<i>In vitro</i> spore germination and gametophyte development of two <i>Cyathea</i> species of South America in response to nutrient media

Marcon, Catiuscia; Lemos, Verônica Kern de; Stein, Isabela Kirch; Droste, Annette

doi:10.1590/2175-7860202374076

Catiuscia Marcon ⁶6Author for correspondence: cati.marcon@hotmail.com

Universidade Feevale, Lab. Biotecnologia Vegetal, Prog. Pós-graduação em Qualidade Ambiental, Novo Hamburgo, RS, Brasil

https://orcid.org/0000-0002-2774-6334

Verônica Kern de Lemos

Universidade Feevale, Lab. Biotecnologia Vegetal, Prog. Pós-graduação em Qualidade Ambiental, Novo Hamburgo, RS, Brasil

https://orcid.org/0009-0004-2686-7574

Isabela Kirch Stein

Universidade Feevale, Lab. Biotecnologia Vegetal, Curso de Ciências Biológicas, Novo Hamburgo, RS, Brasil

https://orcid.org/0000-0003-2449-3500

Annette Droste

Universidade Feevale, Lab. Biotecnologia Vegetal, Prog. Pós-graduação em Qualidade Ambiental, Novo Hamburgo, RS, Brasil

https://orcid.org/0000-0001-8866-1599

6Author for correspondence: cati.marcon@hotmail.com

Area Editor: Dra. Claudine Mynssen

Figures (2)
Tables (4)

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Figure 1
a-c. Location of Cyathea corcovadensis and Cyathea phalerata spore donor populations - a. Brazil, highlighting the state of Rio Grande do Sul in shades of green; b. state of Rio Grande do Sul, highlighting the Atlantic Forest biome in green, the Pampa biome in light green and the location of the municipalities of Três Cachoeiras and Caraá in black; c. in highlight, the municipalities of Três Cachoeiras (1) and Caraá (2) in the Atlantic Forest.

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Figure 2
a-f. Spores and gametophyte development stages of Cyathea corcovadensis - a. non-germinated trilete spore; b. gametophyte with chlorocyte and rhizoid; c. filamentous gametophyte; d-e. laminar gametophyte; f. cordate gametophyte. (c = chlorocyte; r = rhizoid; m = meristematic region). Bars: a-b = 50 µm; c-e = 100 µm; f = 150 µm.

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Table 1
Meyer, Dyer and MS culture media with and without charcoal used for spore germination and gametophyte development of Cyathea corcovadensis and Cyathea phalerata.

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Table 2
Formulations of Meyer, Dyer and MS culture media used for spore germination and gametophyte development of Cyathea corcovadensis and Cyathea phalerata.

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Table 3
Percentage (mean ± standard deviation) of germinated spores (G), laminar gametophytes (LG) and cordate gametophytes (CG) of Cyathea corcovadensis cultivated for up to 90 days in Meyer, Dyer and MS culture media in the absence or presence of activated charcoal. Different letters in the rows and asterisks in the columns indicate that the data differ significantly from each other, according to the Bonferroni test, at 5% significance.

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Table 4
Percentage (mean ± standard deviation) of germinated spores (G), laminar gametophytes (GL) and cordate gametophytes (CG) of Cyathea phalerata cultivated for up to 90 days in Meyer, Dyer and MS culture media in the absence or presence of activated charcoal. Different letters in the rows and asterisks in the columns indicate that the data differ significantly from each other, according to the Bonferroni test at 5% significance.

Culture medium	Activated charcoal	Abbreviation
Meyer	Absence	M
Meyer	Presence	MC
Dyer	Absence	D
Dyer	Presence	DC
MS ¹ 1 MS: Murashige & Skoog (1962) with 50% of the original macronutrient concentration	Absence	50MS
MS with 50% of the original macronutrient concentration	Presence	50MSC
MS with 25% of the original macronutrient concentration	Absence	25MS
MS with 25% of the original macronutrient concentration	Presence	25MSC

