Ocotea porosa: Anatomy and Histochemistry of Leaves and Stems, Chemical Composition, Cytotoxicity and Insecticidal Activities of Essential Oil

Brustulim, Libardone José Ribeiro; Monteiro, Luciane Mendes; Almeida, Valter Paes de; Raman, Vijayasankar; Maia, Beatriz Helena Lameiro de Noronha Sales; Casapula, Inaiara; Paludo, Kátia Sabrina; Bussade, Julia Emília; Rehman, Junaid Ur; Kahn, Ikhlas Ahmed; Farago, Paulo Vitor; Budel, Jane Manfron

doi:10.1590/1678-4324-2020190082

Libardone José Ribeiro Brustulim

State University of Ponta Grossa, Health Science Graduate Program, Ponta Grossa, Paraná, Brazil

http://orcid.org/0000-0002-1020-6948

Luciane Mendes Monteiro

State University of Ponta Grossa, Health Science Graduate Program, Ponta Grossa, Paraná, Brazil

http://orcid.org/0000-0003-2896-8147

Valter Paes de Almeida

State University of Ponta Grossa, Health Science Graduate Program, Ponta Grossa, Paraná, Brazil

http://orcid.org/0000-0003-0603-8184

Vijayasankar Raman

National Center for Natural Products Research, School of Pharmacy, University of Mississippi, University, MS, USA

http://orcid.org/0000-0001-7368-9644

Beatriz Helena Lameiro de Noronha Sales Maia

Federal University of Paraná, Chemistry Department, Curitiba, Paraná, Brazil

http://orcid.org/0000-0001-5896-2892

Inaiara Casapula

Federal University of Paraná, Chemistry Department, Curitiba, Paraná, Brazil

http://orcid.org/0000-0001-9758-3756

Kátia Sabrina Paludo

State University of Ponta Grossa, Health Science Graduate Program, Ponta Grossa, Paraná, Brazil

http://orcid.org/0000-0003-2248-5881

Julia Emília Bussade

University of Mississippi, Department of Modern Languages, University, MS, USA

http://orcid.org/0000-0002-8516-5888

Junaid Ur Rehman

National Center for Natural Products Research, School of Pharmacy, University of Mississippi, University, MS, USA

http://orcid.org/0000-0002-5292-5457

Ikhlas Ahmed Kahn

National Center for Natural Products Research, School of Pharmacy, University of Mississippi, University, MS, USA

http://orcid.org/0000-0001-5464-4643

Paulo Vitor Farago

State University of Ponta Grossa, Health Science Graduate Program, Ponta Grossa, Paraná, Brazil

http://orcid.org/0000-0002-9934-4027

Jane Manfron Budel ^**Correspondence: janemanfron@hotmail.com; Tel.: +55-41-33203124 (J.M.B)

State University of Ponta Grossa, Health Science Graduate Program, Ponta Grossa, Paraná, Brazil

National Center for Natural Products Research, School of Pharmacy, University of Mississippi, University, MS, USA

http://orcid.org/0000-0003-1873-2253

*Correspondence: janemanfron@hotmail.com; Tel.: +55-41-33203124 (J.M.B)

Conflicts of Interest:

“The authors declare no conflict of interest.” “The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results”.

Figures (5)
Tables (2)

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Figure 1
Morpho-anatomy ofOcotea porosa [c, e: Light microscopy; d, f, g, h: FEG-SEM]. a- Plant in habit. b- Leaves. c-h- Leaf in surface view (c- adaxial, e-h- abaxial). [gl-domatia of glands, st- stomata, nt- non-glandular trichome, wa- waxes]. Scale bar: a = 70 cm; b = 4 cm; d = 1 mm; f = 100 µm; c, e = 50 µm; g = 20 µm; h= 5µm.

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Figure 2
Anatomy ofOcotea porosa leaves and stems [a-i: Light microscopy; j, k: FEG-SEM]. a. TS through leaf margin; b. TS of lamina; c. TS of midrib and lamina; d and e. TS of petiole; f-k. TS of stem. [co- collenchyma, cr- crystals, cs- subepidermal layer, ct- cuticle, cx- cortex, ep- epidermis, fi- fibers, nt- non-glandular trichome, pc- phenolic compounds, sc- secretory cell, sg- starch grains, ph- phloem, pp- palisade parenchyma, pi- pith, sp- spongy parenchyma, vb- vascular bundle, xy- xylem]. Scale bar: a, b, e, g, h, i = 50 μm; c, d, f = 200 μm; k =10 μm; j = 5 μm.

