Micropropagation of endemic Corynandra chelidonii var. pallae (Cleomaceae) through nodal explants and validation of their genetic integrity by ISSR markers

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Research Articles | Published:

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Pub Email: contact@vegetosindia.org
Doi: 10.1007/s42535-021-00302-1
First Page: 511
Last Page: 519
Views: 511

Keywords: Corynandra chelidonii var. pallae , Genetic fidelity analysis, ISSR, Organogenesis, Plant regeneration


Micropropagation through nodal explants was attempted for a rare and endemic taxon Corynandra chelidonii var. pallae (Cleomaceae) in view of its medicinal uses, bioactive compounds, declining natural populations due to intermittent sterility, low seed set and erratic seed germination. Multiple shoots were regenerated directly from the nodal explants on Murashige and Skoog’s medium containing different concentrations and combinations of cytokinins and auxins. High-frequency multiple shoots of 8.15 ± 0.249 with 1.63 ± 0.031 shoot length (cm) were obtained on MS medium augmented with the combination of 0.5 mg/L of thidiazuron and 1.0 mg/L of indole-3-acetic acid. Directly regenerated shoots were rooted with 95% root induction on a half-strength MS medium supplemented with 1.0 mg/L of indole-3 butyric acid. The in vitro raised plantlets were hardened in plastic pots and acclimatized initially in the greenhouse conditions and later transferred to the field conditions with a 78% of survival rate. The genetic fidelity of in vitro generated and field transferred plantlets with that of the mother plant was checked by carrying ISSR marker-based polymerase chain reaction method. This protocol on direct organogenesis and genetic fidelity analysis in Corynadra chellidonii var. pallae can be successfully employed for large-scale multiplication and conservation.

                        Corynandra chelidonii var. pallae
                     , Genetic fidelity analysis, ISSR, Organogenesis, Plant regeneration

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Albarello N, Ribeiro IG, Simoes C, de Castro TC, Gianfaldoni MG, Callado CH, Kuster RM, Coelho MGP, Mansur EA (2008) Histological analysis of calluses from in vitro propagated plants of Cleome spinosa Jacq. Revista Brasileira de Biociências 5(S2):699–701

Anburaj J, Singh CR, Sundarraj S, Kannan S (2011) In vitro regeneration of Cleome viscosa—an important medicinal herb. J Cell Mol Biol 6(1):1461–1464

Barakat HH, El-Mousallamy AMD, Souleman AM, Awadalla S (1991) Flavonoids of Ochradenus baccatus. Phytochemistry 30:3777–3779

Begum MM, Kiran KR (2016) Evaluation of methanolic extract of Cleome chelidonii for hepatoprotective activity against paracetamol and ethanol induced hepatotoxicity in rats. Int J Pharma Sci Rev Res 5(1):28–36

Chi VV, Hop T (2002) Cˆay cỏ c´o ´ıch [Vietnam useful plants]. Vietnam Education Publishing House, Hanoi

Chinnappan RS (2020) In vitro conservation of Cleome felina L. J Med Plants 8(4):048–055. https://doi.org/10.15413/ajmp.2020.0107

Devarumath RM, Doule RB, Kawar PG, Naikebawane SB, Nerkar YS (2007) Field performance and RAPD analysis to evaluate genetic fidelity of tissue culture raised plants vis-à-vis conventional sets derived plants of Sugarcane. Sugar Technol 9(1):17–22

Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15

El-Sayed MM, Mahmoud MAA, El-Nahas HAK, El-Toumy SAH, El-Wakil EA, Ghareeb MA (2010) Bio-guided isolation and structure elucidation of antioxidant compounds from the leaves of Ficus sycomorus. Pharmacology 3:317–332

Ethadi SR, Pragada RR, Battu GR (2013) Evolution of anti-inflammatory and hepatoprotective activities of different extracts of Cleome chelidonii root in albino rats. Int J Pharma Biosci 4:111–119

Faisal M, Alatar AA, El-Sheikh MA, Abdel-Salam EM, Qahtan AA (2018) Thidiazuron induced in vitro morphogenesis for sustainable supply of genetically true quality plantlets of Brahmi. Ind Crops Prod 118:173–179

Hassan HMS (2014) Developing an efficient protocol for micropropagation of an endangered medicinal plant, Cleome droserifolia. Middle East J 3(4):1163–1168

Hiregoudar LV, Hosakatte N, Hema BP, Hahn EJ, Paek K (2003) Multiple shoot induction and plant regeneration of Feronia limonia (L.) Swingle. Sci Hortic 98:357–364

