OsHAK4 functions in retrieving sodium from the phloem at the reproductive stage of rice.

Soil salinity significantly limits rice productivity, but it is poorly understood how excess sodium (Na+) is delivered to the grains at the reproductive stage. Here, we functionally characterized OsHAK4, a member of the clade IV HAK/KUP/KT transporter subfamily in rice. OsHAK4 was localized to the plasma membrane and exhibited influx transport activity for Na+, but not for K+. Analysis of organ- and growth stage-dependent expression patterns showed that very low expression levels of OsHAK4 were detected at the vegetative growth stage, but its high expression in uppermost node I, peduncle, and rachis was found at the reproductive stage. Immunostaining indicated OsHAK4 localization in the phloem region of node I, peduncle, and rachis. Knockout of OsHAK4 did not affect the growth and Na+ accumulation at the vegetative stage. However, at the reproductive stage, the hak4 mutants accumulated higher Na+ in the peduncle, rachis, husk, and brown rice compared to the wild-type rice. Element imaging revealed higher Na+ accumulation at the phloem region of the peduncle in the mutants. These results indicate that OsHAK4 plays a crucial role in retrieving Na+ from the phloem in the upper nodes, peduncle, and rachis, thereby preventing Na+ distribution to the grains at the reproductive stage of rice.

Melatonin Increases Root Cell Wall Phosphorus Reutilization via an NO Dependent Pathway in Rice (Oryza sativa).

Melatonin (MT) has been implicated in the plant response to phosphorus (P) stress; however, the precise molecular mechanisms involved remain unclear. This study investigated whether MT controls internal P distribution and root cell wall P remobilization in rice. Rice was treated with varying MT and P levels and analyzed using biochemical and molecular techniques to study phosphorus utilization. The results demonstrated that low P levels lead to a rapid increase in endogenous MT levels in rice roots. Furthermore, the exogenous application of MT significantly improved rice tolerance to P deficiency, as evidenced by the increased biomass and reduced proportion of roots to shoots under P-deficient conditions. MT application also mitigated the decrease in P content regardless in both the roots and shoots. Mechanistically, MT accelerated the reutilization of P, particularly in the root pectin fraction, leading to increased soluble P liberation. In addition, MT enhanced the expression of OsPT8, a gene involved in root-to-shoot P translocation. Furthermore, we observed that MT induced the production of nitric oxide (NO) in P-deficient rice roots and that the mitigating effect of MT on P deficiency was compromised in the presence of the NO inhibitor, c-PTIO, implying that NO is involved in the MT-facilitated mitigation of P deficiency in rice. Overall, our findings highlight the potential of MT as a promising strategy for enhancing rice tolerance to P deficiency and improving P use efficiency in agricultural practices.

A novel OsGST gene encoding 9glutathione reductase negatively regulates cadmium accumulation in rice.

Cadmium (Cd) accumulates in rice and then moves up the food chain, causing serious health problems for humans. Glutathione S-transferase (GST) binds exogenous hazardous compounds to glutathione (GSH), which performs a variety of roles in plant responses to Cd stress. Here, Cd stimulated the transcripts of a novel OsGST gene, and the OsGST protein, which was localized in the nucleus and cytoplasm, was also induced by Cd. In OsGST deletion mutant lines generated by CRISPR/Cas9, more Cd was accumulated, and Cd hypersensitive phenotypes were observed, while transgenic lines overexpressing OsGST exhibited enhanced Cd tolerance and less Cd accumulation. Further analysis indicated that the osgst mutants exhibited considerably greater reactive oxygen species (ROS) and higher GSH level, and the antioxidant activity associated genes' expression were down-regulated, imply that OsGST controlled rice Cd accumulation and resistance through preserving the equilibrium of the GSH and redox in rice.

A NAC transcription factor represses a module associated with xyloglucan content and regulates aluminum tolerance.

