The influence of pre-treatment on germination of three species of <i>Catasetum</i> (Orchidaceae)

Asymbiotic germination techniques are successful in species of the Catasetum genus. The diverse composition of the culture media can be critical; however, viable seeds are required for asymbiotic germination. The reliable method for viability assessment is the tetrazolium test. A decisive step in the efficiency of the tetrazolium test is the seeds preconditioning with sucrose solution, which has the potential to improve germination. Therefore, this work aimed to evaluate the seed germination of three species of the genus Catasetum in culture media, preconditioned or not with sucrose solution. Seeds of the species Catasetum osculatum, Ctsm. galeritum and Ctsm. complanatum were assessed by tetrazolium test and germinated in two different media, pretreated or not with 10% sucrose solution. The statistical analysis showed that the use of sucrose pretreatment significantly increased the germination rates of the species. We conclude that the pretreatment with 10% sucrose for 24 hours, regardless of the species and the culture medium, significantly increases the germination of Ctsm. complanatum, Ctsm. galeritum and Ctsm. osculatum seeds.

Index terms:
orchid; plant culture; seeds; sucrose solution; tetrazolium

Authorship

Milena Cristina Moraes ^**Corresponding author E-mail: milena.bio@uol.com.br

Unoeste Campus II, Agronomy College, Rod. Raposo Tavares Km 572, Bairro Limoeiro, Presidente Prudente, SP, 19067-175, Brasil.UnoesteBrasilPresidente Prudente, SP, BrasilUnoeste Campus II, Agronomy College, Rod. Raposo Tavares Km 572, Bairro Limoeiro, Presidente Prudente, SP, 19067-175, Brasil.

https://orcid.org/0000-0002-1887-385X

Monica Rosa Bertão

Universidade Estadual Paulista (Unesp), School of Sciences, Humanities and Languages, Department of Biotechnology, Avenida Dom Antonio, 2100. Assis, SP, 19806-900. Brasil.Universidade Estadual PaulistaBrazilAssis, SP, BrazilUniversidade Estadual Paulista (Unesp), School of Sciences, Humanities and Languages, Department of Biotechnology, Avenida Dom Antonio, 2100. Assis, SP, 19806-900. Brasil.

https://orcid.org/0000-0002-5925-8829

Ceci Castilho Custódio

https://orcid.org/0000-0002-9611-6731

Nelson Barbosa Machado-Neto

https://orcid.org/0000-0003-0914-0686

*Corresponding author
E-mail: milena.bio@uol.com.br

SCIMAGO INSTITUTIONS RANKINGS

Figures | Tables | Formulas

Figures (3)
Tables (4)
Formulas (1)

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Figure 1
Stained seeds of Catasetum complanatum (A), Catasetum galeritum (B) and Catasetum osculatum (C), with the 2,3,5-triphenyl tetrazolium chloride (TZ). Viable seeds show embryo stained in red (red arrows), empty seed are indicated by black arrows and non-viable seed are indicated by white arrows.

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Figure 2
Germination percentage accumulated of Catasetum complanatum (A), Catasetum galeritum (B) and Catasetum osculatum (C) in MS and MM media, with and without pretreatment.

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Figure 3
Germination images of 12 weeks old seeds of Catasetum complanatum (A), Catasetum galeritum (B) and Catasetum osculatum (C), in MS media without pretreatment (1), MS media with pretreatment (2), MM media without pretreatment (3) and MM media with pretreatment (4).

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Table 1
Catasetum species, their place of origin and conservation status.

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Table 2
Tetrazolium test (TZ) results and empty testae percentages of Catasetum species. Values expressed as means and standard error.

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Table 3
Germination average (%) of Catasetum in MS and MM media, with and without pretreatment and their interactions.

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Table 4
Germination (G%) and germination speed index (GSI) of Catasetum species with and without pretreatment in MS and MM media. Values expressed as means and standard error.

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Figure 1 Stained seeds of Catasetum complanatum (A), Catasetum galeritum (B) and Catasetum osculatum (C), with the 2,3,5-triphenyl tetrazolium chloride (TZ). Viable seeds show embryo stained in red (red arrows), empty seed are indicated by black arrows and non-viable seed are indicated by white arrows.

Figure 2 Germination percentage accumulated of Catasetum complanatum (A), Catasetum galeritum (B) and Catasetum osculatum (C) in MS and MM media, with and without pretreatment.

