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Cited by 19Plant Stress Responses and ToleranceCRISPR and Genetic EngineeringWheat and Barley Genetics and Pathology

Genetic resources and precise gene editing for targeted improvement of barley abiotic stress tolerance

Open access
Karunarathne, Sakura, Walker, Esther, Sharma, Darshan, Li, Chengdao, Han, Yong·2023·Journal of Zhejiang University-SCIENCE B
DOI 10.1631/jzus.b2200552PMC PMC10710907
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Abstract

Abiotic stresses, predominately drought, heat, salinity, cold, and waterlogging, adversely affect cereal crops. They limit barley production worldwide and cause huge economic losses. In barley, functional genes under various stresses have been identified over the years and genetic improvement to stress tolerance has taken a new turn with the introduction of modern gene-editing platforms. In particular, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) is a robust and versatile tool for precise mutation creation and trait improvement. In this review, we highlight the stress-affected regions and the corresponding economic losses among the main barley producers. We collate about 150 key genes associated with stress tolerance and combine them into a single physical map for potential breeding practices. We also overview the applications of precise base editing, prime editing, and multiplexing technologies for targeted trait modification, and discuss current challenges including high-throughput mutant genotyping and genotype dependency in genetic transformation to promote commercial breeding. The listed genes counteract key stresses such as drought, salinity, and nutrient deficiency, and the potential application of the respective gene-editing technologies will provide insight into barley improvement for climate resilience.

AI snapshot

Written from the paper’s full text.

TL;DR

This review compiles more than 150 barley genes associated with abiotic stress tolerance into a single physical map and surveys CRISPR/Cas9, base editing, prime editing, and multiplexing for targeted improvement. It argues that precise gene editing, combined with solutions for transformation genotype dependency and high-throughput mutant genotyping, can accelerate development of climate-resilient barley.

Key findings

  • More than 150 named *Hordeum vulgare* functional and regulatory genes were mapped across all seven barley chromosomes; 44% were induced by drought stress and 23% by salt stress, with several activated under multiple stresses.
  • CRISPR/Cas9 has been used in barley to create insertion/deletion mutations in genes such as eIF4E, HvMPK6, HvNud, HvPAPhy, HvPTST1, HvGBSS1, HvHPT, and HvHGGT, but only three studies—Vlčko and Ohnoutková (2020), Fu et al. (2022), and Karunarathne et al. (2022)—used it for barley germplasm development relevant to abiotic stress.
  • Multiplex editing of two barley cytokinin oxidase/dehydrogenase genes, HvCKX1 and HvCKX3, using a polycistronic tRNA-gRNA construct achieved 21% editing efficiency with heritable mutations.
  • Prime editing has been used in rice and wheat but not yet in barley; it can introduce all base substitutions, small insertions up to 44 bp, and deletions up to 88 bp, and may be combined with multiplex editing for traits such as herbicide resistance.

Methods

  • Narrative review with no primary experiments; the authors performed a literature search and data analysis, focusing on known barley genes as candidates for gene editing and molecular breeding.
  • Candidate genes were compiled from mapping, microarray, and transcriptome studies; gene locations and annotations were obtained by BLASTing NCBI, UniProt, or Morex v1 accession information against the Morex v3 reference sequence, producing Table S1 with accession, function, Morex v3 gene ID, and references.
  • The review also summarizes barley prebreeding resources in Table 1 and discusses CRISPR/Cas9, base editing, prime editing, multiplexing, transformation delivery, and mutant screening methods.

Limitations

  • Stated: current challenges include high-throughput mutant genotyping and genotype dependency in genetic transformation; only a few CRISPR/Cas9 studies have targeted barley abiotic stress, and TALENs or prime editing were unrepresented for these stresses.
  • Stated: many studies examined only seedling-stage responses under controlled laboratory or glasshouse conditions, with only al Abdallat et al. (2014) testing plant response in a field environment; single-gene approaches may not confer acceptable field-level tolerance over a season.
  • Evident from the text: as a review, it provides no new experimental validation, and the compiled gene list reflects the literature’s dominance of drought and salinity studies, so other stresses may be comparatively underrepresented.

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

Integrity signals

Registry

Crossref · 10.1631/jzus.b2200552

Cites

205 of 205 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

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  • Findingnone given

    Several genes were reportedly activated under multiple stresses, represented by transcription factors, reactive oxygen species (ROS) scavengers, and protective proteins, such as late embryogenesis abundant (LEA) proteins.

    “Several genes were reportedly activated under multiple stresses.These are represented by transcription factors, reactive oxygen species (ROS) scavengers, and protective proteins, such as late embryogenesis abundant (LEA) proteins (Table S1).”
    Quote in the paperSubstantial claim
  • Findingnone given

    Numerous overexpression lines have been developed with significant phenotypic improvement reported across a range of abiotic stresses.

    “Numerous overexpression lines have been developed with significant phenotypic improvement reported across a range of abiotic stresses (Table 1).”
    Quote in the paperNot a substantial claim
  • Findingnone given

    Constitutive overexpression yields variable results on plant development and growth depending on the gene targeted.

    “However, constitutive overexpression yielded variable results on plant development and growth, depending on the gene targeted.”
    Quote in the paperSubstantial claim
  • Findingreasoning

    Regulatory genes, such as transcription factors, were a predominant category.

    “Regulatory genes, such as transcription factors, were a predominant category.”
    Quote in the paperSubstantial claim
  • Findingown data

    A modest range of gene-editing tools have been used to develop abiotic stress tolerance in barley.

    “A modest range of gene-editing tools have been used to develop abiotic stress tolerance in barley (Tables 1 and S2).”
    Quote in the paperNot a substantial claim
  • May combine two claims. 46
  • Classified as probably not a substantial claim. 70

  • May combine two claims.