The transcriptional regulation of aluminum (Al) tolerance in plants is largely unknown, although Al toxicity restricts agricultural yields in acidic soils.. Here, we identified a NAM, ATAF1/2, and cup-shaped cotyledon 2 (NAC) transcription factor that participates in Al tolerance in Arabidopsis (Arabidopsis thaliana). Al substantially induced the transcript and protein levels of ANAC070, and loss-of-function anan070 mutants showed remarkably increased Al sensitivity, implying a beneficial role of ANAC070 in plant tolerance to Al toxicity. Further investigation revealed that more Al accumulated in the roots of anac070 mutants, especially in root cell walls, accompanied by a higher hemicellulose and xyloglucan level, implying a possible interaction between ANAC070 and genes that encode proteins responsible for the modification of xyloglucan, including xyloglucan endo-transglycosylases/hydrolase (XTH) or ANAC017. Yeast one hybrid analysis revealed a potential interaction between ANAC070 and ANAC017, but not for other XTHs. Furthermore, dual-luciferase reporter assay, RT-qPCR, and GUS analysis revealed that ANAC070 could directly repress the transcript levels of ANAC017, and knockout of ANAC017 in the anac070 mutant partially restored its Al sensitivity phenotype, indicating that ANAC070 contributes to Al tolerance mechanisms other than suppression of ANAC017 expression. Further analysis revealed that the core transcription factor SENSITIVE TO PROTON RHIZOTOXICITY1 (STOP1) and its target genes, which control Al tolerance in Arabidopsis, may also be involved in ANAC070-regulated Al tolerance. In summary, we identified a transcription factor, ANAC070, that represses the ANAC017-XTH31 module to regulate Al tolerance in Arabidopsis.

A novel aldo-keto reductase gene, OsAKR1, from rice confers higher tolerance to cadmium stress in rice by an in vivo reactive aldehyde detoxification.

Elevated levels of cadmium (Cd) have the ability to impede plant development. Aldo-keto reductases (AKRs) have been demonstrated in a number of plant species to improve tolerance to a variety of abiotic stresses by scavenging cytotoxic aldehydes; however, only a few AKRs have been identified to improve Cd tolerance. The OsAKR1 gene was extracted and identified from rice here. After being exposed to Cd, the expression of OsAKR1 dramatically rose in both roots and shoots, although more pronounced in roots. According to a subcellular localization experiment, the nucleus and cytoplasm are where OsAKR1 is primarily found. Mutants lacking OsAKR1 exhibited Cd sensitive phenotype than that of the wild-type (WT) Nipponbare (Nip), and osakr1 mutants exhibited reduced capacity to scavenge methylglyoxal (MG). Furthermore, osakr1 mutants exhibited considerably greater hydrogen peroxide (H2O2) and malondialdehyde (MDA) levels, and increased catalase (CAT) activity in comparison to Nip. The expression of three isomeric forms of CAT was found to be considerably elevated in osakr1 mutants during Cd stress, as demonstrated by quantitative real-time PCR analysis, when compared to Nip. These results imply that OsAKR1 controlled rice's ability to withstand Cd by scavenging harmful aldehydes and turning on the reactive oxygen species (ROS) scavenging mechanism.

Brassinosteroid decreases cadmium accumulation via regulating gibberellic acid accumulation and Cd fixation capacity of root cell wall in rice (Oryza sativa).

The precise mechanism behind the association between plants' reactions to cadmium (Cd) stress and brassinosteroid (BR) remains unclear. In the current investigation, Cd stress quickly increased the endogenous BR concentration in the rice roots. Exogenous BR also increased the hemicellulose level in the root cell wall, which in turn increased its capacity to bind Cd. Simultaneously, the transcription level of genes responsible for root Cd absorption was decreased, including Natural Resistance-Associated Macrophage Protein 1/5 (OsNRAMP1/5) and a major facilitator superfamily gene called OsCd1. Ultimately, the increased expression of Heavy Metal ATPase 3 (OsHMA3) and the decreased expression of OsHMA2, which was in charge of separating Cd into vacuoles and translocating Cd to the shoots, respectively, led to a decrease in the amount of Cd that accumulated in the rice shoots. In contrast, transgenic rice lines overexpressing OsGSK2 (a negative regulator in BR signaling) accumulated more Cd, while OsGSK2 RNA interference (RNAi) rice line accumulated less Cd. Furthermore, BR increased endogenous Gibberellic acid (GA) level, and applying GA could replicate its alleviative effect. Taken together, BR decreased Cd accumulation in rice by mediating the cell wall's fixation capacity to Cd, which might relied on the buildup of the GA.

