Abstract
Conventional methods for screening for stress-tolerant cereal varieties rely on expensive, labour-intensive field testing and molecular biology techniques. Here, we use the root hair assay (RHA) as a rapid screening tool to identify stress-tolerant varieties at the early seedling stage. Wheat and barley seedlings had stress applied, and the response quantified in terms of programmed cell death (PCD), viability and necrosis. Heat shock experiments of seven barley varieties showed that winter and spring barley varieties could be partitioned into their two distinct seasonal groups based on their PCD susceptibility, allowing quick data-driven evaluation of their thermotolerance at an early seedling stage. In addition, evaluating the response of eight wheat varieties to heat and salt stress allowed identification of their PCD inflection points (35 °C and 150 mM NaCl), where the largest differences in PCD levels arise. Using the PCD inflection points as a reference, we compared different stress effects and found that heat-susceptible wheat varieties displayed similar vulnerabilities to salt stress. Stress-induced PCD levels also facilitated the assessment of the basal, induced and cross-stress tolerance of wheat varieties using single, combined and multiple individual stress exposures by applying concurrent heat and salt stress in a time-course experiment. Two stress-susceptible varieties were found to have low constitutive resistance as illustrated by their high PCD levels in response to single and combined stress exposure. However, both varieties had a fast, adaptive response as PCD levels declined at the other time-points, showing that even with low constitutive resistance, the initial stress cue primes cross-stress tolerance adaptations for enhanced resistance even to a second, different stress type. Here, we demonstrate the RHA's suitability for high-throughput analysis (~4 days from germination to data collection) of multiple cereal varieties and stress treatments. We also showed the versatility of using stress-induced PCD levels to investigate the role of constitutive and adaptive resistance by exploring the temporal progression of cross-stress tolerance. Our results show that by identifying suboptimal PCD levels in vivo in a laboratory setting, we can preliminarily identify stress-susceptible cereal varieties and this information can guide further, more efficiently targeted, field-scale experimental testing.
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Claims 9
Claim · model-written summary
PCD susceptibility separated winter and spring barley varieties after heat shock.
- Taxon
- barley
.jpg?width=60)
Hordeum vulgaree0 - Material
- winter and spring barley varietiese0
- Treatment
- Heat shocke0
- Outcome
- PCD susceptibilitye0
- Value
- seven barley varietiese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
.jpg?width=500)
Verbatim
e0“Heat shock experiments of seven barley varieties showed that winter and spring barley varieties could be partitioned into their two distinct seasonal groups based on their PCD susceptibility”
Claim · model-written summary
Wheat PCD differences under heat were greatest at 35 °C.
- Taxon
- wheat
Triticum aestivume0 - Treatment
- heate0
- Outcome
- PCD inflection pointse0
- Value
- 35 °Ce0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“response of eight wheat varieties to heat and salt stress allowed identification of their PCD inflection points (35 °C and 150 mM NaCl), where the largest differences in PCD levels arise”
Claim · model-written summary
Wheat PCD differences under salt were greatest at 150 mM NaCl.
- Taxon
- wheatTriticum aestivume0
- Treatment
- salt stresse0
- Outcome
- PCD inflection pointse0
- Value
- 150 mM NaCle0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“response of eight wheat varieties to heat and salt stress allowed identification of their PCD inflection points (35 °C and 150 mM NaCl), where the largest differences in PCD levels arise”
Claim · model-written summary
Heat-susceptible wheat varieties were similarly vulnerable to salt stress.
- Taxon
- wheatTriticum aestivume0
- Material
- heat-susceptible wheat varietiese0
- Treatment
- salt stresse0
- Outcome
- vulnerabilities to salt stresse0
- Direction
- similare0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“heat-susceptible wheat varieties displayed similar vulnerabilities to salt stress”
Claim · model-written summary
Stress-induced PCD supported assessment of basal, induced, and cross-stress wheat tolerance.
- Taxon
- wheatTriticum aestivume0
- Outcome
- assessment of the basal, induced and cross-stress tolerancee0
- Comparator
- single, combined and multiple individual stress exposurese0
Not reportedValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Stress-induced PCD levels also facilitated the assessment of the basal, induced and cross-stress tolerance of wheat varieties using single, combined and multiple individual stress exposures”
Claim · model-written summary
Two susceptible wheat varieties had high PCD under single and combined stress exposure.
- Taxon
- wheatTriticum aestivume1
- Material
- Two stress-susceptible varietiese0
- Treatment
- single and combined stress exposuree0
- Outcome
- PCD levelse0
- Direction
- highe0
- Value
- Twoe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Two stress-susceptible varieties were found to have low constitutive resistance as illustrated by their high PCD levels in response to single and combined stress exposure”
e1“eight wheat varieties”
Claim · model-written summary
PCD declined later in both susceptible varieties, indicating rapid adaptation.
- Taxon
- wheatTriticum aestivume1
- Material
- both varietiese0
- Outcome
- PCD levelse0
- Direction
- declinede0
- Qualifier
- other time-pointse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“both varieties had a fast, adaptive response as PCD levels declined at the other time-points”
e1“eight wheat varieties”
Claim · model-written summary
The root hair assay supported high-throughput cereal analysis in about four days.
Verbatim
e0“RHA's suitability for high-throughput analysis (~4 days from germination to data collection) of multiple cereal varieties and stress treatments”
Claim · model-written summary
The authors propose using laboratory PCD screening to target later field trials.
Verbatim
e0“by identifying suboptimal PCD levels in vivo in a laboratory setting, we can preliminarily identify stress-susceptible cereal varieties and this information can guide further, more efficiently targeted, field-scale experimental testing”