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Hydrogen sulfide (H 2 S) is a crucial signaling molecule in plants. Recent studies have shown that H 2 S plays an equally important role as nitric oxide (NO) and hydrogen peroxide (H 2 O 2 ) in plant signaling. Previous studies have demonstrated the involvement of H 2 S in regulating drought and other stressful environmental conditions, but the exact downstream molecular mechanisms activated by the H 2 S signaling molecule remain unclear. In this study, we conducted a comprehensive genome-wide transcriptomic analysis of both wild type (WT) and double mutant ( lcd/des1 ). Arabidopsis thaliana plants were exposed to 40% polyethylene glycol (PEG) to induce drought stress and 20 µM sodium hydrosulfide (NaHS). The resulting transcriptome data were analyzed for differentially significant genes and their statistical enrichments in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The results indicated significant upregulation of genes related to photosynthesis, carbon fixation, plant secondary metabolite biosynthesis, inositol and phosphatidylinositol signaling pathways, and stress-responsive pathways in mutant plants under drought stress. Mutant plants with impaired H 2 S signaling mechanisms displayed greater susceptibility to drought stress compared to wild-type plants. In summary, all findings highlight the pivotal role of H 2 S signaling in stimulating other drought-responsive signaling pathways.

MYB transcription factor is one of the largest families in plants. There are more and more studies on plants responding to abiotic stress through MYB transcription factors, but the mechanism of some family members responding to salt stress is unclear. In this study, physiological and transcriptome techniques were used to analyze the effects of the R2R3-MYB transcription factor AtMYB72 on the growth and development, physiological function, and key gene response of Arabidopsis thaliana . Phenotypic observation showed that the damage of overexpression strain was more serious than that of Col-0 after salt treatment, while the mutant strain showed less salt injury symptoms. Under salt stress, the decrease of chlorophyll content, the degree of photoinhibition of photosystem II (PSII) and photosystem I (PSI) and the degree of oxidative damage of overexpressed lines were significantly higher than those of Col-0. Transcriptome data showed that the number of differentially expressed genes (DEGs) induced by salt stress in overexpressed lines was significantly higher than that in Col-0. GO enrichment analysis showed that the response of AtMYB72 to salt stress was mainly by affecting gene expression in cell wall ectoplast, photosystem I and photosystem II, and other biological processes related to photosynthesis. Compared with Col-0, the overexpression of AtMYB72 under salt stress further inhibited the synthesis of chlorophyll a (Chla) and down-regulated most of the genes related to photosynthesis, which made the photosynthetic system more sensitive to salt stress. AtMYB72 also caused the outbreak of reactive oxygen species and the accumulation of malondialdehyde under salt stress, which decreased the activity and gene expression of key enzymes in SOD, POD, and AsA-GSH cycle, thus destroying the ability of antioxidant system to maintain redox balance. AtMYB72 negatively regulates the accumulation of osmotic regulatory substances such as soluble sugar (SS) and soluble protein (SP) in A. thaliana leaves under salt stress, which enhances the sensitivity of Arabidopsis leaves to salt. To sum up, MYB72 negatively regulates the salt tolerance of A. thaliana by destroying the light energy capture, electron transport, and antioxidant capacity of Arabidopsis.