Auxin acts upstream of nitric oxide to regulate cell wall xyloglucan and its aluminium-binding capacity in Arabidopsis thaliana.

MAIN CONCLUSION: Auxin acts upstream of NO through NOA and XXT5 pathways to regulate the binding capacity of the root cell wall to Al. In our previous study, we identified an unknown mechanism by which 1-naphthaleneacetic acid (NAA) decreased the fixation of aluminum (Al) in the cell wall. Here, we observed that external application of the nitric oxide (NO) donor S-nitrosoglutathion (GSNO) increased the inhibition of Al on root elongation. Further analysis indicated that GSNO could induce Al accumulation in the roots and root cell walls, which is consistent with lower xyloglucan content. In comparison to the Columbia-0 (Col-0) wild type (WT), endogenous NO-reduced mutants noa1 (NOA pathway) and nia1nia2 (NR pathway) were more resistant to Al, with lower root Al content, higher xyloglucan content, and more Al accumulation in the root cell walls. By contrast, the xxt5 mutant with reduced xyloglucan content exhibited an Al-sensitive phenotype. Interestingly, Al treatment increased the endogenous auxin and NO levels, and the auxin levels induced under Al stress further stimulated NO production. Auxin application reduced Al retention in hemicellulose and decreased the xyloglucan content, similar to the effects observed with GSNO. In yucca and aux1-7 mutants, exogenous application of NO resulted in responses similar to those of the WT, whereas exogenous auxin had little effect on the noa1 mutant under Al stress. In addition, as auxin had similar effects on the nia1nia2 mutant and the WT, exogenous auxin and NO had little effect on the xxt5 mutant under Al stress, further confirming that auxin acts upstream of NO through NOA and XXT5 pathways to regulate the binding capacity of the root cell wall to Al.

Auxin is involved in cadmium accumulation in rice through controlling nitric oxide production and the ability of cell walls to bind cadmium.

Although auxin has been linked to plants' responses to cadmium (Cd) stress, the exact mechanism is yet elusive. The objective of the current investigation was to determine the role and the mechanism of auxin in controlling rice's Cd accumulation. Rice roots with Cd stress have higher endogenous auxin levels, and exogenous auxin combined Cd treatment could reduce root cell wall's hemicellulose content when compared with Cd treatment alone, which in turn reduced its fixation of Cd, as well as decreased the expression of OsCd1 (a major facilitator superfamily gene), OsNRAMP1/5 (Natural Resistance-Associated Macrophage Protein 1/5), OsZIP5/9 (Zinc Transporter 5/9), and OsHMA2 (Heavy Metal ATPase 2) that participated in Cd uptake and root to shoot translocation. Furthermore, less Cd accumulated in the shoots as a result of auxin's impact in increasing the expression of OsCAL1 (Cadmium accumulation in Leaf 1), OsABCG36/OsPDR9 (G-type ATP-binding cassette transporter/Pleiotropic drug resistance 9), and OsHMA3, which were in charge of Cd efflux and sequestering into vacuoles, respectively. Additionally, auxin decreased endogenous nitric oxide (NO) levels and antioxidant enzyme activity, while treatment of a NO scavenger-cPTIO-reduced auxin's alleviatory effects. In conclusion, the rice's ability to tolerate Cd toxicity was likely increased by the auxin-accelerated cell wall Cd exclusion mechanism, a pathway that controlled by the buildup of NO.

Salylic acid minimize cadmium accumulation in rice through regulating the fixation capacity of the cell wall to cadmium.

