Plant Genes Research Articles

Mapping an avirulence gene in the sunflower parasitic weed Orobanche cumana and characterization of host selection based on virulence alleles

BackgroundSunflower broomrape (Orobanche cumana Wallr.) is a holoparasitic plant that jeopardizes sunflower production in most areas of Europe and Asia. Recently, populations with increased virulence, classified as race GGV, have been identified in Southern Spain’s Guadalquivir Valley gene pool. These populations overcome resistance genes in hybrids resistant to the predominant race FGV. This study aimed to (i) determine the inheritance and map the avirulence trait segregating in a cross between O. cumana individuals from populations EK23 (FGV) and IN201 (GGV), and (ii) characterize the host effect on the IN201 parental population allelic diversity.ResultsA segregating population consisting of 144 F2:3 families was evaluated for virulence using a differential sunflower genotype (Hybrid 1, resistant to race FGV and susceptible to race GGV) and genotyped with SNP markers. The ratio of avirulent to virulent F2:3 families was not significantly different to 1:3 (χ2 = 0.93; P = 0.34), indicating monogenic control of the avirulence/virulence trait. The AvrG−GV locus was mapped on the upper end of O. cumana chromosome 2, 9.2 cM distal from the SNP markers OS04791 and OS02805. Secretome analysis in the AvrG−GV region revealed a cysteine-rich CAP superfamily- and a glucan 1,3-beta-glucosidase family 3-encoding genes as possible candidates for AvrG−GV. SNP allelic analysis on the IN201 population parasitizing a highly susceptible genotype or the differential genotype Hybrid 1 showed that (i) IN201 structure was shaped towards virulent alleles at SNP loci linked to AvrG−GV (ii) there were significant allelic frequency differences associated with the host genotype at AvrG−GV–linked loci.ConclusionsThis study mapped for the first time an avirulence gene in parasitic plants using a classical genetic approach, confirmed a gene-for-gene model in the O.cumana –sunflower system, and showed the implication of this single avirulence gene in determining the structure of broomrape populations subjected to selection pressure posed by a resistant genotype. The results will contribute to a better understanding of the interaction between crops and weedy parasitic plants, and to effectively manage evolution of virulence by sustainable control strategies based on host genetic resistance.

Medicago truncatula genotype drives the plant nutritional strategy and its associated rhizosphere bacterial communities.

Harnessing the plant microbiome through plant genetics is of increasing interest to those seeking toimproveplant nutrition and health. While genome-wide association studies (GWAS) have been conducted to identify plant genes driving the plant microbiome, more multidisciplinary studies are required to assess the relationships among plant genetics, plant microbiome and plant fitness. Using a metabarcoding approach, we characterized the rhizosphere bacterial communities of a core collection of 155 Medicago truncatula genotypes along with the plant phenotype and investigated the plant genetic effects through GWAS. The different genotypes within the M. truncatula core collection showed contrasting growth and nutritional strategies but few loci were associated with these ecophysiological traits. To go further, we described its associated rhizosphere bacterial communities, dominated by Proteobacteria, Actinobacteria and Bacteroidetes, and defined a core rhizosphere bacterial community. Next, the occurrences of bacterial candidates predicting plant ecophysiological traits of interest were identified using random forest analyses. Some of them were heritable and plant loci were identified, pinpointing genes related to response to hormone stimulus, systemic acquired resistance, response to stress, nutrient starvation or transport, and root development. Together, these results suggest that plant genetics can affect plant growth and nutritional strategies by harnessing keystone bacteria in a well-connected interaction network.

Efficient virus-induced gene silencing in Primulina using two virus vector systems

Virus-induced gene silencing (VIGS) is a post-transcriptional gene silencing method and an effective technique for analyzing plant gene function. However, to date there have been no reports on the establishment of a VIGS system in Primulina, which includes over 200 species of plants, most of which have high ornamental value. In this study, we successfully established a VIGS system in P. 'Alfonso' and P. carinata using TRV vector system with phytoene desaturase (PDS) as marker gene. After being treated with pTRV2-PDS vector, both P. 'Alfonso' and P. carinata plants showed a pronounced photobleaching phenotype. The system was further optimized by exploring different infection conditions. The optimized TRV-based VIGS system was as follows: Agrobacterium OD600 = 0.5, leaf vacuum infiltration, and the silencing efficiency of the system can reach 75 %. Chlorata42 was used as another marker gene and further verified the establishment of TRV VIGS system in P. 'Alfonso'. In addition, the CaLCuV-based VIGS system was also established with PDS as marker gene in P. 'Alfonso'. The establishment of the TRV-based and CaLCuV-based VIGS systems provided a breakthrough in the validation methods for identifying gene function in Primulina genus.

