Cryopreservation of primed tobacco seeds

Priming is a commercial technique used to increase the speed and uniformity of seed germination. However, the physiological quality of primed seeds is usually negatively affected during storage. Tobacco seeds of the cultivar BAT 2101 were used to investigate whether primed seeds could be cryopreserved. The most suitable substances, type of drying, and reheating during priming and cryopreservation of tobacco seeds were studied. Seed priming was performed with water, spermidine, and potassium nitrate, and drying was carried out with silica gel and a saturated saline solution. Seeds were cryopreserved in liquid nitrogen for 24 h and reheated in a water bath for two and five minutes and at ambient temperature. Tobacco seeds primed with spermidine and water can be cryopreserved without loss of physiological quality when quickly dried on silica gel and reheated in a water bath for two minutes.

Index terms:
germination; emergence; Nicotiana tabacum; reheating; drying

Authorship

Camila Aparecida Lopes ^**Corresponding author <camilalopes_sjc@hotmail.com>

Departamento de Agricultura, Universidade Federal de Lavras, Caixa Postal 3037, 37200-000 - Lavras, MG, Brasil.Universidade Federal de LavrasBrazilLavras, MG, BrazilDepartamento de Agricultura, Universidade Federal de Lavras, Caixa Postal 3037, 37200-000 - Lavras, MG, Brasil.

Maria Laene Moreira de Carvalho

Ana Cristina de Souza

João Almir Oliveira

Dayliane Bernardes de Andrade

*Corresponding author <camilalopes_sjc@hotmail.com>

SCIMAGO INSTITUTIONS RANKINGS

Tables

Tables (8)

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Table 1
Water content (%) of tobacco (Nicotiana tabacum) seeds of the cultivar BAT2101 after different forms of priming and drying.

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Table 2
Electrical conductivity (µS. cm⁻¹. g⁻¹) of primed and cryopreserved tobacco (Nicotiana tabacum) seeds of the cultivar BAT 2101 for the factors priming and reheating (A) and drying (B).

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Table 3
First germination count (%) of primed and cryopreserved tobacco (Nicotiana tabacum) seeds of the cultivar BAT 2101 for the factors priming and reheating (A) and drying (B).

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Table 4
Germination (%) of primed and cryopreserved tobacco (Nicotiana tabacum) seeds of the cultivar BAT 2101 for the factors priming and reheating (A) and drying (B).

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Table 5
Germination rate index of primed and cryopreserved tobacco (Nicotiana tabacum) seeds of the cultivar BAT 2101 for the factors priming and reheating (A) and drying (B).

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Table 6
Initial emergence (%) of primed and cryopreserved tobacco (Nicotiana tabacum) seeds of the cultivar BAT 2101 for the factors priming and reheating (A) and drying (B).

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Table 7
Final emergence (%) of primed and cryopreserved tobacco (Nicotiana tabacum) seeds of the cultivar BAT 2101 for the factors priming and reheating (A) and drying (B).

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Table 8
Emergence rate index of primed and cryopreserved tobacco (Nicotiana tabacum) seeds of the cultivar BAT 2101 for the factors priming and reheating (A) and drying (B).

Table 1 Water content (%) of tobacco (Nicotiana tabacum) seeds of the cultivar BAT2101 after different forms of priming and drying.

Priming	Drying	Water content (%)
Priming	Drying	After priming	After drying
Water	Slow	55.49	6.20
Water	Fast	55.49	6.10
SPD	Slow	54.51	6.07
SPD	Fast	54.51	5.93
KNO₃	Slow	55.39	6.31
KNO₃	Fast	55.39	6.11
Control	-	-	6.00

Table 2 Electrical conductivity (µS. cm⁻¹. g⁻¹) of primed and cryopreserved tobacco (Nicotiana tabacum) seeds of the cultivar BAT 2101 for the factors priming and reheating (A) and drying (B).

A
Priming	Drying	Reheating
Priming	Drying	Ambient temp	WB: 2 min	WB: 5 min
Water	Slow	181.68 bC	152.95 aB	147.40 aA
SPD	Slow	148.44 aA	221.02 bC	264.62 bC
KNO₃	Slow	551.40 cC	426.02 cA	485.94 cB
Water	Fast	155.00 aB	147.13 aA	237.70 bC
SPD	Fast	165.19 bB	156.38 bA	186.61 aC
KNO₃	Fast	427.34 cA	510.92 cB	545.81 cC
B
Priming	Reheating		Drying
Priming	Reheating		Slow	Fast
Water	Ambient temp		181.68 B	155.00 A
Water	WB: 2 min		152.95 B	147.13 A
Water	WB: 5 min		147.40 A	237.70 B
SPD	Ambient temp		148.44 A	165.19 B
SPD	WB: 2 min		221.02 B	156.38 A
SPD	WB: 5 min		264.62 B	186.61 A
KNO₃	Ambient temp		551.40 B	427.34 A
KNO₃	WB: 2 min		426.02 A	510.92 B
KNO₃	WB: 5 min		485.94 A	545.81 B
CV (%) = 5.73

Means followed by the same lowercase letter in the column and uppercase letter in the row do not differ by Scott-Knott test at 5% probability level.