Figure 3 Germination images of 12 weeks old seeds of Catasetum complanatum (A), Catasetum galeritum (B) and Catasetum osculatum (C), in MS media without pretreatment (1), MS media with pretreatment (2), MM media without pretreatment (3) and MM media with pretreatment (4).

Table 1 Catasetum species, their place of origin and conservation status.

Catasetum species	Origin¹ 1 Flora do Brasil 2020, Reflora Project (PETINI-BENELLI, 2020).	Conservation Status¹ 1 Flora do Brasil 2020, Reflora Project (PETINI-BENELLI, 2020).
Ctsm. ² 2 Abbreviations of scientific names followed the rules of Royal Horticultural Society for orchid genera and hybrids (RHS, 2017). osculatum	Pará, Rondônia, Mato Grosso do Sul, Mato Grosso	NE³ 3 NE - Not evaluated
Ctsm. galeritum	Amazonas, Pará, Tocantins, Maranhão, Mato Grosso	NE
Ctsm. complanatum	Rondônia, Mato Grosso	NE

¹
Flora do Brasil 2020, Reflora Project (PETINI-BENELLI, 2020PETINI-BENELLI, A. Catasetum in flora e funga do Brasil. Jardim Botânico do Rio de Janeiro. 2022. https://floradobrasil.jbrj.gov.br/FB11312
https://floradobrasil.jbrj.gov.br/FB1131... ).
²
Abbreviations of scientific names followed the rules of Royal Horticultural Society for orchid genera and hybrids (RHS, 2017RHS. Royal Horticultural Society. Alphabetical list of standard abbreviations for natural and hybrid generic names. 2017. https://www.rhs.org.uk/plant-registration-forms
https://www.rhs.org.uk/plant-registratio... ).
³
NE - Not evaluated

Table 2 Tetrazolium test (TZ) results and empty testae percentages of Catasetum species. Values expressed as means and standard error.

Treatments	TZ	Empty testae^* *Empty testae percentages were based in the total count of seeds. (%)
Ctsm. complanatum	44.13 ± 2.66	28.07 ± 2.24
Ctsm. galeritum	21.13 ± 1.56	5.73 ± 0.81
Ctsm. osculatum	21.95 ± 1.64	53.64 ± 11.87

*Empty testae percentages were based in the total count of seeds.

Table 3 Germination average (%) of Catasetum in MS and MM media, with and without pretreatment and their interactions.

Species	Germination (%)
Catasetum complanatum	31.00 a
Catasetum galeritum	15.37 c
Catasetum osculatum	25.02 b
F	21.58*
Culture medium
Murashige and Skoog	22.91 a
Malmgren	25.24 a
F	1.43 ns
Pretreatment
With	42.25 a
Without	4.75 b
F	373.78*
F_int Species x Culture medium	0.48 ns
F_int Species x Pretreatment	24.50*
F_int Culture medium x Pretreatment	9.49*
F_int Species x Culture medium x Pretreatment	0.56ns
CV (%)	23.85

Means followed by equal letters do not differ by Tukey’s test at 5% probability. *Significant at 1% probability by the F test. CV: coefficient of variation.

Table 4 Germination (G%) and germination speed index (GSI) of Catasetum species with and without pretreatment in MS and MM media. Values expressed as means and standard error.

Treatments	G%		GSI
Treatments	Not Pre-treated	Pre-treated	Not Pre-treated	Pre-treated
Ctsm. complanatum	4.70 ± 1.64 aB	57.31 ± 3.39 aA	0.13 ± 0.05 aB	1.72 ± 0.10 aA
Ctsm. galeritum	2.87 ± 0.92 aB	25.80 ± 2.08 cA	0.06 ± 0.02 aB	0.62 ± 0.05 cA
Ctsm. osculatum	6.39 ± 1.78 aB	43.66 ± 3.16 bA	0.11 ± 0.03 aB	0.87 ± 0.07 bA
MS	1.13 ± 0.31 bB	44.70±2,52 aA	0.03±0.01 bB	1.13 ± 0.07 aA
MM	8.84 ± 1.02 aB	39.81±2.42 aA	0.19±0.02 aB	1.01 ± 0.09 aA

Means followed by the same letter, lowercase for column, uppercase for line, do not differ in Tukey’s test at 5% probability.

G S I = G 1 / N 1 + G 2 / N 2 + \dots + G n / N n

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The influence of pre-treatment on germination of three species of <i>Catasetum</i> (Orchidaceae)

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[1] *Corresponding author
E-mail: milena.bio@uol.com.br