Zhi Zheng

The detrimental effects of drought on crop production are strongly influenced by interactions with other biotic and abiotic stresses, including Fusarium crown rot (FCR), a disease known to cause severe yield loss in crops experiencing drought after anthesis. This study aimed to identify loci that simultaneously confer drought tolerance and FCR resistance using a recombinant inbred line (RIL) population. The population derived from a cross between cultivated barley ( Hordeum vulgare L.) and wild barley ( Hordeum spontaneum C. Koch.), was evaluated under drought‐stressed and non‐drought‐stressed conditions. Three loci conferring drought tolerance were identified on chromosome arms 2HL, 4HL and 5HS. Of these, the 2HL locus had the largest effect, influencing leaf wilting, leaf senescence and shoot dry weight, and importantly, also conferred FCR resistance. The RIL population was exposure to FCR infection and drought stress in glasshouse and controlled environment facility, respectively. Genotyping‐by‐sequencing (GBS) was used to develop a high‐density genetic map, which facilitated QTL identification through linkage analysis. Significant correlations were observed among the assessed traits and previously collected agronomic traits. Candidate genes at the 2HL locus were identified based on its location and sequence variations between parental genotypes. This study represents the first identification of such a locus conferring both drought tolerance and FCR resistance, which may open new avenues for leveraging existing drought tolerant genes in enhancing FCR resistance by targeting pathways associated with water stress adaptation.

Keymessage: In this first QTL mapping study of embryo size in barley, novel and stable QTL were identified and candidate genes underlying a significant locus independent of kernel size were identified based on orthologous analysis and comparison of the whole-genome assemblies for both parental genotypes of the mapping population. Embryo, also known as germ, in cereal grains plays a crucial role in plant development. The embryo accounts for only a small portion of grain weight but it is rich in nutrients. Larger embryo translates to a more nutritious grain and larger store of energy reserves, which can benefit seed germination and seedling establishment. However, reports on quantitative trait loci (QTL) affecting embryo size in barley is rare. To understand the genetic basis of embryo size in barley, a population consisting of 201 F9 recombination inbred lines (RILs) was assessed in four environments. Three regions affecting various characteristics of embryo size including embryo length (EL), embryo width (EW) and embryo area (EA) were consistently identified. They located on chromosomes 2H, 4H and 7H, respectively. Among them, the QTL on 7H was not significantly affected by kernel size. Phenotypic variances explained by this QTL for EL, EW and EA were 11.8%, 9.3% and 12.7%, respectively. Taken advantage of the available genomic assemblies of the two parental genotypes, candidate genes for this locus on 7H were identified. In addition, significant correlations between embryo size and early vigour and kernel traits were detected. To our knowledge, the present study is for the first time reporting QTL conferring embryo size by directly measuring the characteristics as quantitative trait in barley, which would broaden our understanding of the genetic basis of barley embryo size and offer valuable targets for future breeding programmes.

Embryo, also known as germ, in cereal grains plays a crucial role in plant development. The embryo accounts for only a small portion of grain weight but it is rich in nutrients. Larger embryo translates to a more nutritious grain and larger store of energy reserves, which can benefit seed germination and seedling establishment. However, reports on quantitative trait loci (QTL) affecting embryo size in barley is rare. To understand the genetic basis of embryo size in barley, a population consisting of 201 F9 recombination inbred lines (RILs) was assessed in four environments. Three regions affecting various characteristics of embryo size including embryo length (EL), embryo width (EW) and embryo area (EA) were consistently identified. They located on chromosomes 2H, 4H and 7H, respectively. Among them, the QTL on 7H was bit affected by kernel size. Phenotypic variances explained by this QTL for EL, EW and EA were 11.8%, 9.3% and 12.7%, respectively. Taken advantage of the available genomic assemblies of the two parental genotypes, candidate genes for this locus on 7H were identified. In addition, significant correlations between embryo size and early vigour and kernel traits were detected. To our knowledge, the present study is for the first time reporting QTL conferring embryo size by directly measuring the characteristics as quantitative trait in barley, which would broaden our understanding of the genetic basis of barley embryo size and offer valuable targets for future breeding programs.

