Green synthesis of NiO NPs for metagenome-derived laccase stabilization: Detoxifying pollutants and wastes.

Laccases play a crucial role in neutralizing environmental pollutants, including antibiotics and phenolic compounds, by converting them into less harmful substances via a unique oxidation process. This study introduces an environmentally sustainable remediation technique, utilizing NiO nanoparticles (NPs) synthesized through green chemistry to immobilize a metagenome-derived laccase, PersiLac1, enhancing its application in pollutant detoxification. Salvadora persica leaf extract was used for the synthesis of NiO nanoparticles, utilizing its phytochemical constituents as reducing and capping agents, followed by characterization through different analyses. Characterization of NiO nanoparticles revealed distinctive FTIR absorption peaks indicating the nanoparticulate structure, while FESEM showed structured NiO with robust interconnections and dimensionality of about 50nm, confirmed by EDX analysis to have a consistent distribution of Ni and O. The immobilized PersiLac1 demonstrated enhanced thermal stability, with 85.55 % activity at 80 °C and reduced enzyme leaching, retaining 67.93 % activity across 15 biocatalytic cycles. It efficiently reduced rice straw (RS) phenol by 67.97 % within 210 min and degraded 70-78 % of tetracycline (TC) across a wide pH range (4.0-8.0), showing superior performance over the free enzyme. Immobilized laccase achieved up to 71 % TC removal at 40-80 °C, significantly outperforming the free enzyme. Notably, 54 % efficiency was achieved at 500 mg/L TC by immobilized laccase at 120 min. This research showed the potential of green-synthesized NiO nanoparticles to effectively immobilize laccase, presenting an eco-friendly approach to purify pollutants such as phenols and antibiotics. The durability and reusability of the immobilized enzyme, coupled with its ability to reduce pollutants, indicates a viable method for cleaning the environment. Nonetheless, the production costs and scalability of NiO nanoparticles for widespread industrial applications pose significant challenges. Future studies should focus on implementation at an industrial level and examine a wider range of pollutants to fully leverage the environmental clean-up capabilities of this innovative technology.

Phylogenomic investigation of safflower (Carthamus tinctorius) and related species using genotyping-by-sequencing (GBS).

Safflower (Carthamus tinctorius, Asteraceae) is a source of high-quality edible oil growing in moisture-limited environments. Despite its economic importance, the relationships to close wild species in Carthamus and the presence and relationships of ecotypes within safflower are still not fully clarified. Here we use genotyping-by-sequencing to identify the wild progenitor of C. tinctorius, infer phylogenetic relationship within the series Carthamus and identify groups of closely related lineages within cultivated safflower. Phylogenetic and population genomic analyses found C. palaestinus to be the closest relative and single progenitor of C. tinctorius, which confirms the Levant as the area of domestication of the crop. Flow cytometry showed all analyzed samples of C. oxyacantha, C. palaestinus and C. tinctorius to be diploid (2n = 2x = 24) with 2C genome sizes of 2.4-2.7 pg. Analyses of a set of 114 worldwide distributed safflower accessions arrived at two to five genetic groups, which showed, however, no correlation with the geographic origins of these accessions. From this, we conclude that the trade of safflower seeds resulted in multiple introductions of genotypes from the Levant into other areas with suitable climate conditions for the plant, as well as exchange of genotypes among these areas.

Evaluation of grain yield stability of tritipyrum as a novel cereal in comparison with triticale lines and bread wheat varieties through univariate and multivariate parametric methods.

Salinity is a major abiotic stress affecting cereal production. Thus, tritipyrum (x. Tritipyrum), a potential novel salt-tolerant cereal, was introduced as an appropriate alternative for cereal production. The purposes of this study were to evaluate agronomic traits, yield, and yield stability of eight primary tritipyrum lines, five promising triticale lines, and four bread wheat varieties and to screen a stable yielding line. The experiments were conducted in randomized complete block designs with three replicates in three locations during four growing seasons. Analysis of variance in each environment and Bartlett's test for the variance homogeneity of experimental errors were made. Subsequently, separate experiments were analyzed as a combined experiment. The stability of grain yield was analyzed according to Eberhart and Russell's regression method, environmental variance, Wrick's ecovalance, Shokla's stability variance, AMMI, and Tai methods. Genotype × environment interactions (GEI) and environments were significant for the agronomic traits. Stability analysis revealed that combined primary tritipyrum line (Ka/b)(Cr/b)-5 and triticale 4115, 4108, and M45 lines had good adaptability in all environments. The results of the AMMI3 model and pattern analysis showed that the new cereal, tritipyrum, had the most stable response in various environments. The tritipyrum line (Ka/b)(Cr/b)-5 had the best yield performance and general adaptability. Based on Tai's method, the contribution of spike number to the stability of grain yield over different environments was higher than that of other yield components. Also, tritipyrum lines demonstrated higher stability compared with wheat and triticale. Totally, M45 triticale and tritipyrum (Ka/b)(Cr/b)-5 lines were the most stable genotypes with high grain yield. Complementary agronomic experiments may then release a new grain crop of triticale and a new pasture line of combined primary tritipyrum for grain and forage. Moreover, the combined tritipyrum line can be used in bread wheat breeding programs for producing salt-tolerant wheat cultivars.