Table 3 First germination count (%) of primed and cryopreserved tobacco (Nicotiana tabacum) seeds of the cultivar BAT 2101 for the factors priming and reheating (A) and drying (B).

A
Priming	Drying	Reheating
Priming	Drying	Ambient temp	WB: 2 min	WB: 5 min
Water	Slow	84 aB	82 aB	90 aA
SPD	Slow	84 aA	83 aA	65 bB
KNO₃	Slow	67 bA	60 bB	59 cB
Water	Fast	83 bB	90 aA	83 cB
SPD	Fast	90 aA	92 aA	87 bB
KNO₃	Fast	89 aA	92 aA	91 aA
B
Priming	Reheating		Drying
Priming	Reheating		Slow	Fast
Water	Ambient temp		84 A	83 A
Water	WB: 2 min		82 B	90 A
Water	WB: 5 min		90 A	83 B
SPD	Ambient temp		84 B	90 A
SPD	WB: 2 min		83 B	92 A
SPD	WB: 5 min		65 B	87 A
KNO₃	Ambient temp		69 B	89 A
KNO₃	WB: 2 min		60 B	92 A
KNO₃	WB: 5 min		59 B	91 A
CV (%) = 12.49

Means followed by the same lowercase letter in the column and uppercase letter in the row do not differ by Scott-Knott test at 5% probability level.

Table 4 Germination (%) of primed and cryopreserved tobacco (Nicotiana tabacum) seeds of the cultivar BAT 2101 for the factors priming and reheating (A) and drying (B).

A
Priming	Drying	Reheating
Priming	Drying	Ambient temp	WB: 2 min	WB: 5 min
Water	Slow	85 bB	82 bC	92 aA
SPD	Slow	89 aA	88 aA	68 bB
KNO₃	Slow	72 cA	61 cB	61 cB
Water	Fast	89 aB	92 aA	89 cB
SPD	Fast	91 aB	88 bC	96 aA
KNO₃	Fast	91 aA	93 aA	92 bA
B
Priming	Reheating		Drying
Priming	Reheating		Slow	Fast
Water	Ambient temp		85 B	89 A
Water	WB: 2 min		82 B	92 A
Water	WB: 5 min		92 A	89 A
SPD	Ambient temp		89 A	91 A
SPD	WB: 2 min		88 A	88 A
SPD	WB: 5 min		68 B	96 A
KNO₃	Ambient temp		72 B	91 A
KNO₃	WB: 2 min		61 B	93 A
KNO₃	WB: 5 min		61 B	92 A
CV (%) = 12.02

Means followed by the same lowercase letter in the column and uppercase letter in the row do not differ by Scott-Knott test at 5% probability level.

Table 5 Germination rate index of primed and cryopreserved tobacco (Nicotiana tabacum) seeds of the cultivar BAT 2101 for the factors priming and reheating (A) and drying (B).

A
Priming	Drying	Reheating
Priming	Drying	Ambient temp	WB: 2 min	WB: 5 min
Water	Slow	14.19 aA	10.33 bB	16.17 aA
SPD	Slow	16.29 aA	16.47 aA	11.42 bB
KNO₃	Slow	11.29 bA	10.49 bA	11.17 bA
Water	Fast	14.42 aA	15.12 aA	14.46 aA
SPD	Fast	15.38 aA	15.09 aA	15.73 aA
KNO₃	Fast	15.20 aA	16.16 aA	15.92 aA
B
Priming	Reheating		Drying
Priming	Reheating		Slow	Fast
Water	Ambient temp		14.19 A	14.42 A
Water	WB: 2 min		10.33 B	15.12 A
Water	WB: 5 min		16.17 A	14.46 A
SPD	Ambient temp		16.29 A	15.38 A
SPD	WB: 2 min		16.47 A	15.09 A
SPD	WB: 5 min		11.42 B	15.73 A
KNO₃	Ambient temp		11.29 B	15.20 A
KNO₃	WB: 2 min		10.49 B	16.16 A
KNO₃	WB: 5 min		11.17 B	15.92 A
CV (%) = 14.97

Means followed by the same lowercase letter in the column and uppercase letter in the row do not differ by Scott-Knott test at 5% probability level.

