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Cited by 8Seed Germination and Physiologythermodynamics and calorimetric analysesPhysiological and biochemical adaptations

Exploring the Applicability of Calorespirometry to Assess Seed Metabolic Stability Upon Temperature Stress Conditions—Pisum sativum L. Used as a Case Study

Open access
Rodrigues, Lénia, Nogales, Amaia, Hansen, Lee D., Santos, Fátima, Rato, Ana Elisa, Cardoso, Hélia·2022·Frontiers in Plant Science
DOI 10.3389/fpls.2022.827117PMC PMC9094064
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Abstract

The availability of phenotyping tools to assist breeding programs in the selection of high-quality crop seeds is of obvious interest with consequences for both seed producers and consumers. Seed germination involves the activation of several metabolic pathways, such as cellular respiration to provide the required ATP and reducing power. This work tested the applicability of calorespirometry, the simultaneous measurement of heat and CO2 rates, as a phenotyping tool to assess seed respiratory properties as a function of temperature. The effect of temperature on seed germination was evaluated after 16 h of seed imbibition by calorespirometric experiments performed in isothermal mode at 15, 20, 25, and 28°C on the seeds of three cultivars of peas (Pisum sativum L.) commonly used in conventional agriculture (cvs. ‘Rondo’, ‘Torta de Quebrar’, and ‘Maravilha d’América’). Significant differences in metabolic heat rate and CO2 production rate (RCO2) as well as in the temperature responses of these parameters were found among the three cultivars. A seed germination trial was conducted during the 6 days of imbibition to evaluate the predictive power of the parameters derived from the calorespirometric measurements. The germination trial showed that the optimal germination temperature was 20°C and low germination rates were observed at extreme temperatures (15 or 28°C). The cv. ‘Torta de Quebrar’ showed significantly higher germination in comparison with the other two cultivars at all three temperatures. In comparison with the other two cultivars, ‘Torta de Quebrar’ has the lowest metabolic heat and CO2 rates and the smallest temperature dependence of these measured parameters. Additionally, ‘Torta de Quebrar’ has the lowest values of growth rate and carbon use efficiency calculated from the measured variables. These data suggest that calorespirometry is a useful tool for phenotyping physiologic efficiency at different temperatures during early germination stages, and can determine the seeds with the highest resilience to temperature variation, in this case ‘Torta de Quebrar’.

AI snapshot

Written from the paper’s full text.

TL;DR

This study tested calorespirometry as a phenotyping tool for pea seed respiratory metabolism under temperature stress, measuring heat and CO₂ rates after 16 h imbibition at 15, 20, 25, and 28 °C in three cultivars. It found cultivar differences in metabolic rates and temperature responses, with ‘Torta de Quebrar’ showing the lowest metabolic heat/CO₂ rates, smallest temperature dependence, and highest/most consistent germination.

Key findings

  • Significant differences among the three cultivars were found in metabolic heat rate (Rq), CO₂ production rate (RCO₂), and their temperature responses; ‘Torta de Quebrar’ had lower Rq than the other two cultivars at all temperatures, and its Rq/RCO₂ ratio was significantly higher at 15 and 28 °C.
  • In the germination trial, ‘Rondo’ had the highest germination at 20 and 25 °C and the lowest at 15 and 28 °C; ‘Torta de Quebrar’ showed significantly higher germination than the other two cultivars at 15, 25, and 28 °C, with almost 100% of seeds germinated.
  • ‘Torta de Quebrar’ had the lowest values of Rq, RCO₂, structural biomass formation rate (Rstruct_biomass), and carbon use efficiency (φ), and the highest Rq/RCO₂ values; it also showed the smallest temperature dependence of Rq, RCO₂, and Rstruct_biomass.
  • Tegument removal improved germination in all cultivars, reaching 100% in ‘Rondo’ and ‘Torta de Quebrar’; physical inhibition was clearly visible in ‘Rondo’, which showed a 30% increase in germination when the tegument was removed or scarified.

Methods

  • Three pea cultivars (‘Rondo’, ‘Torta de Quebrar’, ‘Maravilha d’América’) were used, with two seed lots per cultivar. Calorespirometric measurements were performed in isothermal mode at 15, 20, 25, and 28 °C after 16 h of imbibition; Rq and RCO₂ were measured and used to calculate Rq/RCO₂, Rstruct_biomass, and φ.
  • A germination trial ran for 6 days at 15, 20, 25, and 28 °C in darkness and 75% relative humidity, using 9 replicates of 10 seeds per temperature and cultivar (90 seeds per temperature/cultivar; 360 seeds per cultivar). Root and shoot lengths were measured, and a separate assay compared intact versus scarified/tegument-removed seeds at 25 °C.
  • Statistical analysis used SPSS v22.0; normality and homoscedasticity were checked, and mean comparisons used one-way ANOVA with Tukey’s HSD or the Kruskal-Wallis nonparametric test, with significance at p < 0.05.

