Tetrazolium test for <i>Pinus taeda</i>: preparation, staining, and seed viability classes

The objective of this work was to evaluate preparation and staining methods used in the tetrazolium test for Pinus taeda seeds, with the definition of viability classes. The procedures carried out were those described in the rules for seed testing in Brazil and Association of Official Seed Analysts, as well as an alternative method for the tetrazolium test. For each procedure, different combinations of types of seed cuts, concentrations of tetrazolium salt, and exposure periods were used. The test conducted with the alternative procedure (longitudinal cut adjacent to the embryo and staining of only the half with the embryo) is the most practical. The obtained results allowed classifying seeds into two different viability classes (viable and nonviable), and 0.2% tetrazolium salt for 4 hours is the most effective combination.

Index terms:
germination; Pinaceae; physiological quality

Authorship

Felipe Serrato dos Santos ^**Corresponding author: felipeserrato@hotmail.com

Universidade Federal do Paraná, Departamento de Fitotecnia e Fitossanitarismo, Rua dos Funcionários, no 1.540, Juvevê, CEP 80035-050 Curitiba, PR, Brazil. E-mail: felipeserrato@hotmail.com, maristela@ufpr.brUniversidade Federal do ParanáBrazilCuritiba, PR, Brazil Universidade Federal do Paraná, Departamento de Fitotecnia e Fitossanitarismo, Rua dos Funcionários, no 1.540, Juvevê, CEP 80035-050 Curitiba, PR, Brazil. E-mail: felipeserrato@hotmail.com, maristela@ufpr.br

http://orcid.org/0000-0003-0801-0329

Elisa Serra Negra Vieira

Embrapa Florestas, Estrada da Ribeira, Km 111, Guaraituba, Caixa Postal 319, CEP 83411-000 Colombo, PR, Brazil. E-mail: elisa.vieira@embrapa.brEmbrapa FlorestasBrazilColombo, PR, Brazil Embrapa Florestas, Estrada da Ribeira, Km 111, Guaraituba, Caixa Postal 319, CEP 83411-000 Colombo, PR, Brazil. E-mail: elisa.vieira@embrapa.br

http://orcid.org/0000-0002-0799-7654

Maristela Panobianco

http://orcid.org/0000-0002-9990-2172

*Corresponding author: felipeserrato@hotmail.com

SCIMAGO INSTITUTIONS RANKINGS

Figures | Tables

Figures (2)
Tables (1)

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Figure 1.
Types of cuts performed on Pinus taeda seeds: A and B, transversal cut at the distance of one third from the endosperm distal end; C, longitudinal cut through the endosperm to expose the embryo; D, longitudinal cut adjacent to the embryo, keeping the two halves of the seed together; and E, longitudinal cut adjacent to the embryo, separating half of the seed containing the embryo.

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Figure 2.
Classes of viable Pinus taeda seeds, showing: A, intense red color in the endosperm and embryo; B, carmine red color along the endosperm and embryo; C, carmine red color with a less intense tonality; D, light-red color with transparent spots on the endosperm, not affecting the embryo; E, light-red endosperm and embryo with flaccid tissue; and F, carmine red embryo with small light-red spots on the embryonic axis. Classes of nonviable seeds, showing: G, endosperm and embryo with whitish color; H, seeds that did not color after the process of exposure to tetrazolium salt; I, most of the embryonic axis uncolored and with small colored spots; J, red spots on the endosperm and hypocotyl region; K, uncolored cotyledons; and L, uncolored radicle tip.

Thumbnail

Table 1.
Viability of Pinus taeda seeds when subjected to the germination test and to the tetrazolium test with different combinations of methodologies⁽¹⁾.

Figure 1. Types of cuts performed on Pinus taeda seeds: A and B, transversal cut at the distance of one third from the endosperm distal end; C, longitudinal cut through the endosperm to expose the embryo; D, longitudinal cut adjacent to the embryo, keeping the two halves of the seed together; and E, longitudinal cut adjacent to the embryo, separating half of the seed containing the embryo.

Figure 2. Classes of viable Pinus taeda seeds, showing: A, intense red color in the endosperm and embryo; B, carmine red color along the endosperm and embryo; C, carmine red color with a less intense tonality; D, light-red color with transparent spots on the endosperm, not affecting the embryo; E, light-red endosperm and embryo with flaccid tissue; and F, carmine red embryo with small light-red spots on the embryonic axis. Classes of nonviable seeds, showing: G, endosperm and embryo with whitish color; H, seeds that did not color after the process of exposure to tetrazolium salt; I, most of the embryonic axis uncolored and with small colored spots; J, red spots on the endosperm and hypocotyl region; K, uncolored cotyledons; and L, uncolored radicle tip.

Table 1. Viability of Pinus taeda seeds when subjected to the germination test and to the tetrazolium test with different combinations of methodologies⁽¹⁾.

Assessment		Viability (%)
Germination test		77a
Tetrazolium test
	Procedure/cut/concentration/exposure period/temperature
	Alternative/longitudinal cut discarding one of the halves/0.2%/4 hours/35°C	74a
	Alternative/longitudinal cut discarding one of the halves/1.0%/4 hours/35°C	74a
	Baalbaki et al. (2009)BAALBAKI, R.; ELIAS, S.; MARCOS-FILHO, J.; MCDONALD, M.B. (Ed.). Seed vigor testing handbook. Ithaca: Association of Official Seed Analysts, 2009. 341p. (AOSA. Contribution, 32). /longitudinal cut keeping the two halves together/1.0%/4 hours/35°C	80a
	Alternative/longitudinal cut discarding one of the halves/0.5%/6 hours/35°C	73a
	Alternative/longitudinal cut discarding one of the halves/1.0%/6 hours/35°C	73a
	Brasil (2009)BRASIL. Ministério da Agricultura, Pecuária e Abastecimento. Regras para análise de sementes. Brasília, 2009. 395p./transversal and longitudinal cuts/1.0%/18 hours/30°C	72a
	Alternative/longitudinal cut discarding one of the halves/1.0%/2 hours/35°C	69b
	Alternative/longitudinal cut discarding one of the halves/0.5%/4 hours/35°C	68b
	Alternative/longitudinal cut discarding one of the halves/0.2%/6 hours/35°C	67b
	Alternative/longitudinal cut discarding one of the halves/0.2%/2 hours/35°C	63c
	Alternative/longitudinal cut discarding one of the halves/0.5%/2 hours/35°C	57c
Coefficient of variation (%)		6.10

⁽¹⁾
Means followed by equal letters do not differ by the Scott-Knott test, at 1% probability.

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Tetrazolium test for <i>Pinus taeda</i>: preparation, staining, and seed viability classes

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[1] *Corresponding author: felipeserrato@hotmail.com