Pollinator functional group abundance and floral heterogeneity in an agroecological context affect mating patterns in a self-incompatible wild plant.

Restoration of seminatural field margins can elevate pollinator activity. However, how they support wild plant gene flow through interactions between pollinators and spatiotemporal gradients in floral resources remains largely unknown. Using a farm-scale experiment, we tested how mating outcomes (expected heterozygosity and paternity correlation) of the wild, self-incompatible plant Cyanus segetum transplanted into field margins (sown wildflower or grass-legume strips) were affected by the abundance of different pollinator functional groups (defined by species traits). We also investigated how the maternal plant attractiveness, conspecific pollen donor density, and heterospecific floral richness and density interacted with pollinator functional group abundance to modulate C. segetum mating outcomes. Multiple paternity increased (=lower paternity correlation) with greater local abundance of hoverflies (syrphids) and female medium-sized wild bees (albeit the latter's effect diminished with decreasing maternal plant attractiveness), and the presence of male bumblebees (Bombus) under low local floral richness. Cyanus segetum progeny genetic diversity increased with male Bombus presence but decreased with greater abundance of syrphids and honey bees (Apis mellifera). Overall, field margins supported plant-pollinator interactions ensuring multiple paternity and conservation of allelic diversity in C. segetum progeny. The contribution to plant mating outcomes of different pollinator functional groups was dictated by their local abundance or traits affecting pollen transfer efficiency. The local floral richness or maternal plant attractiveness sometimes modulated these relationships. This complex response of wild plant mating patterns to community interactions has implications for the use of field margins to restore functional pollination systems in farmed landscapes.

Co-option of plant gene regulatory network in nutrient responses during terrestrialization.

Plant responses to nitrate, phosphate and sucrose form a complex molecular network crucial for terrestrial adaptation. However, the origins, functional diversity and evolvability of this network during plant terrestrialization remain scarcely understood. Here we compare the transcriptomic response to these nutrients in the bryophyte Marchantia polymorpha and the streptophyte alga Klebsormidium nitens. We show that the largely species-specific nutrient response pattern is driven by gene regulatory network (GRN) alterations. Intriguingly, while pathways governing the GRNs exhibit modest conservation, M. polymorpha GRNs exhibit more regulatory connections through the redeployment of ancient transcription factor CSD. In M. polymorpha, functional analyses reveal the involvement of pre-existing cytokinin machineries in downstream targets, orchestrating plastic morpho-physiological responses to nutrient status. Our findings implicate the genetic co-option events facilitating successful land plant establishment.

Genome-wide identification and expression analysis of the MORF gene family in celery reveals their potential role in chloroplast development

Chlorophyll is an important nutrient in celery and one of the main indexes of quality evaluation. RNA editing in chloroplasts is an important factor affecting chloroplast development and chlorophyll biosynthesis. Multisite organelle RNA editing factor (MORF) protein is a necessary regulator of chloroplast RNA editing. In this study, a total of 8 MORF genes in celery were identified, which were named AgMORF1a, AgMORF1b, AgMORF2a, AgMORF2b, AgMORF3, AgMORF7, AgMORF8 and AgMORF9 according to their subfamily classification. The physicochemical property, conserved motifs, cis-acting elements and protein interaction were predicted according to the sequences. The phylogenetic relationships and evolutionary selective pressure between MORF genes in celery and other Apiaceae plants were further analyzed. The results showed that AgMORF1b, AgMORF2a, AgMORF2b and AgMORF9 were predicted to be localized in chloroplasts. The evolution of MORF genes in 4 Apiaceae plants including celery, carrot, coriander and water dropwort was influenced by purify selection. Transcriptome data showed that the transcriptional levels of AgMORF2a, AgMORF2b, AgMORF8 and AgMORF9 were relatively higher among all MORF genes in petioles of celery, indicating their major role. RT-qPCR data showed that the expression levels of the above 4 genes were significantly higher in petioles of green celery than those of white celery. This study provided a basis for analyzing the effects of MORF proteins on chloroplast development of celery with different chlorophyll accumulation.