Copper (Cu) is an essential micronutrient for plant growth and development, but environmental Cu pollution has become increasingly severe, adversely affecting both ecosystems and crop productivity. In this study, we identified the AtNAC002 gene as a positive regulator of Cu toxicity in Arabidopsis thaliana. We found that AtNAC002 expression was induced by Cu excess, and the atnac002 mutant was Cu-sensitive, accumulating more Cu than the wild-type. Additionally, atnac002 mutants exhibit reduced activities of antioxidant enzymes (SOD, POD, and CAT), leading to increased levels of reactive oxygen species and malondialdehyde, which decrease Cu resistance. AtNAC002 might play a role in vacuolar and mitochondrial Cu compartmentalization by regulating genes involved in Cu detoxification, specifically COX11 and HCC1. Furthermore, AtNAC002 was implicated in flavone and flavanol biosynthesis, with the atnac002 mutant showing reduced flavonoid content. Our findings suggest that AtNAC002 is integral to the regulation of Cu toxicity tolerance in A. thaliana. This knowledge is critical for advancing our understanding and offers potential molecular breeding targets to enhance plant performance under Cu excess, which is significant for improving global food security and forest restoration., Competing Interests: Declaration of Competing Interest All authors disclosed no relevant relationships., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Although plastic mulch enhances crop yield, its removal and disposal present significant challenges, contributing to macro- and microplastic pollution in agricultural soils. The adverse effects of this pollution on soil and plant health are not fully understood but may stem from the plastic particles or the toxicity of leached chemical additives. This study assessed the impact of macro- and microplastics from nondegradable LDPE-based (LDPEb) and biodegradable PBAT-based (PBATb) mulch films, along with their leachates, on the germination of three plant species. After seven days of incubation, PBAT mulch leached compounds that significantly inhibited Arabidopsis germination, while cotton and tomato exhibited notable tolerance. Notably, PBATb mulch released a higher concentration of compounds, whereas LDPEb mulch exhibited a greater diversity of leached chemicals. Microplastic particles alone did not hinder seed germination, indicating that plastic toxicity primarily arises from the leachates. Many of these leached compounds lack global regulation and hazard information, underscoring the urgent need for further investigation into their environmental impacts and the development of appropriate regulatory frameworks to mitigate the potential toxicity of chemicals from conventional and biodegradable mulches., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Sugar beet (Beta vulgaris L.) shows potential as an energy crop for cadmium (Cd) phytoremediation. To elucidate its in vivo response strategy to Cd exposure, seedlings were treated with 1, 3, and 5 mmol/L CdCl 2 (Cd-1, Cd-3, and Cd-5) for 6 h, using 0 mmol/L CdCl 2 (Cd-0) as the control. The results showed that Cd-3 promoted a unique "hormesis" effect, leading to superior growth performance, increased levels of chlorophyll, soluble protein, and SOD activity, and reduced MDA content in sugar beet, compared to Cd-1, Cd-5, and even Cd-0. GO and KEGG enrichments and PPI networks of transcriptomic analysis revealed that the differentially expressed genes (DEGs) were primarily involved in lipid metabolism, cellular protein catabolism, and photosynthesis. Notably, the MAPK signaling pathway was significantly enriched only under Cd-3, with the up-regulation of ABA-related core gene BvPYL9 and an increase in ABA content after 6 h of Cd exposure. Furthermore, overexpression of BvPYL9 in Arabidopsis thaliana (OE-1 and OE-2) resulted in enhanced growth (fresh weight, dry weight, and root length), as well as higher ABA and soluble protein contents under different Cd treatments. Cd-induced transcriptional responses of BvPYL9 were also evident in OE-1 and OE-2, especially at 10 µmol/L, indicated by qRT-PCR. These findings suggest that ABA-mediated MAPK signaling pathway is activated in response to Cd toxicity, with BvPYL9 being a key factor in the cascade effects for the Cd-induced hormesis in sugar beet., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Nickel (Ni) is an important micronutrient, but excess Ni is toxic to many plant species. Currently, relatively little is known about the genetic basis of the plant responses to Ni toxicity. Here, we demonstrate that NAC32 transcription factor functions as a core genetic hub to regulate the Ni toxicity responses in Arabidopsis. NAC32 negatively regulates root-Ni concentration through the IREG2 (IRON REGULATED2) encoding a transporter. NAC32 also induces local auxin biosynthesis in the root-apex transition zone by upregulating YUCCA 7 (YUC7)/8/9 expression, which results in a local enhancement of auxin signaling in root tips, especially under Ni toxicity, thereby impaired primary root growth. By analyses of various combinations of nac32 and ireg2 mutants, as well as nac32 and yuc7/8/9 triple mutants, including high-order quadruple mutant, we demonstrated that NAC32 negatively regulates Ni stress tolerance by acting upstream of IREG2 and YUC7/8/9 to modulate their function in Ni toxicity responses. ChIPqPCR, EMSA (electrophoretic mobility shift assay) and transient dual-LUC reporter assays showed that NAC32 transcriptionally represses IREG2 expression but activates YUC7/8/9 expression by directly binding to their promoters. Our work demonstrates that NAC32 coordinates Ni compartmentation and developmental plasticity in roots, providing a conceptual framework for understanding Ni toxicity responses in plants., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Cadmium (Cd) pollution poses a significant ecological risk to mangrove ecosystems. Trehalose has excellent potential to mitigate the adverse effects of heavy metals. Unfortunately, the mechanisms related to trehalose-mediated heavy metal tolerance in plants remain elusive. In the present study, we firstly found that Cd induced the accumulation of trehalose and the differential expression of trehalose biosynthesis genes in the roots of mangrove plant Avicennia marina. Then, we found that the application of exogenous trehalose could alleviate the negative effects of Cd on A. marina by phenotypic observation. In addition, photosynthetic parameters and cellular ultrastructure analyses demonstrated that exogenous trehalose could improve the photosynthesis and stabilize the chloroplast and nuclear structure of the leaves of A. marina. Besides, exogenous trehalose could inhibit the Cd 2+ influx from the root to reduce the Cd 2+ content in A. marina. Subsequently, substrate sensitivity assay combined with ion uptake analysis using yeast cells showed that several trehalose biosynthesis genes may have a regulatory function for Cd 2+ transport. Finally, we further identified a positive regulatory factor, AmTPS6, which enhances the Cd tolerance in transgenic Arabidopsis thaliana. Taken together, these findings provide new understanding to the mechanism of Cd tolerance in mangrove A. marina at trehalose aspect and a theoretical basis for the conservation of mangroves in coastal wetlands., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Antimony (Sb) is a toxic pollutant, with Sb(V) being one of its main forms in the environment, but the process and mechanism of plant uptake of Sb(V) remain unclear. To investigate the process of Sb(V) uptake by plants, Arabidopsis thaliana plants were exposed to water culture media supplemented with different Sb(V) concentrations. The distribution, content, and forms of Sb(V) in Arabidopsis roots, and the accumulation and fixation of Sb(V) in Arabidopsis plants were studied. In addition, inhibitor experiments and analyses of gene expression changes were conducted to elucidate the underlying mechanism of its toxicity. Sb(V) entering the roots was mainly adsorbed on the cell wall, and the Sb(V) content in both apoplastic solution and symplastic solution increased with increasing external Sb(V) concentration. Sb(V) concentration in apoplast and symplast were approximately linearly correlated (R 2 =0.980), indicating low affinity of cells for Sb(V) absorption. Moreover, uncouplers significantly inhibited the entry of Sb(V) into the symplast, suggesting that the transmembrane transport of Sb(V) is energy-consuming. Sb(V) entering the cell could be partially reduced to Sb(III), and significant changes in glutathione metabolism gene expression were detected, indicating the important role of glutathione metabolism in the detoxification of Sb(V). From the perspective of the whole plant, although Sb(V) is absorbed by the roots, it is mainly fixed in the leaves and stems. This study revealed the pattern of Sb(V) uptake by plants and elucidated the mechanism of Sb(V) uptake by plants from the perspectives of kinetics, physiology, and genetics., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Peanut (Arachis hypogaea L.) is one of the most important oil crops worldwide. Cadmium (Cd), a heavy metal that is nonessential and toxic, has the potential to significantly impacted the quality and safety of peanut. Despite the known importance of Pleiotropic Drug Resistance (PDR) genes in heavy metal accumulation and transport in plants, there is a lack of comprehensive research on the systematic identification and functional characterization of AhPDRs in peanut. In this study, a total of 38 AhPDR genes were discovered within the peanut genome. Among these, AhPDR24, AhPDR30, and AhPDR33 displayed notable variations in expression levels in response to Cd stress. Particularly noteworthy was the observation that AhPDR33, localized in the plasma membrane, exhibited a significant increase in expression (approximately 3.8-fold) and heightened promoter activity (approximately 4.1-fold) following exposure to Cd (75 μM CdCl2). Furthermore, the study found that the overexpression of AhPDR33 in Arabidopsis resulted in increased root elongation and decreased Cd accumulation (approximately 0.42-fold) compared to wild-type plants. This suggests that AhPDR33 may have a beneficial role in facilitating Cd efflux and tolerance in plants. Additionally, transient silencing of AhPDR33 in peanut demonstrated its positive regulation of Cd tolerance through the promotion of reactive oxygen species (ROS) scavenging and membrane permeability reduction. These findings contribute to the understanding of the molecular mechanisms involved in AhPDR33-mediated Cd tolerance and detoxification in peanut. Furthermore, this study provides comprehensive information to understand the AhPDR gene family, its features, and its expression, which will hold a promising utility as an excellent candidate in the genetic improvement of peanut Cd stress tolerance., Competing Interests: Declaration of Competing Interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The use of nitrogen (N) fertilizers increases crop yield, but the accumulation of residual N in agricultural soils poses significant environmental risks. Improving the N use efficiency (NUE) of crops can help reduce N pollution. While nanomaterials have been shown to enhance crop agronomic traits, more research is needed to clarify the regulatory mechanisms involved. In this study, foliar spraying of carbon dots (CDs, 1 mg mL -1 ) derived from Salvia miltiorrhiza increased the activity of plasma membrane H + -ATPase in Arabidopsis thaliana roots, promoting the uptake, transport, and assimilation of NO 3 - and NH 4 + . The upregulation of N metabolism-related genes, such as AtAMTs and AtNRTs, was also observed in A. thaliana roots. Transcriptome analysis suggested that this regulatory effect is mediated by the shoot-to-root mobile polypeptide CEPD1 (C-terminally encoded peptide DOWNSTREAM 1) signaling pathway. Additionally, foliar application of CDs increased the NUE of sweetpotato (Ipomoea batatas (L.) Lam.) from 2.5% to 8.1%. The upregulation of genes such as CEPD1 in leaves was observed following CDs application under different N conditions. Finally, foliar spraying of CDs significantly increased field yield and enhanced tolerance to low N stress in sweetpotato. Overall, this study demonstrated that foliar application of CDs improved NUE in plants through CEPD1-dependent signaling., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Jian Liu reports financial support was provided by Natural Science Foundation of Shandong Province. Jian Liu reports financial support was provided by National Natural Science Foundation of China., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Pelargonic acid (PA) is a saturated fatty acid commonly found in several organisms, that is known for its phytotoxic effect and its use as bioherbicide for sustainable weed management. Although PA is already commercialised as bioherbicide, its molecular targets and mode of action is unknown according to the Herbicide Resistance Action Committee. Therefore, the aim of this work was focusing on the way this natural active substance impacts the plant metabolism of the model species Arabidopsis thaliana. PA caused increase of secondary and adventitious roots, as well as torsion, loss of gravitropism and phytotoxic effects. Moreover, PA altered the cellular arrangement and the PIN proteins activity. Computational simulations revealed that the intermolecular interactions between PA and the polar auxin transporter protein PIN1 are very similar to those established between the natural auxin IAA and PIN1. However, under intracellular conditions, the PA-PIN1 binding is more energetically stable than the IAA-PIN1. These results suggest that PA could act as an auxin-mimics bioherbicide. The exogenous application of PA would be responsible for the alterations observed both at structural and ultrastructural levels, which would be caused by the alteration on the transport of auxins into the plant, inducing root inhibition and ultimately total stop of root growth., Competing Interests: Declaration of Competing Interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Phloretin is a key secondary metabolite produced by apple trees. Known for its strong antioxidant properties, this dihydrochalcone has been extensively studied in animals but less so in plants. Recently, we identified phloretin as a phytotoxic allelochemical that inhibits growth in the model plant Arabidopsis by disrupting auxin metabolism and distribution in the roots. In this study, we found that phloretin significantly hinders the growth of Arabidopsis seedlings' aerial parts after a short-term treatment (10 days) and causes their decay after long-term exposure (28 days). These effects result from ultrastructural damage in the mesophyll cells of the leaves, including chloroplast displacement and swelling, lesions, and alterations in thylakoid and cell wall organization. Interestingly, phloretin-treated plants showed a decrease in malondialdehyde levels and antioxidant enzyme activities, while hydrogen peroxide and proline levels remained unchanged. This suggests that phloretin-induced chlorosis and seedling decay are not due to oxidative stress but rather to severe chloroplast structural damage, leading to inefficient photosynthesis, starch degradation, starvation, and activation of micro- and macroautophagic processes for self-preservation. Ultimately, these processes result in programmed cell death. These new insights into the phytotoxic effects of phloretin on Arabidopsis shoots could pave the way for future research into phloretin as a potential multitarget bioherbicide and enhance our understanding of autoallelopathy in apple trees., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