High-density linkage mapping for agronomic and physiological traits of rice (Oryza sativa L.) under reproductive-stage salt stress.

Rice is one of the most important cereals of the world, with a substantial amount of genetic variation, and a staple food for more than half of the world's population. Salinity is the second most important abiotic stress after drought that adversely affects rice production globally. Both the seedling and reproductive stages are extremely sensitive to salinity but tolerant at the reproductive stage which is most crucial, as it translates into grain yield. Therefore, it is more important to identify the underlying factors of tolerance at the reproductive stage as a necessary step towards improving varieties for salinity environments. However, because of the difficulties in phenotyping protocols of salinity tolerance screening at the reproductive stage, only a few studies exist on this aspect. In view of this, a study involving 188 F4 rice lines derived from a cross CSR28 × Sadri along with the parents was carried out for phenotyping using a novel screening approach for the reproductive stage in salinity conditions and genotyping by SNP markers (Infinium Illumina 6K SNP chip) to construct a high-saturation linkage map. Quantitative trait loci analysis in an F4 population for physiological traits (chlorophyll a, chlorophyll b and carotenoid) and agronomic traits (plant height, filled grain number, grain yield and spikelet fertility percentage) led to the identification of 14 QTLs with an LOD range of 2.72-4.46 explaining phenotypic variation of 5.29-24.86% on chromosomes 1, 2, 3, 5, 6, 7 and 8. Tolerant alleles were contributed by both CSR28 and Sadri. The results indicated that both physiological and agronomic traits were involved in salinity tolerance at the reproductive stage and majority of the QTLs identified in this study are reported for the first time.

Association analysis between agronomic traits and AFLP markers in a wide germplasm of proso millet (Panicum miliaceum L.) under normal and salinity stress conditions.

BACKGROUND: Proso millet is a highly nutritious cereal considered an essential component of processed foods. It is also recognized with high water-use efficiency as well as short growing seasons. This research was primarily aimed at investigating the genetic diversity among genotypes based on evaluating those important traits proposed in previous researches under both normal and salinity- stress conditions. Use of Amplified fragment length polymorphism (AFLP) molecular markers as well as evaluating the association between markers and the investigated traits under both conditions was also another purpose of this research. RESULTS: According to the phenotypic correlation coefficients, the seed yield had the highest correlation with the forage and biological yields under both conditions. By disintegrating those traits investigated under normal and salinity-stress conditions into principal component analysis, it was found that the first four principal components justified more than 59.94 and 62.48% of the whole variance, respectively. The dendrogram obtained by cluster analysis displayed three groups of genotypes under both normal and salinity- stress conditions. Then, association analyses were conducted on 143 proso millet genotypes and 15 agronomic traits as well as 514 polymorphic AFLP markers (out of 866 created bands) generated by 11 primer combinations (out of the initial 20 primer combinations) EcoRI/MseI. The results obtained by mixed linear model (MLM) indicated that under normal conditions, the M14/E10-45 and M14/E10-60 markers had strong associations with seed yield. A similar trend was also observed for M14/E10-45 and M14/E11-44 markers in relation to forage yield. On the other hand, M14/E10-14, M14/E10-64 markers (for seed yield) and M14/E10-64 marker (for forage yield), had significant and stable association in all environments under salinity-stress conditions. Moreover, a number of markers showed considerable associations and stability under both normal and salinity stress conditions. CONCLUSIONS: According to the analysis of phenotypic data, the wide germplasm of Iranian proso millet has significant variation in terms of measured traits. It can be concluded that markers showing strong associations with traits under salinity-stress conditions are suitable candidates to be used in future marker-assisted selection (MAS) studies to improve salinity-resistance genotypes of Panicum miliaceum in arid and semiarid areas.

Transcriptomic analysis of Aegilops tauschii during long-term salinity stress.

Aegilops tauschii is the diploid progenitor of the bread wheat D-genome. It originated from Iran and is a source of abiotic stress tolerance genes. However, little is known about the molecular events of salinity tolerance in Ae. tauschii. This study investigates the leaf transcriptional changes associated with long-term salt stress. Total RNA extracted from leaf tissues of control and salt-treated samples was sequenced using the Illumina technology, and more than 98 million high-quality reads were assembled into 255,446 unigenes with an average length of 1398 bp and an N50 of 2269 bp. Functional annotation of the unigenes showed that 93,742 (36.69%) had at least a significant BLAST hit in the SwissProt database, while 174,079 (68.14%) showed significant similarity to proteins in the NCBI nr database. Differential expression analysis identified 4506 salt stress-responsive unigenes. Bioinformatic analysis of the differentially expressed unigenes (DEUs) revealed a number of biological processes and pathways involved in the establishment of ion homeostasis, signaling processes, carbohydrate metabolism, and post-translational modifications. Fine regulation of starch and sucrose content may be important features involved in salt tolerance in Ae. tauschii. Moreover, 82% of DEUs mapped to the D-subgenome, including known QTL for salt tolerance, and these DEUs showed similar salt stress responses in other accessions of Ae. tauschii. These results could provide fundamental insight into the regulatory process underlying salt tolerance in Ae. tauschii and wheat and facilitate identification of genes involved in their salt tolerance mechanisms.