Investigation of the effectiveness and molecular mechanisms of thiamin priming to control early blight disease in potato.

In several plant species, thiamin foliar application primes plant immunity and can be effective in controlling various diseases. However, the effectiveness of thiamin against potato pathogens has seldom been investigated. Additionally, the transcriptomics and metabolomics of immune priming by thiamin have not previously been investigated. Here, we tested the effect of thiamin application against Alternaria solani, the causal agent of early blight in potato, and identified associated changes in gene expression and metabolite content. Thiamin applied on foliage at an optimal concentration of 10 mM reduced lesion size by ~33%. However, prevention of lesion growth was temporally limited, as a reduction of lesion size occurred when leaves were inoculated 4 h, but not 24 h, following thiamin treatment. Additionally, the effect of thiamin on lesion size was restricted to the application site and was not systemic. RNA-seq analysis showed that thiamin affected the expression of 308 genes involved in the synthesis of salicylic acid, secondary metabolites, fatty acid, chitin, and primary metabolism, and photosynthesis, which were also amongst the thousands of genes differentially regulated in the response to pathogen alone. Several of these genes and pathways were more differentially expressed and enriched when thiamin and the pathogen were combined. Thiamin also delayed the downregulation of photosynthesis-associated genes in plants inoculated with A. solani. Metabolite analyses revealed that thiamin treatment in the absence of pathogen decreased the amounts of several organic compounds involved in the citric acid cycle. We hypothesize that thiamin primes plant defenses through perturbation of primary metabolism.

Genome-Wide Identification of GATA Family Genes in Potato and Characterization of StGATA12 in Response to Salinity and Osmotic Stress.

GATA factors are evolutionarily conserved transcription regulators that are implicated in the regulation of physiological changes under abiotic stress. Unfortunately, there are few studies investigating the potential role of GATA genes in potato plants responding to salt and osmotic stresses. The physicochemical properties, chromosomal distribution, gene duplication, evolutionary relationships and classification, conserved motifs, gene structure, interspecific collinearity relationship, and cis-regulatory elements were analyzed. Potato plants were treated with NaCl and PEG to induce salinity and osmotic stress responses. qRT-PCR was carried out to characterize the expression pattern of StGATA family genes in potato plants subjected to salinity and osmotic stress. StGATA12 loss-of-function and gain-of-function plants were established. Morphological phenotypes and growth were indicated. Photosynthetic gas exchange was suggested by the net photosynthetic rate, transpiration rate, and stomatal conductance. Physiological indicators and the corresponding genes were indicated by enzyme activity and mRNA expression of genes encoding CAT, SOD, POD, and P5CS, and contents of H2O2, MDA, and proline. The expression patterns of StGATA family genes were altered in response to salinity and osmotic stress. StGATA12 protein is located in the nucleus. StGATA12 is involved in the regulation of potato plant growth in response to salinity and osmotic stress. Overexpression of StGATA12 promoted photosynthesis, transpiration, and stomatal conductance under salinity and osmotic stress. StGATA12 overexpression induced biochemical responses of potato plants to salinity and osmotic stress by regulating the levels of H2O2, MDA, and proline and the activity of CAT, SOD, and POD. StGATA12 overexpression induced the up-regulation of StCAT, StSOD, StPOD, and StP5CS against salinity and osmotic stress. StGATA12 could reinforce the ability of potato plants to resist salinity and osmosis-induced damages, which may provide an effective strategy to engineer potato plants for better adaptability to adverse salinity and osmotic conditions.

