CoChat
Sign Up
Cited by 13Plant tissue culture and regenerationFlowering Plant Growth and CultivationGenetic diversity and population structure

In Vitro Regeneration and ISSR-Based Genetic Fidelity Analysis of Orthosiphon stamineus Benth

Open access
Ali, Hanisah, Musa, Izzah Farhanah, Abu Bakar, Nurul Atikhah, Karsani, Saiful Anuar, Yaacob, Jamilah Syafawati·2019·Agronomy
DOI 10.3390/agronomy9120778
Open this paper in CoChat — free accountOpen PDF

Abstract

Orthosiphon stamineus has been widely used as traditional remedy for various illnesses and diseases, such as cardiovascular diseases and epileptic seizures. In this study, direct regeneration through nodal segment of this species was attempted using Kinetin (6-Furfurylaminopurine) and IAA (indole-3-acetic acid). Optimum regeneration media was identified as MS media supplemented with 2.0 mg L−1 Kin plus 0.5 mg L−1 IAA. This yielded the highest number of shoots (5.57 ± 0.42) and leaves (20.53 ± 1.91) per explant. Acclimatization of the resulting in vitro regenerants was successful in all potting mixtures tested. However, potting mixture PF (1:1:1 ratio of black soil/red soil/compost) was identified as the best medium for acclimatization of this species, as it yielded 100% survival percentage after 90 days of acclimatization. Ten in vitro regenerants of O. stamineus were randomly collected after the third subculture and subjected to genetic variation analysis using inter-simple sequence repeat (ISSR) markers. Out of 20 ISSR markers tested, 10 working primers were observed to produce satisfactory amplification of bands, with an average of 7.11 bands per primer. A total of 610 bands were produced by the 10 primers. The percentage of polymorphism was observed to be very low, yielding only 7.32% polymorphism among all samples. Jaccard dissimilarity analysis was also conducted and very low genetic distance (about 0.1) was found among the in vitro regenerants and between the regenerants with the mother plant, thus ascertaining the clonal nature of the plantlets produced in this study.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.3390/agronomy9120778

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • ISSR markers, applied comparatively between plantlets and mother plant, generally revealed the clonal nature of analyzed regenerants, highlighting genetic fidelity of in vitro propagation, even after several years.

    “However, the ISSR markers, applied comparatively between the plantlets and the mother plant revealed, in general, the clonal nature of the analyzed regenerants, highlighting the genetic fidelity of in vitro propagation, even after several years [70,71,72].”

    From Somaclonal Variation—Advantage or Disadvantage in Micropropagation of the Medicinal Plants (2023) · cites this paper as [70]

    The Introduction says the study compared regenerants with the mother plant using ISSR markers, and the Discussion describes ISSR as a means of assessing clonal fidelity. The Methods (sections 2.2 and 2.4; pages unavailable) describe plantlets examined after the third subculture and regeneration observed for four weeks. This supports the comparison and fidelity assessment, but not the claim that fidelity persisted for several years. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

No citation lists for this paper

No citation lists for this paper yet — checked again daily.

