Atmosphere modification system for respiration analysis of recalcitrant seeds during storage

The modification of atmosphere composition is a strategy to prolong the storage of organisms susceptible to senescence. Atmospheres with high levels of carbon dioxide could be applied to prolong the viability of recalcitrant seeds, which are in a constant process of deterioration due to intense metabolic activity. However, there is a need to develop methods that allow the standardized generation of these modified atmospheres to isolate the effect of low oxygen from high carbon dioxide. In this study, the generation of modified atmospheres was carried out by experiments with insufflated gas (IG) or compensated vacuum (CV), on recalcitrant Inga vera embryos as an experimental model. In IG, CO₂ and N₂ were uninterruptedly injected into a mixing chamber, in which the gases underwent turbulence to be homogenized and subsequently applied to the flasks where the embryos were incubated. In CV, the embryos were placed in flasks, from which a fraction of the air present inside was removed, and subsequently, the gases of interest were injected in defined fractions. The CV method proved to be more efficient and it was possible to restrict the respiratory metabolism of the Inga vera embryos by reducing respiration with the increase in carbon dioxide applied exogenously.

Index terms:
conservation; Inga vera ; metabolic control

Authorship

Gabriel Felipe Manoel

Instituto de Pesquisas Ambientais. Avenida Miguel Stefano, 3687, São Paulo - SP, Brasil.Instituto de Pesquisas AmbientaisBrasilSão Paulo, SP, BrasilInstituto de Pesquisas Ambientais. Avenida Miguel Stefano, 3687, São Paulo - SP, Brasil.

http://orcid.org/0009-0007-3150-1915

Marco Aurelio Tiné

http://orcid.org/0000-0002-2575-5149

Marina Crestana Guardia

http://orcid.org/0000-0002-8676-8648

Catarina Carvalho Nievola

http://orcid.org/0000-0003-0627-1357

Claudio José Barbedo ^* *Corresponding author cbarbedo@sp.gov.br

http://orcid.org/0000-0002-4417-7495

*Corresponding author
cbarbedo@sp.gov.br

SCIMAGO INSTITUTIONS RANKINGS

Figures | Tables

Figures (6)
Tables (3)

Thumbnail

Figure 1
Schematic representation of the process of obtaining carbon dioxide. Initially, a syringe was introduced, through a rubber septum, into a sealed 500 mL Kitasato flask containing 100 g of sodium bicarbonate. Through this syringe, as much air as possible was removed from inside the Kitasato flask (A). Then, acetic acid (20%) was added to the Kitasato flask, using a syringe introduced through the septum (B). The reaction of sodium bicarbonate with acetic acid produced carbon dioxide, which, for the compensated vacuum system, was collected in a latex balloon attached with string or elastic to the lateral outlet of the Kitasato flask (C). After filling the balloon, the acid injection stopped.

Thumbnail

Figure 2
Equipment used in the production of gases. In A, Kitasato flask with sodium bicarbonate, syringe with acetic acid and latex balloon for CO₂ production and capture; in B, PET bottle with liquid nitrogen, in a polystyrene box, with a tube for conducting the gaseous N₂ to the latex balloon; in C, vacuum pump for atmospheric air capture.

Thumbnail

Figure 3
Schematic representation of the process of production of modified atmosphere by the insufflated gas system. CO₂, N₂ and atmospheric air are inflated into the gas mixer, controlling the access valves of each gas, until the desired gas mixture is obtained. From there, the access valve to the respirometer where the embryos to be studied are located is opened. For more details, see the description in Material and Methods.

Thumbnail

Figure 4
Equipment used to obtain the modified atmosphere by the insufflated gas system. In A, a glass bottle with a plastic lid containing silica gel, called a mixer; in B, lid with septa for the insertion of the gas cannulas on the left and the outlet of the gas mixture on the right; in C, complete mixer with detail of the gas inlet control valves and mixture outlet needle in the rectangle; in D, mixer outlet needle inserted into the respirometer for insertion under low pressure and extra needle for atmospheric air outlet and pressure maintenance; in E, Illinois 6600 gas analyzer used to check the percentage of gases.

Thumbnail

Figure 5
Schematic representation of the process of production of modified atmosphere by the compensated vacuum system. In A, the removal of air from the respirometer up to the desired O₂ levels; in B, collection of predetermined volumes of CO₂ and N₂ in syringes; in C, insertion of the gases in the respirometer to compose the modified atmosphere. Removal of air and insertion of CO₂ and N₂ were performed using the same syringe needle coupled to a cannula with a valve (not represented in this diagram). For more details, see the description in Material and Methods.

Thumbnail

Figure 6
Equipment used to obtain modified atmosphere by the compensated vacuum system. In A, glass bottle with metal lid (respirometer), seeds and plastic paraffin film; in B, a respirometer with seeds and the needle that collects a sample to measure gases; in C, Illinois 6600 gas analyzer, which shows the percentages of oxygen and carbon dioxide; in D, respirometer already sealed by plastic paraffin film, before being taken to incubation.

Thumbnail

Table 1
O₂ and CO₂ concentrations obtained after processes of production of modified atmosphere by the insufflated gas and compensated vacuum systems.

Thumbnail

Table 2
O₂ consumption, CO₂ release and respiratory quotient (RQ) of Inga vera embryos incubated at two temperatures, two O₂ concentrations and two CO₂ concentrations.

Thumbnail

Table 3
Germination of Inga vera embryos at the end of incubation treatments at different temperatures and different modified atmospheres.