    Findingnone given

    The review collates about 150 key genes associated with stress tolerance and combines them into a single physical map for potential breeding practices.

    “We collate about 150 key genes associated with stress tolerance and combine them into a single physical map for potential breeding practices.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S4 (based).

    Findingreasoning

    The listed genes counteract key stresses such as drought, salinity, and nutrient deficiency.

    “The listed genes counteract key stresses such as drought, salinity, and nutrient deficiency”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: The identified stress-tolerance genes can be effectively targeted by gene editing to improve barley abiotic stress tolerance.

  • May combine two claims.

    Findingown data

    The list reflects the dominance of drought and salinity studies in the literature, with 44% of identified genes induced by drought stress, followed by 23% induced by salt stress.

    “The list reflects the dominance of drought and salinity studies in the literature, with 44% of identified genes induced by drought stress, followed by 23% induced by salt stress.”
    Quote in the paperNot a substantial claim

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    Findingnone given

    In some cases, plant growth remained healthy under normal and stressed conditions and even exhibited enhanced growth and plant biomass compared to wild type.

    “In some cases, plant growth remained healthy under normal and stressed conditions and even exhibited enhanced growth and plant biomass, compared to the wild type.”
    Quote in the paperNot a substantial claim

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    Findingnone given

    Most of the genes in Table 1 have been overexpressed to achieve drought and frost tolerance, but not through CRISPR/Cas9.

    “Most of the genes in Table 1 have been overexpressed to achieve drought and frost tolerance, but not through CRISPR/Cas9.”
    Quote in the paperNot a substantial claim

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

    Interpretationreasoning

    These genetic resources are minimal, and more research is required to improve the tolerance of barley and other crops to abiotic stresses.

    “However, these resources are minimal, and more research is required to improve the tolerance of barley and other crops to abiotic stresses.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Several barley genes have been exploited to address different stress conditions, including HVA1 for drought, HKT1;1 for salinity, and ARE1 for nitrogen use efficiency.

    “Several barley genes have been exploited to address different stress conditions (Zhou et al., 2016;Hazzouri et al., 2018;Karunarathne et al., 2020;Mwando et al., 2020), including Hordeum vulgare aleurone 1 (HVA1) for drought, high-affinity K + transporter 1;1 (HKT1;1) for salinity, and abnormal cytokinin response 1 repressor 1 (ARE1) for nitrogen use efficiency (NUE) (Sivamani et al., 2000;Han et”
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    Premisecitation

    Demand for cereals both as food and animal feed is likely to be around three billion tonnes per annum by 2050.

    “Demand for cereals both as food and animal feed is likely to be around three billion tonnes per annum by 2050 (FAO, 2009).”
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    • cites FAO, 2009

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    Premisecitation

    Elite stress-tolerant barley genotypes such as 'Golshan' and 'Oxin' for salinity, and 'GrangeR' and 'Bridge' for low-N tolerance have been identified through phenotype screening.

    “Also, elite stresstolerant barley genotypes such as 'Golshan' and 'Oxin' for salinity, and 'GrangeR' and 'Bridge' for low-N tolerance have been identified through phenotype screening (Karunarathne et al., 2020;Bahrani et al., 2023).”
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    • cites Bahrani et al., 2023 source

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    Premisecitation

    Abiotic stresses limit barley production worldwide and cause huge economic losses.

    “They limit barley production worldwide and cause huge economic losses.”
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    • cites Kurnaz, 2014

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    Food demand is expected to increase by 35%-56% to feed a population of nearly ten billion by 2050.

    “food demand which is expected to increase by 35%-56% to feed a population of nearly ten billion by 2050 (van Dijk et al., 2021).”
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    Premisecitation

    Abiotic stresses, predominately drought, heat, salinity, cold, and waterlogging, adversely affect cereal crops.

    “Abiotic stresses, predominately drought, heat, salinity, cold, and waterlogging, adversely affect cereal crops.”
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    • cites Gürel et al., 2016 source

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    • cites Kumar et al., 2021 source

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    Premisecitation

    Drought-related crop losses are reported on all continents, bar Antarctica, and affect all major barley production countries.

    “Drought-related crop losses are reported on all continents, bar Antarctica, and affect all major barley production countries (Kurnaz, 2014;Geng et al., 2016;Stahl et al., 2016;Elliot et al., 2018;Cammarano et al., 2019;Adisa et al., 2020;Kirono et al., 2020;Araneda-Cabrera et al., 2021;Hudzenko et al., 2021;Hunt et al., 2021;Markonis et al., 2021).”
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    Premisecitation

    It is estimated that only 30%-40% of the current year's applied N fertilizer is taken up by crops, with the remainder remaining in the soil or lost out of the cropping system into the air and water.

    “It is estimated that only 30%-40% of the current year's applied N fertilizer is taken up by crops, with the remainder remaining in the soil or lost out of the cropping system into the air and water (Yan et al., 2020;He et al., 2021;Gao et al., 2022).”
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    Barley yield loss is estimated to range from 43% to 78% in Australia.

    “Barley yield loss is estimated to range from 43% to 78% in Australia (Mahajan et al., 2020).”
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    Premisecitation

    Traditional plant breeding techniques have been widely used to improve crop traits, but are labour-intensive and time-consuming.

    “Traditional plant breeding techniques have been widely used to improve crop traits, but are labour-intensive and timeconsuming (Zhu et al., 2020).”
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    Premisecitation

    Increasing temperature may lead to contradictory impacts on different production areas: crop yields are expected to increase in some regions such as Canada and northern Spain, whilst other regions such as France and southern Spain are likely to experience increasing agricultural losses.