Limitations

  • The paper states that no direct link was detected between germination using intact seeds and the calorespirometric parameters; the authors hypothesize a tegument/physical inhibition effect and call for further studies on AOX involvement, including pea genotypes with differences in calorespirometric measurements.
  • The paper states that few calorespirometric studies exist at the seed level under stress conditions, and no similar works were available for comparison at the early germination stage before radicle protrusion.
  • Evident from the text: only three cultivars and two seed lots per cultivar were tested, limiting generalizability; AOX/COX pathway involvement is inferred rather than directly measured; and calorespirometric measurements were taken at a single 16 h imbibition time point.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.3389/fpls.2022.827117

Cites

64 of 66 references verified0 citation errors · 1 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

20 of 48 cited sources checked · 13 supported · 1 flagged · accuracy not yet measured

  • Premisecitation
    § INTRODUCTION · p. 2

    Studies on the effects of adverse environmental conditions on pea have only been done at the plant level.

    “However, studies on the effects of adverse environmental conditions on this plant species have only been done at the plant level (Hernandez et al., 2000; Allen and Ort, 2001).”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites Hernandez et al., 2000 source

      The Introduction says this study examines antioxidant activity in cytosol-enriched fractions and mRNA levels in salt-stressed pea. The Results and discussion (p. 859) report responses in those fractions and in transcripts, so the work was not limited to plant-level measurements. (LLM, conf=0.88)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
    • cites Allen and Ort, 2001 source
  • Premisecitation
    § CONCLUSION · p. 11

    The AOX pathway is a means of relieving stress from temperature and drought.

    “This conclusion agrees with the literature consensus that the AOX pathway is a means of relieving stress from temperature and drought (Wang and Vanlerberghe, 2013; Velada et al., 2014; Dahal and Vanlerberghe, 2017; Campos et al., 2021).”
    Quote in the paperSubstantial claim1 of 3 sources checked
    • cites Wang and Vanlerberghe, 2013 source
    • cites Velada et al., 2014 source

      The Introduction (page unavailable) says AOX limits reactive oxygen species arising from environmental stress. The Discussion (page unavailable) reports evidence that AOX improves cold tolerance and finds AOX1 responsive to cold and heat. The paper does not specifically address drought, so it supports only part of the attributed claim. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites Dahal and Vanlerberghe, 2017 source
  • Findingown data
    § [Exploring the Applicability of Calorespirometry to Assess Seed Metabolic Stability Upon Temperature Stress ConditionsPisum sativum L. Used as a Case Study](https://www.frontiersin.org/articles/10.3389/fpls.2022.827117/full) · p. 1

    cv. 'Torta de Quebrar' showed significantly higher germination than the other two cultivars at all three temperatures.

    “The cv. 'Torta de Quebrar' showed significantly higher germination in comparison with the other two cultivars at all three temperatures.”
    Quote in the paperSubstantial claim
  • Findingown data
    § [Exploring the Applicability of Calorespirometry to Assess Seed Metabolic Stability Upon Temperature Stress ConditionsPisum sativum L. Used as a Case Study](https://www.frontiersin.org/articles/10.3389/fpls.2022.827117/full) · p. 1

    Significant differences in metabolic heat rate and CO2 production rate (RCO2) and in the temperature responses of these parameters were found among the three pea cultivars.

    “Significant differences in metabolic heat rate and CO2 production rate (R CO 2) as well as in the temperature responses of these parameters were found among the three cultivars.”
    Quote in the paperSubstantial claim
  • Findingown data
    § Respiratory Properties of Seeds · p. 4

    The calorespirometric ratio Rq/RCO2 was significantly higher in cv. 'Torta de Quebrar' compared with the other two cultivars at 15 and 28 C.

    “The calorespirometric ratio, R q /R CO 2 was significantly higher in the cv. 'Torta de Quebrar' compared with the other two cultivars at 15 and 28 C ( Figure 2D ).”
    Quote in the paperNot a substantial claim
  • Classified as probably not a substantial claim. 29
  • May combine two claims. 32
  • The quote only partly states this claim. 2

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation
    § INTRODUCTION · p. 2

    Seed performance depends on genetic factors and on the environmental conditions the parent plant was exposed to during seed formation and development, as well as the interaction between genetics and environmental conditions during germination.