Component	Culture medium
Component	Meyer	Dyer	50MS ¹ 1 = with 50% of the macronutrient concentration and without vitamins and amino acids present in the original formulation.	25MS ² 2 = with 25% of the macronutrient concentration and without vitamins and amino acids present in the original formulation.
Macronutrients (mg L^-1)
KH₂PO₄	1000	250	85	42.5
NH₄NO₃	1000	-	825	412.5
MgSO₄ ^. 7H₂O	300	510	185	92.5
CaCl₂ ^. 2H₂O	80	-	220	110
NaCl	100	-	-	-
FeCl₃ ^. 6H₂O	10	-	-	-
KNO₃	-	120	950	475
Ca(NO₃)₂ ^. 4H₂O	-	1,440	-	-
Na₂ EDTA ³ 3 = EDTA: ethylenediamine tetra-acetate.	-	37.3	37.3	37.3
FeSO₄ ^. 7H₂O	-	27.8	27.8	27.8
Micronutrients (mg L^-1)
H₃BO₃	-	-	6.200	6.200
MnSO₄ ^. H₂O	-	-	16.900	16.900
ZnSO₄ ^. 7H₂O	-	-	8.600	8.600
KI	-	-	0.830	0.830
Na₂MoO₄ ^. 2H₂O	-	-	0.250	0.250
CoCl₂ ^. 6H₂O	-	-	0.025	0.025
CuSO₄ ^. 5H₂O	-	-	0.025	0.025

	Day	Activated charcoal ¹ 1 = A: Absence; P: Presence.	Meyer	Dyer	50MS ² 2 = With 50% of the macronutrient concentration and without vitamins and amino acids present in the original formulation.	25MS ³ 3 = With 25% of the macronutrient concentration and without vitamins and amino acids present in the original formulation.	F	p
G	30	A	71.90±5.04^{a^*}	67.20±2.57^{ab^*}	58.10±2.5^{b^*}	59.10±2.71^{b^*}	27.154	<0.001
	30	P	45.40±9.01^a	36.60±10.93^b	16.00±13.82^c	12.20±13.88^c	27.154	<0.001
			F = 357.663				Interaction ⁴ 4 = Indicates interaction between the different culture media and the absence/presence of activated charcoal.
			p < 0.001				3.835	0.013
	60	A	89.70±3.83^{a^*}	87.50±3.63^{a^*}	60.10±5.13^{b^*}	66.90±12.38^{b^*}	42.368	<0.001
	60	P	62.50±9.08^a	50.80±10.96^a	26.90±17.63^b	28.50±15.49^b	42.368	<0.001
			F = 191.078				Interaction
			p < 0.001				1.019	0.389
	90	A	94.00±1.56^{a^*}	86.80±4.13^{ab^*}	74.40±6.45^{bc^*}	70.00±12.23^{c^*}	16.941	<0.001
	90	P	75.80±6.35^a	60.60±9.38^ab	45.10±27.58^b	48.10±18.04^b	16.941	<0.001
			F = 63.304				Interaction
			p < 0.001				0.688	0.562
LG	30	A	59.70±6.78^{a^*}	43.70±3.68^{b^*}	35.90±4.17^{b^*}	37.80±6.14^{b^*}	51.799	<0.001
	30	P	36.60±8.18^a	25.10±7.65^b	9.80±8.21^c	12.80±9.86^c	51.799	<0.001
			F = 212.626				Interaction
			p < 0.001				1.080	0.363
	60	A	76.80±6.30^{a^*}	73.10±5.40^{a^*}	50.60±5.50^{b^*}	58.00±12.60^{b^*}	43.678	<0.001
	60	P	45.70±9.68^a	34.50±11.98^b	14.80±9.61^c	17.20±8.49^c	43.678	<0.001
			F = 324.369				Interaction
			p < 0.001				1.061	0.371
	90	A	64.60±3.02^{ab^*}	70.80±5.59^{a^*}	60.40±3.56^{ab^*}	56.00±11.40^{b^*}	9.612	<0.001
	90	P	43.00±7.36^a	35.90±11.78^ab	22.00±13.30^c	29.60±14.07^bc	9.612	<0.001
			F = 195.901				Interaction
			p < 0.001				3.154	0.030
CG	60	A	7.10±3.1^a	3.70±2.49^ab	0.90±1.28^b	1.70±2.54^b	10.150	<0.001
	60	P	8.40±3.95^a	8.30±3.19^{a^*}	5.10±4.65^a	4.60±3.5^{a^*}	10.150	<0.001
			F = 20.124				Interaction
			p < 0.001				1.056	0.373
	90	A	23.70±2.31^a	10.40±3.20^b	6.30±5.10^b	7.60±3.62^b	33.340	<0.001
	90	P	24.50±7.15^a	17.60±5.50^{b^*}	13.70±9.15^{bc^*}	9.20±4.34^c	33.340	<0.001
			F = 12.100				Interaction
			p = 0.001				2.096	0.108