Plants can sense and respond to non-damaging mechanical stimulation such as touch, rain, or wind. Mechanical stimulation induces an increase of cytosolic calcium ([Ca 2+ ] cyt ), accumulation of phytohormones from the group of jasmonates (JAs) and activation of gene expression, which can be JAs-dependent or JAs-independent. Response to touch shares similar properties with reactions to stresses such as wounding or pathogen attack, and regular mechanical stimulation leads to changes in growth and development called thigmomorphogenesis. Previous studies showed that well-known seismonastic plants such as Venus flytrap (Dionaea muscipula) or sensitive plant (Mimosa pudica) lost their touch-induced motive responses during exposure to general volatile anaesthetic (GVA) diethyl ether. Here, we investigated the effect of diethyl ether anaesthesia on touch response in Arabidopsis thaliana. We monitored [Ca 2+ ] cyt level, accumulation of JAs and expression of touch-responsive genes. Our results showed that none of the investigated responses was affected by diethyl ether. However, diethyl ether alone increased [Ca 2+ ] cyt and modulated JAs-independent touch-responsive genes, thus partially activating touch response non-specifically. Together with our previous studies, we concluded that GVA diethyl ether cannot block the local rise of [Ca 2+ ] cyt but only its systemic propagation dependent on GLUTAMATE LIKE RECEPTOR 3s (GLR3s) channels., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

In vitro plant embryogenesis and microcallus formation are systems which are required for plant regeneration, a process during which cell reprogramming and proliferation are critical. These systems offer many advantages in breeding programmes, such as doubled-haploid production, clonal propagation of selected genotypes, and recovery of successfully gene-edited or transformed plants. However, the low proportion of reprogrammed cells in many plant species makes these processes highly inefficient. Here we report a new strategy to improve in vitro plant cell reprogramming using small molecule inhibitors of mammalian leucine rich repeat kinase 2 (LRRK2), which are used in pharmaceutical applications for cell reprogramming, but never used in plants before. LRRK2 inhibitors increased in vitro embryo production in three different systems and species, microspore embryogenesis of oilseed rape and barley, and somatic embryogenesis in cork oak. These inhibitors also promoted plant cell reprogramming and proliferation in Arabidopsis protoplast cultures. The benzothiazole derivative JZ1.24, a representative compound of the tested molecules, modified the expression of the brassinosteroid (BR)-related genes BIN2, CPD, and BAS1, correlating with an activation of BR signaling. Additionally, the LRRK2 inhibitor JZ1.24 induced the expression of the embryogenesis marker gene SERK1-like. The results suggest that the use of small molecules from the pharmaceutical field could be extended to promote in vitro reprogramming of plant cells towards embryogenesis or microcallus formation in a wider range of plant species and in vitro systems. This technological innovation would help to develop new strategies to improve the efficiency of in vitro plant regeneration, a major bottleneck in plant breeding., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Arsenic poses a global threat to living organisms, compromising crop security and yield. Limited understanding of the transcriptional network integrating arsenic-tolerance mechanisms with plant developmental responses hinders the development of strategies against this toxic metalloid. Here, we conducted a high-throughput yeast one-hybrid assay using as baits the promoter region from the arsenic-inducible genes ARQ1 and ASK18 from Arabidopsis thaliana, coupled with a transcriptomic analysis, to uncover novel transcriptional regulators of the arsenic response. We identified the GLABRA2 (GL2) transcription factor as a novel regulator of arsenic tolerance, revealing a wider regulatory role beyond its established function as a repressor of root hair formation. Furthermore, we found that ANTHOCYANINLESS2 (ANL2), a GL2 subfamily member, acts redundantly with this transcription factor in the regulation of arsenic signaling. Both transcription factors act as repressors of arsenic response. gl2 and anl2 mutants exhibit enhanced tolerance and reduced arsenic accumulation. Transcriptional analysis in the gl2 mutant unveils potential regulators of arsenic tolerance. These findings highlight GL2 and ANL2 as novel integrators of the arsenic response with developmental outcomes, offering insights for developing safer crops with reduced arsenic content and increased tolerance to this hazardous metalloid., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

Genotoxic stress activates the DNA-damage response (DDR) signalling cascades responsible for maintaining genome integrity. Downstream DNA repair pathways include the tyrosyl-DNA phosphodiesterase 1 (TDP1) enzyme that hydrolyses the phosphodiester bond between the tyrosine of topoisomerase I (TopI) and 3'-phosphate of DNA. The plant TDP1 subfamily contains the canonical TDP1α gene and the TDP1β gene whose functions are not fully elucidated. The current study proposes to investigate the involvement of TDP1 genes in DDR-related processes by using Arabidopsis thaliana mutants treated with genotoxic agents. The phenotypic and molecular characterization of tdp1α, tdp1β and tdp1α/β mutants treated with cisplatin (CIS), curcumin (CUR), NSC120686 (NSC), zeocin (ZEO), and camptothecin (CPT), evidenced that while tdp1β was highly sensitive to CIS and CPT, tdp1α was more sensitive to NSC. Gene expression analyses showing upregulation of the TDP2 gene in the double mutant indicate the presence of compensatory mechanisms. The downregulation of POL2A gene in the tdp1β mutant along with the upregulation of the TDP1β gene in pol2a mutants, together with its sensitivity to replication inhibitors (CIS, CTP), point towards a function of this gene in the response to replication stress. Therefore, this study brings novel information relative to the activity of TDP1 genes in plants., (© 2024 The Author(s). Plant, Cell & Environment published by John Wiley & Sons Ltd.)

Phosphorus (P) has crucial roles in plant growth and development. Hydrogen sulphide (H 2 S) has multiple functions in plants, particularly having the ability to promote tolerance to a variety of adversity stresses. However, it is unclear whether H 2 S has a function when plants suffer Pi-deficiency stress. DES1, encoding L-cysteine desulfhydrase1, is a crucial source of H 2 S in Arabidopsis thaliana by catalysing the substrate L-cysteine. Under phosphate starvation, the des1 mutant had a significantly shorter primary root length than the wild-type Col-0, and exogenous application of H 2 S donor NaHS could compensate for the root growth-sensitive phenotype. In contrast, the transgenic lines DES1ox overexpressing DES1 exhibited less sensitivity to phosphate starvation in terms of longer roots compared to the Col-0. These results demonstrate that H 2 S is involved in the regulation of Arabidopsis root growth under phosphate starvation. Moreover, using quantitative real-time polymerase chain reaction experiments to analyse the changes in genes induced by phosphate starvation in des1 mutant and Col-0, we screened to find that the expression of the Sulfoquinovosyl diacylglycerol 1 (SQD1) gene was significantly downregulated in the des1 mutant. Consistently, exogenous H 2 S significantly promoted SQD1 expression levels in roots of Col-0. Taken together, we demonstrate that DES1-mediated H 2 S participates in alleviating root growth inhibition by promoting the expression of SQD1 under Pi starvation., (© 2024 John Wiley & Sons Ltd.)