References 67 · Cited by 13

Research works printed but not linked: 64

  1. Almatar, M.; Rahmat, Z.; Salleh, F.M. Preliminary morphological and anatomical study of Orthosiphon stamineus. Asian J. Pharm. Biol. Res. 2013, 1, 1-6. [CrossRef] · DOI
  2. Akowuah, G.; Ismail, Z.; Norhayati, I.; Sadikun, A. The effects of different extraction solvents of varying polarities on polyphenols of Orthosiphon stamineus and evaluation of the free radical-scavenging activity. Food Chem. 2005, 93, 311-317. [CrossRef] · DOI
  3. Bever, B.O.; Zahnd, G.R. Plants with Oral Hypoglycaemic Action. Q. J. Crude Drug Res. 1979, 17, 139-196. [CrossRef] · DOI
  4. Olennikov, D.N.; Tankhaeva, L.M. Physicochemical characteristics and antioxidant activity of melanoidin pigment from the fermented leaves of Orthosiphon stamineus. Rev. Bras. Farmacogn. 2012, 22, 284-290. [CrossRef] · DOI
  5. Awale, S.; Tezuka, Y.; Banskota, A.H.; Kadota, S. Siphonols A-E: Novel nitric oxide inhibitors from Orthosiphon stamineus of Indonesia. Bioorg. Med. Chem. Lett. 2003, 13, 31-35. [CrossRef] · DOI
  6. Adam, Y.; Somchit, M.N.; Sulaiman, M.R.; Nasaruddin, A.A.; Zuraini, A.; Bustamam, A.A.; Zakaria, Z.A. Diuretic properties of Orthosiphon stamineus Benth. J. Ethnopharmacol. 2009, 124, 154-158. [CrossRef] · DOI
  7. Olah, N.-K.; Radu, L.; Mogoşan, C.; Hanganu, D.; Gocan, S. Phytochemical and pharmacological studies on Orthosiphon stamineus Benth.(Lamiaceae) hydroalcoholic extracts. J. Pharm. Biomed. Anal. 2003, 33, 117-123. [CrossRef] · DOI
  8. Matsubara, T.; Bohgaki, T.; Watarai, M.; Suzuki, H.; Ohashi, K.; Shibuya, H. Antihypertensive actions of methylripariochromene A from Orthosiphon aristatus, an Indonesian traditional medicinal plant. Biol. Pharm. Bull. 1999, 22, 1083-1088. [CrossRef] · DOI
  9. Yehya, A.H.S.; Asif, M.; Kaur, G.; Hassan, L.E.A.; Al-Suede, F.S.R.; Abdul Majid, A.M.S.; Oon, C.E. Toxicological studies of Orthosiphon stamineus (Misai Kucing) standardized ethanol extract in combination with gemcitabine in athymic nude mice model. J. Adv. Res. 2019, 15, 59-68. [CrossRef] · DOI
  10. Robards, K.; Prernzler, P.D.; Tucker, G.; Swatsitang, P.; Glover, W. Phenolic compounds and their role in oxidative processes in fruits. Food Chem. 1999, 66, 401-436. [CrossRef] · DOI · Paper page
  11. Maheswari, C.; Maryammal, R.; Venkatanarayanan, R. Hepatoprotective activity of Orthosiphon stamineus on liver damage caused by paracetamol in rats. Jordan J. Biol. Sci. 2008, 1, 105-108.
  12. Cai, X.; Xiao, C.; Xue, H.; Xiong, H.; Hang, Y.; Xu, J.; Lu, Y. A comparative study of the antioxidant and intestinal protective effects of extracts from different parts of Java tea (Orthosiphon stamineus). Food Sci. Nutr. 2018, 6, 579-584. [CrossRef] [PubMed] · DOI
  13. Choo, B.K.M.; Kundap, U.P.; Kumari, Y.; Hue, S.M.; Othman, I.; Shaikh, M.F. Orthosiphon stamineus Leaf Extract Affects TNF-alpha and Seizures in a Zebrafish Model. Front. Pharmacol. 2018, 9, 139. [CrossRef] · DOI
  14. Seyedan, A.; Alshawsh, M.A.; Alshagga, M.A.; Mohamed, Z. Antiobesity and Lipid Lowering Effects of Orthosiphon stamineus in High-Fat Diet-Induced Obese Mice. Planta Med. 2017, 83, 684-692. [CrossRef] [PubMed] · DOI
  15. Nawi, I.M.; Samad, A.A. Successful plant regeneration of Orthosiphon stamineus from petiole. J. Med. Plants Res. 2012, 6, 4276-4280.
  16. Affendy, H.; Aminuddin, M.; Azmy, M.; Amizi, M.; Assis, K.; Tamer, A. Effect of organic fertilizers application to the growth of Orthosiphon stamineus Benth. intercropped with Hevea brasiliensis Willd. and Durio zibethinus Murr. Int. J. Agric. Res. 2011, 6, 180-187.
  17. Dhoran, V.; Gudadhe, S. Effect of plant growth regulators on seed germination and seedling vigour in Asparagus sprengeri Regelin. Int. Res. J. Biol. Sci. 2012, 1, 6-10.
