Callus induction and bioactive phenolic compounds production from <i>Byrsonima verbascifolia</i> (L.) DC. (Malpighiaceae)

Castro, Ana Hortência Fonsêca; Braga, Karina de Queiroz; Sousa, Fernanda Marinez de; Coimbra, Mairon César; Chagas, Rafael César Russo

doi:10.5935/1806-6690.20160017

Ana Hortência Fonsêca Castro ^** Autor para correspondência

Programa de Pós-Graduação em Ciências Farmacêuticas, Laboratório de Farmacobotânica e Plantas Medicinais, Universidade Federal de São João Del-Rei, Campus Centro-Oeste, Av. Sebastião Gonçalves Coelho, 400, Chanandour, Divinópolis-MG, Brasil, 35.501-296, acastro@ufsj.edu.br

Karina de Queiroz Braga

Laboratório de Farmacobotânica e Plantas Medicinais, Universidade Federal de São João Del-Rei, Campus Centro-Oeste, Av. Sebastião Gonçalves Coelho, 400, Chanandour, Divinópolis-MG, Brasil, 35.501-296, karinaqbraga@yahoo.com.br, fernandamarinez@gmail.com, maironcoimbra@gmail.com

Fernanda Marinez de Sousa

Laboratório de Farmacobotânica e Plantas Medicinais, Universidade Federal de São João Del-Rei, Campus Centro-Oeste, Av. Sebastião Gonçalves Coelho, 400, Chanandour, Divinópolis-MG, Brasil, 35.501-296, karinaqbraga@yahoo.com.br, fernandamarinez@gmail.com, maironcoimbra@gmail.com

Mairon César Coimbra

Laboratório de Farmacobotânica e Plantas Medicinais, Universidade Federal de São João Del-Rei, Campus Centro-Oeste, Av. Sebastião Gonçalves Coelho, 400, Chanandour, Divinópolis-MG, Brasil, 35.501-296, karinaqbraga@yahoo.com.br, fernandamarinez@gmail.com, maironcoimbra@gmail.com

Rafael César Russo Chagas

Programa de Pós-Graduação em Biotecnologia, Universidade Federal de São João Del-Rei, Campus Centro-Oeste, Av. Sebastião Gonçalves Coelho, 400, Chanandour, Divinópolis-MG, Brasil, 35.501-296, rafaelchagas@ufsj.edu.br

* Autor para correspondência

Figures (2)
Tables (3)

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Figure 1
Total phenols (A), flavonoids (B) and tannins (C) contents in B. verbascifolia’s callus. The bar represents the mean􀁲standard deviation of 3 observations. Legend: 1) 4.52 µM 2,4-D; 2) 9.05 µM 2,4-D; 3) 18.10 µM 2,4-D; 4) 4.44 µM BAP; 5) 8.88 µM BAP; 6) 17.75 µM BAP; 7) 4.52 µM 2,4-D + 4.44 µM BAP; 8) 4.52 µM 2,4-D + 8.88 µM BAP; 9) 4.52 µM 2,4-D + 17.75 µM BAP; 10) 9.05 µM 2,4-D + 4.44 µM BAP; 11) 9.05 µM 2,4-D + 8.88 µM BAP; 12) 9.05 µM 2,4-D + 17.75 µM BAP; 13) 18,10 µM 2,4-D + 4.44 µM BAP; 14) 18.10 µM 2,4-D + 8.88 µM BAP; 15) 18.10 µM 2,4-D +17.75 µM BAP; IE = initial explant

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Figure 2
Chromatograms obtained by HPLC-DAD analysis of hydromethanolic extracts by detection at 254 nm. A. Chromatographic profile of the Initial Explant - Peak 1 = benzoic acid derivative, Peaks 2-3 = p-coumaric acid derivative. B. Chromatographic profile of callus from treatment 6 (17.75 µM BAP) in the presence of light - Peaks 1-3 = benzoic acid derivatives, Peaks 4-6 = catechins, Peaks 7-8 = m-coumaric acid derivatives. C. Chromatographic profile of callus from treatment 6 (17.75 µM BAP) in the absence of light - Peaks 1-3 = benzoic acid derivatives, Peaks 4-7 = gallic acid derivatives, Peaks 8-12 = catechins, Peaks 13-15 = m-coumaric acid derivatives. D. Chromatographic profile of callus from treatment 7 (4.52 µM 2,4-D + 4.44 µM BAP) in the absence of light - Peaks 1-2 = cinnamic acid derivatives, Peak 3 = benzoic acid derivative, Peak 4 = gallic acid derivative, Peak 5 = benzoic acid derivative, Peaks 6-7 = gallic acid derivatives, Peaks 8-9 = benzoic acid derivatives, Peaks 10-11 = p-coumaric acid derivatives