    “Seed performance depends on genetic factors and on the environmental conditions the parent plant was exposed to during the formation and the development of seeds as well as the interaction between genetics and environmental conditions during germination (Bettey et al., 2000; Hampton et al., 2013; Li et al., 2017).”
    Quote in the paperNot a substantial claim2 of 3 sources checked
    • cites Bettey et al., 2000 source
    • cites Hampton et al., 2013 source

      Split into 2 atoms — 1 confirmed, 1 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

      Experimental — accuracy not yet measured
    • cites Li et al., 2017

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § INTRODUCTION · p. 2

    An increase of 15-20% yield has been reported if high-quality seeds are used.

    “An increase of 15-20% yield has been reported if high-quality seeds are used (Ambika et al., 2014).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Ambika et al., 2014 source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation
    § INTRODUCTION · p. 2

    The previous use of calorespirometry on seeds is limited, but its applicability for seed phenotyping is known for melon, and more recently for carrot, chickpea, and pea seeds.

    “The previous use of calorespirometry on seeds is limited, but the applicability of calorespirometry for the seed phenotyping of melon is known from Edelstein et al. (2001), and more recently of carrot, chickpea, and pea seeds by Mohanapriya et al. (2019).”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites Edelstein et al. (2001) source
    • cites Mohanapriya et al. (2019) source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § INTRODUCTION · p. 2

    Temperature, water potential, and oxygen are the main environmental factors affecting germination.

    “Temperature, water potential, and oxygen have been recognized as the main environmental factors affecting germination (Finch-Savage and Bassel, 2016).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Finch-Savage and Bassel, 2016 source

  • Classified as probably not a substantial claim.

    Premisecitation
    § INTRODUCTION · p. 2

    Calorespirometry has been used as a screening tool to assess metabolic and respiratory changes associated with cell reprogramming events, including plant plasticity towards different growing temperatures.

    “This approach has been used as a screening tool to assess metabolic and respiratory changes associated with cell reprogramming events (ArnholdtSchmitt et al., 2018) that include plant plasticity towards different growing temperatures (Nogales et al., 2013, 2015, 2020; Campos et al., 2016).”
    Quote in the paperSubstantial claim1 of 3 sources checked
    • cites ArnholdtSchmitt et al., 2018

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites Nogales et al., 2013, 2015, 2020

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites Campos et al., 2016 source

  • May combine two claims.

    Premisecitation
    § DISCUSSION · p. 9

    Seed imbibition immediately activates metabolism linked to germination, accompanied by an increase of respiration rates and heat production, with the embryo being the most responsible for those metabolic changes.

    “In fact, it is known that seed imbibition immediately activates metabolism linked to germination, which is accompanied by an increase of respiration rates and heat production, the embryo being the most responsible for those metabolic changes (Domergue et al., 2019).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites Domergue et al., 2019 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § DISCUSSION · p. 8

    Mohanapriya et al. (2019) highlighted the suitability of calorespirometry to assist pea seed breeding programs focused on the selection of inbred lines.

    “Mohanapriya et al. (2019) highlighted the suitability of calorespirometry to assist pea seed breeding programs focused on the selection of inbred lines.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Mohanapriya et al. (2019) source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § DISCUSSION · p. 9

    Calorespirometry has usually been applied to predict growth responses as a function of temperature in various plant tissues.

    “Calorespirometry has usually been applied to predict growth responses as a function of temperature in various plant tissues (Nogales et al., 2013, 2015, 2020; Campos et al., 2016).”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites Nogales et al., 2013, 2015, 2020

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites Campos et al., 2016 source

      The Introduction says calorespirometry has been used to monitor temperature-dependent growth in carrot in vitro cultures, and the Materials and Methods describe measurements at 21°C and 28°C. Our copy supports that application in carrot cultures, but does not establish that calorespirometry has usually been applied this way across various plant tissues. Page numbers were not provided in the passages. (LLM, conf=0.84)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § DISCUSSION · p. 9

    Plants adapt their growth to their environmental temperature by adapting their respiratory metabolism.

    “Previous reports have shown that plants adapt their growth to their environmental temperature by adapting their respiratory metabolism (Criddle et al., 2005).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Criddle et al., 2005 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § DISCUSSION · p. 9

    Changes in environmental temperatures can strongly influence germination rates through an effect on membrane reorganization during imbibition.

    “The changes in environmental temperatures can strongly influence germination rates as an effect on membrane reorganization during imbibition (Fahad et al., 2017; Doria et al., 2019).”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites Fahad et al., 2017 source

      The Morphological Responses > Growth section says elevated temperatures can reduce seed germination potential, supporting the temperature–germination link. It does not attribute that effect to membrane reorganization during imbibition; the Introduction discusses membrane damage from heat stress but does not connect it to imbibition or germination. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites Doria et al., 2019 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § DISCUSSION · p. 9

    Tegument thickness is the main factor responsible for the physical dormancy and consequent inhibition of seed germination in wild pea (P. sativum subsp. elatius).