Sweet cherry (Prunus avium L.) has become an economically important fruit in China. And its cultivation area has significantly expanded over the last three decades (Wang et al. 2020; Zhao et al. 20...

Key message: This study reported the identification and validation of novel QTL conferring coleoptile length in barley and predicted candidate genes underlying the largest effect QTL based on orthologous analysis and comparison of the whole genome assemblies for both parental genotypes of the mapping population. Coleoptile length (CL) is one of the most important agronomic traits in cereal crops due to its direct influence on the optimal depth for seed sowing which facilitates better seedling establishment. Varieties with longer coleoptiles are preferred in drought-prone areas where less moisture maintains at the top layer of the soil. Compared to wheat, genetic study on coleoptile length is limited in barley. Here, we reported a study on detecting the genomic regions associated with CL in barley by assessing a population consisting of 201 recombinant inbred lines. Four putative QTL conferring CL were consistently identified on chromosomes 1H, 5H, 6H, and 7H in each of the trials conducted. Of these QTL, the two located on chromosomes 5H and 6H (designated as Qcl.caf-5H and Qcl.caf-6H) are likely novel and Qcl.caf-5H showed the most significant effect explaining up to 30.9% of phenotypic variance with a LOD value of 15.1. To further validate the effect of this putative QTL, five pairs of near isogenic lines (NILs) were then developed and assessed. Analysis of the NILs showed an average difference of 21.0% in CL between the two isolines. Notably, none of the other assessed morphological characteristics showed consistent differences between the two isolines for each pair of the NILs. Candidate genes underlying the Qcl.caf-5H locus were also predicted by employing orthologous analysis and comparing the genome assemblies for both parental genotypes of the mapping population in the present study. Taken together, these findings expand our understanding on genetic basis of CL and will be indicative for further gene cloning and functional analysis underly this locus in barley.

Coleoptile length (CL) is one of the most important agronomic traits in cereal crops due to its direct influence on the optimal depth for seed sowing which facilitates better seedling establishment. Varieties with longer coleoptiles are preferred in drought-prone areas where less moisture maintains at the top layer of the soil. Compared to wheat, genetic study on coleoptile length is limited in barley. Here, we reported a study on detecting the genomic regions associated with CL in barley by assessing a population consisting of 201 recombinant inbred lines (RILs). Four putative QTL conferring CL were consistently identified on chromosomes 1H, 5H, 6H, and 7H in each of the trials conducted. Of these QTL, the two located on chromosomes 5H and 6H (designated as Qcl.caf-5H and Qcl.caf-6H) are likely novel and Qcl.caf-5Hshowed the most significant effect explaining up to 30.9% of phenotypic variance with a LOD value of 15.1. To further validate the effect of this putative QTL, five pairs of near isogenic lines (NILs) were then developed and assessed. Analysis of the NILs showed an average difference of 21.0% in CL between the two isolines. Notably, none of the other assessed morphological characteristics showed consistent differences between the two isolines for each pair of the NILs. Candidate genes underlying the Qcl.caf-5H locus were also predicted by employing orthologous analysis and comparing the genome assemblies for both parental genotypes of the mapping population in the present study. Taken together, these findings expand our understanding on genetic basis of CL and will be indicative for further gene cloning and functional analysis underly this locus in barley.

