Morphology and Anatomy of Leaf, Stem and Petiole of Luvunga crassifolia Tanaka (Rutaceae)


  • Sugathini Shunmugam Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor, Malaysia.
  • Nur Syamilah Rosli Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor, Malaysia.
  • Sugumaran Manickam Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia
  • Nur Fatihah Mohd Yusoff Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor, Malaysia.
  • Noorjahan Banu Alitheen Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, UPM Serdang, Selangor, Malaysia.
  • Parameswari Namasivayam Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia



Calcium oxalates, morphology, pellucid dots, secretory cavities


Luvunga crassifolia is an underutilized plant in the Citrus family. Other than brief morphological descriptions, there are no published reports on other identification features of this plant. Thus, the current study was aimed to investigate macroscopic and microscopic diagnostic features of L. crassifolia leaves, stems, and petioles. Macroscopic characterization, optimization of histological procedure, and histochemical analyses of differential stains were carried out on the leaves, stems, and petioles of L. crassifolia. The histological method was optimized by modifying the following parameters: number of fixation days, dehydration duration with degraded series of ethanol or butanol, clearing duration, and infiltration duration. After infiltration, embedding and sectioning of the tissues were performed. Histochemical analyses were carried out using differential stains to identify the cellular components in leaf, stem and, petiole tissue sections. This study showed that L. crassifolia leaves are amphistomatic. Pellucid dots were observed on both adaxial and abaxial leaf surfaces. Secretory cavities, xylem, phloem, and pericyclic fibers were found in the cross-sections of leaf, stem, and petiole. Calcium oxalates were present in the leaf and stem sections, while trichomes were detected in stem and petiole sections. The information obtained from this study will be helpful for the identification and future taxonomic-related studies of this plant species.


D. J. Mabberley, “Mabberley's plant-book: A portable dictionary of plants, their classifications and uses,” 3rd Edition, Cambridge University Press, 2008.

Y. Roskov , T. Kunze , T. Orrell et al., "Species 2000 & ITIS Catalogue of Life, 2014 Annual Checklist,”, Species 2000: Naturalis, Leiden, the Netherlands, 2014. (Accessed: 18 February 2020)

M. Groppo, J. R. Pirani, M. L. Salatino, S. R. Blanco, and J. A. Kallunki, “Phylogeny of Rutaceae based on twononcoding regions from cpDNA,” American Journal of Botany, vol. 95, no. 8, pp. 985−1005, 2008.

N. T. Son, “Notes on the genus Paramignya: Phytochemistry and Biological activity,” Bulletin of Faculty of Pharmacy, Cairo University, vol. 56, no. 1, pp. 1−10, 2018.

N. G. Dengler, “An integral part of botany,” American Journal of Botany, vol. 89, no. 2, pp. 369−373, 2002.

K. A. Babalola,, & A. A. Victoria, “Foliar Epidermal Morphology of Two West African Genera of Haloragaceae R. BR.(Saxifragales),” Journal of Scientific Research and Development, vol. 11, pp. 84−91, 2009.

Tanaka, Meded, Rijks-Herb, 69:8,1931.

W. T. Swingle, & P. C. Reece, “The botany of Citrus and orange relatives in the orange subfamily,” The Citrus Industry, vol. 1, pp. 190−340, 1967.

R. R. Krueger, and L. Navarro, “Citrus germplasm resources,” Citrus genetics, breeding and biotechnology, vol. 45, pp. 140, 2007.

A. Latiff, I. Faridah Hanum, A. Zainudin Ibrahim et al., “On the vegetation and flora of Pulau Tioman, Peninsular Malaysia,” Raffles Bulletin of Zoology, vol. 47, pp. 11−72, 1999.

D. J. Mabberley, “Australian Citreae with notes on other Aurantioideae (Rutaceae),” Telopea, vol. 7, no. 4, pp. 333−344, 1998.

A. Ash, “Manual of leaf architecture: morphological description and categorization of dicotyledonous and net-veined monocotyledonous angiosperms,” Smithsonian Institution, 1999.

B. W. Grant, and I. Vatnick, “Environmental correlates of leaf stomata density,” Teaching issues and Experiments in Ecology, vol. 1, no. 1, pp. 1−24, 2004.

K. Khandelwal, “Practical pharmacognosy techniques and experiments,” 19th edition, Mumbai, Pragati Books Pvt. Ltd., 2008.

D. A. Johansen, “Plant microtechnique,” Plant microtechnique, 1940.

J. E. Kraus, H. C. de Sousa, M. H. Rezende et al., “Astra blue and basic fuchsin double staining of plant materials,” Biotechnic & histochemistry, vol. 73, no. 5, pp. 235-243, 1998.

M. I. Barkatullah, G. Jelani, and I. Ahmad, “Leaf, stem bark and fruit anatomy of Zanthoxylum Armatum dc.(Rutaceae),” Pakistan Journal of Botany, vol. 46, no. 4, pp. 1343−1349, 2014.

S. Singh, L. Manchawal, and M. G. Chauhan, “Pharmacognostic study of male leaves of Trichosanthes dioica Roxb. with special emphasis on microscopic technique,” Journal of Pharmacognosy and Phytotherapy, vol. 2, no. 5, pp. 71–75, 2010.