    “Notably, the increasing temperature may lead to contradictory impacts on different production areas.For example, crop yields are expected to increase in some regions such as in Canada and northern Spain, where warmer winters would improve yields (Masud et al., 2018;Bento et al., 2021), whilst other regions such as France and southern Spain are likely to experience increasing agricultural losses (G”
    Quote in the paperNot a substantial claim0 of 3 sources checked
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    Premisecitation

    Rising sea levels are likely to impact coastal agricultural regions through salt intrusion, with land degradation predicted in Europe.

    “Furthermore, rising sea levels are likely to impact coastal agricultural regions through salt intrusion, with land degradation predicted in Europe (Bosello et al., 2012;Daliakopoulos et al., 2016;Ullah et al., 2021).”
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    Premisecitation

    Excess moisture in the soil limits (hypoxia) or completely depletes (anoxia) oxygen availability, leading to plant oxygen deficiency.

    “Excess moisture in the soil limits (hypoxia) or completely depletes (anoxia) oxygen availability, leading to plant oxygen deficiency (Zahra et al., 2021).”
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    Premisecitation

    A large proportion of barley producing regions are affected worldwide by waterlogging.

    “A large proportion of barley producing regions are affected worldwide (Brisson et al., 2002;Setter and Waters, 2003;Dickin and Wright, 2008;Yavas et al., 2012;Ahmed et al., 2013;Rukhovich et al., 2014;Twining, 2014;Sorokin et al., 2016;Borrego-Benjumea et al., 2019, 2020;Ciancio et al., 2021;Tian et al., 2021;Schmitt et al., 2022).”
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  • Probably not a substantial claim: may not pass S2 (checkable).

    Interpretationreasoning

    More precise gene-editing technologies such as CRISPR/Cas9 gene editing, base editing, and prime editing are imperative to develop new barley lines with superior qualities and better performance under stress conditions.

    “Therefore, more precise gene-editing technologies such as clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated protein 9 (Cas9) gene editing, base editing, and prime editing are imperative to develop new barley lines with superior qualities and better performance under stress conditions (Lin et al., 2021;Karunarathne et al., 2022).”
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    • cites Lin et al., 2021 source

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    • cites Karunarathne et al., 2022 source

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    Premisecitation

    Barley yields have been reduced by from 35% to as much as 70%, dependent on the timing and duration of waterlogging, and are accompanied by a delay in phenology.

    “Barley yields have been reduced by from 35% to as much as 70%, dependent on the timing and duration of waterlogging, and are accompanied by a delay in phenology (Liu et al., 2020a).”
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    • cites Liu et al., 2020a

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    Premisecitation

    Salinity is a constraint in many of the major barley production countries including Australia, Spain, Turkey, Argentina, the USA, and Canada.

    “Salinity is a constraint in many of the major barley production countries including Australia, Spain, Turkey, Argentina, the USA, and Canada (Huffman et al., 2000;Houk et al., 2006;Rengasamy, 2006;Acosta et al., 2011;Gorji et al., 2017;Zaman et al., 2018;Taleisnik and Lavado, 2021).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
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    Premisecitation

    As barley is often the only crop grown in some developing countries, particularly in arid and semi-arid regions, the impacts of drought and heat extremes on barley are of great concern.

    “As barley is often the only crop grown in some developing countries, particularly in arid and semi-arid regions, the impacts of drought and heat extremes on barley are of great concern (Kebede et al., 2019;Visioni et al., 2019).”
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    Premisecitation

    Salinity is one of the leading causes of crop losses worldwide, affecting an estimated 32 million ha of dryland agriculture.

    “Salinity is one of the leading causes of crop losses worldwide, affecting an estimated 32 million ha of dryland agriculture (Wani et al., 2020).”
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    • cites Wani et al., 2020 source

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    Premisecitation

    Increasing NUE in crops is a sustainable solution that not only maximizes yield potential in the face of increasing global food demands, but also would mitigate the economic and environmental impacts of fertilizer use.

    “Increasing NUE in crops is a sustainable solution that not only maximizes yield potential in the face of increasing global food demands, but also would mitigate the economic and environmental impacts of fertilizer use (Houlton et al., 2019;He et al., 2021;Langholtz et al., 2021;Karunarathne et al., 2022).”
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    • cites He et al., 2021

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    • cites Langholtz et al., 2021 source

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    Premisecitation

    Waterlogging has become more frequent and unpredictable due to climate change, with an increased impact on barley cropping regions.

    “Waterlogging has become more frequent and unpredictable due to climate change, with an increased impact on barley cropping regions (Liu et al., 2021).”
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    • cites Liu et al., 2021 source

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    Premisecitation

    Reduced precipitation due to climate change poses increased risks for arid and semi-arid regions as desertification and salinization threaten cropland in the Mediterranean, Africa, parts of Australia, Central America, and parts of the USA.

    “Reduced precipitation due to climate change poses increased risks for arid and semi-arid regions as desertification and salinization threaten cropland in the Mediterranean, Africa, parts of Australia, Central America, and parts of the USA (Corwin, 2021).”
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    Premisecitation

    Nitrogen fertilizers negatively impact aquatic ecosystems through nutrient runoff, and contaminate ground and drinking water.

    “Furthermore, nitrogen fertilizers negatively impact aquatic ecosystems through nutrient runoff, and contaminate ground and drinking water (Houlton et al., 2019;Langholtz et al., 2021).”
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    Premisecitation

    The addition of N fertilizers has significantly increased crop production yields and is essential to maintain food security.

    “The addition of N fertilizers has significantly increased crop production yields and is essential to maintain food security (He et al., 2021;Karunarathne et al., 2022).”
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    Premisecitation

    For barley, the International Barley Genome Sequencing Consortium (2012) published the first reference genome of 'Morex'.