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Table 1
Induction, fresh and dry weight, color and consistency of Byrsonima verbascifolia’s callus, in the presence of light after 45 days cultivation

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Table 2
Induction, fresh and dry weight, color and consistency of B. verbascifolia’s callus, in the absence of light after 45 days cultivation

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Table 3
Phenolic compounds in hydromethanolic extracts in Byrsonima verbascifolia’s callus using HPLC-DAD

Treatment	Growth regulators	Induction (%)	Fresh weight (g)	Dry weight (g)	Color	Consistency
Control	Absence of regulator	-	-	-	-	-
1	4.52 μM 2,4-D	53d	0.71 ± 0.09 b	0.07 ± 0.01 b	Yellow/Red	Friable
2	9.05 μM 2,4-D	87b	0.77 ± 0.08 b	0.04^ Standard deviation values for these data were omitted, since they are less than 0.009. Legend: - (no callus induction) c	Yellow/Red	Friable
3	18.10 μM 2,4-D	68c	0.70 ± 0.07 b	0.04^ Standard deviation values for these data were omitted, since they are less than 0.009. Legend: - (no callus induction) c	Yellow/Red	Friable
4	4.44 μM BAP	7f	0.47 ± 0.09 c	0.03^ Standard deviation values for these data were omitted, since they are less than 0.009. Legend: - (no callus induction) c	Green/Red	Compact
5	8.88 μM BAP	35e	0.48 ± 0.10 c	0.08 ± 0.01 b	Green/Red	Compact
6	17.75 μM BAP	94b	0.21 ± 0.05 d	0.04^ Standard deviation values for these data were omitted, since they are less than 0.009. Legend: - (no callus induction) c	Green/Red	Compact
7	4.52 μM 2,4-D + 4.44 μM BAP	100a	1.77 ± 0.09 a	0.17 ± 0.04 a	Green	Compact
8	4.52 μM 2,4-D + 8.88 μM BAP	100a	1.58 ± 0.12 a	0.15 ± 0.05 a	Green/Red	Compact
9	4.52 μM 2,4-D + 17.75 μM BAP	100a	0.69 ± 0.07 b	0.07 ± 0.01 b	Red	Compact
10	9.05 μM 2,4-D + 4.44 μM BAP	100a	1.55 ± 0.15 a	0.14 ± 0.05 a	Green/Red	Compact
11	9.05 μM 2,4-D + 8.88 μM BAP	100a	0.92 ± 0.20 b	0.07 ± 0.01 b	Green	Compact
12	9.05 μM 2,4-D + 17.75 μM BAP	100a	0.99 ± 0.21 b	0.09 ± 0.02 b	Green	Compact
13	18,10 μM 2,4-D + 4.44 μM BAP	100a	0.77 ± 0.10 b	0.08 ± 0.02 b	Green/Red	Compact
14	18.10 μM 2,4-D + 8.88 μM BAP	100a	1.02 ± 0.25 b	0.08 ± 0.01 b	Green/Red	Compact
15	18.10 μM 2,4-D +17.75 μM BAP	100a	0.28 ± 0.06 d	0.03^ Standard deviation values for these data were omitted, since they are less than 0.009. Legend: - (no callus induction) c	Green/Red	Compact
F		9.456^* * Significant value at 5% level by F test; means in the column followed by the same letter are not significantly different at p ≤ 0.05 by the Tukey test;	5.227^* * Significant value at 5% level by F test; means in the column followed by the same letter are not significantly different at p ≤ 0.05 by the Tukey test;	7.253^* * Significant value at 5% level by F test; means in the column followed by the same letter are not significantly different at p ≤ 0.05 by the Tukey test;

