Resistance In Arabidopsis Research Articles

GhRac9 improves cotton resistance to Verticillium dahliae via regulating ROS production and lignin content.

Rac/Rop proteins, a kind of unique small GTPases in plants, play crucial roles in plant growth and development and in response to abiotic and biotic stresses. However, it is poorly understood whether cotton Rac/Rop protein genes are involved in mediating cotton resistance to Verticillium dahliae. Here, we focused on the function and mechanism of cotton Rac/Rop gene GhRac9 in the defense response to Verticillium dahliae infection. The expression level of GhRac9 peaked at 24 h after V. dahliae infection and remained consistently elevated from 24 to 48 h upon SA treatment. Furthermore, silencing GhRac9 using VIGS (Virus-induced gene silence) method attenuated cotton defense response to V. dahliae by reducing ROS (Reactive Oxygen Species) burst, peroxidase activity and lignin content in cotton plants. On the contrary, heterologous overexpression of GhRac9 enhanced Arabidopsis resistance to V. dahliae and significantly increased ROS production in Arabidopsis plants. Furthemore, transient overexpressing of GhRac9 significantly enhanced ROS burst and POD activity in cotton plants. In addition, GhRac9 positively regulated the expression levels of the genes related to SA signaling pathway in cotton plants. In conclusion, GhRac9 functioned as a positive regulator in the cotton defense response to V. dahliae, which provided important insights for breeding new cotton varieties resistant to V. dahliae.

Immune Priming Promotes Thermotolerance, Whereas Thermopriming Suppresses Systemic Acquired Resistance in Arabidopsis.

Heat stress and pathogens are two serious yield-limiting factors of crop plants. Plants that previously experienced high but sub-lethal temperatures become subsequently tolerant to higher temperatures through the development of acquired thermotolerance (ATT). ATT activation is associated with the elevated expression of heat shock (HS)-related genes such as HSFA2, HSFA3, and HSP101. Similarly, through the development of systemic acquired resistance (SAR), previously experienced plants achieve a higher resistance than naïve plants. SAR activation requires mobile signals and primarily depends on salicylic acid (SA) signaling. Studies to understand the interaction between ATT and SAR are limiting. To investigate the possible interconnection, we studied cross-protection between SAR and ATT on 4-week-old soil-grown Arabidopsis plants. We observed localized pathogen inoculation provides thermotolerance. Pathogens activate the expressions of HSFA2, HSFA3, HSA32, and HSP101 in pathogen-free systemic tissues. Interestingly, pathogen-induced SAR activation is impaired in hsfa2, hsfa3, and hsp101 mutants, suggesting these HS memory genes are essential for SAR induction. In contrast, thermopriming by exposing plants to sublethal temperatures, blocks SAR activation by pathogens. Thermopriming suppresses SAR mobile signal generation, accumulation of SA, and PR1 gene expression in systemic leaves. Altogether, our results demonstrate a complex interaction between SAR and ATT induction pathways in plants.

A chromosome-level reference genome facilitates the discovery of clubroot resistant gene Crr5 in Chinese cabbage

Abstract Brassica rapa includes a variety of important vegetable and oilseed crops, yet it is significantly challenged by clubroot disease. Notably, the majority of genotypes of B. rapa with published genomes exhibit high susceptibility to clubroot disease. The present study presents a high-quality chromosome-level sequence of the genome of the DH40 clubroot-resistant (CR) line, a doubled haploid line derived from the hybrid progeny of a European turnip (ECD01) and two lines of Chinese cabbage. The assembled genome spans 420.92 Mb, with a contig N50 size of 11.97 Mb. Comparative genomics studies revealed that the DH40 line is more closely related to the Chinese cabbage Chiifu than to the turnip ECD04. The DH40 genome provided direct reference and greatly facilitate the map-based cloning of the clubroot resistance gene Crr5, encoding a nucleotide-binding leucine-rich repeat (NLR) protein. Further functional analysis demonstrated that Crr5 confers clubroot resistance in both Chinese cabbage and transgenic Arabidopsis. It responds to inoculation with Plasmodiophora brassicae and is expressed in both roots and leaves. Subcellular localization shows that Crr5 is present in the nucleus. Notably, the TIR domain of Crr5 can autoactivate and trigger cell death. In addition, we developed two Crr5-specific KASP markers and showcased their successful application in breeding CR Chinese cabbage through marker-assisted selection. Overall, our research offers valuable resources for genetic and genomic studies in B. rapa and deepens our understanding of the molecular mechanisms underlying clubroot resistance against Plasmodiophora brassicae.