    “The tegument thickness was highlighted as the main factor responsible for the physical dormancy and the consequent inhibition of seed germination in wild pea ( P. sativum subsp. elatius ) (Hradilová et al., 2019).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Hradilová et al., 2019 source

      The Results section on seed coat thickness reports that dormant wild-pea accessions had thicker seed coats than non-dormant ones, and the Discussion describes a positive correlation between testa thickness and dormancy (page numbers unavailable). These passages support an association, but they do not establish thickness as the main factor responsible for physical dormancy or germination inhibition; some non-dormant accessions also had thick seed coats. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § DISCUSSION · p. 10

    The link between the alternative oxidase (AOX) and structural biomass formation was previously highlighted by Campos et al. (2016) in a carrot primary culture system.

    “The link between the alternative oxidase (AOX), the key enzyme in the alternative respiratory pathway, and structural biomass formation was previously highlighted by Campos et al. (2016) in a carrot primary culture system.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Campos et al. (2016) source

  • Classified as probably not a substantial claim. May combine two claims.

    Premisecitation
    § DISCUSSION · p. 9

    The imbibition period is crucial for seed germination, allowing activation of cellular metabolism including mobilization of food reserves and activation of mitochondria and protein synthesis.

    “Since it provides the required water to hydrate the cellular constituents, the imbibition period is crucial for seed germination, allowing the activation of cellular metabolism which includes the mobilization of food reserves and the activation of mitochondria and protein synthesis (Galland et al., 2014; Paszkiewicz et al., 2017).”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites Galland et al., 2014 source
    • cites Paszkiewicz et al., 2017 source

      The section “Seed Embryo Mitochondria Are Metabolically Reactivated within Minutes of Hydration” reports that mitochondria rapidly regain metabolic activity upon imbibition. The paper mentions a storage-lipid mobilization program and later insertion of new polypeptides, but the passages do not establish that imbibition activates food-reserve mobilization and protein synthesis as the sentence claims. (LLM, conf=0.83)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation
    § DISCUSSION · p. 9

    Seed dormancy is currently observed in the Fabaceae family and has been mainly attributed to physical dormancy.

    “In the Fabaceae family, seed dormancy is currently observed (Smıkal et al., 2014) and has been mainly attributed to the physical dormancy (Martins et al., 2012)”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites Smıkal et al., 2014 source
    • cites Martins et al., 2012 source

  • May combine two claims.

    Findingown data
    § [Exploring the Applicability of Calorespirometry to Assess Seed Metabolic Stability Upon Temperature Stress ConditionsPisum sativum L. Used as a Case Study](https://www.frontiersin.org/articles/10.3389/fpls.2022.827117/full) · p. 1

    The optimal germination temperature for the pea cultivars tested was 20 C, and low germination rates were observed at extreme temperatures (15 or 28 C).

    “The germination trial showed that the optimal germination temperature was 20 Cand low germination rates were observed at extreme temperatures (15 or 28 C).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data
    § [Exploring the Applicability of Calorespirometry to Assess Seed Metabolic Stability Upon Temperature Stress ConditionsPisum sativum L. Used as a Case Study](https://www.frontiersin.org/articles/10.3389/fpls.2022.827117/full) · p. 1

    Compared with the other two cultivars, 'Torta de Quebrar' has the lowest metabolic heat and CO2 rates and the smallest temperature dependence of these measured parameters.

    “In comparison with the other two cultivars, 'Torta de Quebrar' has the lowest metabolic heat and CO2 rates and the smallest temperature dependence of these measured parameters.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data
    § Seed Germination During 6 Days of Imbibition · p. 5

    For cv. 'Rondo', the highest germination rate was achieved at 20 and 25 C, and the lowest germination rates were achieved at the extreme temperatures (15 and 28 C).

    “For the cv. 'Rondo' , the highest germination rate was achieved at 20 and 25 C, and the lowest germination rates were achieved at the extreme temperatures (15 and 28 C) ( Figure 2F ).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data
    § Seed Germination During 6 Days of Imbibition · p. 5

    The decrease in germination at higher temperatures in cvs. 'Rondo' and 'Maravilha d'América' was associated with an increase in the number of dead seeds, associated with mold growth, which reached the maximum at 28 C.

