In vitro Propagation to Conserve the Local Endemic and Endangered Medicinal Plant Helianthemum germanicopolitanum Bornm.

Kapdan, Emine; Sezgin, Mehmet

doi:10.1590/1678-4324-2021200760

Acessibilidade / Reportar erro

Brasil

Brazilian Archives of Biology and Technology

Español English

Brasil

Español English

sumário « anterior atual seguinte »

Sumário

Article - Agriculture, Agribusiness and Biotechnology • Braz. arch. biol. technol. 64 • 2021 • https://doi.org/10.1590/1678-4324-2021200760 copy

In vitro Propagation to Conserve the Local Endemic and Endangered Medicinal Plant Helianthemum germanicopolitanum Bornm.

Authorship SCIMAGO INSTITUTIONS RANKINGS

HIGHLIGHTS

This is the first report of the in vitro propagation of Helianthemum germanicopolitanum Bornm.

It is a comprehensive optimization study in which 3 different media, 64 different PGR combinations and 2 different solidifiers are tested.

The success of acclimatization is 32%.

Abstract

In this study, in vitro propagation and acclimatization of Helianthemum germanicopolitanum Bornm. plant, a local endemic in Çankırı Province (Turkey) with arid and semi-arid lands, and an endangered species taking part among medicinal and aromatic plants were accomplished, which is under-researched. In this study, three basal media [a) Murashige and Skoog b) Gamborg's B5, and c) Nitsch & Nitsch], two gelling agents (agar 7 g/L, and gelrite 2.1 g/L), eight cytokinins and eight auxin doses of plant growth regulators [a) 6-benzyladenin, b) Kinetin-(0, 0.5, 1, and 2 mg/L), c) Indole-3-butyric acid, d) α-napthaleneacetic acid-(0, 0.25, 0.5, and 1 mg/L)] prepared in 64 different combinations with 30 g/L sucrose was added to the basal media and adjusted to pH 5.7 for in vitro propagation of H. germanicopolitanum. During in vitro propagation of the plant, external and internal infections were frequently encountered and this was solved by the developed protocol. The best shoot growth (1.141 cm) and shoot length (0.572 cm) were obtained in the Gamborg’s B5 medium in combination with Kinetin (0.5 mg/L)+Indole-3-butyric acid (0.5 mg/L)+gelrite. The maximum number of shoots (19.50) and the best multiplication rate (94%) were obtained in the media containing benzyladenin (1 mg/L)+Indole-3-butyric acid (0.5 mg/L) plant growth regulator in Murashige and Skoog medium solidified with agar. At the rooting stage, the maximum number of roots (30) was reached in the Murashige and Skoog medium containing gelrite and the best rooting rate (92%) with agar. A hundred plants representing the best shoot and root growth were taken to acclimatization stage, and 32 of these plants adapted to external conditions.

Keywords:
acclimatization; Helianthemum germanicopolitanum; internodes; micropropagation; risk of extinction