VOZ-dependent priming of salicylic acid-dependent defense against Rhizopus stolonifer by β-aminobutyric acid requires the TCP protein TCP2 in peach fruit.

Vascular plant one-zinc finger (VOZ) transcription factors (TFs) play crucial roles in plant immunity. Nevertheless, how VOZs modulate defense signaling in response to elicitor-induced resistance is not fully understood. Here, the defense elicitor β-aminobutyric acid (BABA) resulted in the visible suppression of Rhizopus rot disease of peach fruit caused by Rhizopus stolonifer. Defense priming by BABA was notably associated with increased levels of salicylic acid (SA) and SA-dependent gene expression. Data-independent acquisition proteomic analysis revealed that two VOZ proteins (PpVOZ1 and PpVOZ2) were substantially upregulated in BABA-induced resistance (BABA-IR). Furthermore, the interaction of PpVOZ1 and PpVOZ2 and their potential target of the TEOSINTE-BRANCHED1/CYCLOIDEA/PCF (TCP)-family protein PpTCP2 screened from protein-protein interaction networks was confirmed by yeast two-hybrid (Y2H), luciferase complementation imaging and glutathione S-transferase pull-down assays. Furthermore, subcellular localization, yeast one-hybrid, electrophoretic mobility shift assay and dual-luciferase reporter assays demonstrated that nuclear localization of both PpVOZ1 and PpVOZ2 was critical for their contribution to BABA-IR, as these proteins potentiated the PpTCP2-mediated transcriptional activation of isochorismate synthase genes (ICS1/2). The overexpression of both PpVOZ1 and PpVOZ2 could activate the transcription of SA-dependent genes and provide disease resistance in transgenic Arabidopsis. In contrast, the ppvoz1cas9 and ppvoz2cas9 loss-of-function mutations and the voz1cas9 voz2cas9 double mutation attenuated BABA-IR against R. stolonifer. Therefore, the three identified positive TFs, PpVOZ1, PpVOZ2, and PpTCP2, synergistically contribute to the BABA-activated priming of systemic acquired resistance in postharvest peach fruit by a VOZ-TCP-ICS regulatory module.

Enhanced reactive oxygen species and metabolism are involved in the reduction of tribenuron-methyl residues in Tartary buckwheat mutants

Tartary buckwheat, known for its rapid growth, short growth cycles, and adaptability, is cultivated worldwide, particularly in East Asia and Eastern Europe. However, weed infestations severely impact yield and quality, and the absence of effective herbicides poses a considerable challenge to Tartary buckwheat production. To address this, we used ethyl methane sulfonate (EMS) mutagenesis to create a mutant population of Tartary buckwheat seeds. We applied a targeted screening process using the herbicide tribenuron-methyl (TM) to select mutants with reduced sensitivity to the herbicide. Integrative analysis of transcriptomic and metabolomic data indicated that TM primarily inhibited photosynthesis and amino acid biosynthesis pathways in sensitive plants, leading to toxicity. Conversely, resistant plants could reduce the toxic effects of TM on Tartary buckwheat by increasing antioxidant enzyme activity and enhancing secondary metabolic pathways such as flavonoid biosynthesis. TM also upregulated the expression of a gene encoding uridine diphosphate glucuronic acid transferase-like (FtUGT79L). Overexpression of FtUGT79L in Arabidopsis substantially increased total flavonoid content and improved the resistance of Arabidopsis to TM at the seedling and adult stages. The study provides insights into innovative approaches for breeding herbicide-resistant Tartary buckwheat germplasm and serves as an important reference for the development of herbicide-resistant varieties of other crops.

A Novel Member of miR169 Family Negatively Regulates Maize Resistance Against Bipolaris maydis.

