High temperature (HT) events have negative impact on wheat grains yield and quality. Transcriptome profiles of wheat developing grains of commercial genotypes (Antequera and Bancal) and landraces ...(Ardito and Magueija) submitted to heatwave-like treatments during grain filling were evaluated. Landraces showed significantly more differentially expressed genes (DEGs) and presented more similar responses than commercial genotypes. DEGs were more associated with transcription and RNA and protein synthesis in Antequera and with metabolism alterations in Bancal and landraces. Landraces upregulated genes encoding proteins already described as HT responsive, like heat shock proteins and cupins. Apart from the genes encoding HSP, two other genes were upregulated in all genotypes, one encoding for Adenylate kinase, essential for the cellular homeostasis, and the other for ferritin, recently related with increased tolerance to several abiotic stress in Arabidopsis. Moreover, a NAC transcription factor involved in plant development, known to be a negative regulator of starch synthesis and grain yield, was found to be upregulated in both commercial varieties and downregulated in Magueija landrace. The detected diversity of molecular processes involved in heat response of commercial and traditional genotypes contribute to understand the importance of genetic diversity and relevant pathways to cope with these extreme events.
The genus Avena comprises four distinct genomes organized in diploid (AA or CC), tetraploid (AABB or AACC) and hexaploid species (AACCDD), constituting an interesting model for phylogenetic analysis. ...The aim of this work was to characterize 45S rDNA intergenic spacer (IGS) variability in distinct species representative of Avena genome diversity-A. strigosa (AA), A. ventricosa (CvCv), A. eriantha (CpCp), A. barbata (AABB), A. murphyi (AACC), A. sativa (AACCDD) and A. sterilis (AACCDD) through the assessment of the 5' external transcribed spacer (5'-ETS), a promising IGS region for phylogenetic studies poorly studied in Avena genus. In this work, IGS length polymorphisms were detected mainly due to distinct 5'-ETS sequence types resulting from major differences in the number and organization of repeated motifs. Although species with A genome revealed a 5'-ETS organization (A-organization) similar to the one previously described in A. sativa, a distinct organization was unraveled in C genome diploid species (C-organization). Interestingly, such new organization presents a higher similarity with other Poaceae species than A-genome sequences, supporting the hypothesis of C-genome being the ancestral Avena genome. Additionally, polyploid species with both genomes mainly retain the A-genome 5'-ETS organization, confirming the preferential elimination of C-genome sequences in Avena polyploid species. Moreover, 5'-ETS sequences phylogenetic analysis consistently clustered the species studied according to ploidy and genomic constitution supporting the use of ribosomal genes to highlight Avena species evolutive pathways.
Faba bean (Vicia faba L.) is a facultative cross-pollinating legume crop with a great importance for food and feed due to its high protein content as well as the important role in soil fertility and ...nitrogen fixation. In this work we evaluated genetic diversity and population structure of faba bean accessions from the Western Mediterranean basin and wild related species. For that purpose we screened 53 V. faba, 2 V. johannis and 7 V. narbonensis accessions from Portugal, Spain and Morocco with 28 faba bean Single Sequence Repeats (SSR). SSR genotyping showed that the number of alleles detected per locus for the polymorphic markers ranged between 2 and 10, with Polymorphic Information Content (PIC) values between 0.662 and 0.071, and heterozygosity (HO) between 0-0.467. Heterozygosity and inbreeding coefficient levels indicate a higher level of inbreeding in wild related species than in cultivated Vicia. The analysis of molecular variance (AMOVA) showed a superior genetic diversity within accessions than between accessions even from distant regions. These results are in accordance to population structure analysis showing that individuals from the same accession can be genetically more similar to individuals from far away accessions, than from individuals from the same accession. In all three levels of analysis (whole panel of cultivated and wild accessions, cultivated faba bean accessions and Portuguese accessions) no population structure was observed based on geography or climatic factors. Differences between V. narbonensis and V. johannis are undetectable although these wild taxa are clearly distinct from V. faba accessions. Thus, a limited gene flow occurred between cultivated accessions and wild relatives. Contrastingly, the lack of population structure seems to indicate a high degree of gene flow between V. faba accessions, possibly explained by the partially allogamous habit in association with frequent seed exchange/introduction.