Genome‐Wide Association Study Identifies the Serine/Threonine Kinase ClSIK1 for Low Nitrogen Tolerance in Watermelon Species

ABSTRACTPlants have evolved multiple complex mechanisms enabling them to adapt to low nitrogen (LN) stress via increased nitrogen use efficiency (NUE) as nitrogen deficiency in soil is a major factor limiting plant growth and development. However, the adaptive process and evolutionary roles of LN tolerance‐related genes in plants remain largely unknown. In this study, we resequenced 191 watermelon accessions and examined their phenotypic differences related to LN tolerance. A major gene ClSIK1 encoding a serine/threonine protein kinase involved in the response to LN stress was identified on chromosome 11 using genome‐wide association study and RNA‐Seq analysis. According to a functional analysis, ClSIK1 overexpression can increase the root area, total biomass, NUE and LN tolerance by manipulating multiple nitrogen‐metabolized genes. Interestingly, the desirable LN‐tolerant haplotype ClSIK1HapC was detected in only one wild relative (Citrullus mucosospermus) and likely gradually lost during watermelon domestication and improvement. This study clarified the regulatory effects of ClSIK1 on NUE and adaptations to LN stress, which also identifying valuable haplotypes‐resolved gene variants for molecular design breeding of ‘green’ watermelon varieties highly tolerant to LN stress.

A fungal sRNA silences a host plant transcription factor to promote arbuscular mycorrhizal symbiosis

Summary Cross‐kingdom RNA interference (ckRNAi) is a mechanism of interspecies communication where small RNAs (sRNAs) are transported from one organism to another; these sRNAs silence target genes in trans by loading into host AGO proteins. In this work, we investigated the occurrence of ckRNAi in Arbuscular Mycorrhizal Symbiosis (AMS). We used an in silico prediction analysis to identify a sRNA (Rir2216) from the AM fungus Rhizophagus irregularis and its putative plant gene target, the Medicago truncatula MtWRKY69 transcription factor. Heterologous co‐expression assays in Nicotiana benthamiana, 5′ RACE reactions and AGO1‐immunoprecipitation assays from mycorrhizal roots were used to characterize the Rir2216–MtWRKY69 interaction. We further analyzed MtWRKY69 expression profile and the contribution of constitutive and conditional MtWRKY69 expression to AMS. We show that Rir2216 is loaded into an AGO1 silencing complex from the host plant M. truncatula, leading to cleavage of a host target transcript encoding for the MtWRKY69 transcription factor. MtWRKY69 is specifically downregulated in arbusculated cells in mycorrhizal roots and increased levels of MtWRKY69 expression led to a reduced AM colonization level. Our results indicate that MtWRKY69 silencing, mediated by a fungal sRNA, is relevant for AMS; we thus present the first experimental evidence of fungus to plant ckRNAi in AMS.

Characterization of the Cannabis sativa glandular trichome epigenome

BackgroundThe relationship between epigenomics and plant specialised metabolism remains largely unexplored despite the fundamental importance of epigenomics in gene regulation and, potentially, yield of products of plant specialised metabolic pathways. The glandular trichomes of Cannabis sativa are an emerging model system that produce large quantities of cannabinoid and terpenoid specialised metabolites with known medicinal and commercial value. To address this lack of epigenomic data, we mapped H3K4 trimethylation, H3K56 acetylation, H3K27 trimethylation post-translational modifications and the histone variant H2A.Z, using chromatin immunoprecipitation, in C. sativa glandular trichomes, leaf, and stem tissues. Corresponding transcriptomic (RNA-seq) datasets were integrated, and tissue-specific analyses conducted to relate chromatin states to glandular trichome specific gene expression.ResultsThe promoters of cannabinoid and terpenoid biosynthetic genes, specialised metabolite transporter genes, defence related genes, and starch and sucrose metabolism were enriched specifically in trichomes for histone marks H3K4me3 and H3K56ac, consistent with active transcription. We identified putative trichome-specific enhancer elements by identifying intergenic regions of H3K56ac enrichment, a histone mark that maintains enhancer accessibility, then associated these to putative target genes using the tissue specific gene transcriptomic data. Bi-valent chromatin loci specific to glandular trichomes, marked with H3K4 trimethylation and H3K27 trimethylation, were associated with genes of MAPK signalling pathways and plant specialised metabolism pathways, supporting recent hypotheses that implicate bi-valent chromatin in plant defence. The histone variant H2A.Z was largely found in intergenic regions and enriched in chromatin that contained genes involved in DNA homeostasis.ConclusionWe report the first genome-wide histone post-translational modification maps for C. sativa glandular trichomes, and more broadly for glandular trichomes in plants. Our findings have implications in plant adaptation and stress responses and provide a basis for enhancer-mediated, targeted, gene transformation studies in plant glandular trichomes.

