Plant Cell Reports最新文献

Key message: Multi-omics studies have shown that strigolactone modulates phenolic acid accumulation in the leaves of R. chrysanthum and can enable it to cope with UV-B stress. UV-B stress is an abiotic stress that plants will inevitably suffer during growth and can seriously affect the normal physiological state of plants. Strigolactone, a phytohormone, has been less studied and it is important to investigate its regulation of plant growth under UV-B radiation. In the present study, we investigated the changes in leaves of Rhododendron chrysanthum Pall. (R. chrysanthum) under UV-B radiation. The leaves of R. chrysanthum were collected for widely targeted metabolomics, hormonomics, transcriptomics, proteomics and acetylated proteomics assays. The results showed that the leaves of R. chrysanthum were able to produce a large amount of differential phenolic acids with antioxidant effects under UV-B stress, the content of strigolactone was significantly elevated, and the genes and proteins involved in phenolic acid biosynthesis and strigolactone biosynthesis were significantly altered, and some of the proteins (ASP1, 4CLL7, and CCD1) underwent acetylation modification. Meanwhile, correlation analysis showed that strigolactone was strongly correlated with differential phenolic acids, which might regulate the adaptive responses of the R. chrysanthum under UV-B stress. In this paper, we investigated the effects of strigolactone on the accumulation of phenolic acid compounds and found a strong correlation between strigolactone and elevated phenolic acid levels, which provided insights into the molecular mechanism of plant regulation of phenolic acid accumulation, and facilitated the adoption of measures to mitigate the adverse effects of UV-B stress on plant growth, and to achieve the purpose of protecting plant germplasm resources.

Key message: Transgenic tobacco plant expressed EpCAM-Fc fusion proteins to induce in vivo immune responses producing anti-EpCAM antibodies inhibiting human colorectal cancer cell invasion and migration. Plant is emerging as a promising alternative to produce valuable immunotherapeutic vaccines. In this study, we examined the in vivo anti-cancer efficacy of epidermal cell adhesion molecule (EpCAM)-Fc and EpCAM-FcK fusion proteins produced in transgenic plants as colorectal cancer vaccine candidates. Mice were injected with plant-derived EpCAM-Fc (EpCAM-Fc^P) and EpCAM-Fc^P tagged with KDEL (ER retention signal) (EpCAM-FcK^P), using mammalian-derived EpCAM-Fc (EpCAM-Fc^M) as positive control. Total IgGs from the immunized mice were used to assess immune responses. ELISA tests revealed that IgGs from mice immunized with EpCAM-FcK^P (EpCAM-FcK^P IgG) exhibited the highest absorbance value for binding affinity to recombinant EpCAM-Fc^M compared to IgGs from mice immunized with EpCAM-Fc^P (EpCAM-Fc^P IgG) and EpCAM-Fc^M (EpCAM-Fc^M IgG). Bio-layer interferometry revealed that EpCAM-FcK^P IgG had a higher affinity value than EpCAM-Fc^M IgG and EpCAM-Fc^P IgG. Cell ELISA revealed that EpCAM-FcK^P IgG exhibited the highest binding activity to EpCAM-positive cells SW480 and SW620 compared to EpCAM-Fc^P IgG, EpCAM-Fc^M IgG, and anti-EpCAM mAb. In the transwell invasion assay, EpCAM-FcK^P IgG significantly decreased the numbers of invaded SW480 and SW620 cells compared to EpCAM-Fc^P IgG, whereas EpCAM-Fc^M IgG had similar numbers. In the wound healing assay, EpCAM-FcK^P IgG showed higher migration inhibition compared to EpCAM-Fc^P IgG in both cell types, with similar results to EpCAM-Fc^M IgG in SW620 cells. These results confirm the applicability of plant systems to produce EpCAM-Fc vaccine candidates, inducing the production of anti-EpCAM IgGs against colorectal cancer cells.

Key message: This study identified a gene associated with aluminum stress through GWAS, which regulates aluminum tolerance in alfalfa by contributing to the antioxidant system. Aluminum (Al) ions precipitate in acidic soils with a pH < 5.5, where they are absorbed alongside other nutrients by plants, negatively impacting plant growth. Alfalfa, the most widely grown perennial legume forage in the world, is especially vulnerable to acidic soil conditions. Our research pinpointed MsDUF3700 as a potential gene linked to Al-response traits via genome-wide association analysis in Medicago sativa. MsDUF3700 encodes the domain of unknown function (DUF). We observed higher expression of MsDUF3700 in Al-tolerant alfalfa compared to Al-sensitive ecotypes. MsDUF3700-overexpressing transgenic alfalfa (MsDUF3700-OE) showed shorter root elongation and higher Al accumulation in roots than wild type (WT) under Al conditions. However, the shoots of MsDUF3700-OE lines showed enhanced growth rates under both normal and Al stress conditions. Under Al stress, MsDUF3700-OE lines showed increased H₂O₂ and malondialdehyde (MDA) levels in the roots, alongside reduced catalase activity, In contrast, the shoots showed an inverse trend. In addition, we found that MsDUF3700-OE alfalfa plants had high Al accumulation in the roots and low Al accumulation in the shoots. Transcripts of MsALS3 and MsPALT1, homologs of Al translocation in alfalfa, were downregulated, while MsNrat1, a homolog of transporters absorb Al, was upregulated in the roots of MsDUF3700-OE in alfalfa. Our research indicates that MsDUF3700 plays a role in aluminum stress by participating in antioxidative defense and facilitating aluminum transport from roots to shoots.

