Physiological potential of green bean seeds treated with <i>Bacillus subtilis</i>

Ibanhes Neto, Helio Fernandes; Silva, Ananda Covre da; Sumida, Ciro Hideki; Gouveia, Marjori dos Santos; Pellizzaro, Verônica; Takahashi, Lúcia Sadayo Assari

doi:10.1590/2317-1545v43248603

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ARTICLE • J. Seed Sci. 43 • 2021 • https://doi.org/10.1590/2317-1545v43248603 copy

Physiological potential of green bean seeds treated with Bacillus subtilis

Potencial fisiológico de sementes de feijão-vagem tratadas com Bacillus subtilis

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract:

The association between beneficial microorganisms and green beans can accelerate establishment in the field. The objective of this study was to investigate the influence of seed treatments in different doses of Bacillus subtilis on the physiological potential of green beans. Seeds from the cultivar Macarrão Baixo were used. The study used a completely randomized design in a double factorial scheme with factor A in the treatments: Pelliculation (PC), Pelliculation with Drying (PCD), Pre-Imbibition (PI), Biopriming (BP); factor B, doses of B. subtilis: 0; 7; 14 and 28 mL of a commercial product.kg of seed^-1, and a control. The following treatments were applied: moisture content analysis, first count and final germination, electrical conductivity, length, fresh and dry seedling weight, speed index, and final emergence. Analysis of variance was applied to the data and when significant, it was compared using Tukey’s test and polynomial regression; for the absolute control, orthogonal contrast was applied, with p < 0.05 for all analyses. The treatments did not impair seed viability. Vigor benefited with the use of PI and doses between 16 and 20 mL of p.c.kg of seed^-1. PI alone had the best performance. Biopriming, associated with B. subtilis, favors the vigor of green beans.

Index terms:
Phaseolus vulgaris L.; physiological conditioning; microbiolization; rhizobacteria; emergence of plants

ABRATES - Associação Brasileira de Tecnologia de Sementes Av. Juscelino Kubitschek, 1400 - 3° Andar, sala 31 - Centro,, CEP 86020-000 Londrina/PR - Londrina - PR - Brazil
E-mail: jss@abrates.org.br

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[1] *Corresponding author
E-mail: helioibanhes@hotmail.com

	Initial	After treatment	After drying
	(%)
PC¹ 1 PC: Pelliculation; PCD: Pelliculation with Drying; PI: Pre-imbibition and BP: Biopriming.	11.32	20.08	-
PCD		24.80	11.52
PI		34.24	-
BP		35.61	11.77

	Source of variation	Seed treatment	Dose	Treatment x Dose	Residue	CV (%)	Overall mean
DF		3	3	9	48
MS	NS	120.42	48.25	31.31	51.08	11.92	59.94
	AS	46.75	53.75	45.78	54.46	22.93	32.19
	NGS	36.17	46.17	24.17	24.88	63.33	7.88
	EC	24750.36^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	173.67	154.09	136.61	8.98	130.11
	FGC	231.68^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	75.43	65.15	51.79	12.36	58.23
	RL	2422.86^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	204.02	240.66	126.64	14.23	79.10
	SL	1904.12^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	65.22	303.26^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	62.46	13.87	57.00
	NR	211.38^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	13.08^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	22.21^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	3.74	15.66	12.34
	RFW	2710.45^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	162.61	319.12^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	126.29	14.11	79.64
	SFW	199015.99^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	14777.39	21234.04^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	6615.77	15.38	528.87
	RDW	5.46^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	0.88	0.99^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	0.34	10.32	5.79
	SDW	474.38^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	127.02^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	746.94^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	737.84	15.48	25.33
	ESI	155.62^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	133.59	53.95	31.50	13.39	41.90
	FE	382.06^* * Significant at 5% by Tukey’s test. DF: Degrees of freedom; MS: Mean square; NS: Normal seedlings; AS: Abnormal seedlings; NGS: Non-germinated seeds; EC: Electrical conductivity; FGC: First germination count; RL: Root length; SL: Shoot length; NR: Number of roots; RFW: Root fresh weight; SFW: Shoot fresh weight; RDW: Root dry weight; SDW: Shoot dry weight; ESI: Emergence speed index; FE: Final emergence.	48.40	49.73	60.81	9.93	78.53