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Figure 3
Histochemistry of Ocotea porosa [a, e, f, j: Sudan III; b, g, k: phloroglucinol/HCl; d, i, m, n: ferric chloride (2%); c, h, l: potassium dichromate solution (10%). Transverse sections. a-d: midrib; e-i: petiole; j-n: stem [ct, cuticle; cx, cortex; en, endodermis; ep, epidermis; fi: fibers, gp, ground parenchyma; pc, phenolic compounds; ph, phloem; pi, pith; sc, secretory cavity; sl, sclerenchymatous layer; xy, xylem]. Scale bars: a-d, f-k = 50 μm, e = 200 μm, l, m = 100 μm.

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Figure 4
Cell cytotoxicity was determined using MTT assay. A - MCF-7, B - B16F10 and C - McCoy cells. **p< 0.01 and *** p < 0.001 compared to control. One-way ANOVA with Tukey’s post hoc test. Three independent experiments were performed.

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Figure 5
Morphology of MCF-7and B16F10 cells after 24 h of treatment with O. porosa EO. A and C - Control cells incubated with RPMI only. B and D - cells treated with 77 µg/mL of O. porosa EO. ↑: cell rounding, ➨: bleb formation, ▲: chromatin condensation. Magnification = 1000x, bar = 20 μm.

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Table 1
Chemical composition of Ocotea porosa essential oil.

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Table 2
IC₅₀ values of O. porosa oil for different cell lines and selectivity index (SI). IC₅₀ data were expressed as mean and ± standard error of the mean of three independent experiments.

#	Rt	Al¹	Al²	Chemical compound	%
1	6.080	933	932	α-Pinene	19.71
2	6.544	947	946	Camphene	0.48
3	7.354	973	969	Sabinene	0.60
4	7.464	976	974	β-Pinene	13.86
5	7.954	991	988	Mircene	2.07
6	8.908	1017	1014	α-Terpinene	0.25
7	9.368	1028	1024	Limonene	0.78
8	9.459	1031	1026	1,8-Cineole	0.24
9	10.581	1059	1054	γ-Terpinene	0.36
10	15.640	1177	1174	Terpinen-4-ol	0.43
11	16.248	1191	1186	α-Terpineol	0.35
12	24.286	1376	1374	α-Copaene	0.34
13	24.661	1385	1387	β-Bourbolene	0.53
14	24.994	1392	1389	β-Elemene	0,51
15	26.100	1419	1417	(E)-Caryophyllene	1.70
16	26.905	1439	1440	Aromadendrene	0.86
17	27.491	1453	1452	α-Humulene	0.57
18	27.799	1461	1458	Allo-aromadendrene	1.98
19	28.626	1481	1484	Germacrene D	1.88
20	29.266	1497	1500	Biciclogermacrene	24.62
21	29.959	1515	1513	γ-Cadinene	1,17
22	30.346	1525	1522	δ-Cadinene	0.66
23	31.331	1550	1548	Elemol	0.74
24	31.620	1558	1559	Germacrene B	0.39
25	32.415	1578	1577	Spathulenol	5.34
26	32.650	1584	1590	Globulol	2.09
27	32.951	1592	1592	Viridiflorol	0.99
28	33.207	1599	1600	Guaiol	1.95
29	33.846	1616	1618	1,10-di-epi-cubenol	0.95
30	34.470	1633	1630	γ-Eudesmol	0.42
31	34.716	1640	1639	alloaromadendrene epoxide	0.72
32	35.136	1651	1649	β-Eudesmol	1.77
33	35.251	1654	1652	α-Eudesmol	1.09
34	35.418	1659	1656	Valerianol	1.40
35	35.797	1669	1670	Bulnesol	0.75
Classes of compounds					%
Monoterpenoids hydrocarbons					38.12
Oxygenated Monoterpenoids					1.02
Sesquiterpenoids hydrocarbons					35.21
Oxygenated Sesquiterpenoids					18.20
Total					92.54

Cell lineage	IC₅₀ (mg/mL)	SI
McCoy	9.23 ± 7.04	-
B16F10	8.82 ± 6.27	1.05
MCF-7	185.4 ± 10.91	0.05

[1] *Correspondence: janemanfron@hotmail.com; Tel.: +55-41-33203124 (J.M.B)

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Ocotea porosa: Anatomy and Histochemistry of Leaves and Stems, Chemical Composition, Cytotoxicity and Insecticidal Activities of Essential Oil

Abstract