Abstract
An efficient indirect in vitro plant regeneration protocol for Dianthus caryophyllus L. cv. ‘Master’ was developed using leaf explants. This study revealed the morphogenetic potential of leaf explant as a source for micropropagation. Murashige and Skoog (MS) medium supplemented with 2.0 mg/l 2,4-dichlorophenoxyacetic acid and 0.5 mg/l Naphthalene acetic acid (NAA) resulted in maximum (94.44%) callus induction. MS medium supplemented with 1.5 mg/l thidiazuron, 0.25 mg/l Kinetin and 0.25 mg/l NAA was found to be highest for average shoot regeneration (80.56%), average number of shoots (6.01) and average shoot length (1.93 cm). For in vitro multiplication of shoots, MS medium supplemented with 2.0 mg/l Kinetin and 0.25 mg/l NAA was found to be the best which resulted in 14.64 average number of microshoots. It was observed that the half strength MS basal medium supplemented with 1.5 mg/l Indole-3-butyric acid and 0.02% activated charcoal showed maximum rooting (98.19%) with 9.60 average number of roots per microshoot having 4.24 cm root length. The in vitro rooted plantlets were hardened gradually and were successfully acclimatized under ex vitro conditions. The genetic variation in the in vitro raised plants was confirmed using DNA based markers [Random Amplified Polymorphic DNA and Inter Simple Sequence Repeats] for assessment of genetic stability of plants raised through indirect regeneration.
Access this article
We’re sorry, something doesn't seem to be working properly.
Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.



Similar content being viewed by others
References
Ali A, Afrasiab H, Naz S, Rauf M, Iqhbal J (2008) An efficient protocol for in vitro propagation of carnation (Dianthus caryophyllus L.). Pak J Bot 40:111–121
Kanwar JK, Kumar S (2009) Influence of growth regulators and explants on shoot regeneration in carnation. Hortic Sci 36:140–146
Kharrazi M, Nemati H, Tehranifar A, Bagheri A, Sharifi A (2011) In vitro culture of carnation (Dianthus caryophyllus L.) focusing on the problem of vitrification. J Biol Environ Sci 5(13):1–6
Thakur M, Sharma DR, Sharma SK (2002) In vitro selection and regeneration of carnation (Dianthus caryophyllus L.) plants resistant to culture filtrate of Fusarium oxysporum f. sp. dianthi. Plant Cell Rep 20:825–828
Ahmad N, Srivastava R, Anis M (2006) Improvement in carnation shoot multiplication using thidiazuron. Propag Ornam Plants 6:109–113
Karami O (2008) Induction of embryogenic callus and plant regeneration in carnation (Dianthus caryophyllus L.). J Biol Sci 8(4):68–72
Sayeed Hassan AKM, Munshi JL, Sultana R, Jahan MAA, Khatun R (2011) High frequency in vitro regeneration of Dianthus caryophyllus L., a herbaceous perennial ornamental plant. Bangladesh J Sci Ind Res 46(4):495–498
Esmaiel NM, Doss AA, Barakat MN (2013) An assessment of in vitro culture and plant regeneration from leaf base explants in carnation (Dianthus caryophyllus L.). Food Agric Environ 11(1):1113–1117
Arif M, Rauf S, Din AU, Rauf M, Afrasiab H (2014) High frequency plant regeneration from leaf derived callus of Dianthus caryophyllus L. Am J Plant Sci 5:2454–2463
Ranade R, Kanwar K (2014) Examining synergistic effects of TDZ and TIBA on adventitious shoot induction in Dianthus caryophyllus L. leaf explants. Int J Agric Sci Res 4(2):17–26
Joshi P, Dhawan V (2007) Assessment of genetic fidelity of micropropagated Swertia chirayita plantlets by ISSR marker assay. Biol Plant 51(1):22–26
Gomez KA, Gomez AA (1984) Statistical procedures for agricultural research, 2nd edn. Willy, Hoboken, pp 8–17
Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15