  18. Leng, L.W.; Lai-Keng, C. Plant regeneration from stem nodal segments of Orthosiphon stamineus Benth., a medicinal plant with diuretic activity. In Vitro Cell. Dev. Biol. Plant 2004, 40, 115-118. [CrossRef] · DOI
  19. Zainuddin, Z. In-Vitro Regeneration Of Orthosiphon stamineus (Misai Kucing) Using Axillary Bud. Sci. Herit. J. 2019, 3, 8-10. [CrossRef] · DOI
  20. Sheena, E.; Jeya Jothi, G. In vitro propagation of Orthosiphon stamineus Benth (Lamiaceae) an important medicinal plant using nodal and leaf explants. Pharma Innov. J. 2015, 4, 6-10.
  21. Zarnadze, N.; Diasamidze, I.; Varshanidze, N.; Dolidze, K.; Bolkvadze, T. In vitro reproduction of Kidney Tea (Orthosiphon stamineus Bents). J. Pharm. Pharmacol. 2018, 6, 695-699. [CrossRef] · DOI
  22. Wai-Leng, L.; Lai-Keng, C. Establishment of Orthosiphon stamineus cell suspension culture for cell growth. Plant Cell Tissue Organ Cult. 2004, 78, 101-106. [CrossRef] · DOI
  23. Lim, F.L.; Yam, M.F.; Asmawi, M.Z.; Chan, L.-K. Elicitation of Orthosiphon stamineus cell suspension culture for enhancement of phenolic compounds biosynthesis and antioxidant activity. Ind. Crops Prod. 2013, 50, 436-442. [CrossRef] · DOI
  24. Larkin, P.J.; Scowcroft, W.R. Somaclonal variation-A novel source of variability from cell cultures for plant improvement. Theor. Appl. Genet. 1981, 60, 197-214. [CrossRef] · DOI
  25. Yaacob, J.S.; Loh, H.-S.; Mat Taha, R. Protein Profiling and Histone Deacetylation Activities in Somaclonal Variants of Oil Palm (Elaeis guineensis Jacq.). Sci. World J. 2013, 2013, 613635. [CrossRef] · DOI
  26. Yaacob, J.S.; Mat Taha, R. Genetic stability of in vitro propagated African blue lily (Agapanthus praecox ssp. minimus). Caryologia 2014, 67, 227-233. [CrossRef] · DOI
  27. Murashige, T.; Skoog, F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant. 1962, 15, 473-497. [CrossRef] · DOI
  28. Vujović, T.; Ružić, D.; Cerović, R. In vitro shoot multiplication as influenced by repeated subculturing of shoots of contemporary fruit rootstocks. Hortic. Sci. 2012, 39, 101-107. [CrossRef] · DOI
  29. Viehmannova, I.; Bortlova, Z.; Vitamvas, J.; Cepkova, P.H.; Eliasova, K.; Svobodova, E.; Travnickova, M. Assessment of somaclonal variation in somatic embryo-derived plants of yacon [Smallanthus sonchifolius (Poepp. and Endl.) H. Robinson] using inter simple sequence repeat analysis and flow cytometry. Electron. J. Biotechnol. 2014, 17, 102-106. [CrossRef] · DOI
  30. Reshi, N.A.; Sudarshana, M. In vitro micropropagation of Orthosiphon aristatus (Blume) Miq. J. Med. Plants Res. 2015, 9, 962-967.
  31. Ignacimuthu, S.; Elangomathavan, R.; Prakash, S.; Kathiravan, K.; Seshadri, S. High frequency in vitro propagation of kidney tea plant. Plant Cell Tissue Organ Cult. 2003, 72, 83-86. [CrossRef] · DOI
  32. Al Khateeb, W.; Bahar, E.; Lahham, J.; Schroeder, D.; Hussein, E. Regeneration and assessment of genetic fidelity of the endangered tree Moringa peregrina (Forsk.) Fiori using Inter Simple Sequence Repeat (ISSR). Physiol. Mol. Biol. Plants 2013, 19, 157-164. [CrossRef] [PubMed] · DOI
  33. Veetil, S.E.; Gabriel, J.J. Comparative Phytochemical Screening and Antioxidant Properties of In Vitro and In Vivo Propagated Orthosiphon Stamineus Benth (Lamiaceae). Asian J. Pharm. Clin. Res. 2015, 8, 216-220.
  34. Sridhar, T.M.; Aswath, C.R. Influence of Additives on Enhanced in Vitro Shoot Multiplication of Stevia rebaudiana (Bert.)-An Important Anti Diabetic Medicinal Plant. Am. J. Plant Sci. 2014, 5, 192-199. [CrossRef] · DOI
  35. Ling, A.P.K.; Kok, K.M.; Hussein, S.; Ong, S.L. Effects of plant growth regulators on adventitious roots induction from different explants of Orthosiphon stamineus. Am.-Eurasian J. Sustain. Agric. 2009, 3, 493-501.
  36. Hembrom, M.E.; Martin, K.; Patchathundikandi, S.K.; Madassery, J. Rapid in vitro production of true-to-type plants of Pogostemon heyneaus through dedieferentiated axillary buds. In Vitro Cell. Dev. Biol. Plant 2006, 42, 283-286. [CrossRef] · DOI