PGR Combinations (mg/L)					MS		Gamborg’s B5		N&N
	BA	KIN	IBA	NAA	Agar	Gelrite	Agar	Gelrite	Agar	Gelrite
1	0	0	0	0	0(i)*	0	0	0(i)	0(i)	0(i)
2	0	0	0.25	0	0.30(ghi)	0	0	0	0	0
3	0	0	0.5	0	0	0.01(i)	0	0	0	0
4	0	0	1	0	0	0	0	0	0	0
5	0	0	0	0	0	0.15(i)	0	0	0	0
6	0	0	0	0.25	0	0	0	0	0	0
7	0	0	0	0.5	0	0	0	0.05(hi)	0	0
8	0	0	0	1	0.20(hi)	0	0	0	0	0.05(hi)
9	0.5	0	0	0	0	0	0.15(i)	0	0	0
10	0.5	0	0.25	0	0	0	0	0	0	0
11	0.5	0	0.5	0	0	0	0.16(i)	0.10(i)	0	0
12	0.5	0	1	0	0	0	0	0	0.20(hi)	0
13	0.5	0	0	0	0	0.21(i)	0	0	0	0
14	0.5	0	0	0.25	0	0	0	0	0	0
15	0.5	0	0	0.5	0	0	0	0.03(i)	0	0
16	0.5	0	0	1	0	0	0	0	0	0
17	1	0	0	0	0	0	0	0	0	0
18	1	0	0.25	0	0	0.16(i)	0.18(i)	0	0	0
19	1	0	0.5	0	0.50(cde)	0	0.49(cde)	0	0	0
20	1	0	1	0	0.02(i)	0	0.05(hi)	0	0	0
21	1	0	0	0	0	0	0	0.10(i)	0	0
22	1	0	0	0.25	0.01(i)	0.05(hi)	0	0	0	0
23	1	0	0	0.5	0	0	0	0	0	0
24	1	0	0	1	0	0	0	0	0	0
25	2	0	0	0	0	0	0	0	0	0.03(i)
26	2	0	0.25	0	0	0	0	0	0.05(hi)	0
27	2	0	0.5	0	0.04(hi)	0	0	0	0	0
28	2	0	1	0	0	0	0	0	0	0
29	2	0	0	0	0	0	0	0	0	0
30	2	0	0	0.25	0	0	0	0	0	0
31	2	0	0	0.5	0	0	0	0	0	0
32	2	0	0	1	0	0	0	0	0	0.03(i)
33	0	0	0	0	0	0	0	0	0	0
34	0	0	0.25	0	0	0	0	0	0	0
35	0	0	0.5	0	0	0.25(ghi)	0	0	0	0
36	0	0	1	0	0	0	0	0	0	0
37	0	0	0	0	0	0	0	0	0	0
38	0	0	0	0.25	0	0	0	0	0	0
39	0	0	0	0.5	0	0	0	0	0	0.03(i)
40	0	0	0	1	0	0	0	0	0	0
41	0	0.5	0	0	0	0	0	0	0	0
42	0	0.5	0.25	0	0	0	0	0	0	0
43	0	0.5	0.5	0	0	0	0	1.14(a)	0	0.09(hi)
44	0	0.5	1	0	0	0	0	0	0	0
45	0	0.5	0	0	0.25(ghi)	0	0	0	0	0
46	0	0.5	0	0.25	0	0	0	0	0	0
47	0	0.5	0	0.5	0	0	0	0	0	0
48	0	0.5	0	1	0	0	0	0	0	0
49	0	1	0	0	0	0	0	0	0	0
50	0	1	0.25	0	0	0	0	0.84(ab)	0	0
51	0	1	0.5	0	0	0.65(bc)	0	0.43(def)	0	0
52	0	1	1	0	0.07(hi)	0	0	0.36(fghi)	0	0
53	0	1	0	0	0	0	0	0.25(ghi)	0	0.35(fghi)
54	0	1	0	0.25	0	0	0	0.75(bc)	0	0.05(hi)
55	0	1	0	0.5	0	0	0	0	0	0
56	0	1	0	1	0	0.05(hi)	0	0.14(hi)	0	0.03(i)
57	0	2	0	0	0	0	0	0.22(ghi)	0	0.36(efg)
58	0	2	0.25	0	0.06(hi)	0	0	0	0.45(cde)	0
59	0	2	0.5	0	0	0	0	0	0	0
60	0	2	1	0	0	0	0	0	0	0.05(hi)
61	0	2	0	0	0	0.09(hi)	0	0	0	0
62	0	2	0	0.25	0	0	0	0.47(cde)	0.05(hi)	0.02(i)
63	0	2	0	0.5	0.07(hi)	0	0.52(cd)	0	0	0.05(hi)
64	0	2	0	1	0	0	0.28(ghi)	0	0	0