Comparative assessment of chloroplast transcriptional responses highlights conserved and unique patterns across Triticeae members under salt stress.

Chloroplast functional genomics, in particular understanding the chloroplast transcriptional response is of immense importance mainly due to its role in oxygenic photosynthesis. As a photosynthetic unit, its efficiency and transcriptional activity is directly regulated by reactive oxygen species during abiotic and biotic stress and subsequently affects carbon assimilation, and plant biomass. In crops, understanding photosynthesis is crucial for crop domestication by identifying the traits that could be exploited for crop improvement. Transcriptionally and translationally active chloroplast plays a key role by regulating the PSI and PSII photo-reaction centres, which ubiquitously affects the light harvesting. Using a comparative transcriptomics mapping approach, we identified differential regulation of key chloroplast genes during salt stress across Triticeae members with potential genes involved in photosynthesis and electron transport system such as CytB6f. Apart from differentially regulated genes involved in PSI and PSII, we found widespread evidence of intron splicing events, specifically uniquely spliced petB and petD in Triticum aestivum and high proportion of RNA editing in ndh genes across the Triticeae members during salt stress. We also highlight the role and differential regulation of ATP synthase as member of CF0CF1 and also revealed the effect of salt stress on the water-splitting complex under salt stress. It is worthwhile to mention that the observed conserved down-regulation of psbJ across the Triticeae is limiting the assembly of water-splitting complexes and thus making the BEP clade Triticeae members more vulnerable to high light during the salt stress. Comparative understanding of the chloroplast transcriptional dynamics and photosynthetic regulation will improve the approaches for improved crop domestication.

Identification and validation of functional markers in a global rice collection by association mapping.

Recent results indicate that marker-assisted selection is an effective approach to reduce the cost and to improve the efficacy and accuracy of selection in plant breeding. This study was conducted to identify and validate molecular markers linked to important breeding traits by association mapping. The association was evaluated between 81 molecular markers (STS, SSR, Indel, CAPS, and PCR-based SNP) and 15 morphological traits in a global panel of 100 rice (Oryza sativa) accessions. The population structure analysis identified three main subpopulations. Obvious kinship relationships were also detected between the rice accessions. Association analysis was performed based on the mixed linear model by considering population structure and family relatedness. In addition, the false discovery rate method was used to correct the multiple testing. A total of 47 marker-trait associations were identified, including 22 markers for 14 traits. Among all, the polymorphism at the loci DDR-GL was highly associated with grain characters (grain length, grain width, and length/width ratio). In addition, marker RM3148 was responsible for five important traits simultaneously. Results demonstrated that such informative markers can be very useful for rice breeding programs using marker-assisted selection. Moreover, the diverse populations of rice accessions are a valuable resource for association mapping of morphological traits.

Haplotype analysis of QTLs attributed to salinity tolerance in wheat (Triticum aestivum).

A diverse collection of wheat germplasm, consisting of 100 bread wheat lines with varying levels of salinity tolerance were evaluated based on incomplete block design (lattice) with two replications in field conditions. Plant material was screened for salinity tolerance under normal and saline field conditions. Subsequently in order to assess the haplotype diversity of QTLs attributed to salinity tolerance in wheat (Triticum aestivum), a collection of 30 extremes tolerant and sensitive genotypes among them were selected for genotyping on the basis of morphological, physiological and phenological traits. Genotyping was done using microsatellite markers which had been detected as the flanking regions of large effect QTLs attributed to salinity tolerance on chromosomes 2A, 4D and 3B. Combined analysis of saline and normal conditions revealed that genotypes showed highly significant responses. Association analysis of SSR markers with traits, showed markers Xcfa2121b, Xgwm10 and Xgwm296 on chromosome 2A and markers Xgwm194 and xgwm624 for chromosome 4D, had significant association with most of measured traits. Haplotype diversity analysis showed markers Xgwm10, Xgwm445, Xbarc353.2, Xgwm312, Xgwm515 and Xwmc296 on chromosome 2A as well as markers Xwmc326 and Xgwm345, Xbarc48.4 on chromosomes 3B and 4D were identified as the best markers attributed to salinity tolerance and they can be informative markers for improvement of salinity tolerance through marker-assisted selection programs.