Innate Immunity in Rice: A Defense Against Bacterial Leaf Blight

Rice (Oryza sativa L.) is one of the major staple food crops of the world and sustainable rice production is important for ensuring global food security. Throughout the growing season, a variety of pathogens including fungi, bacteria, viruses, and nematodes, infect different parts of the crop resulting in yield loss. Bacterial leaf blight caused by Xanthomonas oryzae pv. oryzae (Xoo) is one of the major limiting factors in rice production. Being a vascular pathogen, X. oryzae pv. oryzae interferes with a range of physiological and biochemical exchange process in rice. The earlier the pathogen infection, more will be the loss and grain quality is also severely affected due to infection. The spread of the pathogen is too rapid in the favourable climatic conditions. The disease results in 20 to 30 per cent annual loss in rice production and under severe conditions, the yield loss goes up to 80 per cent. Plant pathogens employ different ways to attack host plants and impair plant growth and reproduction. Unlike vertebrates, plants lack mobile immune cells and an adaptive immune system. Plants mainly rely on two interconnected tiers of the innate immune system to perceive and respond to pathogen infection. This innate immune system or basal resistance mediated by a repertoire of Resistance or R genes in plants acts as the first line of pre-formed and inducible defenses that protect the host plants from large number of pathogens. Over the years, extensive investigation on the molecular interactions between rice and Xanthomonas oryzae pv. oryzae has made impressive progress in understanding the molecular basis of rice innate immunity against bacterial leaf blight. Improving plant immunity has been considered as one of the best choices available for achieving economical and sustainable management of bacterial leaf blight in a durable manner. In this review, we summarize the molecular basis of two tiered innate immune system including PAMP triggered immunity (PTI) and Effector triggered immunity (ETI) as well as R genes involved in rice - Xanthomonas oryzae pv. oryzae interaction.

Three endogenous pararetrovirus genomes in Medicago sativa.

The genome sequences of three related endogenous pararetroviruses were obtained by high-throughput genomic sequencing of Medicago sativa. The genomes were found to be integrated within plant genes. The phylogeny revealed that Caulimovirus-MSa3 was closely related to caulimoviruses of petunia, whereas Caulimovirus-MSa1 and Caulimovirus-MSa2 were distinct from constructed clades.

The role of bacteriophages in facilitating the horizontal transfer of antibiotic resistance genes in municipal wastewater treatment plants

Bacteriophages play integral roles in the ecosystem; however, their precise involvement in horizontal gene transfer and the spread of antibiotic resistance genes (ARGs) are not fully understood. In this study, a coculture system involving consortia of bacteriophages and multidrug-resistant bacteria from an aerobic tank in a municipal wastewater treatment plant (WWTP) was established to investigate the functions of bacteriophages in ARG transfer and spread. The results of the cocultivation of the MRB and bacteriophage consortia indicated that the bacterial community remained stable throughout the whole process, but the addition of bacteriophages significantly increased ARG abundance, especially in bacteriophage DNA. Nine out of the 11 identified ARGs significantly increased, indicating that more bacteriophage particles carried ARGs in the system after cocultivation. In addition, 686 plasmids were detected during cocultivation, of which only 3.36 % were identified as conjugative plasmids, which is significantly lower than the proportion found among previously published plasmids (25.2 %, totaling 14,029 plasmids). Our findings revealed that bacteriophages may play important roles in the horizontal transfer of ARGs through both bacteriophage-mediated conduction and an increase in extracellular ARGs; however, conjugative transfer may not be the main mechanism by which multidrug-resistant bacteria acquire and spread ARGs. Unlike in most previous reports, a coculture system of diverse bacteria and bacteriophages was established in this study to assess bacteriophage functions in ARG transfer and dissemination in the environment, overcoming the limitations associated with the isolation of bacteria and bacteriophages, as well as the specificity of bacteriophage hosts.