Key message: Multiple QTLs reveal the polygenic nature of R. rhizogenes-mediated transformation and hairy root formation in roses, with five key regions explaining 12.0-26.9% of trait variability and transformation-related candidate genes identified. Understanding genetic mechanisms of plant transformation remains crucial for biotechnology. This is particularly relevant for roses and other woody ornamentals that exhibit recalcitrant behavior in transformation procedures. Rhizobium rhizogenes-mediated transformation leading to hairy root (HR) formation provides an excellent model system to study transformation processes and host-pathogen interactions. Therefore, this study aimed to identify quantitative trait loci (QTLs) associated with HR formation and explore their relationship with adventitious root (AR) formation in rose as a model for woody ornamentals. A diversity panel of 104 in vitro grown rose genotypes was transformed with R. rhizogenes strain ATCC 15834 carrying a green fluorescent protein reporter gene. Phenotypic data on callus and root formation were collected for laminae and petioles. A genome-wide association study using 23,419 single-nucleotide polymorphism markers revealed significant QTLs on chromosomes one and two for root formation traits. Five key genomic regions explained 12.0-26.9% of trait variability, with some peaks overlapping previously reported QTLs for AR formation. This genetic overlap was supported by weak to moderate correlations between HR and AR formation traits, particularly in petioles. Candidate gene identification through literature review and transcriptomic data analysis revealed ten candidate genes involved in bacterial response, hormone signaling, and stress responses. Our findings provide new insights into the genetic control of HR formation in roses and highlight potential targets for improving transformation efficiency in ornamental crops, thereby facilitating future research and breeding applications.

Key message: A chitinase gene ScChiVII1 which is involved in defense against pathogen stress was characterized in sugarcane. Chitinases, a subclass of pathogenesis-related proteins, catalyze chitin hydrolysis and play a key role in plant defense against chitin-containing pathogens. However, there is little research on disease resistance analysis of chitinase genes in sugarcane, and the systematic identification of their gene families has not been reported. In this study, 85 SsChi and 23 ShChi genes, which were divided into 6 groups, were identified from the wild sugarcane species Saccharum spontaneum and Saccharum hybrid cultivar R570, respectively. Transcriptome analysis and real-time quantitative PCR revealed that SsChi genes responded to smut pathogen stress. The chitinase crude extracted from the leaves of transgenic Nicotiana benthamiana plants overexpressing ScChiVII1 (a homologous gene of SsChi22a) inhibited the hyphal growth of Fusarium solani var. coeruleum and Sporisorium scitamineum. Notably, the chitinase and catalase activities and the jasmonic acid content in the leaves of ScChiVII1 transgenic N. benthamiana increased after inoculation with F solani var. coeruleum, but the salicylic acid, hydrogen peroxide, and malondialdehyde contents decreased. Comprehensive RNA sequencing of leaves before (0 day) and after inoculation (2 days) revealed that ScChiVII1 transgenic tobacco enhanced plant disease resistance by activating transcription factors and disease resistance-related signaling pathways, and modulating the expression of genes involved in the hypersensitive response and ethylene synthesis pathways. Taken together, this study provides comprehensive information on the chitinase gene family and offers potential genetic resources for disease resistance breeding in sugarcane.

Key message: Overexpression of HvbHLH132 from hulless barley impairs in chilling and freezing tolerance at the seedlings stage in Arabidopsis thaliana The basic helix-loop-helix (bHLH) transcription factors (TF) are ubiquitously existed in eukaryote and play crucial roles in numerous biological processes. However, the characterization of their members and functions in hulless barley remains limited. Here, we conducted a genome-wide identification of the HvbHLH gene family and assessed the role of HvbHLH132 in cold stress tolerance. We identified 141 HvbHLH genes, which were categorized into twelve subfamilies. Subcellular localization predictions indicated that the majority of HvbHLH proteins were localized in the nucleus. cis-Acting element analysis revealed that the promoter regions of the HvbHLH family contain diverse elements associated with various biological processes. Expression profiling of the 141 HvbHLH genes in two extreme varieties revealed that HvbHLH132 was significantly induced and exhibited substantial differential expression under cold stress. Analyses of subcellular localization and transactivation activity confirmed that HvbHLH132 specifically localized in the nucleus and contributed to transcriptional activation. Furthermore, overexpression of HvbHLH132 in Arabidopsis resulted in impaired chilling and freezing tolerance at the seedling stage, leading to biochemical changes unfavorable for freezing stress. Additionally, the expression of some cold-responsive genes (COR) genes was significantly less induced compared to wild type under freezing stress. This study provides comprehensive insight into the HvbHLH gene family and reveals a critical role of HvbHLH132 in regulating cold tolerance in plants.