Although salylic acid (SA) has been linked to how plants react to cadmium (Cd) stress, the exact mechanism is still unknown. The endogenous SA concentration in the rice (Oryza sativa L.) roots was enhanced by Cd stress in the current investigation, and exogenous SA reduced the hemicellulose content in root cell wall, which in turn inhibited its Cd binding capacity. What's more, exogenous SA also decreased the transcription level of genes such as Natural Resistance-Associated Macrophage Protein 5 (OsNRAMP5) and a major facilitator superfamily gene-OsCd1 that responsible for root Cd absorption. Finally, less Cd was accumulated in the rice as a result of the higher expression of Heavy Metal ATPase 3 (OsHMA3), Cation/Ca exchanger 2 (OsCCX2) and Pleiotropic Drug Resistance 9 (OsPDR9/OsABCG36) that were responsible for separating Cd into vacuole and getting Cd out of cells, respectively. In contrast, mutant with low SA level accumulated more Cd. Additionally, SA enhanced endogenous nitric oxide (NO) levels, and its alleviatory effects were mimicked by a NO donor, sodium nitroprusside (SNP). In conclusion, SA enhanced rice's Cd resistance through regulating the binding capacity of the cell wall to Cd, a pathway that might dependent on the NO accumulation.

Involvement of the 4-coumarate:coenzyme A ligase 4CL4 in rice phosphorus acquisition and rhizosphere microbe recruitment via root growth enlargement.

MAIN CONCLUSION: The 4-coumarate:coenzyme A ligase 4CL4 is involved in enhancing rice P acquisition and use in acid soil by enlarging root growth and boosting functional rhizosphere microbe recruitment. Rice (Oryza sativa L.) cannot easily acquire phosphorus (P) from acid soil, where root growth is inhibited and soil P is fixed. The combination of roots and rhizosphere microbiota is critical for plant P acquisition and soil P mobilization, but the associated molecular mechanism in rice is unclear. 4CL4/RAL1 encodes a 4-coumarate:coenzyme A ligase related to lignin biosynthesis in rice, and its dysfunction results in a small rice root system. In this study, soil culture and hydroponic experiments were conducted to examine the role of RAL1 in regulating rice P acquisition, fertilizer P use, and rhizosphere microbes in acid soil. Disruption of RAL1 markedly decreased root growth. Mutant rice plants exhibited decreased shoot growth, shoot P accumulation, and fertilizer P use efficiency when grown in soil-but not under hydroponic conditions, where all P is soluble and available for plants. Mutant ral1 and wild-type rice rhizospheres had distinct bacterial and fungal community structures, and wild-type rice recruited some genotype-specific microbial taxa associated with P solubilization. Our results highlight the function of 4CL4/RAL1 in enhancing rice P acquisition and use in acid soil, namely by enlarging root growth and boosting functional rhizosphere microbe recruitment. These findings can inform breeding strategies to improve P use efficiency through host genetic manipulation of root growth and rhizosphere microbiota.

Melatonin reduces cadmium accumulation via mediating the nitric oxide accumulation and increasing the cell wall fixation capacity of cadmium in rice.

Melatonin (MT) is participated in plants' response to cadmium (Cd) tolerance, although its work model remains elusive. Here, the function of MT in adjusting Cd accumulation in rice was investigated. 'Nipponbare' (Nip) was cultured in the -Cd (1/2 Kimura B), -Cd + MT (1/2 Kimura B with 1 µM MT), +Cd (1/2 Kimura B plus 1 µM Cd) and +Cd + MT (1/2 Kimura B with 1 µM Cd and 1 µM MT) nutrient solutions for 7 d. Cd markedly induced the endogenous MT accumulation in rice roots and shoots, even within 1 h. MT applied exogenously elevated the hemicelluloses level, which in turn increased the cell wall's binding capacity to Cd. Furthermore, MT applied exogenously down-regulated the transcription level of Natural Resistance-Associated Macrophage Protein 1 (OsNRAMP1), OsNRAMP5, a major facilitator superfamily gene (OsCd1), and IRON-REGULATED TRANSPORTER 1 (OsIRT1), all of which were responsible for Cd intake, thus less Cd was entered into roots. Moreover, MT applied exogenously also up-regulated transcription level of Cadmium accumulation in Leaf 1 (OsCAL1) and Heavy Metal ATPase 3 (OsHMA3), two genes both attributed to the decreased Cd accumulation in shoots through expelling Cd out of cells and chelating Cd in the vacuoles, respectively. In addition, MT applied exogenously further aggravated the production of nitric oxide (NO) that induced by Cd, while application of a NO donor-SNP mimicked this alleviatory effect of the MT, indicating MT decreased rice Cd accumulation relied on the accumulation of NO.