    “For barley, the International Barley Genome Sequencing Consortium (2012) published the first reference genome of 'Morex'.”
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    • cites International Barley Genome Sequencing Consortium, 2012 source

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    Premisecitation

    Climate change simulation studies predict drought and heat events to worsen over the century with the most severe changes expected to occur in the second half of the century.

    “Climate change simulation studies predict drought and heat events to worsen over the century with the most severe changes expected to occur in the second half of the century.”
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    • cites Challinor et al., 2014 source

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    Premisecitation

    In Russia, the average spring barley yield loss is 13% in plots not treated with herbicides.

    “In Russia, the average spring barley yield loss is 13% in plots not treated with herbicides (Mayerová et al., 2018).”
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    • cites Mayerová et al., 2018 source

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    Premisecitation

    Modelling of USA cropping systems has estimated that a 10% increase in NUE over a 10-year period would increase crop revenue by USD 350 million per annum.

    “Modelling of USA cropping systems has estimated that a 10% increase in NUE over a 10-year period would increase crop revenue by USD 350 million per annum.”
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    • cites Langholtz et al., 2021 source

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    Genome annotation and assembly were subsequently improved.

    “Genome annotation and assembly were subsequently improved (Mascher et al., 2017;Monat et al., 2019).”
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    • cites Mascher et al., 2017 source

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    Premisecitation

    A 20% increase in NUE would increase crop revenue by USD 743 million per annum and reduce the cost of water treatment by about USD 15-136 million per annum.

    “A 20% increase in NUE would increase crop revenue by USD 743 million per annum and reduce the cost of water treatment by about USD 15-136 million per annum (Langholtz et al., 2021).”
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    • cites Langholtz et al., 2021 source

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

    Premisecitation

    Mutagenesis is more acceptable than transgenesis in breeding, yet in some studies its efficiency is reported to be low.

    “Mutagenesis is more acceptable than transgenesis in breeding, yet in some studies its efficiency is reported to be low (Nonaka et al., 2017).”
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    • cites Nonaka et al., 2017 source

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    Premisenone given

    CRISPR is an integral part of the bacterial and archaeal immune systems.

    “It is an integral part of the bacterial and archaeal immune systems, which has now been adapted by scientists in various fields including agriculture.”
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    Premisecitation

    Advancement of the technique to multiplex gene editing allows the simultaneous editing of several targets in a single genome.

    “Advancement of the technique to multiplex gene editing allows the simultaneous editing of several targets in a single genome (Xing et al., 2014).”
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    Interpretationreasoning

    The transient nature of VIGS is of limited benefit to plant breeders.

    “Likewise, virus-induced gene silencing (VIGS) rapidly provides valuable insights into gene function, but its transient nature is of limited benefit to plant breeders.”
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    Premisenone given

    Base editing targets single nucleotide mutations with the use of deactivated Cas9 (dCas9) or nickase Cas9 (nCas9).

    “Base editing targets single nucleotide mutations with the use of deactivated Cas9 (dCas9) or nickase Cas9 (nCas9).”
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  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Gene knockout is the widely used application of CRISPR/Cas9.

    “Gene knockout is the widely used application of CRISPR/Cas9, and single or multiple gene knockouts have been successfully observed in maize, barley, wheat, and rice (Li et al., 2013;Doll et al., 2019;Kim et al., 2019;Lawrenson and Harwood, 2019;Křenek et al., 2021;Zhang et al., 2021;Karunarathne et al., 2022).”
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  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisenone given

    CRISPR/Cas9 is a more precise technique than overexpression.

    “The CRISPR/Cas9 system is a more precise technique than overexpression, yet only Vlčko and Ohnoutková (2020), Fu et al. (2022), and Karunarathne et al. (2022) had used this technology for barley germplasm development (Table 1).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S4 (based).

    Interpretationreasoning

    The development of herbicide resistance in cereals is a cost-effective alternative to avoid crop damage caused by herbicides and to maintain high productivity.

    “Therefore, the development of herbicide resistance in cereals is a cost-effective alternative to avoid crop damage caused by herbicides and to maintain high productivity.”
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  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisereasoning

    CRISPR has been adapted by scientists in various fields including agriculture.

    “It is an integral part of the bacterial and archaeal immune systems, which has now been adapted by scientists in various fields including agriculture.”
    Quote in the paperNot a substantial claim

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

    Premisenone given

    The CRISPR/Cas system is a precise technique in gene-editing.

    “As highlighted under barley prebreeding resources, the CRISPR/Cas system is a precise technique in geneediting.”
    Quote in the paperNot a substantial claim

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

    Premisenone given

    VIGS rapidly provides valuable insights into gene function.

    “Likewise, virus-induced gene silencing (VIGS) rapidly provides valuable insights into gene function, but its transient nature is of limited benefit to plant breeders.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Wild oats, annual ryegrass, and flaxleaf fleabane are the common weeds in barley paddocks.

    “Wild oats, annual ryegrass, and flaxleaf fleabane are the common weeds in barley paddocks (Mahajan et al., 2020), while ryegrass, wild radish, brome grass, and wild oats are reported to be among the costliest to control in Australia (Llewellyn et al., 2016).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Mahajan et al., 2020 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S2 (checkable).

    Interpretationreasoning

    The usefulness of this material for breeding tolerant cultivars is largely dependent on the outcomes of these studies.

    “The usefulness of this material for breeding tolerant cultivars is largely dependent on the outcomes of these studies.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: The outcomes of the gene-editing studies in barley are representative of what can be achieved in breeding tolerant cultivars.

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

    Premisenone given

    Base and prime editing tools are among the most recent advances of CRISPR.

    “Engineered CRISPR systems have become more efficient and flexible, with base and prime editing tools among the most recent advances of CRISPR.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Abiotic stresses activate the abscisic acid (ABA)-independent and -dependent signalling transduction pathways, but also activate signalling pathways typically associated with biotic stresses, such as the jasmonic acid (JA) pathway.