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

Juarez-Vazquez MDC, Jimenez-Arellanes MA, Jimenez-Arellanes MA (2019) Phytochemical investigation, anti-inflammatory and antinociceptive activities from some species of Cleomaceae family: a systematic review. Adv Med Plant Res 7(4):107–128

Kudikala H, Jogam P, Sirikonda A, Mood K, Allini VR (2020) In vitro micropropagation and genetic fidelity studies using SCoT and ISSR primers in Annona reticulata L.: an important medicinal plant. Vegetos 33(3):446–457

Larkin PJ, Scowcroft WR (1981) Somaclonal variation a novel source of variability from cell cultures for plant improvement. Theor Appl Gen 60:197–214

Ludwig-Muller J (2000) Indole-3-butyric acid in plant growth and development. Plant Growth Regul 32:219–230

Minh PN, Tri MD, Phat NT, Dat BT, Hanh NN, Luan NQ, Thanh MT, Huynh CH (2015) Two new flavonol glycosides from the leaves of Cleome chelidonii L.f. J Asian Nat Prod Res 17(4):338–342

Mukherjee AK, Shibani R, Sujaya D, Akhil KD, Pradosh KA, Sudhamoy M, Pratap CP, Ajay KM (2010) Genetic relationships among 22 taxa of bamboo revealed by ISSR and EST-based random primers. Biochem Genet 48:1015–1025

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

Naseem M, Jha KK (1997) Rapid clonal multiplication of Cleome gynandra DC. through tissue culture. Phytomorphology 47(4):405–411

Nguyen PD, Sayagh C, Borie N, Lavaud C (2017) Antiradical flavonol glycosides from the aerial parts of Cleome chelidonii L.f. Phytochemistry 142:30–37

Parimalakrishnan S, Dey A, Smith AA, Manavalan R (2007) Evaluation of antiinflammatory, antinociceptive and antipyretic effects of methanol extract of Cleome chelidonii. Int J Biol Chem Sci 1(3):223–228

Pendli S, Rohela GK, Jogam P, Prasad B, Korra R, Thammidala C (2019) High frequency in vitro plantlet regeneration in Solanum trilobatum L., an important ethno-medicinal plant and confirmation of genetic fidelity of R1 plantlets by using ISSR and RAPD markers. Vegetos 32:508–520. https://doi.org/10.1007/s42535-019-00069-6

Phan NM, Mai DT, Nguyen TP, Bui TD, Nguyen NH, Ngo QL, Ma Thi TT, Chung HH (2015) Two new flavonol glycosides from the leaves of Cleome chelidonii L.f. J Asian Nat Prod Res 17:338e342

Phan NM, Hong TDT, Le Thanh TN, Trong TN, Quoc LN, Trong DT, Quan HN, Bui LCH, Diep XKN, Trong DB, Dinh TM, Tan PN (2021) Hepatoprotection and phytochemistry of the Vietnamese herbs Cleome chelidonii and Cleome viscosa stems. J Chem. https://doi.org/10.1155/2021/5578667

Quang NT, Diet TD, Duong VB, Hieu NT, Hien CM (2011) Preliminary study on chemical composition of Cleome chelidonii L.f., Capparaceae. J Milit Pharm Med 3:40–45

Rathore NS, Rathore N, Shekhawat NS (2013) In vitro propagation and micro morphological studies of Cleome gynandra: A C4 model plant closely related to Arabidopsis thaliana. Acta Physiol Plant 35(9):2691–2698

Reddy CS, Raju VS (2001) A new variety of Cleome chelidonii L.f. (Cleomaceae). J Econ Taxon Bot 25(1):217–219

Rodriguez R, Rey M, Cuozzo L, Ancora G (1990) In vitro propagation of caper (Capparis spinosa L.). Vitro Cell Dev Biol 26(5):531–536

Rohela GK, Jogam P, Shabnam AA, Shukla P, Abbagani S, Ghosh MK (2018) In vitro regeneration and assessment of genetic fdelity of acclimated plantlets by using ISSR markers in PPR-1 (Morus sp.): an economically important plant. Sci Hortic 241:313–321

Rohela GK, Jogam P, Mir MY, Shabnam AA, Shukla P, Abbagani S, Kamili AN (2020) Indirect regeneration and genetic fidelity analysis of acclimated plantlets through SCoT and ISSR markers in Morus alba L. cv. Chinese white. Biotechnol Rep 25:e00417