    “The decrease in germination at higher temperatures in cvs. 'Rondo' and 'Maravilha d'América' was associated with an increase in the number of dead seeds, associated with the development of mold growth, which reached the maximum at 28 C.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data
    § Characterizing Early Imbibition and Metabolic Activation · p. 4

    Water uptake events were completed by 16 h and respiratory heat rates in viable seeds became constant for several hours.

    “These WUevents were completed by 16 h and respiratory heat rates in viable seeds became constant for several hours.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data
    § Seed Germination During 6 Days of Imbibition · p. 5

    The cv. 'Torta de Quebrar' showed significantly higher germination rates at 15, 25, and 28 C in comparison with the other two cultivars, exhibiting almost 100% of the seeds germinated.

    “The cv. 'Torta de Quebrar' showed significantly higher germination rates at 15, 25, and 28 C in comparison with the other two cultivars, exhibiting almost 100% of the seeds germinated.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data
    § Seed Germination During 6 Days of Imbibition · p. 5

    Cumulative germination followed a sigmoid curve showing that seeds of the same cultivar did not germinate at the same time irrespective of thermal conditions.

    “Cumulative germination followed a sigmoid curve showing that seeds of the same cultivar did not germinate at the same time irrespective of thermal conditions.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data
    § DISCUSSION · p. 9

    Cv. 'Torta de Quebrar' exhibited the highest values of seed germination, with slight variation across temperatures.

    “and exhibited the highest values of seed germination, with slight variation across temperatures”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data
    § Effect of Temperature on Respiratory Properties During Seed Germination · p. 5

    The values of Rq/RCO2 for 'Torta de Quebrar' showed a minimum between 20 and 25 C, in 'Rondo' showed a slight decrease with increasing temperature, and in 'Maravilha d'América' was constant between 15-25 C and decreased at 28 C.

    “The values of R q /R CO 2 for 'Torta de Quebrar' showed a minimum between 20 and 25 C, in 'Rondo' showed a slight decrease with increasing temperature, and in 'Maravilha d' America' was constant between 15-25 C and decreased at 28 C.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data
    § Respiratory Properties of Seeds · p. 4

    The Rq values of cvs. 'Rondo' and 'Maravilha d'América' were essentially the same at 15, 20, and 25 C, but differed significantly at 28 C.

    “The R q values of cvs. 'Rondo' and 'Maravilha d'América' were essentially the same when compared at 15, 20, and 25 C, but differed significantly at 28 C.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data
    § DISCUSSION · p. 9

    In cvs. 'Maravilha d'América' and 'Rondo', heat rate stability was reached 8 h after imbibition, coinciding with attainment of the water absorption plateau phase, whereas cv. 'Torta de Quebrar' exhibited slower water uptake and reached the plateau later, at 16 h after imbibition.

    “In cvs. 'Maravilha d'América' and 'Rondo' , the stability of the heat rate was reached 8 h after imbibition, at the same time that the water absorption plateau phase was attained [phase II or middle phase according to Weitbrecht et al. (2011)], while the cv. 'Torta de Quebrar' exhibited a slower WU and reached the plateau at a later time (16 h after imbibition).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data
    § Respiratory Properties of Seeds · p. 4

    The epsilon values were close to the same, between 0.8 and 0.9, for all three cultivars, but in 'Torta de Quebrar' it tended to be slightly lower than the other two cultivars, with significant differences at 15 and 25 C.

    “The + values were close to the same, between 0.8 and 0.9, for all three cultivars, but in 'Torta de Quebrar' it tended to be slightly lower than the other two cultivars, with significant differences at 15 and 25 C ( Figure 2E ).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data
    § Characterizing Early Imbibition and Metabolic Activation · p. 4

    The three pea cultivars started to absorb water immediately on soaking and produced a three-phase water uptake curve.

    “The three cultivars started to absorb water immediately on soaking and produced a three-phase WU curve.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data
    § Seed Germination During 6 Days of Imbibition · p. 5

    The cv. 'Torta de Quebrar' exhibited the greatest root and shoot lengths and more than 70% of seeds with developed shoots at the three higher temperatures.

    “The cv. 'Torta de Quebrar' exhibited the greatest root and shoot lengths and more than 70% of seeds with developed shoots at the three higher temperatures.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data
    § Effect of Tegument on Seed Germination · p. 6

    The removal of tegument improved the germination rate of all cultivars, reaching values of 100% in cv. 'Rondo' and 'Torta de Quebrar'.

    “The removal of tegument improved the germination rate of all cultivars, reaching values of 100% in cv. 'Rondo' and 'Torta de Quebrar' ( Figure 5 ).”
    Quote in the paperNot a substantial claim

  • The quote only partly states this claim.

    Findingnone given
    § Effect of Temperature on Respiratory Properties During Seed Germination · p. 5

    The value of epsilon was invariant with temperature.