Figure 1 Schematic representation of the process of obtaining carbon dioxide. Initially, a syringe was introduced, through a rubber septum, into a sealed 500 mL Kitasato flask containing 100 g of sodium bicarbonate. Through this syringe, as much air as possible was removed from inside the Kitasato flask (A). Then, acetic acid (20%) was added to the Kitasato flask, using a syringe introduced through the septum (B). The reaction of sodium bicarbonate with acetic acid produced carbon dioxide, which, for the compensated vacuum system, was collected in a latex balloon attached with string or elastic to the lateral outlet of the Kitasato flask (C). After filling the balloon, the acid injection stopped.

Figure 2 Equipment used in the production of gases. In A, Kitasato flask with sodium bicarbonate, syringe with acetic acid and latex balloon for CO₂ production and capture; in B, PET bottle with liquid nitrogen, in a polystyrene box, with a tube for conducting the gaseous N₂ to the latex balloon; in C, vacuum pump for atmospheric air capture.

Figure 3 Schematic representation of the process of production of modified atmosphere by the insufflated gas system. CO₂, N₂ and atmospheric air are inflated into the gas mixer, controlling the access valves of each gas, until the desired gas mixture is obtained. From there, the access valve to the respirometer where the embryos to be studied are located is opened. For more details, see the description in Material and Methods.

Figure 4 Equipment used to obtain the modified atmosphere by the insufflated gas system. In A, a glass bottle with a plastic lid containing silica gel, called a mixer; in B, lid with septa for the insertion of the gas cannulas on the left and the outlet of the gas mixture on the right; in C, complete mixer with detail of the gas inlet control valves and mixture outlet needle in the rectangle; in D, mixer outlet needle inserted into the respirometer for insertion under low pressure and extra needle for atmospheric air outlet and pressure maintenance; in E, Illinois 6600 gas analyzer used to check the percentage of gases.

Figure 5 Schematic representation of the process of production of modified atmosphere by the compensated vacuum system. In A, the removal of air from the respirometer up to the desired O₂ levels; in B, collection of predetermined volumes of CO₂ and N₂ in syringes; in C, insertion of the gases in the respirometer to compose the modified atmosphere. Removal of air and insertion of CO₂ and N₂ were performed using the same syringe needle coupled to a cannula with a valve (not represented in this diagram). For more details, see the description in Material and Methods.

Figure 6 Equipment used to obtain modified atmosphere by the compensated vacuum system. In A, glass bottle with metal lid (respirometer), seeds and plastic paraffin film; in B, a respirometer with seeds and the needle that collects a sample to measure gases; in C, Illinois 6600 gas analyzer, which shows the percentages of oxygen and carbon dioxide; in D, respirometer already sealed by plastic paraffin film, before being taken to incubation.

Table 1 O₂ and CO₂ concentrations obtained after processes of production of modified atmosphere by the insufflated gas and compensated vacuum systems.

System	Attempt	Desired concentrations		Obtained concentrations
System	Attempt	O₂	CO₂	O₂	CO₂
Compensated vacuum	1^st	18%	3%	16%	4%
	2^nd	18%	3%	18%	3%
	3^rd	18%	0%	18%	0%
Insufflated gas	1^st	18%	3%	2%	7%
Insufflated gas	2^nd	18%	3%	8%	5%
3^rd	18%	3%	9%	3%

Table 2 O₂ consumption, CO₂ release and respiratory quotient (RQ) of Inga vera embryos incubated at two temperatures, two O₂ concentrations and two CO₂ concentrations.

Gas concentration, temperature (T) and embryo incubation time (t)				O₂ consumption	CO₂ release	RQ
				by the embryos	by the embryos	RQ
[O₂]	[CO₂]	T	t	(μmol.gDM^-1.day^-1)	(μmol.gDM^-1.day^-1)
20.9%	0.0%	25 °C	1 day	123.53 a^* * Means followed by the same letter in the columns do not differ from each other by Tukey test at 5% probability level.	60.18 a	0.48 c
20.9%	0.0%	5 °C	5 days	24.07 c	12.08 bc	0.50 c
18.0%	0.0%	25 °C	1 day	96.34 b	63.47 a	0.66 b
18.0%	0.0%	5 °C	5 days	6.02 c	10.21 c	1.20 a
18.0%	3.0%	25 °C	1 day	87.49 b	23.09 b	0.26 d
18.0%	3.0%	5 °C	5 days	-	-	-

^*
Means followed by the same letter in the columns do not differ from each other by Tukey test at 5% probability level.

Table 3 Germination of Inga vera embryos at the end of incubation treatments at different temperatures and different modified atmospheres.

Temperature	25 °C			5 °C
O₂ concentration	20.9%	18.0%		20.9%	18.0%
CO₂ concentration	0.0%	0.0%	3.0%	0.0%	0.0%	3.0%
Germination	96% a^* * Means followed by the same letter, in the rows, do not differ from each other by Tukey test at 5% probability level.	99% a	99% a	95% a	95% a	94% a

^*
Means followed by the same letter, in the rows, do not differ from each other by Tukey test at 5% probability level.

How to cite

copy

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

Acompanhe os números deste periódico no seu leitor de RSS

Versão para download de PDF

PDF

Inglês

Versões e tradução automática

Versão original do texto

English

Tradução automática

Google Translator
Microsoft Translator

Como citar

RIS

BIBTEX

Outros formatos de citação e exportação:

SciELO - Scientific Electronic Library Online
Rua Dr. Diogo de Faria, 1087 – 9º andar – Vila Clementino 04037-003 São Paulo/SP - Brasil
E-mail: scielo@scielo.org

Leia a Declaração de Acesso Aberto

Métricas

Atmosphere modification system for respiration analysis of recalcitrant seeds during storage

PlumX

Mensagem

[1] *Corresponding author
cbarbedo@sp.gov.br