The predicted world population increase along with climate changes threatens sustainable agricultural supply in the coming decades. It is therefore vital to understand crops diversity associated to ...abiotic stress response. Heat stress is considered one of the major constrains on crops productivity thus it is essential to develop new approaches for a precocious and rigorous evaluation of varietal diversity regarding heat tolerance. Plant cell membrane thermostability (CMS) is a widely used method for wheat thermotolerance assessment although its limitations require complementary solutions. In this work we used CMS assay and explored TdHSP101C genes as an additional tool for durum wheat screening. Genomic and transcriptomic analyses of TdHSP101C genes were performed in varieties with contrasting CMS results and further correlated with heat stress tolerance during fertilization and seed development. Although the durum wheat varieties studied presented a very high homology on TdHSP101C genes (>99%) the transcriptomic assessment allowed the discrimination between varieties with good CMS results and its correlation with differential impacts of heat treatment during inflorescence emergence and seed development on grain yield. The evidences here reported indicate that TdHSP101C transcription levels induced by heat stress in fully expanded leaves may be a promising complementary screening tool to discriminate between durum wheat varieties identified as thermotolerant through CMS.
Wheat is undoubtedly one of the most important crops worldwide and it is essential to study how the distinct varieties answer to heat waves associated with climatic changes, in order to design ...adequate wheat breeding strategies. To assess high temperature (HT) impact in wheat grain characteristics, seven commercial varieties, which have been recommended for production in Portugal, were submitted for one-week HT treatment ten days after anthesis. Firstly, predicted grain technological quality was determined by giving high scores for all varieties studied, based on the allelic compositions of genes encoding high molecular weight glutenins, granule-bound starch synthase and puroindolines. The effects of HT on transcription levels of those genes were, for the first time, evaluated in distinct wheat genotypes, in comparison with control plants. Finally, protein fraction content in mature grains were also estimated in untreated and treated plants. Immature grains from plants, maintained in control conditions, showed significant intervarietal differences in transcription levels of genes associated with grain quality traits, a variability that was significantly reduced in grains from HT treated plants. On the other hand, the influence of HT in mature grain protein-fractions and in gliadin/glutenin ratios revealed intervarietal diversity, even with opposite effects in some varieties. The present study, therefore, discloses marked variability in parameters associated with flour quality between the wheat varieties analyzed, which are differentially affected by HT treatments, similar to heat waves frequently observed in climate change scenarios.
Salinity is an important problem for agriculture in the Mediterranean area, and thus, it is essential to develop mitigation strategies to reduce its impact. The main objective of this study was to ...test the effectiveness of halotolerant plant growth-promoting bacteria (H-PGPB) in improving grapevine salt stress tolerance. Grapevines grafted onto a salt-sensitive rootstock were inoculated with a consortium of H-PGPB. The substrate of half of the plants of each treatment was salinized up to 2 dS m
−1
. Plants grew for six days under these conditions, and afterward, NaCl was removed to assess plant recovery through growth, physiology, and canopy temperature measurements. Inoculation with H-PGPB had a positive effect on plant physiology, but after salt treatment, grapevines stopped their photosynthetic metabolism, leading to severe defoliation. Remarkably, after salt stress removal, inoculated plants re-sprouted faster, demonstrating that H-PGPB inoculation could be a good practice to increase vineyard resilience to salt stress.
Wheat is a dietary staple consumed worldwide strongly responsible for proteins and carbohydrate population intake. However, wheat production and quality will scarcely fulfill forward demands, which ...are compounded by high-temperature (HT) events as heatwaves, increasingly common in Portugal. Thus, landraces assume crucial importance as potential reservoirs of useful traits for wheat breeding and may be pre-adapted to extreme environmental conditions. This work evaluates four Portuguese landrace yield and grain composition through attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, particularly protein content, and their responses to HT treatment mimicking a heatwave. Landraces showed distinct yield traits, especially plant height and first spike grain number, and a similar pattern in FTIR spectra, although revealing differences in grain components' proportions. Comparison between spectra band intensity indicates that Ardito has the highest protein-related peaks, contrary to Magueija, which appears to be the landrace with higher lipid content. In plants submitted to 1 week of HT treatment 10 days after anthesis, the first spike grain size and weight were markedly reduced in all landraces. Additionally, it was observed that a general increase in grain protein content in the four landraces, being the increment observed in Ardito and Grécia, is statistically significant. The comparative assessment of control and HT average FTIR spectra denoted also the occurrence of alterations in grain polysaccharide composition. An integrated assessment of the evaluations performed revealed that Ardito and Magueija landraces presented diverse yield-related characteristics and distinct responses to cope with HT. In fact, the former landrace revealed considerable grain yield diminution along with an increase in grain protein proportion after HT, while the latter showed a significant increase in spikes and grain number, with grain quality detriment. These results reinforce the relevance of scrutinizing old genotype diversity seeking for useful characteristics, particularly considering HT impact on grain production and quality.