The endo-beta mannase MAN7 contributes to cadmium tolerance by modulating root cell wall binding capacity in Arabidopsis thaliana.

The heavy metal cadmium (Cd) is detrimental to crop growth and threatens human health through the food chain. To cope with Cd toxicity, plants employ multiple strategies to decrease Cd uptake and its root-to-shoot translocation. However, genes that participate in the Cd-induced transcriptional regulatory network, including those encoding transcription factors, remain largely unidentified. In this study, we demonstrate that ENDO-BETA-MANNASE 7 (MAN7) is necessary for the response of Arabidopsis thaliana to toxic Cd levels. We show that MAN7 is responsible for mannase activity and modulates mannose content in the cell wall, which plays a role in Cd compartmentalization in the cell wall under Cd toxicity conditions. Additionally, the repression of root growth by Cd was partially reversed via exogenous application of mannose, suggesting that MAN7-mediated cell wall Cd redistribution depends on the mannose pathway. Notably, we identified a basic leucine zipper (bZIP) transcription factor, bZIP44, that acts upstream of MAN7 in response to Cd toxicity. Transient dual-luciferase assays indicated that bZIP44 directly binds to the MAN7 promoter region and activates its transcription. Loss of bZIP44 function was associated with greater sensitivity to Cd treatment and higher accumulation of the heavy metal in roots and shoots. Moreover, MAN7 overexpression relieved the inhibition of root elongation seen in the bzip44 mutant under Cd toxicity conditions. This study thus reveals a pathway showing that MAN7-associated Cd tolerance in Arabidopsis is controlled by bZIP44 upon Cd exposure.

Boron supply restores aluminum-blocked auxin transport by the modulation of PIN2 trafficking in the root apical transition zone.

The supply of boron (B) alleviates the toxic effects of aluminum (Al) on root growth; however, the mechanistic basis of this process remains elusive. This study filled this knowledge gap, demonstrating that boron modifies auxin distribution and transport in Al-exposed Arabidopsis roots. In B-deprived roots, treatment with Al induced an increase in auxin content in the root apical meristem zone (MZ) and transition zone (TZ), whereas in the elongation zone (EZ) the auxin content was decreased beyond the level required for adequate growth. These distribution patterns are explained by the fact that basipetal auxin transport from the TZ to the EZ was disrupted by Al-inhibited PIN-FORMED 2 (PIN2) endocytosis. Experiments involving the modulation of protein biosynthesis by cycloheximide (CHX) and transcriptional regulation by cordycepin (COR) demonstrated that the Al-induced increase of PIN2 membrane proteins was dependent upon the inhibition of PIN2 endocytosis, rather than on the transcriptional regulation of the PIN2 gene. Experiments reporting on the profiling of Al3+ and PIN2 proteins revealed that the inhibition of endocytosis of PIN2 proteins was the result of Al-induced limitation of the fluidity of the plasma membrane. The supply of B mediated the turnover of PIN2 endosomes conjugated with indole-3-acetic acid (IAA), and thus restored the Al-induced inhibition of IAA transport through the TZ to the EZ. Overall, the reported results demonstrate that boron supply mediates PIN2 endosome-based auxin transport to alleviate Al toxicity in plant roots.