Key message: BnaPAP17s associated with root-secreted APases activity were identified by genome-wide association study, and those were induced by Pi-deficiency. BnaPAP17s were involved in improving exogenous organophosphorus utilization as secreted APases. Deficiency of available phosphorus (P) in soil has become an important limiting factor for yield and quality in oilseed rape (Brassica napus). In many soils, organic P (Po) is the main component of the soil P pool. Po must be hydrolyzed to inorganic P (Pi) through acid Phosphatase (APases), and then taken up by plants. However, root-secreted APases (SAP) activity, as a quantitative trait, plays an important role in soil Po utilization; those genetic loci are not clear in B. napus. In this study, we performed a genome-wide association study for SAP activity under Pi-deficiency using a panel of 350 accessions of B. napus and more than 4.5 million polymorphic single nucleotide polymorphisms (SNPs). Thirty-five significant SNPs associated with SAP activity were identified. BnaA01.PAP17 (BnaA01g27810D) was a candidate gene underlying lead SNP (ChrA01_19576615). We experimentally verified that both BnaA01.PAP17 and its three homologous genes had similar expression pattern in response to Pi-deficiency. The dynamic changes in BnaPAP17s expression level were opposite to those of Pi concentration in both roots and leaves, suggesting their potential utility as Pi marker genes in B. napus. Transient expression of BnaPAP17s in tobacco leaves proved that BnaPAP17s were located in the apoplast as secreted APases. The overexpression of BnaPAP17s enhanced SAP activity in response to Pi-deficiency and resulting in increased P content in plants when ATP was supplied as the sole P resource. Taken together, these results suggest that BnaPAP17s contributed to SAP activity, thus having a function in extracellular Po utilization in B. napus.

Key message: GmABCI5 and GmABCI13 enhance Al tolerance through regulating the composition of root cell wall, and in this process, GmABCI5 and GmABCI13 may act in the form of a complex. Aluminum (Al) toxicity is a major factor limiting plant growth in acidic soils. ATP-binding cassette (ABC) transporters are involved in plant tolerance to various environmental stresses. However, there are few reports on the ABC transporters implicated in soybean tolerance to Al toxicity. Here, we reported that two genes, GmABCI5 and GmABCI13, were involved in Al tolerance in soybean (Glycine max). GmABCI5 and GmABCI13 encode a nucleotide-binding domain and a transmembrane domain of a bacterial-type ABC transporter, respectively. The expression of both GmABCI5 and GmABCI13 was mainly induced by Al in the roots. GmABCI5 was localized at the plasma membrane and also in the cytoplasm and nucleus, while GmABCI13 was only localized at the plasma membrane. Furthermore, GmABCI5 could physically interact with GmABCI13. Overexpression of GmABCI5 or GmABCI13 in Arabidopsis reduced Al accumulation in roots and enhanced Al tolerance. However, expression of GmABCI5 and/or GmABCI13 in yeast cells did not affect Al uptake. Under Al stress, transgenic Arabidopsis lines expressing GmABCI5 or GmABCI13 had lower Al content in root cell walls than wild-type plants. Further analysis showed that Al content in cell wall fractions (pectin and hemicellulose 1) of transgenic lines was significantly lower than that of wild-type plants, which was coincident with the changes of pectin and hemicellulose 1 content under Al stress. These results indicate that GmABCI5 and GmABCI13 form an ABC transporter complex to regulate Al tolerance by affecting the modification of cell wall.

Single-cell transcriptomic techniques have ushered in a new era in plant biology, enabling detailed analysis of gene expression at the resolution of individual cells. This review delves into the transformative impact of these technologies on our understanding of plant development and their far-reaching implications for plant biotechnology. We present a comprehensive overview of the latest advancements in single-cell transcriptomics, emphasizing their application in elucidating complex cellular processes and developmental pathways in plants. By dissecting the heterogeneity of cell populations, single-cell technologies offer unparalleled insights into the intricate regulatory networks governing plant growth, differentiation, and response to environmental stimuli. This review covers the spectrum of single-cell approaches, from pioneering techniques such as single-cell RNA sequencing (scRNA-seq) to emerging methodologies that enhance resolution and accuracy. In addition to showcasing the technological innovations, we address the challenges and limitations associated with single-cell transcriptomics in plants. These include issues related to sample preparation, cell isolation, data complexity, and computational analysis. We propose strategies to mitigate these challenges, such as optimizing protocols for protoplast isolation, improving computational tools for data integration, and developing robust pipelines for data interpretation. Furthermore, we explore the practical applications of single-cell transcriptomics in plant biotechnology. These applications span from improving crop traits through precise genetic modifications to enhancing our understanding of plant-microbe interactions. The review also touches on the potential for single-cell approaches to accelerate breeding programs and contribute to sustainable agriculture. This review concludes with a forward-looking perspective on the future impact of single-cell technologies in plant research. We foresee these tools becoming essential in plant biotechnology, spurring innovations that tackle global challenges in food security and environmental sustainability. This review serves as a valuable resource for researchers, providing a roadmap from sample preparation to data analysis and highlighting the transformative potential of single-cell transcriptomics in plant biotechnology.