Doses (mL cp.kg seed^-1)
		0	7	14	28			0	7	14	28
SL	PC¹ 1 PC: Pelliculation; PCD: pelliculation with drying; PI: Pre-imbibition and BP: Biopriming.	53.5 bc^* * Means followed by different letters in the column are significant at 5% probability by Tukey’s test.	62.3 ab	55.2 a	47.0 b	SFW	PC	469.9 a	305.9 c	429.4 c	353.8 b
	PCD	39.0 c	45.7 c	37.1 b	83.5 a		PCD	555.8 a	504.5 b	535.6 bc	530.2 a
	PI	71.6 a	71.6 a	56.3 a	58.2 b		PI	531.1 a	744.6 a	719.8 a	650.4 a
	BP	54.7 b	54.5 bc	60.1 a	61.3 b		BP	440.3 a	523.7 b	597.2 ab	574.9 a
NR	PC	13.2 a	10.4 b	11.9 b	9.3 b	RDW	PC	5.5 a	4.4 b	5.6 a	4.3 b
	PCD	9.5 b	8.5 b	11.0 b	9.6 b		PCD	5.5 a	5.4 b	5.9 a	5.6 a
	PI	12.8 ab	22.1 a	18.4 a	17.5 a		PI	5.6 a	6.6 a	6.2 a	6.1 a
	BP	9.2 b	11.5 b	10.7 b	11.9 b		BP	5.4 a	6.6 a	6.6 a	6.4 a
RFW	PC	73.8 a	51.5 b	65.9 b	50.9 b	SDW	PC	22.3 ab	15.4 c	21.8 b	16.5 b
	PCD	82.6 a	79.0 a	86.4 ab	81.6 a		PCD	25.5 a	20.3 bc	22.8 b	28.4 a
	PI	72.7 a	90.4 a	91.1 a	90.2 a		PI	25.2 a	33.7 a	36.0 a	33.9 a
	BP	78.0 a	93.5 a	93.4 a	93.2 a		BP	17.2 b	23.8 b	31.5 a	31.3 a

		Variables	R²	Maximum dose (mL.kg seed^-1)	Maximum Effect
SL	PC	$Ŷ = - 0.46 x + 49.51$ ^* * Significant at 5% by Tukey’s test.	54.38	0.0	49.5
	PCD	$Ŷ = 57.9$	-	-	57.9
	PI	$Ŷ = - 0.08 x 2 + 2.51 x + 59.14$ ^* * Significant at 5% by Tukey’s test.	53.33	16.7	80.0
	BP	$Ŷ = 55.71$	-	-	55.7
NR	PC	$Ŷ = - 0.11 x + 12.58$ ^* * Significant at 5% by Tukey’s test.	63.24	0.0	12.6
	PCD	$Ŷ = 9.7$	-	-	9.7
	PI	$Ŷ = - 0.03 x 2 + 0.87 x + 14.03$ ^* * Significant at 5% by Tukey’s test.	53.33	16.0	21.0
	BP	$Ŷ = 10.8$	-	-	10.8
RFW	PC	$Ŷ = 60.5$	-	-	60.5
	PCD	$Ŷ = 82.4$	-	-	82.4
	PI	$Ŷ = 0.61 x + 79.35$ ^* * Significant at 5% by Tukey’s test.	59.34	28.0	93.2
	BP	$Ŷ = 88.8$	-	-	88.8
SFW	PC	$Ŷ = 388.5$	-	-	388.5
	PCD	$Ŷ = 531.5$	-	-	531.5
	PI	$Ŷ = - 0.82 x 2 + 26.30 x + 550.08$ ^* * Significant at 5% by Tukey’s test.	83.98	16.1	760.8
	BP	$Ŷ = - 0.42 x 2 + 16.89 x + 436.45$ ^* * Significant at 5% by Tukey’s test.	98.79	19.7	602.7
RDW	PC	$Ŷ = 5.0$	-	-	5.0
	PCD	$Ŷ = 5.7$	-	-	5.7
	PI	$Ŷ = 6.2$	-	-	6.2
	BP	$Ŷ = - 0.01 x 2 + 0.14 x + 5.56$ ^* * Significant at 5% by Tukey’s test.	89.87	17.9	6.9
SDW	PC	$Ŷ = 18.0$	-	-	18.0
	PCD	$Ŷ = 0.03 x 2 + 0.60 x + 24.86$ ^* * Significant at 5% by Tukey’s test.	87.32	28.0	28.4
	PI	$Ŷ = - 0.04 x 2 + 1.30 x + 25.53$ ^* * Significant at 5% by Tukey’s test.	98.24	18.1	37.2
	BP	$Ŷ = - 0.033 x 2 + 1.44 x + 16.67$ ^* * Significant at 5% by Tukey’s test.	97.68	22.1	32.6