MicroRNAs (miRNAs) have been confirmed to play important roles in plant defense response. However, the key maize miRNAs involved in the defense response against Bipolaris maydis are very limited. In this study, a novel member of the miR169 family in response to B. maydis, named zma-miR169s, was discovered and investigated. The expression levels of pre-miR169s and zma-miR169s were significantly repressed during B. maydis infection. The CRISPR/Cas9-induced zma-miR169s mutant exhibited more resistance against B. maydis, whereas overexpression of zma-miR169s enhanced susceptibility, supporting that zma-miR169s might play a negative role in maize resistance. Moreover, RNA-seq and Gene Ontology analysis showed that differentially expressed genes were highly enriched in the oxidation-reduction process and plant hormone pathway. Hence, reactive oxygen species (ROS) and plant hormone levels were further investigated. ROS detection confirmed that the zma-miR169s mutant accumulated more ROS, while less ROS was detected in transgenic maize OE-miR169s. Furthermore, more remarkable changes in PR-1 expression levels and salicylic acid (SA) contents were detected in the zma-miR169s mutant compared with wild-type and transgenic maize during B. maydis infection. Additionally, nuclear transcription factors (NF-YA1 and NF-YA13) were identified as targets regulated by zma-miR169s through the agrobacterium-mediated transient expression method. Overexpression of ZmNF-YA13 enhanced Arabidopsis resistance to Pseudomonas syringae pv. tomato DC3000. Taken together, our results suggest that zma-miR169s negatively regulates maize defense responses by influencing ROS accumulation and the SA-dependent signaling pathway.

Molecular identification and expression patterns of sweet cherry HIPPs and functional analysis of PavHIPP16 in cold stress.

The HIPP proteins are involved in low-temperature stress, the growth of sweet cherry, and may be potential targets for genetic improvement. PavHIPP16 improved cold resistance in Arabidopsis. In response to abiotic stressors, the heavy metal-associated isoprenylated plant protein (HIPP) proteins play a crucial regulatory role. Although the function of HIPP has been identified in some plants, there have been fewer systematic studies conducted on sweet cherry (Prunus avium L.). Therefore, we performed a comprehensive analysis and expression profiling of PavHIPPs using bioinformatics, RT-PCR, and qRT-PCR techniques. Our findings revealed that 28 PavHIPP genes were unevenly distributed across eight chromosomes. We predicted nine motifs in PavHIPP proteins and observed similar gene structures among highly homologous proteins. The promoter sequences of PavHIPPs contained numerous regulatory elements associated with an adversity of stress. The expression levels of some members showed varying degrees of change under low-temperature treatment. These genes were differentially expressed during flower and fruit development. Arabidopsis overexpressing the PavHIPP16 (OE) gene showed significantly lower relative conductivity and malondialdehyde (MDA) content compared with the wild-type (WT) plants under cold environment. Conversely, peroxidase (POD) activity, superoxide dismutase (SOD) activity, and proline content were significantly higher in OE Arabidopsis than in WT plants. Overall, our results suggest that PavHIPP16 OE Arabidopsis thaliana exhibited enhanced adaptability compared to WT plants under cold conditions. This study provides a foundation for future investigations of the pathways regulating sweet cherry growth and development mediated by the HIPP genes.

The Jahn–Teller distortion‐induced electronic structure regulation of Mn‐doped Co3O4 for enhanced acetone detection

AbstractThe modulation of the electronic structure of metal oxides is crucial to enhance their gas‐sensing performance. However, there is lacking in profound study on the effect of electronic structure regulation on sensing performance. Herein, we propose an innovative strategy of Jahn–Teller distortion‐induced electronic configuration regulation of Co3O4 to improve acetone sensing performance. After the introduction of Mn3+ into Co3O4 (Mn‐Co3O4), the Jahn–Teller distortion of high‐spin Mn3+ (t2g3eg1) conversed to low‐spin Mn4+ (t2g3eg0), resulting in conversion of Co3+ (t2g6eg0) into Co2+ (t2g6eg1). As expected, Mn‐Co3O4 exhibits a high response value of 46.7 toward 100 ppm acetone, low limit of detection of 0.75 ppb, high selectivity, and high stability, which are overwhelmingly superior to previous Co3O4‐based acetone sensors. The dynamics and thermodynamics analysis demonstrate that the Mn doping improves sensing reaction rate, reduces reaction barrier, and promotes the charge transfer. The theoretical calculations further prove the charge transfer from Mn to Co derived from Jahn–Teller distortion and support promoting the adsorption of acetone on Co3O4 by Mn dopant. Moreover, we demonstrated the substantial potential application of Mn‐Co3O4 sensor as a monitoring gas sensor in pest resistance of Arabidopsis. This work provides a new strategy to design sensing materials from electronic configuration perspective.image