  37. Nordine, A.; Bousta, D.; El Khanchoufi, A.; El Meskaoui, A. An efficient and rapid in vitro propagation system of Thymus hyemalis Lange, a wild medicinal and aromatic plant of Mediterranean region. Int. J. Pharma Biosci. Technol. 2013, 1, 118-129.
  38. Saha, S.; Kader, A.; Sengupta, C.; Ghosh, P. In vitro propagation of Ocimum gratissimum L.(Lamiaceae) and its evaluation of genetic fidelity using RAPD marker. Am. J. Plant Sci. 2012, 3, 64. [CrossRef] · DOI
  39. Marin, J. High survival rates during acclimatization of micropropagated fruit tree rootstocks by increasing exposures to low relative humidity. In Proceedings of the I International Symposium on Acclimatization and Establishment of Micropropagated Plants, Sani-Halkidiki, Macedonia, Greece, 19 September 2001; pp. 139-142.
  40. Chandra, S.; Bandopadhyay, R.; Kumar, V.; Chandra, R. Acclimatization of tissue cultured plantlets: From laboratory to land. Biotechnol. Lett. 2010, 32, 1199-1205. [CrossRef] · DOI · Paper page
  41. Hoque, M.; Ali, M.; Karim, N. Embryogenic callus induction and regeneration of elite Bangladeshi Indica rice cultivars. Plant Tissue Cult. Biotechnol. 2007, 17, 65-70. [CrossRef] · DOI
  42. Mishra, R.; Wang, H.-Y.; Yadav, N.R.; Wilkins, T.A. Development of a highly regenerable elite Acala cotton (Gossypium hirsutum cv. Maxxa)-A step towards genotype-independent regeneration. Plant Cell Tissue Organ Cult. 2003, 73, 21-35. [CrossRef] · DOI
  43. Rashid, H.; Khan, S.A.; Zia, M.; Chaudhary, M.F.; Hanif, Z.; Chaudary, Z. Callus induction and regeneration in elite sugarcane cultivar HSF-240. Pak. J. Bot. 2009, 41, 1645-1649.
  44. Dang, J.C.; Kumaria, S.; Kumar, S.; Tandon, P. Micropropagation of Ilex khasiana, a critically endangered and endemic holly of Northeast India. AoB Plants 2011, 2011. [CrossRef] · DOI
  45. Cui, Y.; Deng, Y.; Zheng, K.; Hu, X.; Zhu, M.; Deng, X.; Xi, R. An efficient micropropagation protocol for an endangered ornamental tree species (Magnolia sirindhorniae Noot. & Chalermglin) and assessment of genetic uniformity through DNA markers. Sci. Rep. 2019, 9, 9634.
  46. Markovic, M.; Grbic, M.; Djukic, M. Micropropagation of Endangered and Decorative Species Dianthus pinifolius Sibth. et Sm. Braz. Arch. Biol. Technol. 2016, 59, e16150320. [CrossRef] · DOI
  47. Ramirez-Estrada, K.; Vidal-Limon, H.; Hidalgo, D.; Moyano, E.; Golenioswki, M.; Cusidó, R.M.; Palazon, J. Elicitation, an effective strategy for the biotechnological production of bioactive high-added value compounds in plant cell factories. Molecules 2016, 21, 182. [CrossRef] · DOI
  48. Dias, M.I.; Sousa, M.J.; Alves, R.C.; Ferreira, I.C. Exploring plant tissue culture to improve the production of phenolic compounds: A review. Ind. Crops Prod. 2016, 82, 9-22. [CrossRef] · DOI
  49. Manimekalai, R.; Nagarajan, P.; Kumaran, P. Comparison of effectiveness of RAPD, ISSR and SSR markers for analysis of coconut (Cocos nucifera L.) germplasm accessions. Trop. Agric. Res. 2006, 18, 217.
  50. Sornakili, A.; Rathinam, P.K.; Thiruvengadum, R.; Kuppusamy, P. Comparative assessment of RAPD and ISSR markers to study genetic polymorphism in Colletotrichum gloeosporioides isolates of mango. Asian J. Plant Pathol. 2017, 11, 130-138. [CrossRef] · DOI
  51. Rayar, J.K.; Arif, M.; Singh, U.S. Relative efficiency of RAPD and ISSR markers in assessment of DNA polymorphism and genetic diversity among Pseudomonas strains. Afr. J. Biotechnol. 2015, 14, 1097-1106.
  52. Fernandez, M.; Figueiras, A.; Benito, C. The use of ISSR and RAPD markers for detecting DNA polymorphism, genotype identification and genetic diversity among barley cultivars with known origin. Theor. Appl. Genet. 2002, 104, 845-851. [CrossRef] [PubMed] · DOI