Background: The widespread evolution of pesticide resistance poses a significant challenge to current agriculture, necessitating the discovery of molecules with new modes of action. Despite extensive efforts, no major molecules with new modes of action have been commercialized for decades. Most pesticides function by binding to specific pockets on target enzymes, enabling a single target site mutation to confer resistance. An alternative approach is the disruption of protein-protein interactions (PPI), which require complementary mutations on both interacting partners for resistance to occur. Thus, our aim is the discovery and design of small-molecule inhibitors that target the interface of the PPI complex of O-acetylserine sulfhydrylase (OASS) and serine acetyltransferase (SAT), key obligatory interacting plant enzymes involved in the biosynthesis of the amino acid cysteine., Results: By employing in silico filtering techniques on a virtual library of 30 million small molecules, we identified initial hits capable of binding OASS and interfering with its interaction with a peptide derived from SAT with a half-maximal inhibitory concentration (IC 50 ) of 34 μm. Subsequently, we conducted molecular chemical optimizations, generating an early lead molecule (PJ4) with an IC 50  value of 4 μm. PJ4 successfully inhibited the germination of Arabidopsis thaliana seedlings and inhibited clover growth in a pre-emergence application at an effective concentration of 4.6 kg ha -1 ., Conclusion: These new compounds described herein can serve as promising leads for further optimization as herbicides with a new mode-of-action. This technology can be used for discovering new modes of action chemicals inhibiting all pest groups. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Salt stress has a detrimental impact on both plant growth and global crop yields. B-box proteins have emerged as pivotal players in plant growth and development regulation. Although the precise role of B-box proteins orchestrating salt stress responses in B. napus (Brassica napus) is not well understood in the current literature, further research and molecular explorations are required. Here, we isolated the B-box protein BnBBX22.A07 from B. napus. The overexpression of BnBBX22.A07 significantly improved the salt tolerance of Arabidopsis (Arabidopsis thaliana) and B. napus. Transcriptomic and histological analysis showed that BnBBX22.A07 enhanced the salt tolerance of B. napus by activating the expression of reactive oxygen species (ROS) scavenging-related genes and decreasing salt-induced superoxide anions and hydrogen peroxide. Moreover, BnBBX22.A07 interacted with BnHY5.C09, which specifically bound to and activated the promoter of BnWRKY33.C03. The presence of BnBBX22.A07 enhanced the activation of BnHY5.C09 on BnWRKY33.C03. Overexpression of BnHY5.C09 and BnWRKY33.C03 improved the salt tolerance of Arabidopsis. Functional analyses revealed that BnBBX22.A07-mediated salt tolerance was partly dependent on WRKY33. Taken together, we demonstrate that BnBBX22.A07 functions positively in salt responses not only by activating ROS scavenging-related genes but also by indirectly activating BnWRKY33.C03. Notably, our study offers a promising avenue for the identification of candidate genes that could be harnessed in breeding endeavours to develop salt-resistant transgenic crops., (© 2024 John Wiley & Sons Ltd.)

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., Competing Interests: Declarations. Conflict of interest: The authors declare no conflict of interest., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Key Message: Constitutive expression of cucumber CsACS2 in Arabidopsis disrupts anther dehiscence and male fertility via ethylene signaling and DNA methylation, revealing new avenues for enhancing crop reproductive traits. The cucumber gene CsACS2, encoding ACC (1-aminocyclopropane-1-carboxylic acid) synthase, plays a pivotal role in ethylene biosynthesis and sex determination. This study investigates the effects of constitutive CsACS2 expression in Arabidopsis thaliana on anther development and male fertility. Transgenic Arabidopsis plants overexpressing CsACS2 exhibited male sterility due to inhibited anther dehiscence, which was linked to suppressed secondary cell wall thickening. RNA-Seq analysis revealed upregulation of ethylene signaling pathway genes and downregulation of secondary cell wall biosynthesis genes, with gene set enrichment analysis indicating the involvement of DNA methylation. Rescue experiments demonstrated that silver nitrate (AgNO₃) effectively restored fertility, while 5-azacytidine (5-az) partially restored it, highlighting the roles of ethylene signaling and DNA methylation in this process. Constitutive CsACS2 expression in Arabidopsis disrupts anther development through ethylene signaling and DNA methylation pathways, providing new insights into the role of ethylene in plant reproductive development and potential applications in crop improvement., Competing Interests: Declarations. Conflict of interests: The authors state that they have no known financial conflicts of interest or personal relationships that may have influenced the research presented in this paper., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The role of molecular hydrogen (H 2 ) in autophagy during inflammatory response is controversial in mammalian cells. Although the stimulation of H 2 production in response to osmotic stress was observed in plants, its synthetic pathway and the interrelationship between its induction and plant autophagy remain unclear. Here, the induction of autophagy was observed in Arabidopsis upon osmotic stress, assessing by the autophagosome formation and autophagy-related genes expression. Above responses were intensified by H 2 fumigation. Meanwhile, the reduction in seedling growth and roots vigor was obviously abolished, accompanied by reestablishing redox balance. These H 2 responses were markedly impaired in T-DNA knockout lines atg2, atg5, and atg18. Further evidence showed that the increased endogenous H 2 synthesis by genetic manipulation, not only stimulated autophagosome formation, but also triggered various plant responses toward osmotic stress. By contrast, these responses were obviously abolished by the disruption of endogenous H 2 synthesis with the addition of 2,6-dichloroindophenol sodium salt. Together, the integrated genetic and molecular evidence clearly illustrated the requirement of autophagy activation in H 2 control of plant osmotic tolerance., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Cadmium (Cd) represents a hazardous heavy metal, prevalent in agricultural soil due to industrial and agricultural expansion. Its propensity for being absorbed by edible plants, even at minimal concentrations, and subsequently transferred along the food chain poses significant risks to human health. Accordingly, it is imperative to investigate novel genes and mechanisms that govern Cd tolerance and detoxification in plants. Here, we discovered that the transcription factor MYC2 directly binds to the promoters of HMA2 and HMA4 to repress their expression, thereby altering the distribution of Cd in plant tissues and negatively regulating Cd stress tolerance. Additionally, molecular, biochemical, and genetic analyses revealed that MYC2 interacts and cooperates with MYB43 to negatively regulate the expression of HMA2 and HMA4 and Cd stress tolerance. Notably, under Cd stress conditions, MYC2 undergoes degradation, thereby alleviating its inhibitory effect on HMA2 and HMA4 expression and plant tolerance to Cd stress. Thus, our study highlights the dynamic regulatory role of MYC2, in concert with MYB43, in regulating the expression of HMA2 and HMA4 under both normal and Cd stress conditions. These findings present MYC2 as a promising target for directed breeding efforts aimed at mitigating Cd accumulation in edible plant roots., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Heavy metals, including Hg 2+ , Cr 6+ and Cd 2+ , have always been a major issue in environmental pollution, leading to abnormal changes in the levels of biologically active molecules including Cys in plants, seriously affecting all aspects of the growth and development of plants. This makes it essential to develop a simple and practical method to study the potential impact of heavy metals on plants. In this paper, our research group has developed near-infrared fluorescent probe WRM-S, which has the advantages of fast response, sensitivity to Cys, and successfully applying it to cells and zebrafish. Moreover, it combined the close relationship between heavy metal stress on plants and Cys, using Cys as the detection target, monitoring the internal environment changes of two plants under Hg 2+ , Cr 6+ , and Cd 2+ stress in the environment, and then conducting 3D imaging. The results indicated that the probe has strong penetration ability in plant tissues, and revealed abnormal changes in plant Cys levels caused by heavy metal stress-induced cellular oxidative stress or cytotoxicity. Thus, the in-situ imaging detection of this probe provides a direction for the physiological dynamics research of plant environmental stress., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

S-nitrosoglutathione reductase (GSNOR) is involved in the response to cadmium (Cd) exposure. In this study, the plants of mutant (gsnor1-3) with lossing-function of- and over-expression (GSNOR OE 5) of GSNOR were used to clear the role of GSNOR in Cd tolerance. GSNOR activity increased through upregulating the expression of the AtGSNOR gene and protein in Arabidopsis thaliana under Cd stress, which attenuated Cd tolerance. Oxidative damage was more serious in GSNOR OE 5 and was alleviated in gsnor1-3 under Cd stress, compared with Col-0. Induction of GSNOR facilitated H 2 O 2 accumulation but inhibited catalase (CAT) activity in shoots under Cd stress. This phenotype was eliminated by 3-amino-1,2,4-triazole (3-AT), a CAT inhibitor. In addition, the expressions of AtCAT1 and AtCAT2 were down-regulated with increasing GSNOR activity under Cd stress. This suggested that GSNOR was involved in the accumulation of hydrogen peroxide (H 2 O 2 ) through regulating CAT expression and activity under Cd exposure. Furthermore, Cd tolerance and CAT activity were improved by spraying S-nitrosoglutathione (GSNO) onto the surface of the leaves. The in vitro activity of CAT increased with GSNO concentration until a GSNO/CAT ratio of 2 was reached. Thus, CAT activity was relative to GSNOR through regulating the expression and S-nitrosylation level of proteins. In summary, the Cd-induced promotion of GSNOR activity aggravated Cd toxicity in plants by mediating H 2 O 2 accumulation controlled by CAT., (© 2024. The Author(s).)

