Histocytological Study of Somatic Embryogenesis in the Tree Cinnamomum camphora L. (Lauraceae)

  • Ruyue JING Central South Forestry University of Science and Technology, Changsha 410004
  • Peilan WANG Central South Forestry University of Science and Technology, Changsha 410004
  • Zhen HUANG Sichuan Academy of Forestry Sciences, Chengdu 610000
  • Zhihui LI Central South Forestry University of Science and Technology, Changsha 410004
Keywords: Cinnamomum camphora L.; histocytology; malformed embryos; primary somatic embryos; secondary somatic embryos; somatic embryogenesis


Histocytological studies were conducted on primary, secondary, and malformed embryos produced during somatic embryogenesis of Cinnamomum camphora L. to better understand its development. Exploring its callus types and structures provided a theoretical basis for clarifying the mechanism of somatic embryogenesis, which may shed light on the mechanism of zygotic embryogenesis. We used immature zygotic embryos as explants to induce somatic embryos, forming many embryogenic calli that differentiated into mature somatic embryos. Our results showed that somatic embryogenesis of C. camphora was similar to that of zygotic embryos. We have been dedifferentiated four types of callus. Compared with non-embryogenic cells, embryogenic cells had a closer arrangement, larger nucleus, thicker cytoplasm, more starch granules and easier to stain into black. Somatic embryogenesis had two pathways: direct (predominate) and indirect (rare). Embryogenic cells of C. camphora could have either an internal or external origin, the latter being primary, for which occurrence sites include epidermis and near-epidermis (little internally). Mostly arising from single cells, C. camphora follows two developmental pathways: single-cell equal as opposed to unequal, wherein both divide to form multi-cell proembryos. However, multicellular origins can occasionally occur and feature physiological isolation during somatic embryo development. This development has four embryo stages: globular, heart-shaped, torpedo, and cotyledon, with procambium cells apparent in globular embryos and late cotyledons forming “Y-shaped” vascular bundles. Secondary embryos were present in all stages, directly occurring on primary embryo’s germ and radicle end surfaces. We conclude that secondary and primary embryos of C. camphora undergo similar developmental processes. At the same time, conjoined cotyledon embryos and morphological abnormal embryos were found, with an internal origin more likely to generate abnormal embryos.


Metrics Loading ...


Al Gabbiesh AH, Ghabeish M, Kleinwächter M, Selmar D (2014). Plant regeneration through somatic embryogenesis from calli derived from leaf bases of Laurus nobilis L. (Lauraceae). Plant Tissue Culture and Biotechnology 24(2):213-221.

Alemanno L, Berthouly M, Michaux-Ferrière N (1996). Histology of somatic embryogenesis from floral tissues cocoa. Plant Cell, Tissue and Organ Culture 46(3):187-194.

Babu KN, Sajina A, Minoo D, John CZ, Mini PM, Tushar KV (2003) Micropropagation of camphor tree (Cinnamomum camphora). Plant Cell, Tissue and Organ Culture 74(2):179-183.

Benelli C, Fabbri A, Grassi S, Lambardi M, Rugini E (2001). Histology of somatic embryogenesis in mature tissues of olive (Olea europaea L.). The Journal of Horticultural Science and Biotechnology 76(1):112-119.

Button J, Botha CEJ (1975). Enzymicmaceration of citrus callus and the regeneration of plant from single cells. Journal of Experimental Botany 26(5):723-729.

Canhoto JM, Lopes ML, Cruz GS (1999). Somatic embryogenesis induction in Bay Laurel (Laurus nobilis L.). In: Somatic embryogenesis in woody plants. Springer, Dordrecht pp 341-367.

Catarina C, Moser JR, Bouzon Z, Floh E, Maraschin M, Viana AM (2005). Protocol of somatic embryogenesis: Ocotea catharinensis Mez. (Lauraceae). In: Protocol for somatic embryogenesis in woody plants. Springer, Dordrecht pp 427-443.

Catarina CS, Olmedo AD, Meyer GD, Macedo J, Amorim WD, Viana AM (2004). Repetitive somatic embryogenesis of Ocotea catharinensis Mez. (Lauraceae): effect of somatic embryo developmental stage and dehydration. Plant Cell, Tissue and Organ Culture 78(1):55-62.

Chen J, Shi J, Zhu Q, Huang M (2003). Studies on the somatic embryogenesis of Liriodendron hybrids (L. chinese × L. tulipifera). Forestry Science 4:49-53.

Chen J, Zhang Y, Li T, Wang P, Wang G, Shi J (2012), Study on origin and development of somatic embryos of Liriodendron hybrids. Journal of Nanjing Forestry University (Natural Science Edition) 36 (1):16-20.