  53. Parveen, S.; Saharan, M.; Verma, A.; Sharma, I. Comparative analysis of RAPD and ISSR marker assays for detecting genetic polymorphism in Tilletia indica. Eur. J. Exp. Biol. 2013, 3, 380-387.
  54. Pathak, H.; Dhawan, V. ISSR assay for ascertaining genetic fidelity of micropropagated plants of apple rootstock Merton 793. In Vitro Cell. Dev. Biol. Plant 2012, 48, 137-143. [CrossRef] · DOI · Paper page
  55. Muthusamy, S.; Kanagarajan, S.; Ponnusamy, S. Efficiency of RAPD and ISSR markers system in accessing genetic variation of rice bean (Vigna umbellata) landraces. Electron. J. Biotechnol. 2008, 11, 32-41. [CrossRef] · DOI
  56. Seth, S.; Panigrahi, J. In vitro organogenesis of Abutilon indicum (L.) Sweet from leaf derived callus and assessment of genetic fidelity using ISSR markers. J. Hortic. Sci. Biotechnol. 2019, 94, 70-79. [CrossRef] · DOI
  57. Dinara, S.M.; Elena, V.K.; Tatyana, I.N. Assessment of Genetic Fidelity of Fritillaria dagana (Liliaceae) Regenerated Plants Using ISSR Markers. In Proceedings of the BIO Web of Conferences, Novosibirsk, Russia, 8-12 October 2018; p. 00029.
  58. Babu, G.A.; Vinoth, A.; Ravindhran, R. Direct shoot regeneration and genetic fidelity analysis in finger millet using ISSR markers. Plant Cell Tissue Organ Cult. 2018, 132, 157-164. [CrossRef] · DOI
  59. Rohela, G.K.; Jogam, P.; Shabnam, A.A.; Shukla, P.; Abbagani, S.; Ghosh, M.K. In vitro regeneration and assessment of genetic fidelity of acclimated plantlets by using ISSR markers in PPR-1 (Morus sp.): An economically important plant. Sci. Hortic. 2018, 241, 313-321. [CrossRef] · DOI
  60. Mudoi, K.D.; Saikia, S.P.; Goswami, A.; Gogoi, A.; Bora, D.; Borthakur, M. Micropropagation of important bamboos: A review. Afr. J. Biotechnol. 2013, 12, 2770-2785.
  61. Zakaria, M.H. Review of Policies and Issues in the Malaysian Herbal Industry; FFTC Agricultural Policy Articles; Food and Fertilizer Technology Centre (FTTC): Taiper, Taiwan, 2015.
  62. Gonçalves, S.; Romano, A. Production of plant secondary metabolites by using biotechnological tools. In Secondary Metabolites: Sources and Applications; INTECH Open Access Publisher: London, UK, 2018; pp. 81-99.
  63. Sampaio, B.L.; Edrada-Ebel, R.; Da Costa, F.B. Effect of the environment on the secondary metabolic profile of Tithonia diversifolia: A model for environmental metabolomics of plants. Sci. Rep. 2016, 6, 29265. [CrossRef] · DOI
  64. Ncube, B.; Finnie, J.; Van Staden, J. Quality from the field: The impact of environmental factors as quality determinants in medicinal plants. S. Afr. J. Bot. 2012, 82, 11-20. [CrossRef] · DOI

Authors
Ali, H., Musa, I. F., Abu Bakar, N. A., Karsani, S. A., & Yaacob, J. S.
Affiliations
  • University of Malaya (MY)
  • Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur 50603, Malaysia
Corresponding author
Yaacob, Jamilah Syafawati
ORCID
Venue
Agronomy
Publisher
MDPI AG
Published
2019-11-20
Volume
9
Issue
12
Pages
778
ISSN
2073-4395 (ISSN-L)
DOI
10.3390/agronomy9120778
OpenAlex
W2991046082
Cited by
13
References
67
Citations by year
2026: 1 · 2025: 2 · 2024: 1 · 2023: 3 · 2022: 3 · 2021: 1 · 2020: 2
FWCI
0.50
Type
journal article
Language
en
Open access
Licence
Locations
  • Agronomy (publishedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
Universiti Malaya (RG297-14AFR, PG156-2014B, CEBAR RU006-2018)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant tissue culture and regeneration
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Flowering Plant Growth and Cultivation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic diversity and population structure
Keywords
Orthosiphon stamineus, in vitro regeneration, ISSR markers, genetic fidelity, clonality, acclimatization, potting mix, kinetin, indole-3-acetic acid, micropropagation, genetic variation, medicinal plants

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review