Guard Cell Activity of PIF4 Represses Disease Resistance in Arabidopsis.

Phytochrome Interacting Factor 4 (PIF4) plays a central role in coordinating plant growth regulation by integrating multiple environmental cues. However, studies on whether and how PIF4 regulates plant immunity have inconsistent findings. In this study, we investigated the role of PIF4 in disease resistance against Pst DC3000 by characterizing its loss-of-function mutants using different inoculation strategies. Our findings reveal that pif4 mutants exhibit enhanced disease resistance with spray inoculation but not with infiltration inoculation compared to wild-type plants, and that mutants displayed more closed stomata apertures, indicating that PIF4 promotes stomatal opening. Importantly, expression of PIF4 by a guard-cell-specific promoter was sufficient to restore disease resistance to the wild-type level in the pif4 mutant. Additionally, PIF4 overexpression enhances disease symptom development independent of disease resistance and chlorophyll degradation, while the loss of PIF4 function leads to higher chlorophyll accumulation. Thus, our findings highlight a crucial function of PIF4 in regulating stomata-mediated disease resistance and chlorophyll accumulation, providing new insights into the connection of growth and defense in plants.

Border-like cell formation mediated by SgPG1 confers aluminum resistance in Stylosanthes guianensis.

Stylosanthes is an important forage legume in tropical areas with strong resistance to aluminum (Al) toxicity, though knowledge of mechanisms underlying this resistance remains fragmentary. We found that border-like cells (BLCs) were constitutively produced surrounding the root tips of all 54 examined Stylosanthes guianensis genotypes, but not the Stylosanthes viscose genotype TF0140. In genotypic comparisons under Al conditions, the S. guianensis genotype RY#2 retained significantly more Al in BLCs and thereby showed higher relative root growth than TF0140. Formation of BLCs accompanied changes in cell wall pectin epitopes and differential expression of genes involved in pectin metabolism, including a polygalacturonase (SgPG1). The expression pattern of SgPG1 was consistent with the formation of BLCs in both RY#2 and TF0140. SgPG1 was localized in cell walls and exhibited high activities mediating demethyl-esterified homogalacturonan degradation. Overexpressing SgPG1 changed cell wall pectin epitopes, enhanced BLCs production, and Al resistance in both Arabidopsis and Stylosanthes hairy roots. Furthermore, combining protein-DNA binding assays in vitro and in vivo, a bHLH transcription factor SgbHLH19 was demonstrated to be the upstream regulator of SgPG1. Our study demonstrates that S. guianensis Al resistance mainly relies on BLCs, whose formation involves cell wall pectin epitope modification by SgPG1.

HIP is involved in NaCl and endoplasmic reticulum stress resistance in Arabidopsis

Heat shock protein 70 (HSP70)-interacting proteins (HIPs) have been studied in animals. HIPs perform diverse cellular functions, ranging from alleviating stress to suppressing the formation of insoluble protein, but how their orthologs function in plants is less known. This study shows that Arabidopsis HIP is a cytosolic and nuclear protein associated with HSP70. The hip mutants showed compromised tolerance to NaCl and endoplasmic reticulum (ER) stress, although they grew normally under standard growth conditions. Furthermore, hip mutants presented a decreased HSP70 level compared with the wild type under NaCl and ER stress conditions. These findings suggest the involvement of HIP in NaCl and ER stress tolerance.