Seashore paspalum (Paspalum vaginatum) is a warm-season and perennial turfgrass and is known for its cadmium (Cd)-stress tolerance. Here, a Phosphoproteomics analysis was performed to examine the key proteins relating to Cd tolerance in seashore paspalum. Fructose 1,6-biphosphate aldolase, PvFBA1, was identified for its phosphorylated state after exposure to Cd stress. Specifically, the phosphorylation of PvFBA1 was enhanced in several metabolic pathways, including pentose phosphate pathway (PPP), carbon fixation and biosynthesis of amino acids under Cd stress. By transforming PvFBA1 into Arabidopsis, the PvFBA1-OE plants exhibited longer roots, greater FBA activity and higher soluble sugar content than WT under 100 µM CdCl 2 treatment. By expressing the PvFBA1 in yeast, a serine 50 phosphorylation site was identified as functional site. By microscale thermophoresis experiment, we indicted that PvFBA1can bind Cd directly enhancing its phosphorylation level to alleviate the damage of Cd. This finding may provide new insights into the molecular mechanisms of plants Cd tolerance., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2024. Published by Elsevier Inc.)

Echinochloa phyllopogon is a noxious weed that can harm rice over prolonged periods. Recently, a penoxsulam-resistant variant of E. phyllopogon with a mutation in the acetolactate synthase (ALS) gene was collected in Northeastern China. In the present study, the molecular mechanism underlying herbicide resistance in mutant populations was evaluated. The GR 50 and IC 50 values of the herbicide-resistant mutant 1-11 were 27.0- and 21.4-fold higher than those of the susceptible population 2-31, respectively. In addition, pre-application of malathion reduced the GR 50 value of the resistant population. Additionally, mutant populations developed cross-resistance to other ALS inhibitors. E. phyllopogon ALS sequencing showed a Trp-574-Leu mutation in ALS2 variant 1-11. Molecular docking showed that the Trp-574-Leu substitution reduced the number of hydrogen bonds and altered the interaction between penoxsulam and ALS2. Transgenic Arabidopsis plants harboring the ALS2 mutant gene also showed resistance to penoxsulam and other ALS inhibitors. Overall, our study demonstrated that the Trp-574-Leu mutation and P450-mediated metabolic resistance lead to the cross-resistance of E. phyllopogon to ALS inhibitors., Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Cytokinin is central to coordinating plant adaptation to environmental stresses. Here, we first demonstrated the involvement of cytokinin in Arabidopsis responses to arsenite [As(III)] stress. As(III) treatment reduced cytokinin contents, while cytokinin treatment repressed further primary root growth in Arabidopsis plants under As(III) stress. Subsequently, we revealed that the cytokinin signaling members ARR1 and ARR12, the type-B ARABIDOPSIS RESPONSE REGULATORs, participate in cytokinin signaling-mediated As(III) responses in plants as negative regulators. A comprehensive transcriptome analysis of the arr1 and arr12 single and arr1,12 double mutants was then performed to decipher the cytokinin signaling-mediated mechanisms underlying plant As(III) stress adaptation. Results revealed important roles for ARR1 and ARR12 in ion transport, nutrient responses, and secondary metabolite accumulation. Furthermore, using hierarchical clustering and regulatory network analyses, we identified two NODULIN 26-LIKE INTRINSIC PROTEIN (NIP)-encoding genes, NIP1;1 and NIP6;1, potentially involved in ARR1/12-mediated As(III) uptake and transport in Arabidopsis. By analyzing various combinations of arr and nip mutants, including high-order triple and quadruple mutants, we demonstrated that ARR1 and ARR12 redundantly function as negative regulators of As(III) tolerance by acting upstream of NIP1;1 and NIP6;1 to modulate their function in arsenic accumulation. ChIP-qPCR, EMSA, and transient dual-LUC reporter assays revealed that ARR1 and ARR12 transcriptionally activate the expression of NIP1;1 and NIP6;1 by directly binding to their promoters and upregulating their expression, leading to increased arsenic accumulation under As(III) stress. These findings collectively provide insights into cytokinin signaling-mediated plant adaptation to excessive As(III), contributing to the development of crops with low arsenic accumulation., (© 2024 Society for Experimental Biology and John Wiley & Sons Ltd.)

Copper (Cu) and zinc (Zn) are both oxidation-reducing metal elements that are necessary for plant growth, and their effect often depends on their concentration. However, there are few studies that have investigated how plants are stressed and affected when the two ions are present simultaneously, especially when one ion is beneficial due to a low concentration and the other is detrimental due to a high concentration. To address this question, we treated Arabidopsis plants with either high or/and low concentrations of the two ions and investigated their mutual effects and the underlying molecular mechanism, focusing on photosynthetic function. The results showed that the photosynthetic pigment content and the performance of photosynthetic systems were most affected when both metal ions were present at detrimental concentrations (60 μM Cu for Cu 60 and 350 μM Zn for Zn 350 ). These include the effective openness of the photoreaction center, the electron transport rate and efficiency of photosystem II (PSII), the NPQ-dependent energy dissipation and the activity of photosystem I (PSI). However, when the harmful concentration of one of the two metals is combined with the beneficial concentration of the other metal (Cu 5 +Zn 350 or Zn 50 +Cu 60 ), these photosynthetic indicators are compensated to different degrees but the negative effects of copper ions at high dose are more difficult to eliminate than zinc ions. These results were also confirmed by gene expression analysis, which provides a clue to understanding the interaction between heavy metal ions, reducing metal toxicity and improving the tolerance of plants to heavy metals in practice., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Salvia miltiorrhiza Bunge, a model plant for medicinal research, is extensively utilized for its dried roots and rhizomes for treatment of various diseases. Soil salinization hinders the large-scale cultivation and industrial production of S. miltiorrhiza by affecting its active compounds. Methyl jasmonate (MeJA) is a crucial plant hormone that regulates plant responses under salt stress. Jasmonate zim domain (JAZ) proteins function as transcriptional repressors in jasmonic acid (JA) signaling pathways. This study explores the interaction between JA and salt stress by using transgenic Arabidopsis thaliana to elucidate the roles of SmJAZ3, SmJAZ4, and SmJAZ8. We found that 2.5 μM MeJA reduced the inhibitory effect of 150 mM NaCl on wild-type seed germination, and this effect was reversed by 15 μM dihydroxyindole-2-carboxylic acid (DIECA). Similar results were observed in transgenic A. thaliana lines overexpressing SmJAZ3/4/8. Inclusion of SmJAZ3/4 enhanced salt resistance by increasing antioxidant enzyme activity, chlorophyll content, proline content, and Na + /K + content, while SmJAZ8 had the opposite effect. These findings suggest that appropriate concentrations of MeJA can alleviate the negative effect of salt stress on plant growth and development. Investigating the salt tolerance of SmJAZ3/4/8 is significant for cultivating high-quality salt-tolerant S. miltiorrhiza., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Salvia miltiorrhiza holds significant importance in traditional Chinese medicine. Stress-associated proteins (SAP), identified by A20/AN1 zinc finger structural domains, play crucial roles in regulating plant growth, development, resistance to biotic and abiotic stress, and hormone responses. Herein, we conducted a genome-wide identification of the SAP gene family in S. miltiorrhiza. The expression analysis revealed a significant upregulation of SmSAP4 under methyl jasmonate (MeJA) and salt stress. Overexpressing SmSAP4 in S. miltiorrhiza hairy roots increased tanshinones content while decreasing salvianolic acids content, while RNAi-silencing SmSAP4 had the opposite effect. SmSAP4 overexpression in both Arabidopsis thaliana and S. miltiorrhiza hairy roots decreased their salt stress tolerance, accompanied by increased activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), and a hindered ability to maintain the Na +  : K + ratio. Further investigations demonstrated that MeJA alleviated the inhibitory effect of SmJAZ3 on SmSAP4 activation by SmbHLH37 and SmERF73. However, MeJA did not affect the inhibition of SmSAP4 activation by SmJAZ8 through SmbHLH37. In summary, our research reveals that SmSAP4 negatively regulates the accumulation of salvianic acid through the SmJAZs-SmbHLH37/SmERF73-SmSAP4 module and positively impacting the accumulation of tanshinones. Additionally, it functions as a negative regulator under salt stress., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