Treatment	Growth regulators	Induction (%)	Fresh weight (g)	Dry weight (g)	Color	Consistency
Control	Absence of regulator	-	-	-	-	-
1	4.52 μM 2,4-D	71 c	0.68 ± 0.16 b	0.06 ± 0.01 b	Yellow	Friable
2	9.05 μM 2,4-D	73 c	0.85 b ± 0.11	0.04^ Standard deviation values for these data were omitted, since they are less than 0.009. Legend: - (no callus induction) c	Yellow/Red	Friable
3	18.10 μM 2,4-D	82 b	0.34 ± 0.05 c	0.03^ Standard deviation values for these data were omitted, since they are less than 0.009. Legend: - (no callus induction) c	Yellow	Friable
4	4.44 μM BAP	53 d	0.15 ± 0.05 d	0.02^ Standard deviation values for these data were omitted, since they are less than 0.009. Legend: - (no callus induction) c	Yellow/Red	Compact
5	8.88 μM BAP	74 c	0.19 ± 0.04 d	0.03^ Standard deviation values for these data were omitted, since they are less than 0.009. Legend: - (no callus induction) c	Yellow/Red	Compact
6	17.75 μM BAP	59 d	0.32 ± 0.07 c	0.04^ Standard deviation values for these data were omitted, since they are less than 0.009. Legend: - (no callus induction) c	Yellow	Compact
7	4.52 μM 2,4-D + 4.44 μM BAP	100 a	1.66 ± 0.18 a	0.15 ± 0.05 a	Yellow	Friable
8	4.52 μM 2,4-D + 8.88 μM BAP	100 a	1.49 ± 0.15 a	0.14 ± 0.05 a	Yellow/Red	Compact
9	4.52 μM 2,4-D + 17.75 μM BAP	100 a	0.88 ± 0.10 b	0.06 ± 0.01 b	Yellow/Red	Compact
10	9.05 μM 2,4-D + 4.44 μM BAP	100 a	0.81 ± 0.09 b	0.04^ Standard deviation values for these data were omitted, since they are less than 0.009. Legend: - (no callus induction) c	Yellow/Red	Compact
11	9.05 μM 2,4-D + 8.88 μM BAP	100 a	1.58 ± 0.10 a	0.15 ± 0.05 a	Yellow	Compact
12	9.05 μM 2,4-D + 17.75 μM BAP	100 a	1.43 ± 0.10 a	0.13 ± 0.02 a	Yellow	Compact
13	18,10 μM 2,4-D + 4.44 μM BAP	100 a	0.84 ± 0.05 b	0.07 ± 0.01 b	Yellow	Compact
14	18.10 μM 2,4-D + 8.88 μM BAP	100 a	0.87 ± 0.09 b	0.05 ± 0.01 c	Yellow/Red	Compact
15	18.10 μM 2,4-D +17.75 μM BAP	100 a	0.34 ± 0.05 c	0.04^ Standard deviation values for these data were omitted, since they are less than 0.009. Legend: - (no callus induction) c	Yellow/Red	Compact
F		10.778^* * Significant value at 5% level by F test; means in the column followed by the same letter are not significantly different at p ≤ 0.05 by the Tukey test;	6.534^* * Significant value at 5% level by F test; means in the column followed by the same letter are not significantly different at p ≤ 0.05 by the Tukey test;	8.123^* * Significant value at 5% level by F test; means in the column followed by the same letter are not significantly different at p ≤ 0.05 by the Tukey test;