Table 6 Initial emergence (%) of primed and cryopreserved tobacco (Nicotiana tabacum) seeds of the cultivar BAT 2101 for the factors priming and reheating (A) and drying (B).

A
Priming	Drying	Reheating
Priming	Drying	Ambient temp	WB: 2 min	WB: 5 min
Water	Slow	85 aB	83 bB	88 aA
SPD	Slow	84 aB	88 aA	66 bC
KNO₃	Slow	56 bB	57 cB	59 cA
Water	Fast	88 aA	91 aA	89 bA
SPD	Fast	89 aB	92 aA	89 bB
KNO₃	Fast	88 aB	83 bC	93 aA
B
Priming	Reheating		Drying
Priming	Reheating		Slow	Fast
Water	Ambient temp		85 B	88 A
Water	WB: 2 min		83 B	91 A
Water	WB: 5 min		88 A	89 A
SPD	Ambient temp		84 B	89 A
SPD	WB: 2 min		88 B	92 A
SPD	WB: 5 min		66 B	89 A
KNO₃	Ambient temp		56 B	88 A
KNO₃	WB: 2 min		57 B	83 A
KNO₃	WB: 5 min		59 B	93 A
CV (%) = 11.44

Means followed by the same lowercase letter in the column and uppercase letter in the row do not differ by Scott-Knott test at 5% probability level.

Table 7 Final emergence (%) of primed and cryopreserved tobacco (Nicotiana tabacum) seeds of the cultivar BAT 2101 for the factors priming and reheating (A) and drying (B).

A
Priming	Drying	Reheating
Priming	Drying	Ambient temp	WB: 2 min	WB: 5 min
Water	Slow	87 aB	84 bC	90 aA
SPD	Slow	85 bB	89 aA	67 bC
KNO₃	Slow	59 cB	59 cB	62 cA
Water	Fast	92 aA	92 aA	90 cA
SPD	Fast	92 aA	93 aA	93 aA
KNO₃	Fast	91 aA	84 bC	94 aA
B
Priming	Reheating		Drying
Priming	Reheating		Slow	Fast
Water	Ambient temp		87 B	92 A
Water	WB: 2 min		84 B	92 A
Water	WB: 5 min		90 A	90 A
SPD	Ambient temp		85 B	92 A
SPD	WB: 2 min		89 B	93 A
SPD	WB: 5 min		67 B	93 A
KNO₃	Ambient temp		59 B	91 A
KNO₃	WB: 2 min		59 B	84 A
KNO₃	WB: 5 min		62 B	94 A
CV (%) = 11.26

Means followed by the same lowercase letter in the column and uppercase letter in the row do not differ by Scott-Knott test at 5% probability level.

Table 8 Emergence rate index of primed and cryopreserved tobacco (Nicotiana tabacum) seeds of the cultivar BAT 2101 for the factors priming and reheating (A) and drying (B).

A
Priming	Drying	Reheating
Priming	Drying	Ambient temp	WB: 2 min	WB: 5 min
Water	Slow	16.70 aA	16.47 aA	17.33 aA
SPD	Slow	16.16 aA	17.13 aA	12.14 bB
KNO₃	Slow	11.32 bA	12.87 bA	11.21 bA
Water	Fast	16.78 aA	17.07 aA	17.30 aA
SPD	Fast	17.15 aA	17.64 aA	16.39 aA
KNO₃	Fast	16.54 aA	14.59 bB	16.94 aA
B
Priming	Reheating		Drying
Priming	Reheating		Slow	Fast
Water	Ambient temp		16.70 A	16.78 A
Water	WB: 2 min		16.47 A	17.06 A
Water	WB: 5 min		17.32 A	17.30 A
SPD	Ambient temp		16.16 A	17.15 A
SPD	WB: 2 min		17.13 A	17.64 A
SPD	WB: 5 min		12.14 B	16.39 A
KNO₃	Ambient temp		11.32 B	16.54 A
KNO₃	WB: 2 min		12.87 B	14.59 A
KNO₃	WB: 5 min		11.21 B	16.94 A
CV (%) = 6.50

Means followed by the same lowercase letter in the column and uppercase letter in the row do not differ by Scott-Knott test at 5% probability level.

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Cryopreservation of primed tobacco seeds

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[1] *Corresponding author <camilalopes_sjc@hotmail.com>