    “The value of + was invariant with temperature.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data
    § DISCUSSION · p. 10

    The lower + and higher Rq/RCO2 values were inversely correlated with germination rates, cv. 'Torta de Quebrar' showing a high germination rate at all temperatures tested.

    “The lower + and higher R q /R CO 2 values were inversely correlated with germination rates, as cv. 'Torta de Quebrar' showed a high germination rate at all temperatures tested.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingcitation
    § DISCUSSION · p. 9

    Physical dormancy was clearly seen in cv. 'Rondo'.

    “which was clearly seen in cv. 'Rondo'”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation
    § INTRODUCTION · p. 2

    Seed vigor, comprised of germination and seedling performance, affects vegetative growth and is frequently related to crop yield.

    “Seed vigor, comprised of germination and seedling performance, affects vegetative growth and is frequently related to crop yield (TeKrony and Egli, 1991; Ellis, 1992).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites TeKrony and Egli, 1991 source
    • cites Ellis, 1992 source

  • May combine two claims.

    Interpretationown data
    § INTRODUCTION · p. 2

    Calorespirometry is a useful tool for phenotyping physiologic efficiency at different temperatures during early germination stages and can determine the seeds with the highest resilience to temperature variation, in this case 'Torta de Quebrar'.

    “These data suggest that calorespirometry is a useful tool for phenotyping physiologic efficiency at different temperatures during early germination stages, and can determine the seeds with the highest resilience to temperature variation, in this case 'Torta de Quebrar'.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data
    § Seed Germination During 6 Days of Imbibition · p. 5

    High resilience under extreme temperatures, here associated with high seed vigor, was seen in cv. 'Torta de Quebrar', which exhibited seedling development at all tested temperatures.

    “High resilience under extreme temperatures, here associated with high seed vigor, was seen in cv. 'Torta de Quebrar' , that exhibited seedling development at all tested temperatures.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisenone given
    § DISCUSSION · p. 8

    The use of calorespirometry to evaluate metabolic changes upon different growing temperatures was not considered.

    “However, the use of this technique to evaluate metabolic changes upon different growing temperatures was not considered.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation
    § INTRODUCTION · p. 2

    During germination, progressive depletion of oxygen creates anaerobic conditions, leading to activation of fermentative metabolism as the main source of ATP production.

    “During germination, the progressive depletion of oxygen creates anaerobic conditions consequently leading to the activation of fermentative metabolism, working as the main source of ATP production (Ma et al., 2020).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Ma et al., 2020

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Classified as probably not a substantial claim.

    Premisecitation
    § Evaluation of Temperature Effect on Seed Germination and Seedling Development · p. 4

    The agronomically acceptable minimum (aam) germination is 70% for pea seeds.

    “The agronomically acceptable minimum (aam) germination is 70% for pea seeds (Miguel, 1983).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Miguel, 1983

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Classified as probably not a substantial claim.

    Premisecitation
    § INTRODUCTION · p. 2

    Calorespirometry is a rapid method for simultaneously measuring the metabolic heat rate and CO2 emission rate of biological samples at different environmental temperatures.

    “Calorespirometry is a rapid method for simultaneously measuring the metabolic heat rate and CO2 emission rate of biological samples at different environmental temperatures (Nogales et al., 2013, 2015, 2020).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Nogales et al., 2013, 2015, 2020

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Classified as probably not a substantial claim. May combine two claims.

    Premisecitation
    § INTRODUCTION · p. 2

    Combining respiratory heat and CO2 rates as a function of temperature enables calculation of carbon use efficiency and growth rate as functions of temperature to select the temperature range to which plants are well-adapted.

    “Combining respiratory heat and CO2 rates as a function of temperature enables the calculation of carbon use efficiency (the fraction of carbon substrate incorporated into structural biomass, + ) and growth rate as functions of temperature to select the temperature range to which plants are well-adapted (Hansen et al., 2009).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Hansen et al., 2009 source

  • May combine two claims.

    Premisecitation
    § DISCUSSION · p. 8

    The advantage of the calorespirometric method over methods currently used for phenotyping seed viability is that in addition to measuring CO2 production rates, the production of metabolic heat is also measured, with both rates known to be correlated with plant growth rates.

    “The advantage of the calorespirometric method over the methods currently used for phenotyping seed viability is that in addition to measuring CO2 production rates, the production of metabolic heat is also measured, with both rates known to be correlated with plant growth rates (Criddle et al., 1991).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Criddle et al., 1991 source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation
    § INTRODUCTION · p. 2

    Pea is a widespread crop across Europe and plays a preponderant role in human nutrition as one of the main sources of dietary plant protein.