PGR Combinations (mg/L)					MS		Gamborg’s B5		N&N
	BA	KIN	IBA	NAA	Agar	Gelrite	Agar	Gelrite	Agar	Gelrite
1	0	0	0	0	0	0	0	0	0(i)	0(i)
2	0	0	0.25	0	0.50(hi)*	0	0	0	0	0
3	0	0	0.5	0	0	0.50(hi)	0	0	0	0
4	0	0	1	0	0	0	0	0	0	0
5	0	0	0	0	0	1.50(ghi)	0	0	0	0
6	0	0	0	0.25	0	0	0	0	0	0
7	0	0	0	0.5	0	0	0	0.30(hi)	0	0
8	0	0	0	1	2.40(ghi)	0	0	0	0	0.10(i)
9	0.5	0	0	0	0	0	3.00(fgh)	0	0	0
10	0.5	0	0.25	0	0	0	0	0	0	0
11	0.5	0	0.5	0	0	0	3.10(fgh)	0.50(hi)	0	0
12	0.5	0	1	0	0	0	0	0	10.00(defg)	0
13	0.5	0	0	0	0	18.10(ab)	0	0	0	0
14	0.5	0	0	0.25	0	0	0	0	0	0
15	0.5	0	0	0.5	0	0	0	0.10(i)	0	0
16	0.5	0	0	1	0	0	0	0	0	0
17	1	0	0	0	0	0	0	0	0	0
18	1	0	0.25	0	0	11.80(def)	0.80(hi)	0	0	0
19	1	0	0.5	0	19.50(a)	0	2.00(gh)	0	0	0
20	1	0	1	0	0.10(i)	0	0.40(hi)	0	0	0
21	1	0	0	0	0	0	0	0.20(i)	0	0
22	1	0	0	0.25	1.50(ghi)	0.10(i)	0	0	0	0
23	1	0	0	0.5	0	0	0	0	0	0
24	1	0	0	1	0	0	0	0	0	0
25	2	0	0	0	0	0	0	0	0	0.50(hi)
26	2	0	0.25	0	0	0	0	0	1.00(hi)	0
27	2	0	0.5	0	0.40(hi)	0	0	0	0	0
28	2	0	1	0	0	0	0	0	0	0
29	2	0	0	0	0	0	0	0	0	0
30	2	0	0	0.25	0	0	0	0	0	0
31	2	0	0	0.5	0	0	0	0	0	0
32	2	0	0	1	0	0	0	0	0	0.10(i)
33	0	0	0	0	0	0	0	0	0	0
34	0	0	0.25	0	0	0	0	0	0	0
35	0	0	0.5	0	0	1.70(ghi)	0	0	0	0
36	0	0	1	0	0	0	0	0	0	0
37	0	0	0	0	0	0	0	0	0	0
38	0	0	0	0.25	0	1.00(hi)	0	0	0	0
39	0	0	0	0.5	0	0	0	0	0	0.20(i)
40	0	0	0	1	0	0	0	0	0	0
41	0	0.5	0	0	0	0	0	0	0	0
42	0	0.5	0.25	0	0	0	0	0	0	0
43	0	0.5	0.5	0	0	0	0	2.35(ghi)	0	15.38(bc)
44	0	0.5	1	0	0	0	0	0	0	0
45	0	0.5	0	0	13.50(cdef)	0	0	0	0	0
46	0	0.5	0	0.25	0	0	0	0	0	0
47	0	0.5	0	0.5	0	0	0	0	0	0
48	0	0.5	0	1	0	0	0	0	0	0
49	0	1	0	0	0	0	0	0	0	0
50	0	1	0.25	0	0	0	0	3.84(fgh)	0	0
51	0	1	0.5	0	0	9.27(de)	0	8.30(efgh)	0	0
52	0	1	1	0	0	0	0	1.78(ghi)	0	0
53	0	1	0	0	0	0	0	0.50(hi)	0	14.50(cde)
54	0	1	0	0.25	0	0	0	1.40(ghi)	0	16.50(bc)
55	0	1	0	0.5	0	0	0	0	0	0
56	0	1	0	1	0	0.10(i)	0	0.70(hi)	0	14.00(cde)
57	0	2	0	0	0	0	0	0.20(i)	0	19.06(a)
58	0	2	0.25	0	13.50(cdef)	0	0	0	18.00(ab)	0
59	0	2	0.5	0	0	0	0	0	0	0
60	0	2	1	0	0	0.40(hi)	0	0	0	0.20(i)
61	0	2	0	0	0	4.50(fghi)	0	0	0	0
62	0	2	0	0.25	0	0	0	3.27(fgh)	8.00(efgh)	0.10(i)
63	0	2	0	0.5	0.30(hi)	0	1.90(gh)	0	0	0.20(i)
64	0	2	0	1	0	0	3.40(fg)	0	0	0