PbrATG6 modulates reactive oxygen species metabolism and interacts with PbrTLP15 synergistic enhancement of pear resistance to Botryosphaeria dothidea

Autophagy is vital for plant defense against pathogens, with ATG6 being a key gene in this process. At present, little has been reported on the potential function and molecular mechanisms of ATG6 mediated pathogen resistance in pear. This study investigates the function of the pear homolog of ATG6 (PbrATG6) in resistance to Botryosphaeria dothidea. PbrATG6 is expressed differentially in pear tissues and its expression increases upon infection. Overexpression of PbrATG6 enhances resistance in Arabidopsis and pear calli, while silencing it increases susceptibility. PbrTLP15, a pathogenesis-related protein belonging to the PR5 family, was found that interacts with PbrATG6 by a yeast two-hybrid screening. Yeast two-hybrid, luciferase complementation imaging, bimolecular fluorescence complementation assays and pull-down assays showed that PbrATG6 interacts with PbrTLP15. The transient silencing transgenic assays of PbrATG6 and PbrTLP15 revealed that PbrATG6 could cooperate with PbrTLP15 to regulate pear B. dothidea resistance. In addition, transcriptional analyses of autophagy key genes in pTRV-PbrTLP15 and transmission electron microscopy (TEM) assays also implied that PbrTLP15 does affect autophagy. Hence, PbrATG6 and PbrTLP15 may synergistically enhance pear B. dothidea disease resistance. It provides a new strategy for the study of autophagy in pear disease resistance and enriches the research on pear disease resistance mechanism.

Genome-wide identification of Aux/IAA gene family members in grape and functional analysis of VaIAA3 in response to cold stress.

Twenty-five VvIAA genes and eighteen VaIAA genes were identified from Pinot Noir and Shanputao, respectively. The overexpression of VaIAA3 in transgenic Arabidopsis increased cold tolerance by regulating auxin, ABA and ethylene signaling. Aux/IAA genes are key genes involved in regulating auxin signal transduction in plants. Although IAA genes have been characterized in various plant species, the role of IAA genes in grape cold resistance is unclear. To further explore the members of the Aux/IAA gene family in grape and their functions, in this study, using genomic data for Pinot Noir (Vitis vinifera cv. 'Pinot Noir') and Shanputao (Vitis amurensis), 25 VvIAA genes and 18 VaIAA genes were identified. The VaIAA genes presented different expression patterns at five different temperatures (28 ± 1 °C, 5 ± 1 °C, 0 ± 1 °C, -5 ± 1 °C, and -10 ± 1 °C) according to qRT‑PCR results. VaIAA3 was selected as a candidate gene for further functional analysis because of its high expression level under low-temperature stress. Subcellular localization experiments revealed that VaIAA3 was localized in the nucleus. Additionally, under 4 °C treatment for 24 h, relative expression level of VaIAA3, antioxidant enzyme activity, survival rate, and cold-responsive gene expression in three transgenic lines (OE-1, OE-2, OE-3) were greater, whereas relative electrolytic conductivity (REC), malondialdehyde (MDA) content and hydrogen peroxide (H2O2) content were lower than those of the wild type (WT). Transcriptome sequencing analysis revealed that VaIAA3 regulated cold stress resistance in Arabidopsis thaliana (Arabidopsis) through pathways involving auxin, ABA, JA, or ethylene. Importantly, heterologous overexpression of VaIAA3 increased the resistance of Arabidopsis to cold stress, which provides a theoretical basis for the further use of VaIAA3 to improve cold resistance in grape.

The pathogen-induced peptide CEP14 is perceived by the receptor-like kinase CEPR2 to promote systemic disease resistance in Arabidopsis.