    “This species is a widespread crop across Europe, playing a preponderant role in human nutrition as one of the main sources of dietary plant protein (Dhall, 2017).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Dhall, 2017

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation
    § INTRODUCTION · p. 2

    Pea is adapted to a wide range of climatic regimes, from semiarid to temperate maritime, with a germination temperature ranging from 20 to 1.1 C.

    “Pea is adapted to a wide range of climatic regimes, from semiarid to temperate maritime, with a germination temperature ranging from 20 to 1.1 C (Raveneau et al., 2011).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Raveneau et al., 2011 source

  • Classified as probably not a substantial claim.

    Premisecitation
    § INTRODUCTION · p. 2

    Plant growth rates have been correlated with heat rates.

    “In addition to CO2 production rates, plant respiration is linked to the production of metabolic heat, and plant growth rates have been correlated with heat rates (Criddle et al., 1991).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Criddle et al., 1991 source

  • Classified as probably not a substantial claim.

    Premisecitation
    § INTRODUCTION · p. 2

    A linear relationship between seed viability and metabolic rate determined with flow-through respirometry has been demonstrated in seeds of several plant species.

    “Dalzielli and Tomlinson (2017) have recently demonstrated a linear relationship between seed viability and metabolic rate determined with flow-through respirometry in the seeds of several plant species.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Dalzielli and Tomlinson (2017)

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Interpretationown data
    § CONCLUSION · p. 11

    Calorespirometry could identify the seeds with the highest resilience to temperature variation, which in this case were 'Torta de Quebrar'.

    “Moreover, it could identify the seeds with the highest resilience to temperature variation, which were in this case 'Torta de Quebrar'.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisenone given
    § DISCUSSION · p. 9

    Respiratory heat rate Rq is a measure of the total rate of catabolic oxidation by both the alternative oxidase (AOX) and cytochrome oxidase (COX) pathways.

    “Respiratory heat rate, R q is thus a measure of the total rate of catabolic oxidation by both the alternative oxidase (AOX) and cytochrome oxidase (COX) pathways.”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Interpretationown data
    § CONCLUSION · p. 11

    There is a potential existence of physical inhibition of germination in cv. 'Rondo'.

    “revealing the potential existence of physical inhibition”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation
    § DISCUSSION · p. 10

    Stressful conditions such as extreme temperatures lead to changes in metabolic pathways that are reflected in high values of Rq/RCO2 and low values of +.

    “Stressful conditions, due for example, to extreme temperatures, lead to the changes in metabolic pathways, which are reflected in high values of R q /R CO 2 and low values of + (Rank et al., 1991; Hansen et al., 2009).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Rank et al., 1991 source
    • cites Hansen et al., 2009 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § DISCUSSION · p. 9

    The substrate carbon conversion efficiency (+) varies between 0 and 0.85 in non-growing plants/tissues and rapidly growing vegetative plants and tissues, respectively.

    “According to previous studies, the value of + varies between 0 and 0.85 in non-growing plants/tissues and rapidly growing vegetative plants and tissues, respectively (Amthor, 2000; Hansen et al., 2005).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Amthor, 2000 source
    • cites Hansen et al., 2005 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § DISCUSSION · p. 10

    When a plant's environment reaches the temperature limits of growth, the efficiency of respiration goes to zero.

    “When a plant's environment reaches these temperatures, the efficiency of respiration goes to zero (McCarlie et al., 2003).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites McCarlie et al., 2003 source

  • May combine two claims.

    Interpretationreasoning
    § DISCUSSION · p. 9

    Because ΔHCO2 and ΔHB are constant and ΔHCO2 is much greater than ΔHB, Rq/RCO2 varies inversely with +, decreasing as + increases.

    “Since 1 H CO 2 and 1 H B are constant and 1 H CO 2 &gt; &gt; 1 H B , R q /R CO 2 varies inversely with + , i.e., R q /R CO 2 decreases as + increases.”
    Quote in the paperNot a substantial claim
    • The authors state this as an assumption: The temperature limits of plant growth correspond to the genetically determined biochemical limits.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § DISCUSSION · p. 9

    The optimum germination temperature is near 20 °C in most cases.

    “where other authors observed that the optimum germination temperature was near 20 C in most cases (Raveneau et al., 2011).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Raveneau et al., 2011 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § DISCUSSION · p. 9

    The water uptake plateau phase is characterized by a decrease in respiration prior to germination.

    “According to Bewley et al. (2013), the WU plateau phase is characterized by a decrease in respiration prior to germination.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Bewley et al. (2013) source
54 of 54

  • The paper relies on this without support.

