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Although the functions of carotenogenic genes are well documented, little is known about the mechanisms that regulate their expression, especially those genes involved in α- and β-branch carotenoid metabolism. In this study, an R2R3-MYB transcriptional factor (CrMYB68) that directly regulates the transformation of α- and β-branch carotenoids was identified using Green Ougan (MT), a stay-green mutant of Citrus reticulata cv Suavissima. A comprehensive analysis of developing and harvested fruits indicated that reduced expression of β-carotene hydroxylases 2 (CrBCH2) and 9-cis-epoxycarotenoid dioxygenase 5 (CrNCED5) was responsible for the delay in the transformation of α- and β-carotene and the biosynthesis of ABA. Additionally, the expression of these genes was negatively correlated with the expression of CrMYB68 in MT. Further, electrophoretic mobility shift assays (EMSAs) and dual luciferase assays indicated that CrMYB68 can directly and negatively regulate CrBCH2 and CrNCED5. Moreover, transient overexpression experiments using leaves of Nicotiana benthamiana indicated that CrMYB68 can also negatively regulate NbBCH2 and NbNCED5. To overcome the difficulty of transgenic validation, we quantified the concentrations of carotenoids and ABA, and gene expression in a revertant of MT. The results of these experiments provide more evidence that CrMYB68 is an important regulator of carotenoid metabolism.

Chemical design of SiO2-based glasses with high elastic moduli and low weight is of great interest. However, it is difficult to find a universal expression to predict the elastic moduli according to the glass composition before synthesis since the elastic moduli are a complex function of interatomic bonds and their ordering at different length scales. Here we show that the densities and elastic moduli of SiO2-based glasses can be efficiently predicted by machine learning (ML) techniques across a complex compositional space with multiple (>10) types of additive oxides besides SiO2. Our machine learning approach relies on a training set generated by high-throughput molecular dynamic (MD) simulations, a set of elaborately constructed descriptors that bridges the empirical statistical modeling with the fundamental physics of interatomic bonding, and a statistical learning/predicting model developed by implementing least absolute shrinkage and selection operator with a gradient boost machine (GBM-LASSO). The predictions of the ML model are comprehensively compared and validated with a large amount of both simulation and experimental data. By just training with a dataset only composed of binary and ternary glass samples, our model shows very promising capabilities to predict the density and elastic moduli for k-nary SiO2-based glasses beyond the training set. As an example of its potential applications, our GBM-LASSO model was used to perform a rapid and low-cost screening of many (~105) compositions of a multicomponent glass system to construct a compositional-property database that allows for a fruitful overview on the glass density and elastic properties.

Key messageCitWRKY28 and CitNAC029 are involved in cuticular wax synthesis as indicated by the comparative analysis of fruit aliphatic wax content between Citrus reticulata and Citrus trifoliata and gene co-expression analysis.Cuticular wax covers the fruit surface, playing important roles in reduction of fruit water loss and resistance to pathogen invasion. However, there is limited research on the synthesis and transcriptional regulation of cuticular wax in citrus fruit. In this study, we characterized the variations of aliphatic wax in HJ (Citrus reticulata) and ZK (Citrus trifoliata) from young fruit to mature fruit, as well as performed transcriptome sequencing on 27 samples at different fruit developmental stages. The results revealed that the ZK fruit always had a higher aliphatic wax content than the HJ fruit during development. qRT-PCR analysis demonstrated that two KCS genes, CitKCS1 and CitKCS12, had the most significant difference in expression between HJ and ZK. Furthermore, a heterologous expression assay in Arabidopsis indicated that CitKCS1 and CitKCS12 are involved in cuticular wax synthesis. Subsequently, gene co-expression network analysis screened CitWRKY28 and CitNAC029. Dual luciferase and EMSA assays indicated that CitWRKY28 might bind to the promoter of CitKCS1 and CitKCS12 and CitNAC029 might bind to that of CitKCS1 to activate their expression. Moreover, CitWRKY28 and CitNAC029 could promote the accumulation of cuticular wax in Arabidopsis leaves. Our findings provide new insights into the synthesis and regulation of cuticular wax and valuable information for further mining of wax-related genes in citrus fruit.