Secreted plant peptides that trigger cellular signaling are crucial for plant growth, development, and adaptive responses to environmental stresses. In Arabidopsis (Arabidopsis thaliana), the C-TERMINALLY ENCODED PEPTIDE (CEP) family is a class of secreted signaling peptides that is phylogenetically divided into 2 groups: group I (CEP1-CEP12) and group II (CEP13-CEP15). Several group I CEP peptides regulate root architecture and nitrogen starvation responses, whereas the biological activity and roles of group II CEPs remain unknown. Here, we report that a group II CEP peptide, CEP14, functions as a pathogen-induced elicitor of Arabidopsis immunity. In response to infection by the bacterial pathogen Pseudomonas syringae, CEP14 expression was highly induced via the salicylic acid pathway in Arabidopsis leaves and roots. In the absence of a pathogen attack, treatment of Arabidopsis plants with synthetic CEP14 peptides was sufficient to trigger immune responses. Genetic and biochemical analyses demonstrated that the receptor-like kinase CEP RECEPTOR 2 (CEPR2) perceives CEP14 to trigger plant immunity. The SOMATIC EMBRYOGENESIS RECEPTOR KINASES (SERKs) BRASSINOSTEROID INSENSITIVE 1-ASSOCIATED RECEPTOR KINASE 1 (BAK1) and SERK4 also participated in CEP14 perception by forming CEP14-induced complexes with CEPR2. Overexpression of CEP14 largely enhanced Arabidopsis resistance to P. syringae, while CEP14 or CEPR2 mutation significantly attenuated Arabidopsis systemic resistance to P. syringae. Taken together, our data reveal that the pathogen-induced CEP14 peptide, which is perceived by the CEPR2-BAK1/SERK4 receptor complexes, acts as an endogenous elicitor to promote systemic disease resistance in Arabidopsis.

Comprehensive studies of the serine carboxypeptidase-like (SCPL) gene family in Carya cathayensis revealed the roles of SCPL4 in epigallocatechin-3-gallate (EGCG) synthesis and drought tolerance

Hickory (Carya cathayensis) nuts are rich in epigallocatechin-3-gallate (EGCG) with multiple health functions. EGCG also regulates plant growth, development and stress responses. However, research on the synthesis mechanism of EGCG and its function in hickory is currently limited. Herein, 44 serine carboxypeptidase-like (SCPL) members were identified from the hickory genome and classified into three major categories: SCPL-I, SCPL-II, and SCPL-III. In the CcSCPLs-IA branch, CcSCPL3/4/5/8/9/11/13 showed differential expression patterns in various tissues, especially with relatively high expression levels in plant roots, female flowers and seed coat. These proteins have a catalytic triad composed of serine (Ser), aspartic acid (Asp) and histidine (His). Ser-His in the triad and arginine (Arg) mediated the docking of CcSCPL3/4/5/11 with 1-O-galloyl-β-d-glucose (βG) and epigallocatechin (EGC), whereas the Asp of the triad did not. CcSCPL4 was further confirmed to promote the synthesis of EGCG in tobacco leaves. CcSCPL4 may function as monomer and be mainly localized within cellular structures outside the nucleus. Notably, the expression level of CcSCPL4 significantly changed after drought, cold, and salt stress, with the highest expression level under drought stress. Meanwhile CcSCPL4 over-expression could enhance the drought resistance of Saccharomyces cerevisiae and Arabidopsis. This study elucidates key enzymes for EGCG synthesis and their role in drought resistance, providing insights into the EGCG synthesis pathway and molecular breeding of hickory in future.

Callose and Salicylic Acid Are Key Determinants of Strigolactone-Mediated Disease Resistance in Arabidopsis

Research has demonstrated that strigolactones (SLs) mediate plant disease resistance; however, the basal mechanism is unclear. Here, we provide key genetic evidence supporting how SLs mediate plant disease resistance. Exogenous application of the SL analog, rac-GR24, increased Arabidopsis thaliana resistance to virulent Pseudomonas syringae. SL-biosynthetic mutants and overexpression lines of more axillary growth 1 (MAX1, an SL-biosynthetic gene) enhanced and reduced bacterial susceptibility, respectively. In addition, rac-GR24 promoted bacterial pattern flg22-induced callose deposition and hydrogen peroxide production. SL-biosynthetic mutants displayed reduced callose deposition but not hydrogen peroxide production under flg22 treatment. Moreover, rac-GR24 did not affect avirulent effector-induced cell death between Col-0 and SL-biosynthetic mutants. Furthermore, rac-GR24 increased the free salicylic acid (SA) content and significantly promoted the expression of pathogenesis-related gene 1 related to SA signaling. Importantly, rac-GR24- and MAX1-induced bacterial resistance disappeared completely in Arabidopsis plants lacking both callose synthase and SA. Taken together, our data revealed that callose and SA are two important determinants in SL-mediated plant disease resistance, at least in Arabidopsis.