    Differences among the three pea cultivars in calorespirometric and germination parameters reflect genetic differences in seed metabolic stability rather than seed lot, maternal environment, or other non-genetic factors.

    “Three cultivars of pea ( P. sativum ) provided by flora Lusitana R , commonly used in conventional agriculture were selected for calorespirometric measurements and germination trials: cvs. 'Rondo' (lots YF001522 and G2003378), 'Torta de Quebrar' (lots FL5581 and FL5719), and 'Maravilha d'América' (lots 04528CIT07X and AN3815-7251).”

  • The authors state this as an assumption.

    The temperature limits of plant growth correspond to the genetically determined biochemical limits.

    “The model that relates these two calorespirometric parameters assumes that the temperature limits of plant growth correspond to the genetically determined biochemical limits.”

  • The paper relies on this without support.

    None of the calorespirometric parameters are likely to differ between seeds and other growing plant tissues.

    “Since none of these parameters are likely to differ between seeds and other growing plant tissues, we must look to the inefficiencies in respiratory metabolism”

  • The authors state this as an assumption.

    The time required to achieve stable and reproducible metabolic heat and CO2 rates is related to mitochondria biogenesis processes, and by the end of this time all mitochondria are functional, producing stable respiratory rates.

    “we hypothesize that the required time to achieve stable and reproducible metabolic heat and CO2 rates is related to the mitochondria biogenesis processes and that by the end of this time, all mitochondria were functional, producing stable respiratory rates.”
4 of 4

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Two incompatible definitions of carbon use efficiency

    Minor

    A fraction of substrate carbon incorporated into biomass uses total substrate carbon as its denominator. A ratio of biomass-formation rate to CO2-production rate instead uses respired carbon as its denominator, so the two stated definitions are not equivalent.

  • Three temperatures stated where four were tested

    Minor

    The summary describes the cultivar comparison as covering three temperatures, while the methods specify four and the results report comparisons at all four.

2 of 2

Cites

Flagged first, then by citations

Research works printed but not linked: 3

  1. Dhall, 2017, Pea cultivation. Centre for Communication and International Linkages.
  2. 2013, Seed and Seed Quality: Technical Information for FAO Emergency Staff.
  3. Miguel, 1983, Métodos de Germinação de Sementes das Espécies Mais Utilizadas pela Agricultura Portuguesa.

Cited by

Still being checked.

Related

Authors
Rodrigues, L., Nogales, A., Hansen, L. D., Santos, F., Rato, A. E., & Cardoso, H.
Affiliations
  • University of Évora (PT)
  • University of Lisbon (PT)
  • Brigham Young University (US)
  • Consejo Superior de Investigaciones Científicas (ES)
Corresponding author
Cardoso, Hélia
ORCID
Venue
Frontiers in Plant Science
Publisher
Frontiers Media SA
Published
2022-04-27
Volume
13
Pages
827117-827117
ISSN
1664-462X (ISSN-L)
DOI
10.3389/fpls.2022.827117
PMC
PMC9094064
OpenAlex
W4224930873
Semantic Scholar
7bfa7e917e5f13137d0896445aa4eee7289c86e0
Cited by
8
References
66
Citations by year
2026: 1 · 2025: 2 · 2024: 2 · 2023: 2 · 2022: 1
FWCI
0.86
Type
journal article
Language
en
Open access
Licence
Locations
  • Frontiers in Plant Science (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Repositório da Universidade de Lisboa (University of Lisbon) (publishedVersion, other-oa)
  • PubMed Central (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Portuguese National Funding Agency for Science, Research and Technology (RCAAP Project by FCT) (submittedVersion, cc-by)
Funders
  • European Commission (s HORIZON 2020, 727230, HORIZON2020, UIDB/04129/2020, UIDB/05183/2020)
  • Fundação para a Ciência e a Tecnologia (57/2016, UIDB/05183/2020, 04129, /2017, UIDB/04129/2020, CEECIND/01769/2017, UIDB/05183)
  • Wageningen University and Research
  • Instituto Superior de Agronomia (UIDB/05183/2020, UIDB/04129/2020)
  • Staatssekretariat für Bildung, Forschung und Innovation (17.00090)
Topics
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Seed Germination and Physiology
  • Physical Sciences › Chemistry › Physical and Theoretical Chemistry › thermodynamics and calorimetric analyses
  • Physical Sciences › Environmental Science › Ecology › Physiological and biochemical adaptations
Keywords
seed germination, temperature stress, Pisum sativum, pea cultivars, seed respiration, CO2 production rate, seed imbibition, carbon use efficiency, germination temperature, phenotyping, cultivar screening

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-25.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-25.

Cited sources are checked by gpt-6-sol.

Statistics and arithmetic are recomputed, not judged.

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