    “Abiotic stresses activate the abscisic acid (ABA)-independent and -dependent signalling transduction pathways, but also activate signalling pathways typically associated with biotic stresses, such as the jasmonic acid (JA) pathway (Visioni et al., 2019).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Visioni et al., 2019 source

      Not checked in the free run.

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

    Premisenone given

    Engineered CRISPR systems have become more efficient and flexible.

    “Engineered CRISPR systems have become more efficient and flexible, with base and prime editing tools among the most recent advances of CRISPR.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Modifying regulatory genes can alter the expression of multiple downstream stress response genes, and therefore has the potential to produce a more significant and durable phenotypic change than targeting a single functional gene.

    “Modifying regulatory genes can alter the expression of multiple downstream stress response genes, and therefore has the potential to produce a more significant and durable phenotypic change than targeting a single functional gene (Umezawa et al., 2006).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Umezawa et al., 2006 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Plant stress response is interactive and non-additive, with some genes acting synergistically and others antagonistically.

    “Plant stress response is interactive and non-additive, with some genes acting synergistically and others antagonistically.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Mittler, 2006 source

      Not checked in the free run.

    • cites Atkinson and Urwin, 2012 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Some weeds have developed resistance to herbicides that inhibit acetyl-coenzyme A carboxylase (ACCase), acetolactate synthase (ALS), and enol pyruvyl shiquimate phosphate synthase (EPSPS), suggesting that herbicides with new modes of action are needed.

    “Some of these weeds have developed resistance to herbicides that inhibit acetyl-coenzyme A carboxylase (ACCase), acetolactate synthase (ALS), and enol pyruvyl shiquimate phosphate synthase (EPSPS), suggesting that herbicides with new modes of action are needed (Galon et al., 2022).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Galon et al., 2022 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S4 (based).

    Premisenone given

    HvHVP10 is known to be upregulated in response to salt stress in barley roots.

    “This might have been because HvHVP10 is known to be upregulated in response to salt stress in barley roots.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Prime editing is the latest technique that can induce precise mutations.

    “Prime editing is the latest technique that can induce precise mutations (Lin et al., 2021).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Lin et al., 2021 source

      Not checked in the free run.

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

    Premisecitation

    Researchers have sometimes used gene expression studies with reverse transcription-polymerase chain reaction (RT-PCR) or differential gene expression to determine potential genes for any given trait.

    “Researchers have sometimes used gene expression studies with reverse transcription-polymerase chain reaction (RT-PCR) or differential gene expression to determine potential genes for any given trait (Feng et al., 2020a;Karunarathne et al., 2022).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Feng et al., 2020a

      Not checked in the free run.

    • cites Karunarathne et al., 2022 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisenone given

    CRISPR/Cas9 can be reprogrammed for transcriptional regulation by fusing transcriptional activator domains to a dCas9 in barley.

    “CRISPR/Cas9 can be reprogrammed for transcriptional regulation by fusing transcriptional activator domains to a dCas9 in barley.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    New systems such as Cas12, Cas13a, and Cas13b are in the pipeline.

    “CRISPR/Cas9 is the most widely used system, but new systems such as Cas12, Cas13a, and Cas13b are in the pipeline (Smargon et al., 2017;Zaidi et al., 2017;Aman et al., 2018).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Smargon et al., 2017 source

      Not checked in the free run.

    • cites Zaidi et al., 2017 source

      Not checked in the free run.

    • cites Aman et al., 2018 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Plant prime-editing systems have been used in rice and wheat, but not in barley.

    “Plant prime-editing systems have been used in rice and wheat, but not in barley (Lin et al., 2020;Xu et al., 2020).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Lin et al., 2020 source

      Not checked in the free run.

    • cites Xu et al., 2020

      Not checked in the free run.

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

    Interpretationcitation

    CRISPRa may be a promising application in other monocots too, including barley, to study the transcriptional network underlying abiotic stresses.

    “Therefore, this may be a promising application in other monocots too, including barley, to study the transcriptional network underlying abiotic stresses.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Base editing can be repurposed as a gene loss of function tool by creating a stop codon or a gene knockdown by creating an upstream open reading frame (uORF).

    “It can also be repurposed as a gene loss of function tool by creating a stop codon or a gene knockdown by creating an upstream open reading frame (uORF) (Billon et al., 2017;Kuscu et al., 2017;Xiong et al., 2022).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Billon et al., 2017 source

      Not checked in the free run.

    • cites Kuscu et al., 2017 source

      Not checked in the free run.

    • cites Xiong et al., 2022 source

      Not checked in the free run.

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

    Premisenone given

    CRISPR/Cas9 is the most widely used system.

    “CRISPR/Cas9 is the most widely used system, but new systems such as Cas12, Cas13a, and Cas13b are in the pipeline (Smargon et al., 2017;Zaidi et al., 2017;Aman et al., 2018).”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Where gene knockouts in the coding region cause unfavourable pleiotropic effects, modification of the promoters to induce gene activation is desirable in developing stress tolerance in crops.

    “Where gene knockouts in the coding region cause unfavourable pleiotropic effects, modification of the promoters to induce gene activation is desirable in developing stress tolerance in crops (Ren C et al., 2022).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Ren C et al., 2022

      Not checked in the free run.

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

    Premisecitation

    Introduction of a stop codon to the Solanum lycopersicum L. glutamate decarboxylase SlGAD2 and SlGAD3 genes through CRISPR/Cas9 technology increased the GABA content in the tomatoes by 7-fold to 15-fold.