Bioinformatics Analysis of the Panax ginseng Cyclophilin Gene and Its Anti-Phytophthora cactorum Activity.

In this paper, Panax ginseng cyclophilin (PgCyP) was successfully obtained through a genetic engineering technique. A bioinformatics method was used to analyze the physicochemical properties and structure of PgCyP. The results showed that PgCyP belongs to the cyclophilin gene family. The protein encoded by the PgCyP gene contains the active site of PPIase (R62, F67, and H133) and a binding site for cyclosporine A (W128). The relative molecular weight of PgCyP is 187.11 bp; its theoretical isoelectric point is 7.67, and it encodes 174 amino acids. The promoter region of PgCyP mainly contains the low-temperature environmental stress response (LTR) element, abscisic acid-responsive cis-acting element (ABRE), and light-responsive cis-acting element (G-Box). PgCyP includes a total of nine phosphorylation sites, comprising four serine phosphorylation sites, three threonine phosphorylation sites, and two tyrosine phosphorylation sites. PgCyP was recombined and expressed in vitro, and its recombinant expression was investigated. Furthermore, it was found that the recombinant PgCyP protein could effectively inhibit the germination of Phytophthora cactorum spores and the normal growth of Phytophthora cactorum mycelia in vitro. Further experiments on the roots of susceptible Arabidopsis thaliana showed that the PgCyP protein could improve the resistance of arabidopsis to Phytophthora cactorum. The findings of this study provide a basis for the use of the PgCyP protein as a new type of green biopesticide.

Functional characterization of Arabidopsis hydroxynitrile lyase in response to abiotic stress and the regulation of flowering time.

Hydroxynitrile lyases (HNLs) are a class of hydrolytic enzymes from a wide range of sources, which play crucial roles in the catalysis of the reversible conversion of carbonyl compounds derived from cyanide and free cyanide in cyanogenic plant species. HNLs were also discovered in non-cyanogenic plants, such as Arabidopsis thaliana, and their roles remain unclear even during plant growth and reproduction. The pattern of expression of the HNL in A. thaliana (AtHNL) in different tissues, as well as under abiotic stresses and hormone treatments, was examined by real-time quantitative reverse transcription PCR (qRT-PCR) and an AtHNL promoter-driven histochemical β-glucuronidase (GUS) assay. AtHNL is highly expressed in flowers and siliques, and the expression of AtHNL was dramatically affected by abiotic stresses and hormone treatments. The overexpression of AtHNL resulted in transgenic A. thaliana seedlings that were more tolerance to mannitol and salinity. Moreover, transgenic lines of A. thaliana that overexpressed this gene were less sensitive to abscisic acid (ABA). Altered expression of ABA/stress responsive genes was also observed in hnl mutant and AtHNL-overexpressing plants, suggesting AtHNL may play functional roles on regulating Arabidopsis resistance to ABA and abiotic stresses by affecting ABA/stress responsive gene expression. In addition, the overexpression of AtHNL resulted in earlier flowering, whereas the AtHNL mutant flowered later than the wild type (WT) plants. The expression of the floral stimulators CONSTANS (CO), SUPPRESSOR OF OVER EXPRESSION OF CO 1 (SOC1) and FLOWERING LOCUS T (FT) was upregulated in plants that overexpressed AtHNL when compared with the WT plants. In contrast, expression of the floral repressor FLOWERING LOCUS C (FLC) was upregulated in AtHNL mutants and downregulated in plants that overexpressed AtHNL compared to the WT plants. This study revealed that AtHNL can be induced under abiotic stresses and ABA treatment, and genetic analysis showed that AtHNL could also act as a positive regulator of abiotic stress and ABA tolerance, as well as flowering time.

A GDSL motif-containing lipase modulates Sclerotinia sclerotiorum resistance in Brassica napus.