Decrease in hemicellulose content and its retention of iron contributes to phosphorus deficiency alleviated iron deficiency in Arabidopsis thaliana.

The physiological and molecular mechanisms between phosphorus (P) and iron (Fe) interactions are still elusive although they have been extensively investigated. In this study, we uncovered that limiting P supply could alleviate Fe deficiency in Arabidopsis (Col-0). Under Fe deficiency, P deficiency (-Fe-P) decreased cell wall Fe accumulation in root, but elevated Fe accumulation in the shoot, implying that the reduced Fe retention in the root cell wall may contribute to the P-deficiency-alleviated Fe deficiency in the shoot. On the other hand, increasing P supply could mimic the degree of Fe deficiency in terms of the expressions of genes induced after Fe deficient treatment. The components of the root cell wall showed that there was no distinction in the pectin content and the Fe retention in pectin between -Fe and -Fe-P treatments, while hemicellulose 1 content and Fe retained in it were decreased significantly in -Fe-P treatment as compared with -Fe treatment. The time-course experiment showed that decreasing cell wall retained Fe was mainly from the corresponding decrease in hemicellulose 1 retained Fe. Furthermore, the up-regulation of IRT1 expression in -Fe-P was obviously lower than -Fe. All these suggest that the P deficiency-induced decrease of hemicellulose 1 component leads to reutilization of root cell wall Fe and improvement of Fe nutrition in shoot in Fe deficient Arabidopsis. Our results provide a novel explanation of the interplay between Fe and P in Arabidopsis.

Salicylic Acid Acts Upstream of Auxin and Nitric Oxide (NO) in Cell Wall Phosphorus Remobilization in Phosphorus Deficient Rice.

Salicylic acid (SA) is thought to be involved in phosphorus (P) stress response in plants, but the underlying molecular mechanisms are poorly understood. Here, we showed that P deficiency significantly increased the endogenous SA content by inducing the SA synthesis pathway, especially for up-regulating the expression of PAL3. Furthermore, rice SA synthetic mutants pal3 exhibited the decreased root and shoot soluble P content, indicating that SA is involved in P homeostasis in plants. Subsequently, application of exogenous SA could increase the root and shoot soluble P content through regulating the root and shoot cell wall P reutilization. In addition, - P + SA treatment highly upregulated the expression of P transporters such as OsPT2 and OsPT6, together with the increased xylem P content, suggesting that SA also participates in the translocation of the P from the root to the shoot. Moreover, both signal molecular nitric oxide (NO) and auxin (IAA) production were enhanced when SA is applied while the addition of respective inhibitor c-PTIO (NO scavenger) and NPA (IAA transport inhibitor) significantly decreased the root and shoot cell wall P remobilization in response to P starvation. Taken together, here SA-IAA-NO-cell wall P reutilization pathway has been discovered in P-starved rice.

Gibberellic acid alleviates cadmium toxicity in rice by regulating NO accumulation and cell wall fixation capacity of cadmium.

Gibberellic acid (GA) has been implicated in the response of plants to cadmium (Cd) stress, but the underlying mechanism remains unclear. In the present study, our aim was to confirm the role of GA in regulating the accumulation of Cd in rice. We found that Cd stress elevated the endogenous GA level in the rice roots. Exogenous GA application not only decreased the fixation of Cd in the root cell wall through reducing the hemicelluloses content, but also decreased the expression of OsNRAMP5 (Natural Resistance-Associated Macrophage Protein 5) and OsCd1 (a major facilitator superfamily gene). Both OsNRAMP5 and OsCd1 are related to Cd absorption, therefore, less Cd was accumulated in the roots. Furthermore, GA increased the expression of OsHMA3 (Heavy Metal ATPase 3) and OsCAL1 (Cadmium accumulation in Leaf 1), which are responsible for sequestering the Cd to the vacuoles and effluxing the Cd outside the cell, respectively, as a result, less Cd was accumulated in the shoots. In contrast, more Cd was accumulated in GA deficient lines. Furthermore, GA decreased the endogenous NO levels and the activity of antioxidant enzymes, while application of a NO scavenger-cPTIO diminished the alleviatory role of GA. In summary, the GA accelerated cell wall Cd exclusion mechanism probably improved rice tolerance to Cd toxicity via regulating the accumulation of NO.