Stressful high temperatures on plants can limit whole-plant function and decrease crop productivity. However, little is known regarding heat stress effects on microtubule cytoskeleton and chromatin ...in roots from intact plants. Here we studied high temperature effects on cell division, microtubule and chromatin organization patterns in rye root tips from intact plants subjected to 40ºC for 4 h and after different recovery periods (0RT, 7RT, 24 RT). We showed that heat stress induced changes in nuclear morphology as detected by the unusual presence of interphase cells with irregularly shaped nuclei, probably associated with changes in chromosome segregation at anaphase, leading to micronuclei formation as well as changes in the mitotic index. These alterations were associated to differential effects in microtubules organization in both heat-stressed interphase and mitotic cells at 0RT and 7RT. Although no changes in the distribution of H3 phosphorylation of Ser 10 residues on chromatin were found in cells from heat-stressed plants, marked alterations in chromatin DNA methylation patterns were detected. These effects included higher agglutination of 5-methylcytosine domains in both interphase and metaphase cells compared to controls. Taken together these results seem to suggest that alterations in microtubule conformation upon heat stress influences nuclear chromatin organization and cell cycle progression. However, when seedlings recovered from stress (24RT), root tip cells presented microtubule configurations and chromatin organization patterns similar to controls. We conclude that in spite of heat stress markedly altered cell cycle progression and distribution of epigenetic marks, these responses are transient to cope with such stress conditions in the roots.
Wheat is one of the most important cereals for food and feed, and it is, therefore, necessary to determine the effects of short-term high temperature events (heatwaves) during grain filling. These ...heatwave events are increasingly common, especially in Portugal. In this work, seven commercial varieties recommended for production in Portugal were submitted to one-week high temperature (HT) treatment ten days after anthesis to evaluate heat effects on grain yield and quality. Grain yield parameters, such as grain number and weight, were evaluated as well as grain composition through attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy. Variation in HT response between varieties was detected. Grain number and weight tended to decrease in most varieties analyzed. However, two varieties proved to be more resilient since grain number and weight remain unaltered in the Bancal variety, which is the one with better yield results, and even increased in the Pata Negra variety. Regarding grain composition, the comparison between ATR-FTIR spectra of milled grains from control and HT plants revealed alterations in peaks assigned to polysaccharides and proteins. Additionally, a model was built based on nitrogen elemental analysis to predict protein content in flour samples through spectral data that corroborated the differences identified by spectra profile comparison. Moreover, both analyses showed that the intervarietal diversity observed in control conditions was significantly reduced in HT treated plants. The results obtained highlight the intervarietal diversity of wheat response to HT, regarding grain yield parameters, grain composition, and particularly, protein content.
Allopolyploidy is a significant evolutionary process, resulting in new species with diploid or greater chromosome complements derived from two or more progenitor species. We examined the chromosomal ...consequences of genomic merger in Arabidopsis suecica, the allotetraploid hybrid of Arabidopsis thaliana and Arabidopsis arenosa. Fluorescence in situ hybridization with centromere, nucleolus organizer region (NOR), and 5S rRNA gene probes reveals the expected numbers of progenitor chromosomes in natural A. suecica, but one pair of A. thaliana NORs and one pair of A. arenosa-derived 5S gene loci are missing. Similarly, in newly formed synthetic A. suecica-like allotetraploids, pairs of A. thaliana NORs are gained de novo, lost, and/or transposed to A. arenosa chromosomes, with genotypic differences apparent between F3 siblings of the same F2 parent and between independent lines. Likewise, pairs of A. arenosa 5S genes are lost and novel linkages between 5S loci and NORs arise in synthetic allotetraploids. By contrast, the expected numbers of A. arenosa-derived NORs and A. thaliana-derived 5S loci are found in both natural and synthetic A. suecica. Collectively, these observations suggest that some, but not all, loci are unstable in newly formed A. suecica allotetraploids and can participate in a variety of alternative rearrangements, some of which resemble chromosomal changes found in nature.
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