Cadmium (Cd) is a significant heavy metal contaminant within the environment, carrying a notable level of toxicity that presents a substantial hazard to both plant and human. Carrot (Daucus carota), a significant root vegetable crop globally, have evolved multiple transcriptional regulatory mechanisms to cope with Cd stress, with a crucial involvement of the myeloblastosis (MYB) transcription factor. In this study, the DcMYB62 gene encoding 288 amino acids, localized in the nucleus and demonstrated transcription activation property, was isolated from carrot (cv. 'Kuroda'). There was a positive relationship observed between the levels of DcMYB62 expression and the accumulation patterns of carotenoids in two distinct carrot cultivars. Further investigation revealed that the expression of DcMYB62 improved Cd tolerance of Arabidopsis by increasing seed germination rate, root length, and overall survival rate. The levels of carotenoids in DcMYB62 transgenic Arabidopsis surpassed those in wild type, accompanied by elevated expression levels of 15-cis-phytoene desaturase, zeta-carotene desaturase, and carotenoid isomerase. Meanwhile, the heterologous expression of DcMYB62 promoted the biosynthesis of abscisic acid (ABA) and hydrogen sulfide (H 2 S), which in turn suppressed the formation of hydrogen peroxide and superoxide anion, while also stimulating stomatal closure. Furthermore, the heterologous expression of DcMYB62 increased the transcription of genes associated with heavy metal resistance in Arabidopsis, notably nicotianamine synthase. Overall, this study contributes to understanding how DcMYB62 promote Cd stress resistance of plants by regulating the biosynthesis pathways of carotenoids, ABA, and H 2 S, which offers valuable insights into the regulatory mechanism connecting DcMYBs with Cd stress response of carrot., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

The phytohormone abscisic acid (ABA) is an important regulator of plant growth, but its potential participation in the process of in vitro shoot regeneration has not to date been reported. Here, we found that ABA appeared to inhibit in vitro shoot regeneration. ABA represses the formation of stem cell niches, thereby reducing the shoot regeneration by localizing the expression of WUSCHEL (WUS). During in vitro shoot regeneration, enrichment of H3K9ac in the specific region of WUS is a necessary event for its activation which could be inhibited by exogenous ABA. These findings reveal the potential function, as well as the possible way of ABA in regulating de novo shoot regeneration in Arabidopsis., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Background: Developing herbicide-resistant (HR) crop cultivars is an efficient way to control weeds and minimize crop yield losses. However, widespread and long-term herbicide application has led to the evolution of resistant weeds. Here, we established a resistant (R) E. indica population, collected from imidazolinone-resistant rice cultivar fields., Results: The R population evolved 4.5-fold resistance to imazamox. Acetolactate synthase (ALS) gene sequencing and ALS activity assays excluded the effect of target-site resistance in this population. P450 inhibitor malathion pretreatment significantly reversed resistance to imazamox. RNA sequencing showed that a P450 gene CYP81A104 was expressed higher in R versus susceptible (S) plants. Arabidopsis overexpressing CYP81A104 showed resistance to ALS inhibitors (imazamox, tribenuron-methyl, penoxsulam and flucarbazone-sodium), PSII inhibitor (bentazone), hydroxyphenyl pyruvate dioxygenase inhibitor (mesotrione) and auxin mimics (MCPA), which was generally consistent with the results presented in the R population., Conclusion: This study confirmed that the CYP81A104 gene endowed resistance to multiherbicides with different modes-of-action. Our findings provide an insight into the molecular characteristics of resistance and contribute to formulating an appropriate strategy for weed management in HR crops. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

NHX5 and NHX6, Arabidopsis endosomal antiporters, play a vital role in facilitating ion and pH homeostasis in endosomal compartments. Studies have found that NHX5 and NHX6 are essential for protein trafficking, auxin homeostasis, and plant growth and development. Here, we report the role of NHX5 and NHX6 in brassinosteroid (BR) signaling. We found that hypocotyl growth was enhanced in nhx5 nhx6 under epibrassinolide (eBR) treatment. nhx5 nhx6 bri1 was insensitive to eBR treatment, indicating that NHX5 and NHX6 are downstream of the BRI1 receptor in BR signaling. Moreover, confocal observation with both hypocotyls and root tips showed that BRI1-YFP localization in the plasma membrane (PM) was reduced in nhx5 nhx6. Interestingly, brefeldin A (BFA) treatment showed that formation of the BFA bodies containing BRI1 and their disassembling were disrupted in nhx5 nhx6. Further genetic analysis showed that NHX5/NHX6 and SYP22 may act coordinately in BR signaling. NHX5 and NHX6 may regulate SYP22 function by modulating cellular K + and pH homeostasis. Importantly, NHX5 and NHX6 colocalize and interact with SYP22, but do not interact with BRI1. In summary, our findings indicate that NHX5/NHX6/SYP22 complex is essential for the regulation of BRI1 recycling and PM localization. The H + -leak facilitated by NHX5 and NHX6 offers a means of controlling BR signaling in plants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

Isoxaben is a pre-emergent herbicide used to control broadleaf weeds. While the phytotoxic mechanism is not completely understood, isoxaben interferes with cellulose synthesis. Certain mutations in cellulose synthase complex proteins can confer isoxaben tolerance; however, these mutations can cause compromised cellulose synthesis and perturbed plant growth, rendering them unsuitable as herbicide tolerance traits. We conducted a genetic screen to identify new genes associated with isoxaben tolerance by screening a selection of Arabidopsis thaliana T-DNA mutants. We found that mutations in a FERREDOXIN-NADP(+) OXIDOREDUCTASE-LIKE (FNRL) gene enhanced tolerance to isoxaben, exhibited as a reduction in primary root stunting, reactive oxygen species accumulation and ectopic lignification. The fnrl mutant did not exhibit a reduction in cellulose levels following exposure to isoxaben, indicating that FNRL operates upstream of isoxaben-induced cellulose inhibition. In line with these results, transcriptomic analysis revealed a highly reduced response to isoxaben treatment in fnrl mutant roots. The fnrl mutants displayed constitutively induced mitochondrial retrograde signalling, and the observed isoxaben tolerance is partially dependent on the transcription factor ANAC017, a key regulator of mitochondrial retrograde signalling. Moreover, FNRL is highly conserved across all plant lineages, implying conservation of its function. Notably, fnrl mutants did not show a growth penalty in shoots, making FNRL a promising target for biotechnological applications in breeding isoxaben tolerance in crops., (© 2024 The Author(s). Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.)

Anchorene, identified as an endogenous bioactive carotenoid-derived dialdehyde and diapocarotenoid, affects root development by modulating auxin homeostasis. However, the precise interaction between anchorene and auxin, as well as the mechanisms by which anchorene modulates auxin levels, remain largely elusive. In this study, we conducted a comparative analysis of anchorene's bioactivities alongside auxin and observed that anchorene induces multifaceted auxin-like effects. Through genetic and pharmacological examinations, we revealed that anchorene's auxin-like activities depend on the indole-3-pyruvate-dependent auxin biosynthesis pathway, as well as the auxin inactivation pathway mediated by Group II Gretchen Hagen 3 (GH3) proteins that mainly facilitate the conjugation of indole-3-acetic acid (IAA) to amino acids, leading to the formation of inactivated storage forms. Our measurements indicated that anchorene treatment elevates IAA levels while reducing the quantities of inactivated IAA-amino acid conjugates and oxIAA. RNA sequencing further revealed that anchorene triggers the expression of numerous auxin-responsive genes in a manner reliant on Group II GH3s. Additionally, our in vitro enzymatic assays and biolayer interferometry (BLI) assay demonstrated anchorene's robust suppression of GH3.17-mediated IAA conjugation with glutamate. Collectively, our findings highlight the significant role of carotenoid-derived metabolite anchorene in modulating auxin homeostasis, primarily through the repression of GH3-mediated IAA conjugation and inactivation pathways, offering novel insights into the regulatory mechanisms of plant bioactive apocarotenoids., (© 2024 The Author(s). Journal of Integrative Plant Biology published by John Wiley & Sons Australia, Ltd on behalf of Institute of Botany, Chinese Academy of Sciences.)