Key message: This study reported validation and fine mapping of a Fusarium crown rot resistant locus on chromosome arm 6HL in barley using near isogenic lines, transcriptome sequences, and a large near isogenic line-derived population. Fusarium crown rot (FCR), caused by Fusarium pseudograminearum, is a chronic and serious disease affecting cereal production in semi-arid regions globally. The increasing prevalence of this disease in recent years is attributed to the widespread adoption of minimum tillage and stubble retention practices. In the study reported here, we generated eight pairs of near isogenic lines (NILs) targeting a putative QTL (Qcrs.caf-6H) conferring FCR resistance in barley. Assessing the NILs confirmed the large effect of this locus. Aimed to develop markers that can be reliably used in incorporating this resistant allele into breeding programs and identify candidate genes, transcriptomic analyses were conducted against three of the NIL pairs and a large NIL-derived population consisting of 1085 F7 recombinant inbred lines generated. By analyzing the transcriptomic data and the fine mapping population, Qcrs.caf-6H was delineated into an interval of 0.9 cM covering a physical distance of ~ 547 kb. Six markers co-segregating with this locus were developed. Based on differential gene expression and SNP variations between the two isolines among the three NIL pairs, candidate genes underlying the resistance at this locus were detected. These results would improve the efficiency of incorporating the targeted locus into barley breeding programs and facilitate the cloning of causal gene(s) responsible for the resistance.

Background: The thermotolerant yeast is beneficial in terms of efficiency improvement of processes and reduction of costs, while Saccharomyces cerevisiae does not efficiently grow and ferment at high-temperature conditions. The sterol composition alteration from ergosterol to fecosterol in the cell membrane of S. cerevisiae affects the thermotolerant capability. Results: In this study, S. cerevisiae ERG5, ERG4, and ERG3 were knocked out using the CRISPR–Cas9 approach to impact the gene expression involved in ergosterol synthesis. The highest thermotolerant strain was S. cerevisiae ERG5ΔERG4ΔERG3Δ, which produced 22.1 g/L ethanol at 37 °C using the initial glucose concentration of 50 g/L with an increase by 9.4% compared with the wild type (20.2 g/L). The ethanol concentration of 9.4 g/L was produced at 42 ℃, which was 2.85-fold of the wild-type strain (3.3 g/L). The molecular mechanism of engineered S. cerevisiae at the RNA level was analyzed using the transcriptomics method. The simultaneous deletion of S. cerevisiae ERG5, ERG4, and ERG3 caused 278 up-regulated genes and 1892 down-regulated genes in comparison with the wild-type strain. KEGG pathway analysis indicated that the up-regulated genes relevant to ergosterol metabolism were ERG1, ERG11, and ERG5, while the down-regulated genes were ERG9 and ERG26. S. cerevisiae ERG5ΔERG4ΔERG3Δ produced 41.6 g/L of ethanol at 37 °C with 107.7 g/L of corn liquefied glucose as carbon source. Conclusion: Simultaneous deletion of ERG5, ERG4, and ERG3 resulted in the thermotolerance improvement of S. cerevisiae ERG5ΔERG4ΔERG3Δ with cell viability improvement by 1.19-fold at 42 °C via modification of steroid metabolic pathway. S. cerevisiae ERG5ΔERG4ΔERG3Δ could effectively produce ethanol at 37 °C using corn liquefied glucose as carbon source. Therefore, S. cerevisiae ERG5ΔERG4ΔERG3Δ had potential in ethanol production at a large scale under supra-optimal temperature.

Fusarium crown rot (FCR), caused by Fusarium pseudograminearum, is a chronic and serious disease affecting semi-arid regions globally. The increasing prevalence of this disease in recent years is attributed to the widespread adoption of minimum tillage and stubble retention practices. In the study reported here, we generated eight pairs of near isogenic lines (NILs) targeting a putative QTL (Qcrs.caf-6H) conferring FCR resistance in barley. Assessing the NILs confirmed the large effect of this locus. Aimed to develop markers that can be reliably used in incorporating this resistant locus into breeding programs and identify candidate genes, transcriptomic analyses were conducted against three of the NIL pairs and a large NIL-derived population consisting of 1,085 F7 recombinant inbred lines (RILs) was generated. By analysing the transcriptomic data and the fine mapping population, Qcrs.caf-6Hwas delineated into an interval of 0.9 cM covering a physical distance of ~547 kb. Six markers co-segregating with this locus were developed. Based on differential gene expression and SNP variations between the two isolines among the three NIL pairs, candidate genes underlying the resistance at this locus were detected. These results would improve the efficiency of incorporating the targeted locus into barley breeding programs and facilitate the cloning of causal gene(s) responsible for the resistance.