    “Introduction of a stop codon to the Solanum lycopersicum L. glutamate decarboxylase SlGAD2 and SlGAD3 genes through CRISPR/ Cas9 technology increased the GABA content in the tomatoes by 7-fold to 15-fold (Nonaka et al., 2017).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Nonaka et al., 2017 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Compared to other gene-editing methods such as TALEN or zinc finger nuclease (ZFN), CRISPR/Cas9 has advantages in simple target design, multiplexed mutation, versatility, and efficiency.

    “Compared to other gene-editing methods such as TALEN or zinc finger nuclease (ZFN), CRISPR/Cas9 has advantages in simple target design, multiplexed mutation, versatility, and efficiency (Leong et al., 2018).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Leong et al., 2018 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationcitation

    Base-editing studies conducted in Arabidopsis and rice provide confidence in using this technique in barley to improve useful traits, for instance, inducing early or late flowering to escape stress windows such as those for frost or drought.

    “Base-editing studies conducted in Arabidopsis and rice provide confidence in using this technique in barley to improve useful traits, for instance, inducing early or late flowering to escape stress windows such as those for frost or drought (Li et al., 2020;Xiong et al., 2022).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Li et al., 2020

      Not checked in the free run.

    • cites Xiong et al., 2022 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    Prime editing provides another platform for generating herbicide resistance in barley, especially as it can introduce pre-determined single base substitutions precisely.

    “They provide another platform for generating herbicide resistance in barley, especially as they can introduce pre-determined single base substitutions precisely.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    A γ-aminobutyric acid (GABA)-enriched tomato from Japan was the first CRISPR-edited food to enter the market.

    “A γ-aminobutyric acid (GABA)-enriched tomato from Japan was the first CRISPR-edited food to enter the market.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    These mutants are currently being screened under different stress conditions.

    “A number of mutants were obtained with improved agronomic and plant architecture traits such as seed number, spike length, tiller number, and root structure, and are currently being screened under different stress conditions (Nejat, 2022).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Nejat, 2022

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisenone given

    Some agronomic traits are controlled by a few quantitative trait loci (QTL).

    “Some agronomic traits are controlled by a few quantitative trait loci (QTL), and in such cases editing one gene is less likely to give a favourable phenotype.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    SDN-2 and SDN-3 rely on homology-directed repair (HDR) in which a template DNA sequence is used.

    “SDN-2 and SDN-3 rely on homology-directed repair (HDR) in which a template DNA sequence is used (Grohmann et al., 2019).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Grohmann et al., 2019 source

      Not checked in the free run.

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

    Premisecitation

    SDN-1 products rely on a non-homologous end joining (NHEJ) pathway to introduce a few base insertions or deletions.

    “SDN-1 products rely on a non-homologous end joining (NHEJ) pathway to introduce a few base insertions or deletions.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Grohmann et al., 2019 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    dPCR is not suitable for large amplicons and cannot detect multiple targets in one sample.

    “Third generation digital PCR (dPCR) enables rare mutation detection and trace DNA detection but is not suitable for large amplicons and cannot detect multiple targets in one sample (Mao et al., 2019).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Mao et al., 2019

      Not checked in the free run.

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

    Premisecitation

    Altered conformation due to a single base change in the DNA can cause different band patterns on non-denaturing gels which separate mutant and wild-type DNA.

    “Altered conformation due to a single base change in the DNA can cause different band patterns on non-denaturing gels which separate mutant and wild-type DNA (Zhu, 2005).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Zhu, 2005 source

      Not checked in the free run.

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

    Premisecitation

    Stable transformation of barley from immature embryos, microspore-derived embryos, and callus from young embryos was first reported by Wan and Lemaux (1994).

    “Stable transformation of barley from immature embryos, microspore-derived embryos, and callus from young embryos was first reported by Wan and Lemaux (1994).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Wan and Lemaux (1994) source

      Not checked in the free run.

  • May combine two claims.

    Interpretationnone given

    Nanoparticles can avoid genotype dependency in plants but require further insight in terms of application.

    “It can avoid genotype dependency in plants but requires further insight in terms of application.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Next generation sequencing (NGS) is useful for sequencing large mutant populations at lower cost.

    “The more advanced next generation sequencing (NGS) is useful for sequencing large mutant populations at lower cost (Peterson et al., 2016;Yang et al., 2022).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Peterson et al., 2016 source

      Not checked in the free run.

    • cites Yang et al., 2022 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Third generation digital PCR (dPCR) enables rare mutation detection and trace DNA detection.

    “Third generation digital PCR (dPCR) enables rare mutation detection and trace DNA detection but is not suitable for large amplicons and cannot detect multiple targets in one sample (Mao et al., 2019).”
    Quote in the paperNot a substantial claim

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

    Premisereasoning

    Single-strand conformation polymorphism (SSCP) can be used to detect SNPs due to its simplicity and low cost.

    “Single-strand conformation polymorphism (SSCP) on the other hand can be used to detect SNPs due to its simplicity and low cost.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    The transformation capacity of pan-genome accessions is yet to be tested.

    “A barley pan-genome is now available that provides more sequence information for gene editing, but the transformation capacity of pan-genome accessions is yet to be tested (Jayakodi et al., 2020).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Jayakodi et al., 2020 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    The availability of barley pan-genome sequences facilitates the identification of structural variations and conserved sequences within the genes of interest and pinpoints the most plausible gene for editing.

    “Furthermore, the availability of barley pan-genome sequences facilitates the identification of structural variations and conserved sequences within the genes of interest and pinpoints the most plausible gene for editing (Jayakodi et al., 2020).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Jayakodi et al., 2020 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    HRM generates sequence-related melting profiles and reveals the genotype at the level of a single nucleotide.

    “It generates sequence-related melting profiles and reveals the genotype at the level of a single nucleotide (Chatzidimopoulos et al., 2019).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Chatzidimopoulos et al., 2019 source

      Not checked in the free run.