Aluminum-dependent stoppage of root growth requires the DNA damage response (DDR) pathway including the p53-like transcription factor SUPPRESSOR OF GAMMA RADIATION 1 (SOG1), which promotes terminal differentiation of the root tip in response to Al dependent cell death. Transcriptomic analyses identified Al-induced SOG1-regulated targets as candidate mediators of this growth arrest. Analysis of these factors either as loss-of-function mutants or by overexpression in the als3-1 background shows ERF115, which is a key transcription factor that in other scenarios is rate-limiting for damaged stem cell replenishment, instead participates in transition from an actively growing root to one that has terminally differentiated in response to Al toxicity. This is supported by a loss-of-function erf115 mutant raising the threshold of Al required to promote terminal differentiation of Al hypersensitive als3-1. Consistent with its key role in stoppage of root growth, a putative ERF115 barley ortholog is also upregulated following Al exposure, suggesting a conserved role for this ATR-dependent pathway in Al response. In contrast to other DNA damage agents, these results show that ERF115 and likely related family members are important determinants of terminal differentiation of the root tip following Al exposure and central outputs of the SOG1-mediated pathway in Al response., (© 2024 The Author(s). Plant, Cell & Environment published by John Wiley & Sons Ltd.)

Hydrogen peroxide (H 2 O 2 ) is naturally produced by plant cells during normal development and serves as a messenger that regulates cell metabolism. Despite its importance, the relationship between hydrogen peroxide and the target of rapamycin (TOR) pathway, as well as its impact on cell division, has been poorly analyzed. In this study, we explore the interaction of H 2 O 2 with TOR, a serine/threonine protein kinase that plays a central role in controlling cell growth, size, and metabolism in Arabidopsis thaliana. By applying two concentrations of H 2 O 2 exogenously (0.5 and 1 mM), we could correlate developmental traits, such as primary root growth, lateral root formation, and fresh weight, with the expression of the cell cycle gene CYCB1;1, as well as TOR expression. When assessing the expression of the ribosome biogenesis-related gene RPS27B, an increase of 94.34% was noted following exposure to 1 mM H 2 O 2 treatment. This increase was suppressed by the TOR inhibitor torin 2. The elimination of H 2 O 2 accumulation with ascorbic acid (AA) resulted in decreased cell division as well as TOR expression. The potential molecular mechanisms associated with the effects of H 2 O 2 on the cell cycle and TOR expression in roots are discussed in the context of the results., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Plant pathogenic fungi pose a substantial challenge to agricultural production, but the conventional fungicide-based approaches are losing importance. As agents with broad-spectrum antibacterial effects, gold nanoparticles (Au NPs) are found to have antifungal effects; however, no study has examined their application in agriculture as fungicides. Accordingly, this study investigates the activity of 2-mercaptoimidazole-capped Au NPs (MI-Au NPs) against the 'top' plant pathogenic fungi, finding that they could inhibit Magnaporthe oryzae , Botrytis cinerea , Fusarium pseudograminearum and Colletotrichum destructivum by inducing cytoplasmic leakage. Moreover, MI-Au NPs are found to protect plants from infection by B. cinerea . Specifically, pot experiments demonstrate that MI-Au NPs decrease the incidence rate of B. cinerea infection in Arabidopsis thaliana from 74.6% to 6.2% and in Solanum lycopersicum from 100% to 10.9%, outperforming those achieved by imazalil. Furthermore, the biosafety assays reveal that MI-Au NPs cannot penetrate the cuticle of plant cells or negatively influence plant growth, and it is safe to mammalian cells. In summary, the findings of this study will support the development of NP-based antifungal agents for use in agriculture.

Background: Drought stress markedly constrains plant growth and diminishes crop productivity. Strigolactones (SLs) exert a beneficial influence on plant resilience to drought conditions. Nevertheless, the specific function of SLs in modulating cotton's response to drought stress remains to be elucidated., Results: In this study, we assess the impact of exogenous SL (rac-GR24) administration at various concentrations (0, 1, 5, 10, 20 µM) on cotton growth during drought stress. The findings reveal that cotton seedlings treated with 5 µM exogenous SL exhibit optimal mitigation of growth suppression induced by drought stress. Treatment with 5 µM exogenous SL under drought stress conditions enhances drought tolerance in cotton seedlings by augmenting photosynthetic efficiency, facilitating stomatal closure, diminishing reactive oxygen species (ROS) generation, alleviating membrane lipid peroxidation, enhancing the activity of antioxidant enzymes, elevating the levels of osmoregulatory compounds, and upregulating the expression of drought-responsive genes. The suppression of cotton SL biosynthesis genes, MORE AXILLARY GROWTH 3 (GhMAX3) and GhMAX4b, impairs the drought tolerance of cotton. Conversely, overexpression of GhMAX3 and GhMAX4b in respective Arabidopsis mutants ameliorates the drought-sensitive phenotype in these mutants., Conclusion: These observations underscore that SLs significantly bolster cotton's resistance to drought stress., (© 2024. The Author(s).)

Selective perturbation of protein interactions with chemical compounds enables dissection and control of developmental processes. Differentiation of stomata, cellular valves vital for plant growth and survival, is specified by the basic-helix-loop-helix (bHLH) heterodimers. Harnessing a new amination reaction, we here report a synthesis, derivatization, target identification, and mode of action of an atypical doubly-sulfonylated imidazolone, Stomidazolone, which triggers stomatal stem cell arrest. Our forward chemical genetics followed by biophysical analyses elucidates that Stomidazolone directly binds to the C-terminal ACT-Like (ACTL) domain of MUTE, a master regulator of stomatal differentiation, and perturbs its heterodimerization with a partner bHLH, SCREAM in vitro and in plant cells. On the other hand, Stomidazolone analogs that are biologically inactive do not bind to MUTE or disrupt the SCREAM-MUTE heterodimers. Guided by structural docking modeling, we rationally design MUTE with reduced Stomidazolone binding. These engineered MUTE proteins are fully functional and confer Stomidazolone resistance in vivo. Our study identifies doubly-sulfonylated imidazolone as a direct inhibitor of the stomatal master regulator, further expanding the chemical space for perturbing bHLH-ACTL proteins to manipulate plant development., (© 2024. The Author(s).)

Main Conclusion: AtPLC1 plays a critical role in plant growth, development, and response to drought stress. Phosphoinositide-specific phospholipase C (PI-PLC) hydrolyzes substrates to generate secondary messengers crucial for plant growth, development, and stress responses. Drought escape (DE) response is an adaptive strategy that plants employ under drought conditions. The expression levels of the flower meristem-specific gene APETALA 1 and flowering regulatory genes FLOWERING LOCUS T and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 were downregulated in plc1, and FLOWERING LOCUS C was upregulated. The flowering time of the plc1flc double mutant was earlier than that of the wild type. Transcriptome analysis revealed that the Gene Ontology of differentially expressed genes (DEGs) was enriched in abscisic acid (ABA) response signaling, and Kyoto Encyclopedia of Genes and Genomes analysis revealed differential gene expression annotated to plant hormone signaling pathways. Our experiments show that AtPLC1 is upregulated by ABA in Arabidopsis. Under ABA induction and water stress, wild-type plants exhibit a DE response, and the DE response in plc1 disappears. Expression levels of ABA signaling pathway transcription factors ABA-responsive element-binding factors 3 (ABF3) and ABF4 were downregulated in plc1. In conclusion, our study suggests that AtPLC1 participates in regulating plant growth and development and participates in the DE response through the regulation of ABA signaling pathway transcription factors ABF3/ABF4. The study enhances our comprehension of the role of AtPLC1 in plant development and drought stress, providing a theoretical foundation for further investigation into DE responses., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Methylglyoxal (MG), a highly reactive and cytotoxic α-oxoaldehyde compound, can over-accumulate under abiotic stress, consequently injuring plants or even causing death. Glyoxalase I (GLYI), the first enzyme of the glyoxalase pathway, plays multiple roles in the detoxification of MG and in abiotic stress responses. However, the GLY1 gene in maize has been little studied in response to abiotic stress. In this study, we screened a glyoxalase I gene ( ZmGLYI-8 ) and overexpressed in Arabidopsis . This gene was localized in the cytoplasm and can be induced in maize seedlings under multiple stress treatments, including salt, drought, MG, ABA, H 2 O 2 and high temperature stress. Phenotypic analysis revealed that after MG, salt and drought stress treatments, overexpression of ZmGLYI-8 increased the tolerance of transgenic Arabidopsis to MG, salt and drought stress. Furthermore, we demonstrated that the overexpression of ZmGLYI-8 scavenges accumulated reactive oxygen species, detoxifies MG and enhances the activity of antioxidant enzymes to improve the resistance of transgenic Arabidopsis plants to salt and drought stress. In summary, this study preliminarily elucidates the molecular mechanism of the maize ZmGLYI-8 gene in transgenic Arabidopsis and provides new insight into the breeding of salt- and drought-tolerant maize varieties.

