Reference-quality genomes of both sexes are essential for studying sex determination and sex-chromosome evolution, as their gene contents and expression profiles differ. Here, we present independent chromosome-level genome assemblies for the female (XX) and male (XY) genomes of desert poplar (Populus euphratica), resolving a 22.7-Mb X and 24.8-Mb Y chromosome. We also identified a relatively complete 761-kb sex-linked region (SLR) in the peritelomeric region on chromosome 14 (Y). Within the SLR, recombination around the partial repeats for the feminizing factor ARR17 (ARABIDOPSIS RESPONSE REGULATOR 17) was potentially suppressed by flanking palindromic arms and the dense accumulation of retrotransposons. The inverted small segments S1 and S2 of ARR17 exhibited relaxed selective pressure and triggered sex determination by generating 24-nt small interfering RNAs that induce male-specific hyper-methylation at the promoter of the autosomal targeted ARR17. We also detected two male-specific fusion genes encoding proteins with NB-ARC domains at the breakpoint region of an inversion in the SLR that may be responsible for the observed sexual dimorphism in immune responses. Our results show that the SLR appears to follow proposed evolutionary dynamics for sex chromosomes and advance our understanding of sex determination and the evolution of sex chromosomes in Populus.

Background: Zygophyllumis an important medicinal plant, with notable properties such as resistance to salt, alkali, and drought, as well as tolerance of poor soils and shifting sand. However, the response mechanism ofZygophyllumspp. to abiotic stess were rarely studied.Results: Here, we aimed to explore the salt-tolerance genes ofZygophyllumplants by transcriptomic and metabolic approaches. We choseZ. brachypterum,Z. obliquumandZ. fabagoto screen for salt tolerant and sensitive species. Cytological observation showed that both the stem and leaf ofZ. brachypterumwere significantly thicker than those ofZ. fabago.Then, we treated these three species with different concentrations of NaCl, and found thatZ. brachypterumexhibited the highest salt tolerance (ST), whileZ. fabagowas the most sensitive to salt (SS). With the increase of salt concentration, the CAT, SOD and POD activity, as well as proline and chlorophyll content in SS decreased significantly more than in ST. After salt treatment, the proportion of open stomata in ST decreased significantly more than in SS, although there was no significant difference in stomatal number between the two species. Transcriptomic analysis identified a total of 11 overlapping differentially expressed genes (DEGs) in the leaves and roots of the ST and SS species after salt stress. Two branched-chain-amino-acid aminotransferase (BCAT) genes among the 11 DEGs, which were significantly enriched in pantothenate and CoA biosynthesis, as well as the valine, leucine and isoleucine biosynthesis pathways, were confirmed to be significantly induced by salt stress through qRT-PCR. Furthermore, overlapping differentially abundant metabolites showed that the pantothenate and CoA biosynthesis pathways were significantly enriched after salt stress, which was consistent with the KEGG pathways enriched according to transcriptomics.Conclusions: In our study, transcriptomic and metabolomic analysis revealed that BCAT genes may affect the pantothenate and CoA biosynthesis pathway to regulate the salt tolerance ofZygophyllumspecies, which may constitute a newly identified signaling pathway through which plants respond to salt stress.

Drought stress severely affects global plant growth and production. The enhancement of plant water-use efficiency (WUE) and drought tolerance by the manipulation of the stomata is an effective strategy to deal with water shortage. However, increasing the WUE and drought tolerance by manipulation on the stomata has rarely been tested in Brassica napus. Here, we isolated Bna.EPF2, an epidermal patterning factor (EPF) in Brassica napus (ecotype Westar), and identified its role in drought performance. Bna.EPF2 overexpression lines had a reduction average of 19.02% in abaxial stomatal density and smaller stomatal pore size, leading to approximately 25% lower transpiration, which finally resulted in greater instantaneous WUE and enhanced drought tolerance. Interestingly, the reduction in stomatal density did not affect the CO₂ assimilation or yield-related agronomic traits in Bna.EPF2 overexpression plants. Together with the complementation of Bna.EPF2 significantly decreasing the stomatal density of Arabidopsis epf2, and Bna.EPF2 being expressed in mature guard cells, these results suggest that Bna.EPF2 not only functions in stomatal density development, but also in stomatal dimension in Brassicas. Taken together, our results suggest that Bna.EPF2 improves WUE and drought tolerance by the regulation of stomatal density and stomatal size in Brassica without growth and yield penalty, and provide insight into the manipulation of this gene in the breeding of drought tolerant plants with increased production under water deficit conditions.