Classical theoretical approaches to evaluate CPTU-based hydraulic conductivity gave the expression of a bilinear relation defined by KD-BqQt, for undrained and drained soils. In fact, a dividing line may be selected with a certain degree of subjectivity, and several undrained/drained points occur in opposite sites, which may make us doubt the necessity of two intersecting lines to distinguish the undrained and drained soils. And a uniform and relatively simple curve including an arc, parabola, or ellipse correlation may be more suitable and accessible from a practical point of view. A database in the Yangtze Delta region has been collated to assess the three curves compared with the bilinear line. With the graphical and statistical analyses, the results indicate that the arc, parabola, and ellipse simplified curves could give better performance than the bilinear line. Through the statistical analyses and the number of variables, the overall best curve was the ellipse with only two variables, which is expected to improve the application scope and simplicity of hydraulic conductivity.

Typical thioredoxin (TRX) plays an important role in maintaining redox balance in plants. However, the typical TRX genes in wheat still need to be comprehensively and deeply studied. In this research, a total of 48 typical TaTRX genes belonging to eight subtypes were identified via a genome-wide search in wheat, and the gene structures, protein conserved motifs, and protein 3D structures of the same subtype were very similar. Evolutionary analysis showed that there are two pairs of tandem duplication genes and 14 clusters of segmental duplication genes in typical TaTRX family members; TaTRX15 , TaTRX36 , and TaTRX42 had positive selection compared with the orthologs of their ancestral species; rice and maize have 11 and 13 orthologous typical TRXs with wheat, respectively. Gene Ontology (GO) analysis indicated that typical TaTRXs were involved in maintaining redox homeostasis in wheat cells. Estimation of ROS content, determination of antioxidant enzyme activity, and gene expression analysis in a line overexpressing one typical TaTRX confirmed that TRX plays an important role in maintaining redox balance in wheat. A predictive analysis of cis -acting elements in the promoter region showed that typical TaTRXs were extensively involved in various hormone metabolism and response processes to stress. The results predicted using public databases or verified using RT-qPCR show that typical TaTRXs were able to respond to biotic and abiotic stresses, and their expression in wheat was spatiotemporal. A total of 16 wheat proteins belonging to four different families interacting with typical TaTRXs were predicted. The above comprehensive analysis of typical TaTRX genes can enrich our understanding of this gene family in wheat and provide valuable insights for further gene function research.

Key messageThe ER or donut-like structures localized aquaporin NIP5;1, which interacts with PIPs and alters their localization from plasma membrane to donut-like structures, regulates water permeability.NOD26-like intrinsic proteins (NIPs) play important roles in nutrient uptake and response to various stresses. However, there have been few studies of their functions in water transportation in citrus. Here, we demonstrate the functions of a novel citrus NIP aquaporin (CsNIP5;1) via multiple physiological and biochemical experiments. CsNIP5;1 showed high water permeability when expressed in Xenopus laevis oocytes and yeast. However, subcellular localization assays showed that this protein was localized in the endoplasmic reticulum (ER) or donut-like structures in citrus callus and tobacco leaf. Meanwhile, overexpression of CsNIP5;1 led to a reduction in the water permeability of citrus callus. Protein–protein interaction experiments and subcellular localization assays further revealed that CsNIP5;1 physically interacted with PIPs (CsPIP1;1 and AtPIP2;1), which altered their subcellular localization from the plasma membrane to donut-like structures. Together, CsNIP5;1 was identified as a good water channel when expressed in oocytes and yeast. Meanwhile, CsNIP5;1 participated in the regulation of water permeability of citrus callus, which may be associated with CsNIP5;1-induced re-localization of water channels PIPs. In summary, these results provide new insights into the regulatory mechanism of AQPs-mediated water diffusion.

The interactions between solute atoms and crystalline defects such as vacancies, dislocations, and grain boundaries are essential in determining alloy properties. Here we present a general linear correlation between two descriptors of local electronic structures and the solute-defect interaction energies in binary alloys of body-centered-cubic (bcc) refractory metals (such as W and Ta) with transition-metal substitutional solutes. One electronic descriptor is the bimodality of the d -orbital local density of states for a matrix atom at the substitutional site, and the other is related to the hybridization strength between the valance sp- and d- bands for the same matrix atom. For a particular pair of solute-matrix elements, this linear correlation is valid independent of types of defects and the locations of substitutional sites. These results provide the possibility to apply local electronic descriptors for quantitative and efficient predictions on the solute-defect interactions and defect properties in alloys. Understanding the interactions between solute atoms and crystalline defects is essential for determining alloy properties. Here the authors use a linear regression model to propose a quantitative correlation between local electronic structure descriptors and the solute-defect interaction energies in bcc refractory alloys.