Key Message: Hydrogen peroxide promoted leaf senescence by sulfenylating the magnesium chelating protease I subunit (CHLI1) in the chlorophyll synthesis pathway, and inhibited its activity to reduce chlorophyll synthesis. Leaf senescence is the final and crucial stage of plant growth and development, during which chlorophyll experiences varying degrees of destruction. It is well-known that the higher ROS accumulation is a key factor for leaf senescence, but whether and how ROS regulates chlorophyll synthesis in the process are unknown. Here, we report that H 2 O 2 inhibits chlorophyll synthesis during leaf senescence via the I subunit of magnesium-chelatase (CHLI1). During leaf senescence, the decrease of chlorophyll content is accompanied by the increase of H 2 O 2 accumulation, as well as the inhibition of catalase (CAT) genes expression. The mutant cat2-1, with increased H 2 O 2 shows an accelerated senescence phenotype and decreased CHLI1 activity compared with the wild type. H 2 O 2 inhibits CHLI1 activity by sulfenylating CHLI1 during leaf senescence. Consistent with this, the chli1 knockout mutant displays the same premature leaf senescence symptom as cat2-1, while overexpression of CHLI1 in cat2-1 can partially restore its early senescence phenotype. Taken together, these results illustrate that CAT2-mediated H 2 O 2 accumulation during leaf senescence represses chlorophyll synthesis through sulfenylating CHLI1, and thus inhibits its activity, providing a new insight into the pivotal role of chlorophyll synthesis as a participant in orchestrating the leaf senescence., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Fusicoccin-A (FC-A) is a diterpene glucoside produced by a pathogenic fungus. Since its discovery, FC-A has been widely recognized as a phytotoxin that induces stomatal opening and leaf wilting, eventually leading to plant death. In this study, we present the first evidence that FC-A enhances plant growth by stabilizing the protein-protein interaction between plasma membrane (PM) H + -ATPase and 14-3-3 in guard cells. Long-term treatment of Arabidopsis plants with FC-A resulted in ~ 30% growth enhancement. Structurally similar fusicoccin-J (FC-J) showed a similar degree of growth-promotion activity as FC-A, whereas the more hydrophilic fusicoccin-H (FC-H) exhibited no effect on plant growth, indicating that the enhancement of plant growth observed with FC-A and FC-J involves upregulation of the protein-protein interaction between PM H + -ATPase and 14-3-3 in guard cells, which promotes stomatal opening and photosynthesis., (© 2024. The Author(s).)

Indole-3-carbinol (I3C), a hydrolysis product of indole-3-methylglucosinolate, is toxic to herbivorous insects and pathogens. In mammals, I3C is extensively studied for its properties in cancer prevention and treatment. Produced in Brassicaceae, I3C reversibly inhibits root elongation in a concentration-dependent manner. This inhibition is partially explained by the antagonistic action of I3C on auxin signaling through TIR1. To further elucidate the mode of action of I3C in plants, we have employed a forward-genetic amiRNA screen that circumvents functional redundancy. We identified and characterized two amiRNA lines with impaired I3C response. The first line, ICT2 , targets the phosphatidylinositol 4-phosphate 5-kinase family (PIP5K), exhibiting tolerance to I3C, while the second line, ICS1 , targets the Wall-Associated Kinases (WAK1-3) family, showing susceptibility to I3C. Both lines maintain I3C-induced antagonism of auxin signaling, indicating that their phenotypes are due to auxin-independent mechanisms. Transcript profiling experiments reveal that both lines are transcriptionally primed to respond to I3C treatment. Physiological, metabolomic, and transcriptomic analysis reveal that these kinases mediate numerous developmental processes and are involved in abiotic and biotic stress responses.

Roots play an important role in plant growth, including providing essential mechanical support, water uptake, and nutrient absorption. Nanomaterials play a positive role in improving plant root development, but there is limited knowledge of how nanomaterials affect lateral root (LR) formation. Poly (acrylic) acid coated nanoceria (cerium oxide nanoparticles, PNC) are commonly used to improve plant stress tolerance due to their ability to scavenge reactive oxygen species (ROS). However, its impact on LR formation remains unclear. In this study, we investigated the effects of PNC on LR formation in Arabidopsis thaliana by monitoring ROS levels and Ca2+ distribution in roots. Our results demonstrate that PNC significantly promote LR formation, increasing LR numbers by 26.2%. Compared to controls, PNC-treated Arabidopsis seedlings exhibited reduced H2 O2 levels by 18.9% in primary roots (PRs) and 40.6% in LRs, as well as decreased O 2 · - levels by 47.7% in PRs and 88.5% in LRs. When compared with control plants, Ca2+ levels were reduced by 35.7% in PRs and 22.7% in LRs of PNC-treated plants. Overall, these results indicate that PNC could enhance LR development by modulating ROS and Ca2+ levels in roots.

Indole-3-acetic acid (IAA) can regulate plant growth and thus modulate the accumulation of polycyclic aromatic hydrocarbons (PAHs). However, the effect of endogenous IAA on PAHs accumulation and its influencing factors remains unclear. To unravel this, two different IAA expression genotypes of Arabidopsis thaliana, i.e., IAA-underproducing yucca1D [YUC1] mutant and wild type [WT]) were selected and treated with different fluoranthene (Flu) concentrations (0 mg/L [CK], 5 mg/L [Flu5], and 20 mg/L [Flu20]) to reveal the impact mechanism of endogenous IAA on Flu uptake by plants. The results indicated that under Flu5 treatment, the bioconcentration factors (BCF) and translocation factors (TF) of Flu in WT were 41.4 % and 14.3 % higher than those in YUC1. Similarly, under Flu20 treatment, the BCF and TF of Flu in WT were also 42.2 % and 8.2 % higher than those in YUC1. In addition, the BCF and TF were 72.5 % and 35.8 % higher under Flu5 treatment compared to Flu20 treatment for WT, and 73.4 % and 28.6 % higher respectively for YUC1. Moreover, WT exhibited higher plant growth (biomass, root morphology indicators [root length, root area and number of tips]) and IAA content compared to YUC1 under identical Flu treatments. Plant growth and IAA content declined with the increase of Flu concentration in both YUC1 and WT leaves compared with CK treatment. Conversely, in WT roots, root biomass and morphology indicators promoted followed by a decrease as the concentration of Flu increased. Additionally, the antioxidant enzyme activities (SOD, POD, and CAT) of WT were 11.1 %, 16.7 %, and 28.9 % higher than those of YUC1 under Flu5 treatment, and 13.6 %, 12.9 %, and 26.5 % higher under Flu20 treatment. Compared with CK treatment, SOD and POD activities promoted with increasing Flu concentration, whereas CAT activities decreased. Variability separation analysis revealed that level of IAA primarily influenced Flu accumulation in WT or under Flu5 treatments, whereas antioxidant enzyme activity primarily affected Flu accumulation in YUC1 or under Flu20 treatments. Exploring the relationship between the IAA synthesis gene YUCCA and IAA levels, alongside Flu accumulation, could yield novel insights into the regulation of PAH accumulation in plants., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)