PGR Combinations (mg/L)					MS		Gamborg’s B5		N&N
	BA	KIN	IBA	NAA	Agar	Gelrite	Agar	Gelrite	Agar	Gelrite
1	0	0	0	0	0(g)	0(g)	0	0	0	0
2	0	0	0.25	0	0	0	0	0	0	0
3	0	0	0.5	0	0	0	0	0	0	0
4	0	0	1	0	0	0	0	0	0	0
5	0	0	0	0	0	0	0	0	0	0
6	0	0	0	0.25	0	0	0	0	0	0
7	0	0	0	0.5	0	0	0	0	0	0
8	0	0	0	1	0	0	0	0	0	0.50(a)
9	0.5	0	0	0	0	0	0.40(ab)	0	0	0
10	0.5	0	0.25	0	0	0	0	0	0	0
11	0.5	0	0.5	0	0.10(cef)	0	0	0	0	0
12	0.5	0	1	0	0	0	0	0	0	0
13	0.5	0	0	0	0	0	0	0	0	0
14	0.5	0	0	0.25	0	0	0	0	0	0
15	0.5	0	0	0.5	0	0	0	0	0	0
16	0.5	0	0	1	0	0	0.10(cef)	0	0	0
17	1	0	0	0	0	0	0	0	0	0
18	1	0	0.25	0	0	0	0	0.10(cef)	0	0
19	1	0	0.5	0	0	0	0	0	0	0
20	1	0	1	0	0	0	0	0	0	0
21	1	0	0	0	0	0	0	0	0	0
22	1	0	0	0.25	0	0	0	0	0	0
23	1	0	0	0.5	0	0	0	0	0	0
24	1	0	0	1	0	0	0	0	0	0
25	2	0	0	0	0	0	0	0	0	0
26	2	0	0.25	0	0	0	0	0	0.20(bce)	0
27	2	0	0.5	0	0	0	0	0	0	0
28	2	0	1	0	0	0	0	0	0	0
29	2	0	0	0	0	0	0	0	0	0
30	2	0	0	0.25	0	0	0	0	0	0
31	2	0	0	0.5	0	0	0	0	0	0
32	2	0	0	1	0	0	0	0	0	0
33	0	0	0	0	0	0	0	0	0	0
34	0	0	0.25	0	0	0	0	0	0	0
35	0	0	0.5	0	0	0	0	0	0	0
36	0	0	1	0	0	0	0	0	0	0
37	0	0	0	0	0	0	0	0	0	0
38	0	0	0	0.25	0	0	0	0	0	0
39	0	0	0	0.5	0	0	0	0	0	0
40	0	0	0	1	0	0	0	0	0	0
41	0	0.5	0	0	0	0	0	0	0	0
42	0	0.5	0.25	0	0	0	0	0	0	0
43	0	0.5	0.5	0	0	0	0	0	0	0.07(cef)
44	0	0.5	1	0	0	0	0	0	0	0
45	0	0.5	0	0	0	0	0	0	0	0
46	0	0.5	0	0.25	0	0	0	0	0	0
47	0	0.5	0	0.5	0	0	0	0	0	0
48	0	0.5	0	1	0	0	0	0	0	0
49	0	1	0	0	0	0	0	0	0	0
50	0	1	0.25	0	0	0	0	0	0	0
51	0	1	0.5	0	0	0	0	0	0	0
52	0	1	1	0	0	0	0	0	0	0
53	0	1	0	0	0	0	0	0	0	0
54	0	1	0	0.25	0	0	0	0	0	0
55	0	1	0	0.5	0	0	0	0	0	0
56	0	1	0	1	0	0	0	0	0	0
57	0	2	0	0	0	0	0	0	0	0
58	0	2	0.25	0	0	0	0	0	0	0
59	0	2	0.5	0	0	0	0	0	0	0
60	0	2	1	0	0	0	0	0	0	0
61	0	2	0	0	0	0	0	0	0	0.30(bc)
62	0	2	0	0.25	0	0	0	0	0	0
63	0	2	0	0.5	0	0	0	0	0	0
64	0	2	0	1	0	0	0	0	0	0