The response of DNA methyltransferase and demethylase genes to abiotic stresses in tomato seedling

DNA methylation plays an important role in regulating plant growth, development and gene expression. However, less is known about the response of DNA methyltransferase and demethylase genes to various stresses. In this study, the effects of abiotic stresses on DNA methylation gene expression patterns in tomato seedlings were investigated. Results showed that most tomato DNA methyltransferase and demethylase genes contained stress-related elements. The expression of SlDML1 was significantly induced by cadmium (Cd) and sodium chloride (NaCl) stresses. SlDML2 was more sensitive and reached its maximum value under polyethylene (PEG) stress at 24 h. The expression of SlMET3L was repressed to varying degrees under Cd, NaCl and PEG stresses at 48 h. However, 5-aza-2′-deoxycytidine (5-azadC) treatment decreased the Cd and PEG stress tolerance by down-regulating the expression of DNA methyltransferase except for the SlMET3L, and up-regulating the expression levels of SlDML2, SlDML3 and SlDML4, cadmium transporters (SlHMA5, SlCAX3, and SlACC3) and osmoregulators (SlDREB, SlLEA and SlHSP70). Whereas 5-azadC treatment alleviated the salt stress through up-regulating DNA methyltransferase gene expression, and down-regulating the expression level of SlDML1, SlDML3, and SlDML4, SlHKT1, SlNHX1, and SlSOS1. Collectively, 5-azadC impaired Cd and PEG stress tolerance and enhanced salt stress tolerance by regulating the expression of methylation-related and stress-related genes in tomato seedlings. These results may provide useful information for further analysing function and evolution of DNA methylation methyltransferase and demethylase genes in tomato under stress conditions.

OsPIPK-FAB, A Negative Regulator in Rice Immunity Unveiled by OsMBL1 Inhibition

Phosphatidylinositol signaling system plays a crucial role in plant physiology and development, phosphatidylinositol phosphate kinases (PIPKs) are one of the essential enzymes responsible for catalyzing the synthesis of phosphatidylinositol bisphosphate (PIP2) within this signaling pathway. However, its mechanism of signal transduction remains poorly exploited in plants. OsMBL1, a jacalin-related mannose-binding lectin in rice, plays a crucial role in plant defense mechanisms, acting as a key component of the pattern-triggered immunity (PTI) pathway. Here, a rice phosphatidylinositol-phosphate kinase FAB (OsPIPK-FAB), a member of the rice PIPKs family, as an interacting protein of OsMBL1 through yeast-two-hybrid (Y2H) screening assay. And this interaction was confirmed by using co-immunoprecipitation (Co-IP) and pull-down assay techniques. Furthermore, we demonstrated that the deletion of OsPIPK-FAB gene in plant enhanced resistance against rice blast while overexpression of OsPIPK-FAB increases sensitivity to the fungal infection. Additionally, through determination and measurement of the plant inositol 1,4,5-trisphosphate (IP3) contents and the plant phosphatidylinositol 4-phosphate 5-kinase (PIP5K) activity, we revealed that OsMBL1 inhibits the PIP5K kinase activity of OsPIPK-FAB as well as the plant IP3 contents in rice. Conclusively, these findings indicated that OsPIPK-FAB serves as a novel and critical component that is negatively involved in PTI activation and was inhibited by OsMBL1.

Effects of water deficit on two cultivars of Hibiscus mutabilis: A comprehensive study on morphological, physiological, and metabolic responses

Hibiscus mutabilis, commonly known as the cotton rose, is a widely cultivated ornamental and has been acclaimed as the representative flower of the 2024 World Horticultural Exposition. The growth and ornamental characteristics of Hibiscus mutabilis can be affected by drought stress. Therefore, we investigated the physiological and metabolic responses of drought-sensitive Hibiscus mutabilis JRX-1 and drought-tolerant Hibiscus mutabilis CDS-4 under drought stress. The results of the physiological analyses revealed that, compared to JRX-1,CDS-4 maintained good growth and greater water use efficiency through stronger antioxidant defences, osmoregulatory capacity and stomatal regulation. A total of 3277 metabolites were identified in positive and negative ion modes, of which 663 metabolites presented changes in expression under drought conditions, including 306 upregulated metabolites and 357 downregulated metabolites. Secondary metabolites, such as flavonoids and diterpenoids, are crucial in the plant response to drought stress. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that the differentially aboundant metabolites were significantly enriched in the pathways valine, leucine and isoleucine degradation; linoleic acid metabolism; one carbon pool by folate; and folate biosynthesis. The results of this study will not only help to elucidate and apply the physiological and metabolic regulatory strategies of Hibiscus mutabilis to improve its adaptation to water deficit conditions, but will also provide valuable guidance to breeders and molecular biologists in the screening and use of drought resistant genes in ornamental plants.