Sclerotinia stem rot (SSR) caused by Sclerotinia sclerotiorum (Lib.) De Bary is a devastating disease infecting hundreds of plant species. It also restricts the yield, quality, and safe production of rapeseed (Brassica napus) worldwide. However, the lack of resistance sources and genes to S. sclerotiorum has greatly restricted rapeseed SSR-resistance breeding. In this study, a previously identified GDSL motif-containing lipase gene, B. napus GDSL LIPASE-LIKE 1 (BnaC07.GLIP1), encoding a protein localized to the intercellular space, was characterized as functioning in plant immunity to S. sclerotiorum. The BnaC07.GLIP1 promoter is S. sclerotiorum-inducible and the expression of BnaC07.GLIP1 is substantially enhanced after S. sclerotiorum infection. Arabidopsis (Arabidopsis thaliana) heterologously expressing and rapeseed lines overexpressing BnaC07.GLIP1 showed enhanced resistance to S. sclerotiorum, whereas RNAi suppression and CRISPR/Cas9 knockout B. napus lines were hyper-susceptible to S. sclerotiorum. Moreover, BnaC07.GLIP1 affected the lipid composition and induced the production of phospholipid molecules, such as phosphatidylethanolamine, phosphatidylcholine, and phosphatidic acid, which were correlated with decreased levels of reactive oxygen species (ROS) and enhanced expression of defense-related genes. A B. napus bZIP44 transcription factor specifically binds the CGTCA motif of the BnaC07.GLIP1 promoter to positively regulate its expression. BnbZIP44 responded to S. sclerotiorum infection, and its heterologous expression inhibited ROS accumulation, thereby enhancing S. sclerotiorum resistance in Arabidopsis. Thus, BnaC07.GLIP1 functions downstream of BnbZIP44 and is involved in S. sclerotiorum resistance by modulating the production of phospholipid molecules and ROS homeostasis in B. napus, providing insights into the potential roles and functional mechanisms of BnaC07.GLIP1 in plant immunity and for improving rapeseed SSR disease-resistance breeding.

A 3R-MYB transcription factor is involved in Methyl Jasmonate-Induced disease resistance in Agaricus bisporus and has implications for disease resistance in Arabidopsis

IntroductionMethyl jasmonate (MeJA) and MYB transcription factors (TFs) play important roles in pathogen resistance in several plants, but MYB TFs in conjunction with MeJA-induced defense against Pseudomonas tolaasii in edible mushrooms remain unknown. ObjectivesTo investigate the role of a novel 3R-MYB transcription factor (AbMYB11) in MeJA-induced disease resistance of Agaricus bisporus and in the resistance of transgenic Arabidopsis to P. tolaasii. MethodsMushrooms were treated with MeJA alone or in combination with phenylpropanoid pathway inhibitors, and the effects of the treatments on the disease-related and physiological indicators of the mushrooms were determined to assess the role of MeJA in inducing resistance and the importance of the phenylpropanoid pathway involved. Subcellular localization, gene expression analysis, dual-luciferase reporter assay, electrophoretic mobility shift assay, and transgenic Arabidopsis experiments were performed to elucidate the molecular mechanism of AbMYB11 in regulating disease resistance. ResultsMeJA application greatly improved mushroom resistance to P. tolaasii infection, and suppression of the phenylpropanoid pathway significantly weakened this effect. MeJA treatment stimulated the accumulation of phenylpropanoid metabolites, which was accompanied by increased the activities of biosynthetic enzymes and the expression of phenylpropanoid pathway-related genes (AbPAL1, Ab4CL1, AbC4H1) and an AbPR-like gene, further confirming the critical role of the phenylpropanoid pathway in MeJA-induced responses to P. tolaasii. Importantly, AbMYB11, localized in the nucleus, was rapidly induced by MeJA treatment under P. tolaasii infection; it transcriptionally activated the phenylpropanoid pathway-related and AbPR-like genes, and AbMYB11 overexpression in Arabidopsis significantly increased the transcription of phenylpropanoid-related genes, the accumulation of total phenolics and flavonoids, and improved resistance to P. tolaasii. ConclusionThis study clarified the pivotal role of AbMYB11 as a regulator in disease resistance by modulating the phenylpropanoid pathway, providing a novel idea for the breeding of highly disease-resistant edible mushrooms and plants.