9_12


Moras B, Loffredo L, Rey S (2018) Quality assessment of saffron (Crocus sativus L.) extracts via UHPLC-DAD-MS analysis and detection of adulteration using gardenia fruit extract (Gardenia jasminoides Ellis). Food Chem 257:325–332


Mousa GT, Abdul-Hafeez EY, Ibrahim OHM (2016) Response of Gardenia plants grown under various growth media and ferrous sulfate application. Pak J Agric Sci 52:651–658


Moyo M, Aremu AO, Van Staden J (2015) Insights into the multifaceted application of microscopic techniques in plant tissue culture systems. Planta 242(4):773–790


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


Nower AA (2013) Hamza EM (2013) Production of Gardenia jasminoides scions via tissue culture for grafting on Gardenia thunbergia under greenhouse conditions. J Appl Sci Res 9(4):3118–3128


Pereira MP, Rodrigues LCA, Corrêa FF, Castro EM, Ribeiro VE, Pereira FJ (2016) Cadmium tolerance in Schinus molle trees is modulated by enhanced leaf anatomy and photosynthesis. Trees 30:807–814


Phatak RS (2015) Phytochemistry, pharmacological activities and intellectual property landscape of Gardenia jasminoides Ellis: a review. Pharmacogn J 7:254–265


Pontikis CA (1983) In vitro propagation of Gardenia jasminoides. Plant Propag 29:13–14


Qin FM, Liu BL, Zhang Y, Zhou GX (2015) A new triterpenoid from the fruits of Gardenia jasminoides var. radicans Makino. Nat Prod Res 29(7):633–637


Rajput BS, Jani M, Ramesh K, Manokari M, Jogam P, Allini VR, Kher MM, Shekhawat MS (2020) Large-scale propagation of Bambusa balcooa Roxb.: an industrially important bamboo species. Ind Crops Prod. https://doi.org/10.1016/j.indcrop.2020.112905


Rashotte AM, Poupart J, Waddell CS, Muday GK (2003) Transport of the two natural auxins, indole-3-butyric acid and indole-3-acetic acid, in Arabidopsis. Plant Physiol 133:761–772


Revathi J, Manokari M, Shekhawat MS (2018) Optimization of factors affecting in vitro regeneration, flowering, ex vitro rooting and foliar micromorphological studies of Oldenlandia corymbosa L.—a multipotent herb. Plant Cell Tissue Organ Cult 134(1):1–13


Salim SAA, Hamza SY (2017) An efficient protocol for micropropagation of Gardenia jasminoides Ellis. Biosci Biotechnol Res Asia 14:757–766


Scaramuzzi F, D’Elia C (1984) Clonal multiplication of Gardenia grandiflora Lour. C R Acad Sci Paris 299:657–662


Serret M, Trillas M (2000) Effects of light and sucrose levels on the anatomy, ultrastructure, and photosynthesis of Gardenia jasminoides Ellis leaflets cultured in vitro. Int J Plant Sci 161(2):281–289


Serret M, Trillas M, Araus J (2001) The effect of in vitro culture conditions on the pattern of photoinhibition during acclimation of Gardenia plantlets to ex vitro conditions. Photosynthetica 39:67–73


Shekhawat MS, Manokari M (2017) Comparative foliar micromorphological studies of in vitro and field transferred plants of Morinda citrifolia L. Acta Bot Hung 59(3–4):427–438


Shekhawat MS, Manokari M (2018) Micromorpho-anatomical evaluation of in vitro and field transferred plants of Coccinia indica Wight & Arn. Agric Res 7(2):135–144


Shekhawat MS, Kannan N, Manokari M (2015a) In vitro propagation of traditional medicinal and dye yielding plant Morinda coreia Buch.-Ham. S Afr J Bot 100:43–50


Shekhawat MS, Kannan N, Manokari M, Ravindran CP (2015b) Enhanced micropropagation protocol of Morinda citrifolia L. through nodal explants. J Appl Res Med Arom Plants 2(4):174–181


Sung YY, Lee AY, Kim HK (2014) The Gardenia 455 jasminoides extract and its constituent, geniposide, elicit anti-allergic effects on atopic dermatitis by inhibiting histamine in vitro and in vivo. J Ethnopharmacol 156:33–40


Wafa SN, Taha RM, Mohajer S, Mahmad N, Abdul BAA (2016) Organogenesis and ultrastructural features of in vitro grown Canna indica L. BioMed Res Int. https://doi.org/10.1155/2016/2820454


Wang J, Lu J, Lv C, Xu T, Jia L (2012) Three new triterpenoid saponins from root of Gardenia jasminoides Ellis. Fitoterapia 83(8):1396–1401


Wu X, Zhou Y, Yin F, Mao C, Li L, Cai B, Lu T (2014) Quality control and producing areas differentiation of Gardeniae fructus for eight bioactive constituents by HPLCDAD465 ESI/MS. Phytomedicine 21(4):551–559


Zhou Y, Zhang J, Tang R, Zhang J (2015) Simultaneous dyeing and functionalization of silk with three natural yellow dyes. Ind Crops Prod 64:224–232




 


Acknowledgements


The work was financially supported by the Researchers Supporting Project (RSP-2023R86), King Saud University, Riyadh, Saudi Arabia.


Author Information


Manokari M.
Biotechnology Unit, Kanchi Mamunivar Government Institute for Postgraduate Studies and Research, Puducherry, India