G. L. Nesom, “Citrus trifoliata (Rutaceae): Review of biology and distribution in the USA,” Phytoneuron, vol. 46, pp. 1-14, 2014.

A. Padaki, and N. Parthasarathy, “Abundance and distribution of lianas in tropical lowland evergreen forest of Agumbe, Central Western Ghats, India,” Tropical Ecology, vol. 41, no. 2, pp. 143−154, 2000.

Luvunga monophylla (DC.) Mabb, Australian Tropical Rainforest Plants, 8th Edition, CSIRO Publication,, 2020, (Accessed: 23 April 2020).

D. J. Mabberley, “Citrus (Rutaceae): A review of recent advances in etymology, systematics and medical applications,” Blumea-Biodiversity, Evolution and Biogeography of Plants, vol. 49, no. 2−3, pp. 481−498, 2004.

I. Hameed, F. Hussain, and G. Dastagir, “Stomatal studies of some selected medicinal plants of Polygonaceae,” Pakistan Journal of Botany, vol. 40, no. 6, pp. 2273−2280, 2008.

M. Prabhakar, “Structure, delimitation, nomenclature and classification of stomata,” Acta Botanica Sinica-English Edition, vol. 46, no. 2, pp. 242−252, 2004.

N. A. Igboabuchi, and C. V. Ilodibia, “A Study on the Anatomy of Zanthoxylum macrophylla (L.) Sarg.(Rutaceae),” Asian Journal of Biology, pp. 1−6, 2017.

D. Kumar, K. Kumar, S. Kumar et al., “Pharmacognostic evaluation of leaf and root bark of Holoptelea integrifolia Roxb, ”Asian Pacific journal of tropical biomedicine, vol. 2, no. 3, pp. 169-175, 2012.

S. Nabin, S. C. Nath, and D. Simanta, “Foliar micromorphological characters of few taxa of the genus Aquilaria Lamk growing in North-East India,” Advances in Plant Sciences, vol. 13, no. 2, pp. 551−558, 2000.

K. Kubitzki, J. A. Kallunki, M. Duretto, and P. G. Wilson, “Rutaceae,” In Flowering Plants. Eudicots, The Families and Genera of Vascular Plants, Springer, Berlin, Heidelberg, vol. 10, pp. 276−356, 2010.

G. W. Turner, “A brief history of the lysigenous gland hypothesis,” The Botanical Review, vol. 65, no. 1, pp. 76−88, 1999.

G. W. Turner, A. M. Berry, and E. M. Gifford, “Schizogenous secretory cavities of Citrus limon (L.) Burm. F. and a reevaluation of the lysigenous gland concept,” International Journal of Plant Sciences, vol. 159, no. 1, pp. 75−88, 1998.

K. Soundappan, R. Bajaj, B. N. Vaidya, and N. Joshee, “Anatomical and biochemical investigations on medicinal tree Murraya koenigii Spreng,” Journal of Biotech Research, vol. 9, pp. 70−78, 2018.

S. R. Machado, Y. Canaveze, and T. M.Rodrigues, “Structure and functioning of oil cavities in the shoot apex of Metrodorea nigra A. St.-Hil.(Rutaceae),” Protoplasma, vol. 254, no. 4, pp. 1661−1674, 2017.

M. Tekin and N. Eruygur, “The structural studies on the medicinal plant Haplophyllum telephioides,” Revista Brasileira de Farmacognosia, vol. 26, no. 5, pp. 544−552, 2016.

R. Charoensup, T. Duangyod, P. Phuneerub, and C. Singharachai, “Pharmacognostic specification of Zanthoxylum limonella (Dennst.) Alston: Fruits and seeds in Thailand,” Journal of Advanced Pharmaceutical Technology & Research, vol. 7, no. 4, pp. 134, 2016.

D. G. Hall, E. D. Ammar, and K. D. Bowman, “Epifluorescence and stereomicroscopy of trichomes associated with resistant and susceptible host plant genotypes of the Asian citrus psyllid (Hemiptera: Liviidae), vector of citrus greening disease bacterium,” Journal of Microscopy and Ultrastructure, vol. 6, no. 1, pp. 56, 2018.

M. S. Sedeek, N. F. Kirollos, C. G. Michel, and M. A. Abdel-Kawy, “Botanical and genetic characterization of Citrus maxima (BURM.) Merrill. F. Rutaceae,” International Journal of Pharmacy and Pharmaceutical Sciences, vol. 9, no. 1, pp. 260−272, 2017.

E. E. Jarald, E. Sheeja, S. Parial, H. Arya, and A. Bajpai, “Morphoanatomy of stems of Murraya koenigii Spreng,” Journal of Biological Sciences, vol. 8, no. 3, pp. 654−658, 2008.

T. G. Muntoreanu, R. da Silva Cruz, and G. F. Melo-de-Pinna, “Comparative leaf anatomy and morphology of some neotropical Rutaceae: Pilocarpus Vahl and related genera,” Plant Systematics and Evolution, vol. 296, no. 1−2, pp. 87−99, 2011.