	Day	Activated charcoal ¹ 1 = A: Absence; P: Presence.	Meyer	Dyer	50MS ² 2 = With 50% of the macronutrient concentration and without vitamins and amino acids present in the original formulation.	25MS ³ 3 = With 25% of the macronutrient concentration and without vitamins and amino acids present in the original formulation.	F	p
G	30	A	6.50±3.89^b	12.20±4.34^a	8.40±3.40^{ab^*}	9.80±2.70^{ab^*}	13.793	<0.001
	30	P	6.30±3.68^b	11.50±3.10^a	2.10±1.19^b	5.80±4.31^b	13.793	<0.001
			F = 13.049				Interaction ⁴ 4 = Indicates interaction between the different culture media and the absence/presence of activated charcoal.
			p = 0.001				3.448	0.021
	60	A	46.60±7.06^a	33.50±11.99^b	38.60±4.06^{ab^*}	35.80±5.24^{b^*}	17.932	<0.001
	60	P	42.60±4.81^a	38.80±8.47^a	19.20±9.25^b	23.80±7.07^b	17.932	<0.001
			F = 19.322				Interaction
			p < 0.001				9.610	<0.001
	90	A	51.30±6.24^{a^*}	46.10±4.56^ab	44.70±3.95^{b^*}	41.90±3.14^{b^*}	25.865	<0.001
	90	P	42.90±2.13^a	44.30±4.64^a	28.30±6.41^b	32.30±5.65^b	25.865	<0.001
			F = 71.082				Interaction
			p < 0.001				7.761	<0.001
LG	30	A	3.10±1.59^b	5.80±2.78^ab	5.00±3.39^{ab^*}	7.40±2.67^{a^*}	9.418	<0.001
	30	P	3.50±2.37^b	7.30±2.36^a	0.90±1.37^b	3.40±2.83^b	9.418	<0.001
			F = 7.674				Interaction
			p = 0.007				6.817	<0.001
	60	A	19.80±6.89^a	21.50±4.72^a	24.30±5.33^{a^*}	19.40±6.38^{a^*}	4.523	0.006
	60	P	15.80±4.86^ab	17.30±6.65^a	4.10±3.14^c	9.60±3.68^bc	4.523	0.006
			F = 63.828				Interaction
			p < 0.001				10.084	<0.001
	90	A	17.90±5.74^{b^*}	23.40±3.56^{a^*}	25.60±3.63^{a^*}	22.70±4.37^{a^*}	10.124	<0.001
	90	P	13.40±2.30^b	19.40±3.53^a	6.80±2.15^c	12.20±2.44^b	10.124	<0.001
			F = 134.261				Interaction
			p < 0.001				17.892	<0.001
CG	60	A	20.90±4.43^a	6.30±3.02^b	8.60±3.06^b	10.60±3.69^b	30.510	<0.001
	60	P	19.30±3.33^a	12.90±3.81^b	9.60±7.18^bc	6.60±5.52^c	30.510	<0.001
			F = 0.251				Interaction
			p = 0.618				5.193	0.003
	90	A	28.10±1.72^a	14.50±5.12^b	14.40±3.09^b	14.90±4.12^b	28.957	<0.001
	90	P	23.90±4.04^a	17.10±5.70^b	13.40±5.10^b	13.90±7.91^b	28.957	<0.001
			F = 0.670				Interaction
			p = 0.416				1.597	0.198

[1] 6Author for correspondence: cati.marcon@hotmail.com

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In vitro spore germination and gametophyte development of two Cyathea species of South America in response to nutrient media

Abstract