Sample	Peak number	Rt (min.)	λmax (nm)	Compound	Reference
Initial explant	1	3.491	264	Benzoic acid derivative	Maatta; Kamal-Eldin; Torronen (2003)MÄÄTTÄ, K. R.; KAMAL-ELDIN, A.; TÖRRÖNEN, A. R. High-performance liquid chromatography (HPLC) analysis of phenolic compounds in berries with diode array and electrospray ionization mass spectrometric (MS) detection: ribes species. Journal of Agricultural and Food Chemistry, v. 51, n. 23, p. 6736-6744, 2003.
	2	21.037	326	p-cumaric acid derivative	*Sakakibara et al.* (2003)**SAKAKIBARA, H. et al. Simultaneous determination of all polyphenols in vegetables, fruits, and teas. Journal of Agricultural and Food Chemistry, v. 51, n. 3, p. 571-581, 2003.
	3	23.264	326	p-cumaric acid derivative
Treatment 6 (presence of light)	1	3.506	265	Benzoic acid derivative	*Sakakibara et al.* (2003)**SAKAKIBARA, H. et al. Simultaneous determination of all polyphenols in vegetables, fruits, and teas. Journal of Agricultural and Food Chemistry, v. 51, n. 3, p. 571-581, 2003.
	2	10.433	258
	3	13.146	258
	4	17.780	279	Cathechin	Reference substance; *Sakakibara et al.* (2003)**SAKAKIBARA, H. et al. Simultaneous determination of all polyphenols in vegetables, fruits, and teas. Journal of Agricultural and Food Chemistry, v. 51, n. 3, p. 571-581, 2003.
	5	18.393	279
	6	19.125	279
	7	22.149	270	m-cumaric acid derivative	*Sakakibara et al.* (2003)**SAKAKIBARA, H. et al. Simultaneous determination of all polyphenols in vegetables, fruits, and teas. Journal of Agricultural and Food Chemistry, v. 51, n. 3, p. 571-581, 2003.
	8	23.226	270	m-cumaric acid derivative
Treatment 6 (absence of light)	1	2.163	274	Benzoic acid derivative	*Sakakibara et al.* (2003)**SAKAKIBARA, H. et al. Simultaneous determination of all polyphenols in vegetables, fruits, and teas. Journal of Agricultural and Food Chemistry, v. 51, n. 3, p. 571-581, 2003.
	2	3.509	272
	3	4.096	259
	4	4.780	262	Gallic acid derivative	*Abad-Garcia et al.* (2009)**ABAD-GARCÍA, B. et al.A general analytical strategy for the characterization of phenolic compounds in fruit juices by high-performance liquid chromatography with diode array detection coupled to electrospray ionization and triple quadrupole mass spectrometry. Journal of Chromatography A, v. 1216, n. 28, p. 5398-5415, 2009.
	5	5.055	262
	6	10.462	261
	7	13.164	257
	8	14.721	278	Cathechin	Reference substance; *Sakakibara et al.* (2003)**SAKAKIBARA, H. et al. Simultaneous determination of all polyphenols in vegetables, fruits, and teas. Journal of Agricultural and Food Chemistry, v. 51, n. 3, p. 571-581, 2003.
	9	17.351	279
	10	17.791	279
	11	18.384	279
	12	19.122	279
	13	22.127	273	m-cumaric acid derivative	*Sakakibara et al.* (2003)**SAKAKIBARA, H. et al. Simultaneous determination of all polyphenols in vegetables, fruits, and teas. Journal of Agricultural and Food Chemistry, v. 51, n. 3, p. 571-581, 2003.
	14	22.313	273
	15	23.195	273
Treatment 7 (absence of light)	1	2.225	277	Cinamic acid derivative	*Sakakibara et al.* (2003)**SAKAKIBARA, H. et al. Simultaneous determination of all polyphenols in vegetables, fruits, and teas. Journal of Agricultural and Food Chemistry, v. 51, n. 3, p. 571-581, 2003.
	2	2.878	278	Cinamic acid derivative
	3	3.477	269	Benzoic acid derivative
	4	5.051	261	Gallic acid derivative
	5	5.761	271	Benzoic acid derivative
	6	8.303	260	Gallic acid derivative	*Sakakibara et al.* (2003)**SAKAKIBARA, H. et al. Simultaneous determination of all polyphenols in vegetables, fruits, and teas. Journal of Agricultural and Food Chemistry, v. 51, n. 3, p. 571-581, 2003.
	7	10.426	261	Gallic acid derivative
	8	12.899	255	Benzoic acid derivative	*Sakakibara et al.* (2003)**SAKAKIBARA, H. et al. Simultaneous determination of all polyphenols in vegetables, fruits, and teas. Journal of Agricultural and Food Chemistry, v. 51, n. 3, p. 571-581, 2003.
	9	13.092	258	Benzoic acid derivative
	10	23.256	327	p-cumaric acid derivative
	11	29.490	308	p-cumaric acid derivative

[1] * Autor para correspondência

Brasil

Brasil

Callus induction and bioactive phenolic compounds production from Byrsonima verbascifolia (L.) DC. (Malpighiaceae)¹ Pesquisa financiada pela Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) (CRA APQ 000683-11)

Indução de calos e produção de compostos fenólicos bioativos em Byrsonima verbascifolia (L.) DC. (Malpighiaceae)

ABSTRACT

Brasil

Brasil

Callus induction and bioactive phenolic compounds production from Byrsonima verbascifolia (L.) DC. (Malpighiaceae)1 Pesquisa financiada pela Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) (CRA APQ 000683-11)

Indução de calos e produção de compostos fenólicos bioativos em Byrsonima verbascifolia (L.) DC. (Malpighiaceae)

ABSTRACT

Callus induction and bioactive phenolic compounds production from Byrsonima verbascifolia (L.) DC. (Malpighiaceae)¹ Pesquisa financiada pela Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) (CRA APQ 000683-11)