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

    Premisereasoning

    Enzyme recognition sites are usually chosen close to the protospacer adjacent motif (PAM), and thus any mutations away from PAM/recognition site can give false negative results, limiting the application of this method.

    “Enzyme recognition sites are usually chosen close to the protospacer adjacent motif (PAM), and thus any mutations away from PAM/recognition site can give false negative results, limiting the application of this method.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    De novo induction of gene-edited meristems avoids the tissue culture step and can be induced in aseptically grown seedlings and soil-grown plants.

    “De novo induction of geneedited meristems also avoids the tissue culture step and can be induced in aseptically grown seedlings and soil-grown plants (Maher et al., 2020).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Maher et al., 2020 source

      Not checked in the free run.

  • May combine two claims.

    Premisenone given

    Regardless of the delivery method, the process involves tissue culture, which is tedious, time-consuming, and genotype-dependent.

    “Regardless of the delivery method, the process involves tissue culture, which is tedious, time-consuming, and genotype-dependent.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    SYNV delivery does not require the isolation of plant cells or tissues and can be delivered directly into intact plants.

    “This does not require the isolation of plant cells or tissues and can be delivered directly into intact plants.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Sanger sequencing is widely used and useful for low-volume high-quality DNA sequencing.

    “Sanger sequencing is widely used and useful for low-volume high-quality DNA sequencing (Nonaka et al., 2017;Doll et al., 2019).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Nonaka et al., 2017 source

      Not checked in the free run.

    • cites Doll et al., 2019 source

      Not checked in the free run.

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

    Premisenone given

    The spring barley cultivar 'Golden Promise' has been used for decades with a successful transformation and regeneration rate.

    “The spring barley cultivar 'Golden Promise' has been used for decades with a successful transformation and regeneration rate.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    TA-cloning can be used to clone multiple DNA fragments for sequencing, which can detect heterozygous or chimeric mutations.

    “TA-cloning can be used to clone multiple DNA fragments for sequencing, which can detect heterozygous or chimeric mutations (Ma et al., 2020).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Ma et al., 2020

      Not checked in the free run.

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

    Premisecitation

    Several molecular techniques from agarose gel electrophoresis to sequencing can be used for genotyping mutants.

    “Several molecular techniques from agarose gel electrophoresis to sequencing can be used for this task (Zhang et al., 2021;Karunarathne et al., 2022).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Zhang et al., 2021 source

      Not checked in the free run.

    • cites Karunarathne et al., 2022 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Transgene-free homozygous mutants are desirable and in barley are feasible in the T1 generation.

    “Transgene-free homozygous mutants are desirable and in barley are feasible in the T1 generation (Zeng et al., 2020).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Zeng et al., 2020 source

      Not checked in the free run.

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

    Premisenone given

    Both Sanger sequencing and NGS enable the identification of the exact order of a gene sequence, and the alignment of this DNA or protein sequence helps detect the respective amino acid change in mutants.

    “Both techniques enable the identification of the exact order of a gene sequence, and the alignment of this DNA or protein sequence helps us to detect the respective amino acid change in mutants.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    PCR-restriction enzyme digestion has proved to be useful in genotyping mutants.

    “PCR-restriction enzyme digestion has proved to be useful in genotyping mutants.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    All the above techniques must be followed by sequencing, which is the gold standard for genotyping mutants.

    “Regardless, all the above techniques must be followed by sequencing, which is the gold standard for genotyping mutants (Xu et al., 2020;Zhang et al., 2021).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Xu et al., 2020

      Not checked in the free run.

    • cites Zhang et al., 2021 source

      Not checked in the free run.

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

    Premisecitation

    Tracking of InDels by Decomposition (TIDE) is an effective web tool that can be used to analyse sequencing data, which estimates the spectra and frequencies of small insertions and deletions generated from gene editing.

    “Tracking of InDels by Decomposition (TIDE) is an effective web tool (https://tide.nki.nl) that can be used to analyse sequencing data, which estimates the spectra and frequencies of small insertions and deletions generated from gene editing (Brinkman et al., 2014).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Brinkman et al., 2014 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisenone given

    CRISPR/Cas9 has already been applied in both monocots and dicots to improve numerous agronomic traits.

    “CRISPR/Cas9 gene-editing provides an ideal platform with high efficiency and versatility, which has already been applied in both monocots and dicots to improve numerous agronomic traits, and some products have already been commercialised.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S4 (based).

    Interpretationnone given

    A combination of gene-editing with speed breeding techniques can accelerate crop breeding to develop new cultivars with desirable traits.

    “A combination of gene-editing with speed breeding techniques can accelerate crop breeding to develop new cultivars with desirable traits.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based). May combine two claims.

    Premisenone given

    Further applications of CRISPR/Cas9 not discussed in this review include haploid induction and generating male sterile lines to support hybrid vigour.

    “Further applications of CRISPR/Cas9 not discussed in this review include haploid induction and generating male sterile lines to support hybrid vigour.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisenone given

    Some CRISPR/Cas9 products have already been commercialised.

    “CRISPR/Cas9 gene-editing provides an ideal platform with high efficiency and versatility, which has already been applied in both monocots and dicots to improve numerous agronomic traits, and some products have already been commercialised.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S4 (based).

    Interpretationnone given

    CRISPR/Cas9-aided domestication for climate resilience and other desirable traits takes only a short time compared to traditional domestication.

    “Barley wild species have huge genetic diversity and CRISPR/Cas9-aided domestication for climate resilience and other desirable traits takes only a short time compared to traditional domestication.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S4 (based).

    Premisenone given

    Barley wild species have huge genetic diversity.

    “Barley wild species have huge genetic diversity and CRISPR/Cas9-aided domestication for climate resilience and other desirable traits takes only a short time compared to traditional domestication.”
    Quote in the paperNot a substantial claim
103 of 103

  • The paper relies on this without support.