Kumar A, Verma A, Singh SK, Raghava SPS, Kumar PA (2006) In vitro shoot regeneration from leaf segments of carnation (Dianthus caryophyllus L.) via indirect organogenesis. Plant Cell Biotechnol Mol Biol 7:65–68
Radojevic L, Dragosavac DC, Spiric J, Stevanovic B, Stevanovic V (2010) In vitro propagation of Dianthus ciliatus ssp. dalmaticus and D. giganteus ssp. croaticus (Caryophyllaceae) from stem segment cultures. Bot Serbica 34(2):153–161
Mizory L, Yaseen S, Hassan AM (2014) Effect of different explant and different concentrations of plant growth regulators on micropropagation of (Dianthus caryophyllus L.). J Agric Vet Sci 7(1):1–9
Halperin W (1986) Attainment and retention of morphogenetic capability in vitro. In: Vasil K (ed) Cell culture and somatic cell genetics of plants, vol 3. Academic Press, Orlendo, pp 3–47
Nehra NS, Stushnoff C, Kartha KK (1990) Regeneration of plant from immature leaf derived callus of strawberry (Fragaria x annanosa). Plant Sci 66:119–126
Sahoo Y, Pattnaik SK, Chand PK (1997) Plant regeneration from callus cultures of Morus indica L. derived from seedlings and mature plants. Sci Hortic 69:85–98
Salehi H (2006) Can a general shoot proliferation and rooting medium be used for a number of carnation cultivars. Afr J Biotechnol 5(1):25–30
Bora A, Singh S, Talukdar MC, Hazarika BN (2007) An efficient method for in vitro plant regeneration in carnation. Indian J Hortic 64(4):439–443
Bora A, Talukdar MC, Hazarika BN (2006) Direct organogenesis in Dianthus caryophyllus Linn. J Ornam Hortic 9(3):164–168
Kanwar K, Deshmukh AJ, Deepika R, Kaushal B (2007) Efficient in vitro rapid axillary and proliferation from mature Terminalia chebula Retz.: medicinal tree. Tree For Sci 1(1):92–94
Markovic M, Popovic M, Vilotic D (2013) Micropropagation of Dianthus deltoides L. through shoot tip and nodal cuttings culture. Arch Biol Sci 65(1):17–22
Uddin I, Rashid H, Khan N, Perveen F, Tai TH, Tanaka K (2007) Selection of promising salt tolerant rice mutants derived from cultivar ‘Drew’ and their antioxidant enzyme activity under salt stress. J Breed Genet 39(2):89–98
Kanwar K, Joseph J, Deepika R (2010) Comparison of in vitro regeneration pathways in Punica granatum L. Plant Cell Tissue and Organ Cult 100:199–207
Rahman M, Ahmed B, Islam R, Mandal A, Hossain M (2014) A biotechnological approach for the production of red gerbera (Gerbera Jamesonii Bolus). Nova J Med Biol Sci 2(1):1–6
Robert AV, Smith EF (1990) Preparation of in vitro chrysanthemum for transplantation to soil: protection of roots by cellulose plugs. Plant Cell Tissue Organ Cult 21:129–132
Martyr RF (1981) New developments in the use of graded horticultural perlite. Acta Horticu 126:143–146
Bhatia R, Singh KP, Sharma TR, Jhang T (2011) Evaluation of the genetic fidelity of in vitro propagated Gerbera (Gerbera jamesonii Bolus) using DNA based marker. Plant Cell Tissue and Organ Cult 104:131–135
Kaushal B, Kanwar K (2006) Assessment of genetic stability of micropropagated plants of Morus alba L. using RAPD markers. Physiol Mol Biol Plants 12(3):387–392
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
Acknowledegment
The authors would like to thank Dr. Y. S. Parmar University of Horticulture and Forestry, Solan, Himachal Pradesh, India for providing laboratory and field facilities.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
Both authors declare no conflict of interest.
Rights and permissions
About this article
Cite this article
Thakur, K., Kanwar, K. In Vitro Plant Regeneration by Organogenesis from Leaf Callus of Carnation, Dianthus caryophyllus L. cv. ‘Master’. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 88, 1147–1155 (2018). https://doi.org/10.1007/s40011-017-0851-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s40011-017-0851-2