Cui K, Wang J, Xing G, Wang Y (1997). Ultrastructural and cytochemical localization of ATPase activity studies of embryonic cell differentiation in Lycium barbarum L. Journal of Northwest Botany 17(6):106-110.

Cui R, Fan T, Qu F, Zhai F, Liang L, Gong X (2008). Study on somatic embryogenesis of Cyclamen persicum Mill. Journal of Sichuan University (Natural Science Edition) 45(6):1477-1484.

Denchev P, Velcheva M, Atanassov A (1991). A new approach to direct somatic embryogenesis in Medicago. Plant Cell Reports 10(6-7):338-341.

Du L (2005). Preliminary studies on plant regeneration via somatic embryogenesis and Agrobacterium-mediated transformation of campor tree (Cinnamomum camphora L.). Huazhong Agricultural University.

Du L, Ye Y, Bao M (2006). Study on somatic embryogenesis and plant regeneration of immature zygotic embryo of Cinnamomum camphora. Scientia Silvae Sinicae 42(6):37-39 (in Chinese).

Du L, Zhou S, Bao M (2007). Effect of plant growth regulators on direct somatic embryogenesis in camphor tree (Cinnamomum camphora L.) from immature zygotic embryos and embryogenic calli induction. Forestry Studies in China 9(4):267-271.

Du K, Cao H, Zhang H, Zhao N, Li T (1997). Establishment of a system with high synchronous frequency of somatic embryogenesis and embryo seedling formation in Camellia sinensis var. assamica. Acta Botanica Sinica 39:1126-1130.

Grieve M (1979). A modern herbal. In: Leyel CF (Ed). A modern herbal. Jonathan Cape, Thirty Bedford Square, London pp 155-156.

Guan Z, Guo B, Wei Y (2011). Morphological and cytological observation of somatic embryogenesis and development in Lycopersivon esculentum Mill. Journal of Chinese Electron Microscopy Society 30(2):158-165.

Guo S, Geng Z, Zhang W, Liang J, Wang C, Deng Z, Du S (2016). The chemical composition of essential oils from Cinnamomum camphora and their insecticidal activity against the stored product pests. International Journal of Molecular Sciences 17(11):1836.

Häggman H, Vuosku J, Sarjala T, Jokela A, Niemi K (2006). Somatic embryogenesis of pine species: from functional genomics to plantation forestry. In: Somatic embryogenesis. Springer, Berlin, Heidelberg pp 119-140.

Hao J, Zhou X, Li B, Guo S, Chen R, Zhang J (1994). Observation on somatic embryogenesis in vitro culture of Cnidium monnieri young stem. Wuhan Botanical Research 3:247-250.

Hao J, Zhou X, Li S (1995). Morphological and cytological observation of somatic embryogenesis and development in Foeniculum vulgare. Journal of Experimental Biology 3:339-347.

Hayta-Smedley S, Özbek N, Ansiz A, Bayraktar M, Gurel A (2018). The effect of different plant growth regulators on callus induction from hypocotyl explants and plantlet regeneration through somatic embryo in cotton (Gossypium hirsutum L.) genotype Nazilli-143. Anadolu Ege Tarımsal Araştırma Enstitüsü Dergisi 28(2):55-61.

Huang LC, Huang B, Murashige T (1998). A micropropagation protocol for Cinnamomum camphora. In Vitro Cellular and Developmental Biology-Plant 34(2):141-146.

Hemant S, Vashistha BD (2010). In vitro propagation of Cinnamomum camphora (L.) Nees & Eberm using shoot tip explants. Annals of Biology 26(2):109-114.

Kong L, Dai D, Shang M, Li K, Zhang CX (2009). Thidiazuron-induced somatic embryos, their multiplication, maturation, and conversion in Cinnamomum pauciflorum Nees (Lauraceae). New Forests 38(2):131-142.

Kumar PR, Tsunoda S (1980). Variation in oil content and fatty acid composition among seeds from Cruciferae. In: Tsunoda S (Ed). Brassica crops and wild allies. Tokyo: Japan Scientific Societies Press pp 235-252.

Lattoo SK, Bamotra S, Dhar RS, Khan S, Dhar AK (2006). Rapid plant regeneration and analysis of genetic fidelity of in vitro derived plants of Chlorophytum arundinaceum Baker-an endangered medicinal herb. Plant Cell Reports 25(6):499-506.

Liang Z, Yuan C, Zhang B (2018). Experience in making paraffin sections of minute tissues. Chinese Journal of Histochemistry and Cytochemistry 4:368-373.

Lili S, Ying C, Nanyang W, Caiping X, Fuliang C (2012). Histological observation of somatic embryogenesis and adventitious buds induction from Ginkgo biloba L. different explants in vitro culture. Chinese Forestry Science and Technology 3:107.

Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15(3):473-479.

Pelegrini LL, Ribas LLF, Amano E, Quoirin M (2013). Somatic embryogenesis and morphoanatomy of Ocotea porosa somatic embryos. Ciência Florestal 23(4):595-605.

Pelegrini LL, Ribas LLF, Zanette F, Koehler HS (2011). Micropropagation of Ocotea porosa (Nees & Martius) Barroso. African Journal of Biotechnology 10(9):1527-1523.

Pliego-Alfaro F, Murashige T (1988). Somatic embryogenesis in avocado (Persea americana Mill.) in vitro. Plant Cell, Tissue and Organ Culture 12(1):61-66.

Quoirin M, Pelegrini L, Ribas L (2011). Induction and repetitive embryogenesis of Ocotea porosa. In: BMC Proceedings 5(7):148.

Sánchez-Romero C, Márquez-Martín B, Pliego-Alfaro F (2006). Somatic and zygotic embryogenesis in avocado. In: Somatic embryogenesis. Springer, Berlin, Heidelberg pp 271-284.

Santa-Catarina C, Moser JR, Bouzon Z, Floh E, Maraschin M, Viana AM (2005). Protocol of somatic embryogenesis: Ocotea catharinensis Mez.(Lauraceae). In: Protocol for somatic embryogenesis in woody plants. Springer, Dordrecht pp 427-443.

Schwendiman J, Pannetier C, Michaux-Ferrière N (1988). Histology of somatic embryogenesis from leaf explants of the oil palm Elaeis guineensis. Annals of Botany 62(1):43-52.

Shekhawat MS, Manokari M (2018). Micromorphological and anatomical evaluation of in vitro and field transferred plants of Coccinia indica. Agricultural Research 7(2):135-144.

Shi X, Dai X, Liu G, Zhang J, Ning G, Bao M (2010). Cyclic secondary somatic embryogenesis and efficient plant regeneration in camphor tree (Cinnamomum camphora L.). In Vitro Cellular and Developmental Biology-Plant 46(2):117-125.

Shi XP (2009). Enhancement of somatic embryogenesis and genetic transformation with Barnase and PaFT genes in Cinnamomum camphora L. PhD Dissertation, Huazhong Agricultural University, Wuhan, Hubei, China (in Chinese).

Steward FC, Mapes MO, Hears K (1958). Growth and organized development of cultured cells. II. Growth and division of freely suspended cells. American Journal of Botany 45(10):705-708.

Wang Y (2004). Histocytological study on somatic embryogenesis of Hevea brasiliensis. South China Tropical Agriculture University.

Wang Y, Cui K, Chen K, Gao Z, Zhang D, Jiao C (1993). Study on cytoembryology and starch growth and decline dynamics of somatic embryogenesis in Triticum acstivum tissue culture. Journal of Experimental Biology 3:259-267.

Williams EG, Maheswaran G (1986). Somatic embryogenesis: factors influencing coordinated behaviour of cells as an embryogenic group. Annals of Botany 57(4):443-462.

Xiang W, Zhao J, Wu J, Huang J, Yu W, Wu L, Zhang Q (2015). Morphological and cytological observation of somatic embryogenesis and development in Torreya grandis ‘Merrillii’. Acta Horticulturae Sinica 42 (8):1477-1486.

Xinju Z (2009). Research overview on the breeding and cultivation of Cinnamomum camphora. Guangdong Forestry Science and Technology 2009-01.

Xue B (2017). Histological research of somatic embryo genesis and development in two Lily species native to China. Shenyang Agricultural University.

Yang L, Li Z, Lin L (2018). Effects of hormon on the somatic embryogenesis from different provenances of Cinnamomum camphora. Journal of Central South Forestry University 38(12):115-120.

Zhang C, Yao Z, Zhao Z, Qi J (2007). Histological observation of somatic embryogenesis from cultured embryos of Quercus variabilis BI. Journal of Plant Physiology and Molecular Biology 33(1):33-38.

Zhang W (1982). Morphological study on embryogenesis in leaf culture of Saintpaulia ionantha Wendl. Acta Botanica Sinica 24(8):282-284.

Zou H, Yong K, Wang W (2009). Research progress on tissue culture and rapid propagation technology of Cinnamomum camphora L. Fujian Agricultural Science and Technology 1:71-72.

How to Cite
JING, R., WANG, P., HUANG, Z., & LI, Z. (2019). Histocytological Study of Somatic Embryogenesis in the Tree Cinnamomum camphora L. (Lauraceae). Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 47(4), 1348-1358. https://doi.org/10.15835/nbha47411655
Research Articles