PGR Doses	Basal Media
	MS		Gamborg’s B5		N&N
	Agar	Gelrite	Agar	Gelrite	Agar	Gelrite
0 mg/L	2.50(fghi)*	0(i)	0(i)	0(i)	0(i)	0(i)
0.25 mg/L	5.38(fg)	1.83(ghi)	0(i)	0(i)	4.25(fgh)	1.15(hi)
0.5 mg/L	1.25(hi)	30.08(a)	4.00(fgh)	17.25(b)	9.40(cde)	8.80(cde)
1 mg/L	0(i)	1.80(ghi)	4.27(fg)	10.83(cd)	11.65(cd)	12.41(cd)

PGR Doses	Basal Media
	MS		Gamborg’s B5		N&N
	NAA	IBA	NAA	IBA	NAA	IBA
0 mg/L	5.00(de)*	2.25(efg)	0(g)	0(g)	0(g)	0(g)
0.25 mg/L	0.83(fg)	3.66(efg)	0(g)	0(g)	1.40(fg)	4.00(efg)
0.5 mg/L	3.42(efg)	15.00(a)	6.00(de)	16.25(a)	12.33(bc)	5.87(de)
1 mg/L	0(g)	1.80(fg)	5.06(de)	10.04(bc)	12.83(bc)	11.23(bc)

Basal media	Plant Growth Regulator (PGR)
Basal media	NAA	IBA
MS	0.223(e)*	0.685(cde)
Gamborg’s B5	0.538(de)	1.914(a)
N&N	0.749(bcd)	0.972(bc)

PGR	Basal Media
	MS		Gamborg’s B5		N&N
	Agar	Gelrite	Agar	Gelrite	Agar	Gelrite
IBA	0.312(def)*	0.513(bc)	0.046(f)	0.572(ab)	0.325(cdef)	0.416(bcd)
NAA	0.204(ef)	0.405(bcd)	0.599(ab)	0.367(cde)	0.313(def)	0.636(a)

Basal Media	Solidifier
Basal Media	Agar	Gelrite
MS	11.296(cd)*	6.950(e)
Gamborg’s B5	29.842(a)	11.696(cd)
N&N	15.993(b)	10.224(de)

Instituto de Tecnologia do Paraná - Tecpar Rua Prof. Algacyr Munhoz Mader, 3775 - CIC, 81350-010 Curitiba PR Brazil, Tel.: +55 41 3316-3052/3054, Fax: +55 41 3346-2872 - Curitiba - PR - Brazil
E-mail: babt@tecpar.br

Acompanhe os números deste periódico no seu leitor de RSS

[1] *Correspondence: sezgin@karatekin.edu.tr; Tel.: +90 3762189537 (M.S.).