Sugar is a primary determinant of citrus fruit flavour, but undergoes varied accumulation processes across different citrus varieties owing to high genetic variability. Sucrose phosphate synthase (SPS), a key enzyme in glucose metabolism, plays a crucial role in this context. Despite its significance, there is limited research on sugar component quality and the expression and regulatory prediction of SPS genes during citrus fruit development. Therefore, we analysed the sugar quality formation process in ‘Kiyomi’ and ‘Succosa’, two citrus varieties, and performed a comprehensive genome-wide analysis of citrus CsSPSs . We observed that the accumulation of sugar components significantly differs between the two varieties, with the identification of four CsSPSs in citrus. CsSPS sequences were highly conserved, featuring typical SPS protein domains. Expression analysis revealed a positive correlation between CsSPS expression and sugar accumulation in citrus fruits. However, CsSPS expression displays specificity to different citrus tissues and varieties. Transcriptome co-expression network analysis suggests the involvement of multiple transcription factors in shaping citrus fruit sugar quality through the regulation of CsSPSs . Notably, the expression levels of four CsWRKYs ( CsWRKY2 , CsWRKY20 , CsWRKY28 , CsWRKY32 ), were significantly positively correlated with CsSPSs and CsWRKY20 might can activate sugar accumulation in citrus fruit through CsSPS2 . Collectively, we further emphasize the potential importance of CsWRKYs in citrus sugar metabolism, our findings serve as a reference for understanding sugar component formation and predicting CsSPS expression and regulation during citrus fruit development.

During citrus fruit development, exogenous gibberellin (GA) and 6-benzylaminopurine (6-BA, a synthetic cytokinin (CTK)) are both known to promote citrus peel thickness; however, the differences in their regulatory mechanisms on cell wall metabolism in citrus peels remain unclear. In this study, we found that GA treatment significantly increased cell wall polysaccharides in citrus peels, such as pectin and cellulose, whereas 6-BA treatment led to a notable accumulation of lignin. RNA-sequencing data revealed that several fruit ripening-related cell wall degradation genes, such as PME3 , PL18 , and EXPA1 / 8 , exhibited decreased expression levels in both GA and 6-BA treatments. Additionally, a set of cell wall polysaccharide synthesis genes was upregulated in response to GA treatment but was largely downregulated in 6-BA-treated peels. Conversely, a group of lignin biosynthesis genes was upregulated in 6-BA-treated peels. GA treatment inhibited DELLA proteins (encoded by RGA and GAI ) in the GA signaling pathway, whereas 6-BA treatment increased the expression of B-ARRs ( ARR1 and ARR2 ) in the CTK signaling pathway. Furthermore, GA treatment elevated endogenous CTK levels, while 6-BA treatment also enhanced endogenous GA content, suggesting a reciprocal interaction between these two hormonal pathways.

Selenium stress can adversely affect plants by inhibiting growth, impairing oxidative stress resistance, and inducing toxicity. In this experiment, we investigated the effect of exogenous 24-epibrassinolide (24-EBL; 2.0 mg/L), a brassinosteroid (BR), on alleviating selenium stress in peach trees by analyzing its impact on biomass, selenium accumulation, and the expression of selenium metabolism-related genes in peach seedlings. The results demonstrated that 24-EBL could effectively mitigate biomass loss in peach seedlings exposed to selenium stress. Compared to the Se treatment alone, the 24-EBL+Se treatment resulted in a significant 16.55% increase in root selenium content and a more pronounced 30.39% increase in selenium content in the aboveground parts. Regarding the subcellular distribution, the cell wall was the primary site of Se deposition, accounting for 42.3% and 49.8% in the root and aboveground parts, respectively, in the Se treatment. 24-EBL further enhanced Se distribution at this site, reaching 42.9% and 63.2% in root and aboveground parts, respectively, in the 24-EBL+Se treatment. The 24-EBL+Se treatment significantly increased the contents of different chemical forms of Se, including ethanol-soluble, water-soluble, and salt-soluble Se. The quantitative real-time PCR (qRT-PCR) results indicated that the Se treatment promoted the expression of organic Se assimilation genes (SATs, OAS-TL B, and OAS-TL C), and 24-EBL application further increased their expression. Meanwhile, the Se-only treatment up-regulated the organic Se metabolism gene CGS1. Consequently, we propose that 24-EBL alleviates Se stress in peach seedlings by enhancing Se uptake and assimilation, and by adjusting subcellular distribution and chemical forms.