Savitikadi P, Jogam P, Rohela GK, Ellendula R, Dulam S, Rao VA, Abbagani S (2020) Direct regeneration and genetic fidelity analysis of regenerated plants of Andrographis echioides (L.)—an important medicinal plant. Ind Crops Prod 155:112766

Sahoo Y, Pattnaik SK, Chand PK (1997) In vitro clonal propagation of an aromatic medicinal herb Ocimum basilicum (L.) (Sweet Basil) by axillary shoot proliferation. Vitro Cell Dev Biol 33:293–296

Sandhya D, Jogam P, Manokari M, Shekhawat MS, Jadaun JS, Allini VR, Abbagani S (2021) High-frequency in vitro propagation and assessment of genetic uniformity and micro-morphological characterization of Origanum majorana L.—a highly traded aromatic herb. Biocatal Agric Biotechnol 34:102024

Sheeba H, Syed Ali M, Anuradha V (2019) Bioactive compounds and antimicrobial activity of fungal crude extract from medicinal plants. J Pharma Sci Res 11(5):1826–1833

Simoes C, Santos A, Albarello N, Figueiredo S (2004) Shoot organogenesis and plantlet regeneration from stem explants of Cleome rosea Vahl (Capparaceae). J Plant Biotechnol 6:199–204

Sirangi S, Jogam P, Nemali G, Ragan A, Abbagani S, Raju VS (2020) Intraspecific genetic variation in Corynandra chelidonii (Angiosperms: Cleomaceae) as revealed by SCoT, ISSR and RAPD analyses. J Plant Biotechnol 47:289–297

Songsak T, Lockwood GB (2002) Glucosinolates of seven medicinal plants from Thailand. Fitoterapia 73(3):209–216

Songsak T, Lockwood GB (2004) Production of two volatile glucosinolate hydrolysis compounds in Nasturtium montanum and Cleome chelidonii plant cell cultures. Fitoterapia 75:296–301

Sridhar N, Kiran BVS, Sasidhar DT, Kanthal LK (2014) In vitro antimicrobial screening of methanolic extracts of Cleome chelidonii and Cleome gynandra. Bangladesh J Pharmacol 9(2):161–166

Sujatha D, Ravi C, Raghuvardhan L, Prasad B, Gulab KR, Sadanandam A, Christopher RT (2013) In vitro plantlet regeneration and genetic transformation of sponge gourd (Luffa cylindrica L.). Afr J Plant Sci 7(6):244–252

Sumitha V, Gurulakshmi M (2015) Antioxidant and free radical scavenging activity of leaf extracts of Cleome chelidonii. Int J Innov Pharm 2(3):228–236

Supriya D, Manabendra DC, Pranab BMa (2013) Micropropagation of Dioscorea alata L. through nodal segments. Afr J Biotechnol 12(47):6611–6617

Trilochana Y, Babu DJM, Rao PR (2017) The study of antihyperglycaemic activity of aqueous extract of root of Cleome chelidonii herb in rats. Indian J Res Pharm Biotechnol 5(2):88–93

Vemula S, Koppula T, Jogam P, Mohammed M (2020) In vitro high frequency multiplication and assessment of genetic fidelity of Corallocarpus epigaeus: an endangered medicinal plant. Vegetos 33(1):63–73

Rekha NV, Godasu SK, Jyothi GSVD, Hemanth K (2019) Phytochemical and pharmacological activities of Polygala chainensis, Cleome chelidonii. Int J Pharmacogn 6(7):253–258



SS is grateful to the University Grants Commission, New Delhi, for the award of the BSR-RFSMS fellowship. The authors are thankful to Prof. Sadanandam Abbagani, Department of Biotechnology, Kakatiya University, for his valuable suggestions and encouragement. The authors thank the Head of the Departments of Botany and Biotechnology, Kakatiya University, Warangal, India, for providing the facilities.

Author Information

Sirangi Subhash
Plant Systematics Laboratory, Department of Botany, Kakatiya University, Warangal, India

Jogam Phanikanth
Department of Biotechnology, Kakatiya University, Warangal, India

Rohela Gulab Khan
Biotechnology Section, Central Sericultural Research and Training Institute, Central Silk Board, Jammu and Kashmir, India

Ragan Ajmeera
Department of Biotechnology, Kakatiya University, Warangal, India

Raju Vatsavaya S.
Plant Systematics Laboratory, Department of Botany, Kakatiya University, Warangal, India