    The identified stress-tolerance genes can be effectively targeted by gene editing to improve barley abiotic stress tolerance.

    “The listed genes counteract key stresses such as drought, salinity, and nutrient deficiency, and the potential application of the respective gene-editing technologies will provide insight into barley improvement for climate resilience.”

  • The paper relies on this without support.

    The Morex v3 reference sequence is adequate for mapping genes across barley genotypes.

    “Gene locations and annotations were obtained by blasting the National Center for Biotechnology Information (NCBI), UniProt, or Morex v1 accession information provided in publications against the Morex v3 reference sequence (Colmsee et al., 2015;Afgan et al., 2018;Tello-Ruiz et al., 2022).”

  • The paper relies on this without support.

    The outcomes of the gene-editing studies in barley are representative of what can be achieved in breeding tolerant cultivars.

    “The usefulness of this material for breeding tolerant cultivars is largely dependent on the outcomes of these studies.”

  • The paper relies on this without support.

    Cereal crops including barley suffer significant yield and economic losses due to increased abiotic stresses from climate change.

    “Cereal crops including barley suffer significant yield and economic losses in the face of increased abiotic stresses due to climate change.”
4 of 4

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Using CRISPR/Cas9 gene editing technologies, barley abiotic stress tolerance has been enhanced by targeting specific genes associated with drought and salinity resistance.

    “Using CRISPR/Cas9 gene editing technologies, barley abiotic stress tolerance has been enhanced by targeting specific genes associated with drought and salinity resistance (Karunarathne et al., 2023; Yadav et al., 2023).”

    From Emerging applications of gene editing technologies for the development of climate-resilient crops (2025) · cites this paper as Karunarathne et al., 2023

    Section 4.2 says the barley genes used to achieve drought tolerance were overexpressed, but not through CRISPR/Cas9. It also reports that CRISPR-generated HvHVP10 knockout lines had decreased, not enhanced, salt tolerance. The review discusses CRISPR as a potential strategy, but these passages contradict the claimed improvements. (LLM, conf=0.88)

    Checked by a reasoning model — may be wrong.

    Experimental — accuracy not yet measured

  • Gene pyramiding has been effectively demonstrated in several crops, including rice, maize, and barley.

    “Gene pyramiding has been effectively demonstrated in several crops, including rice, maize, and barley.”

    From Emerging applications of gene editing technologies for the development of climate-resilient crops (2025) · cites this paper as Karunarathne et al., 2023

    Section 4.2 reports heritable mutations from multiplex editing of two barley genes and simultaneous multigene activation in rice. These are related demonstrations, but the paper does not establish that gene pyramiding was effectively demonstrated in all three named crops; its discussion of maize in Section 4.4 concerns genetic transformation, not pyramiding. Page numbers were not available in our copy. (LLM, conf=0.77)

    Experimental — accuracy not yet measured

  • A significant technical hurdle is the high-throughput mutant genotyping required to identify and confirm the presence of multiple edited genes in a single plant.

    “A significant technical hurdle is the high-throughput mutant genotyping required to identify and confirm the presence of multiple edited genes in a single plant (Karunarathne et al., 2023).”

    From Emerging applications of gene editing technologies for the development of climate-resilient crops (2025) · cites this paper as Karunarathne et al., 2023

Cites

1 retracted

Flagged first, then by citations

20 of the 225 references couldn’t be matched to a known paper.

Research works printed but not linked: 4

  1. Heffer P, Prud’homme M, 2016. Global nitrogen fertiliser demand and supply: trend, current level and outlook. Proceedings of 2016 International Nitrogen Initiative Conference, Melbourne, Australia.
  2. Hudzenko VM, Demydov OA, Polishchuk TP, et al., 2021. Comprehensive evaluation of spring barley yield and tolerance to abiotic and biotic stresses. Ukr J Ecol, 11(8):48–55. https://doi.org/10.15421/2021_267 · DOI
  3. Llewellyn R, Ronning D, Ouzman J, et al., 2016. Impact of Weeds on Australian Grain Production: the Cost of Weeds to Australian Grain Growers and the Adoption of Weed Management and Tillage Practices. Technical Report No. EP161334, Grains Research and Development Corporation, Canberra, Australia. https://grdc.com.au/__data/assets/pdf_file/0027/75843/grdc_weeds_review_r8.pdf.pdf
  4. Nejat N, 2022. Gene Editing of Elite Malting Barley Cultivar RGT Planet Using Agrobacterium-Mediated Delivery of CRISPR/Cas9. PhD Thesis, Murdoch University, Perth, Australia.

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Authors
Karunarathne, S., Walker, E., Sharma, D., Li, C., & Han, Y.
Affiliations
  • Murdoch University (AU)
  • Department of Primary Industries and Regional Development (AU)
Corresponding author
Li, Chengdao
ORCID
Venue
Journal of Zhejiang University-SCIENCE B
Publisher
Zhejiang University Press
Published
2023-07-03
Volume
24
Issue
12
Pages
1069-1092
ISSN
  • 1673-1581 (ISSN-L)
  • 1862-1783
DOI
10.1631/jzus.b2200552
PMC
PMC10710907
OpenAlex
W4382932510
Cited by
18
References
225
Citations by year
2026: 3 · 2025: 9 · 2024: 4 · 2023: 2
FWCI
2.84
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Zhejiang University SCIENCE B (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Stress Responses and Tolerance
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Wheat and Barley Genetics and Pathology
Keywords
barley, abiotic stress tolerance, CRISPR/Cas9 genome editing, drought tolerance, salt tolerance, heat stress, cold stress, waterlogging, nutritional deficiency, base editing, prime editing, multiplex genome editing, genetic resources, climate resilience, crop breeding

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