Despite the high economic and ecological importance of forests, our knowledge of the adaptive evolution of leaf traits remains very limited. Euphrates poplar (Populus euphratica), which has high tolerance to arid environment, has evolved four heteromorphic leaf forms, including narrow (linear and lanceolate) and broad (ovate and broad-ovate) leaves on different crowns. Here, we revealed the significant functional divergence of four P. euphratica heteromorphic leaves at physiological and cytological levels. Through global analysis of transcriptome and DNA methylation across tree and leaf developmental stages, we revealed that gene expression and DNA epigenetics differentially regulated key processes involving development and functional adaptation of heteromorphic leaves, such as hormone signaling pathways, cell division, and photosynthesis. Combined analysis of gene expression, methylation, ATAC-seq, and Hi-C-seq revealed longer interaction of 3D genome, hypomethylation, and open chromatin state upregulates IAA-related genes (such as PIN-FORMED1 and ANGUSTIFOLIA3) and promotes the occurrence of broad leaves while narrow leaves were associated with highly concentrated heterochromatin, hypermethylation, and upregulated abscisic acid pathway genes (such as Pyrabactin Resistance1-like10). Therefore, development of P. euphratica heteromorphic leaves along with functional divergence was regulated by differentially expressed genes, DNA methylation, chromatin accessibility, and 3D genome remodeling to adapt to the arid desert. This study advances our understanding of differential regulation on development and functional divergence of heteromorphic leaves in P. euphratica at the multi-omics level and provides a valuable resource for investigating the adaptive evolution of heteromorphic leaves in Populus.

Populus euphratica, a key tree species in the ecologically fragile area of the Tarim River Basin, plays a crucial role in maintaining the ecological balance of the desert. In order to explore the response of Populus euphratica leaves to river water resources and drought climate change, the complex topological relationship between the leaf traits of Populus euphratica was comprehensively analyzed by establishing the leaf trait network of the mainstream of the Tarim River, and the central traits that play a key role in regulating the phenotype of Populus euphratica were identified. The adaptation strategies of Populus euphratica to different environments were clarified from the perspective of the overall leaf trait network. The results show that, with a decrease in river water flow, the internal coordination of the leaf traits weakened, forming a looser network structure. In the hyper-arid region, the leaf traits were more independent, the network was simpler, and it was easier for the leaf to coordinate the aggregation of specific traits. Leaf thickness and the related photosynthetic traits are the key to adaptive strategies, particularly the cuticle thickness and midvein vascular bundle area. Leaf anatomical traits are more critical than stoichiometry traits in drought resistance. Leaves tend to give priority to the relationship between structural traits. Populus euphratica leaves adjust their trait modularization through a trait substitution strategy to manage different drought conditions.

Oxytropis glabra DC. is a perennial poisonous plant to livestock belonging to the genus Oxytropis, Leguminosae, mainly distributed in Northwestern China. As a poisonous grass, this species protects plant diversity in degraded grasslands by sheltering adjacent plants. In this study, the complete chloroplast genome with a total size of 122,094 bp was reported. Our annotations showed that the chloroplast genome contains 109 genes, including 76 protein-coding genes, 29 tRNA genes, and four rRNA genes. This work presents complete chloroplast genome information, which will be valuable for studying the evolution and genetic diversity of O. glabra.