Treatment	PC	PCD	PI	BP
NS	58.25	57.38	63.50	60.63
AS	32.00	34.50	30.38	31.38
NGS	9.75	8.13	6.13	7.50
EC	137.52 b	174.27 a	78.77 c	129.88 b
FGC	58.00 ab	53.38 b	62.63 a	58.94 ab
RL	68.68 b	72.23 b	96.38 a	79.12 b
SL	43.88 c	57.91 b	70.50 a	55.71 b
NR	11.19 b	9.66 b	17.71 a	10.81 b
RFW	60.53 b	82.40 a	86.86 a	88.76 a
SFW	388.51 c	531.5 b	661.46 a	534.02 b
RDW	5.01 c	5.67 b	6.19 ab	6.28 a
SDW	18.98 c	24.22 b	32.19 a	25.93 b
ESI	37.58 b	44.67 a	41.77 ab	43.60 a
FE	73.25 b	82.63 a	75.50 ab	82.75 a

	EC	FGC	RL	SL	NR	RFW	SFW	RDW	SDW	ESI	FE
Treat./Dose	Means
AC	129.92	61.5	64.88	49.45	10.4	57.7	446.45	4.97	23.17	39.73	75
	Estimates
PC/0.0	-3.15	-7	10.6	4.1	2.78^* * Significant at 5%.	16.11	18.45	0.62	-0.9	0.16	2.5
PC/7.0	2.01	1	5.1	12.9^* * Significant at 5%.	-0.03	-6.21	-140.54^* * Significant at 5%.	-0.5	-7.77^* * Significant at 5%.	-0.19	0.5
PC/14.0	9.64	-7	16.8	5.8	1.53	8.2	-17.06	0.64	-1.43	-4.39	-9
PC/28.0	21.89^* * Significant at 5%.	-1	6.6	-2.5	-1.1	-6.79	-92.62	-0.58	-6.66^* * Significant at 5%.	-4.17	-1
PCD/0.0	41.84^* * Significant at 5%.	-3	-7.1	-10.4	-0.88	24.87^* * Significant at 5%.	109.32	0.59	2.3	0.02	6
PCD/7.0	48.59^* * Significant at 5%.	-13	9.7	-3.7	-1.9	21.34^* * Significant at 5%.	58.02	0.5	-2.89	10.05^* * Significant at 5%.	11.50^* * Significant at 5%.
PCD/14.0	44.97^* * Significant at 5%.	-7	2	-12.4^* * Significant at 5%.	0.6	28.66^* * Significant at 5%.	89.11	1.00^* * Significant at 5%.	-0.39	1.08	4.5
PCD/28.0	41.97^* * Significant at 5%.	-10	43.3^* * Significant at 5%.	34.1^* * Significant at 5%.	-0.78	23.93^* * Significant at 5%.	83.76	0.71	5.18	8.60^* * Significant at 5%.	8.5
PI/0.0	-47.42^* * Significant at 5%.	2	35.0^* * Significant at 5%.	22.1^* * Significant at 5%.	2.4	15.02	84.67	0.71	2.05	-0.16	-2
PI/7.0	-57.25^* * Significant at 5%.	3	31.3^* * Significant at 5%.	22.2^* * Significant at 5%.	11.68^* * Significant at 5%.	32.67^* * Significant at 5%.	298.15^* * Significant at 5%.	1.71^* * Significant at 5%.	10.50^* * Significant at 5%.	2.87	-1
PI/14.0	-52.89^* * Significant at 5%.	-1	8	6.9	8.03^* * Significant at 5%.	33.41^* * Significant at 5%.	273.33^* * Significant at 5%.	1.29^* * Significant at 5%.	12.85^* * Significant at 5%.	2.3	4
PI/28.0	-47.06^* * Significant at 5%.	1	9.2	8.8	7.13^* * Significant at 5%.	35.53^* * Significant at 5%.	203.90^* * Significant at 5%.	1.21^* * Significant at 5%.	10.69^* * Significant at 5%.	3.15	1
BP/0.0	-0.65	2	7.2	5.3	-1.2	20.28^* * Significant at 5%.	-6.21	0.51	-5.99^* * Significant at 5%.	-3.1	4.5
BP/7.0	-3.35	-3	11.9	5.1	1.05	35.77^* * Significant at 5%.	77.26	1.63^* * Significant at 5%.	0.62	3.81	10.5
BP/14.0	2.91	-11	20.7^* * Significant at 5%.	10.6	0.3	35.70^* * Significant at 5%.	150.74^* * Significant at 5%.	1.64^* * Significant at 5%.	8.32^* * Significant at 5%.	3.53	6.5
BP/28.0	0.94	2	17.8^* * Significant at 5%.	11.8	1.5	32.49^* * Significant at 5%.	128.48^* * Significant at 5%.	1.48^* * Significant at 5%.	8.08^* * Significant at 5%.	11.23*	9.5