Citric acid-driven cadmium uptake and growth promotion mechanisms in Brassica napus

Citric acid (CA) is well-known for mitigating cadmium (Cd) toxicity in plants. Yet, the underlying mechanisms driving growth promotion, Cd detoxification/tolerance, and enhanced phytoremediation processes remain incompletely understood. This study investigated the effects of CA application (2.5 mM) on Brassica napus grown in Cd-contaminated (30 mg kg−1) growth medium through a controlled pot experiment. Cd exposure alone significantly impaired various plant physiological parameters in B. napus. Whereas CA application significantly (p < 0.05) enhanced physiological attributes, Cd detoxification and tolerance by modulating key genes involved in photosynthesis and Cd transport, including the metal-transporting P1B-type ATPases (Cd/zinc heavy metal-transporting ATPase 1; HMA1) and light-harvesting chlorophyll a/b-binding 3 (LHCB3). Notably, CA application increased Cd accumulation in stems and leaves by 4% and 35%, respectively, enhancing bioconcentration factors (BCF) by 12% in stems and 40% in leaves while reducing root BCF by 10%. This translocation was facilitated by the upregulation of HMA4, HMA2, and plant Cd resistance (PCR2) genes in plant leaves, improving Cd mobility within the plant. Furthermore, CA induced a 34% increase in phytochelatins and a 32% upregulation in metallothioneins, accompanied by a significant reduction in oxidative stress markers, including a 40% decrease in hydrogen peroxide and a 44% decline in malondialdehyde levels in leaves. Enhanced antioxidant enzyme activity and osmolyte accumulation further contributed to improved Cd detoxification/sequestration in leaves, reduced oxidative stress, and improved photosynthetic efficiency, resulting in enhanced plant biomass production and Cd tolerance. Transcriptomic analysis showed that CA treatment substantially influenced the expression of 12,291 differentially expressed genes (DEGs), with 750 common genes consistently downregulated in CK vs Cd treatment group but upregulated in Cd vs Cd-CA treatment group. Additionally, CA modulated 11 DEGs associated with 32 gene ontologies in the citrate pathway under Cd stress, highlighting its targeted regulatory effect on metabolic pathways involved in Cd stress response. This study offers novel insights into the synergistic role of CA in promoting plant growth and regulating Cd uptake in B. napus, highlighting its potential to enhance phytoremediation strategies.

Mapping of dwarfing gene and identification of mutant allele on plant height in wheat

Plant height is one of the most critical factors influencing wheat plant architecture, and the application of Green Revolution genes has led to a reduction in plant height and an increase in yield. Discovering new dwarfing genes and alleles can contribute to enhance the genetic diversity of wheat. Here we obtained an EMS induced dwarf wheat mutant je0166 with increased grain weight, which exhibited a reduction in plant height ranging from 46.47% to 49.40%, and its cell length was shorter. The mutant je0166 was sensitive to exogenous gibberellin, but its sensitivity was lower than that of its wild type. Genetic analysis on plant height and gene mapping located the target region to a 4.07 cM interval on chr. 4AL. Within this interval, we identified a co-segregated mutation in Rht-A1h, which is a novel allele of the Green Revolution gene Rht-A1. We also found large fragment inversions in the genetic map of the mutant. The novel allele diversifies natural allelic variations and could be utilized in future wheat improvement. Furthermore, we demonstrated that chemical mutagen treatment led to large fragment inversion.

A history-dependent integrase recorder of plant gene expression with single-cell resolution

During development, most cells experience a progressive restriction of fate that ultimately results in a fully differentiated mature state. Understanding more about the gene expression patterns that underlie developmental programs can inform engineering efforts for new or optimized forms. Here, we present a four-state integrase-based recorder of gene expression history and demonstrate its use in tracking gene expression events in Arabidopsis thaliana in two developmental contexts: lateral root initiation and stomatal differentiation. The recorder uses two serine integrases to mediate sequential DNA recombination events, resulting in step-wise, history-dependent switching between expression of fluorescent reporters. By using promoters that express at different times along each of the two differentiation pathways to drive integrase expression, we tie fluorescent status to an ordered progression of gene expression along the developmental trajectory. In one snapshot of a mature tissue, our recorder is able to reveal past gene expression with single cell resolution. In this way, we are able to capture heterogeneity in stomatal development, confirming the existence of two alternate paths of differentiation.