Auxin facilitates cell wall phosphorus reutilization in a nitric oxide-ethylene dependent manner in phosphorus deficient rice (Oryza sativa L.).

Auxin is involved in stress responses of plants, such as phosphorus (P) deficiency in rice. Studies on whether auxin participates in cell-wall inorganic phosphorous (Pi) reutilization in Pi-starved rice are scarce. This study explored the mechanisms underlying auxin-facilitated cell-wall Pi-reutilization in rice roots. Pi deficiency rapidly induced auxin accumulation in roots; exogenous auxin [α-naphthaleneacetic acid (NAA), a permeable analog of auxin] elevated soluble Pi content in roots and shoots by increasing pectin content by enhancing activity of pectin methylesterase, and upregulating the transcript level of PHOSPHORUS-TRANSPORTER-2, such that more Pi was translocated to the shoot. Irrespective of the Pi status, exogenous auxin induced nitric oxide (NO) and ethylene production, while exogenous sodium nitroprusside (an NO donor) and 1-aminocyclopropane-1-carboxylic acid (a precursor of ethylene) had no effect on auxin content, suggesting that auxin may act upstream of NO and ethylene. The beneficial effect of NAA in increasing soluble Pi content in roots and shoots disappeared when 2-(4-carboxyphenyl)- 4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (a scavenger of NO) or aminoethoxyvinylglycine (an inhibitor of ethylene) were applied, suggesting that auxin facilitates cell-wall Pi-reutilization in a NO-ethylene-dependent manner in Pi-deficient rice. Our study results suggest auxin application as an effective agronomic practice for improving plant Pi nutrition in P-deficient conditions.

Distinct Patterns of Rhizosphere Microbiota Associated With Rice Genotypes Differing in Aluminum Tolerance in an Acid Sulfate Soil.

Rhizosphere microbes are important for plant tolerance to various soil stresses. Rice is the most aluminum (Al)-tolerant small grain cereal crop species, but the link between rice Al tolerance and rhizosphere microbiota remains unclear. This study aimed to investigate the microbial community structure of aluminum-sensitive and Al-tolerant rice varieties in acid sulfate soil under liming and non-liming conditions. We analyzed the rice biomass and mineral element contents of rice plants as well as the chemical properties and microbial (archaea, bacteria, and fungi) communities of rhizosphere and bulk soil samples. The results showed that the Al-tolerant rice genotype grew better and was able to take up more phosphorus from the acid sulfate soil than the Al-sensitive genotype. Liming was the main factor altering the microbial diversity and community structure, followed by rhizosphere effects. In the absence of liming effects, the rice genotypes shifted the community structure of bacteria and fungi, which accounted for the observed variation in the rice biomass. The Al-tolerant rice genotype recruited specific bacterial and fungal taxa (Bacillus, Pseudomonas, Aspergillus, and Rhizopus) associated with phosphorus solubilization and plant growth promotion. The soil microbial co-occurrence network of the Al-tolerant rice genotype was more complex than that of the Al-sensitive rice genotype. In conclusion, the bacterial and fungal community in the rhizosphere has genotype-dependent effects on rice Al tolerance. Aluminum-tolerant rice genotypes recruit specific microbial taxa, especially phosphorus-solubilizing microorganisms, and are associated with complex microbial co-occurrence networks, which may enhance rice growth in acid sulfate soil.

Transcription factor ANAC004 enhances Cd tolerance in Arabidopsis thaliana by regulating cell wall fixation, translocation and vacuolar detoxification of Cd, ABA accumulation and antioxidant capacity.

Cadmium (Cd) is toxic to plants, which have evolved multiple strategies to cope with Cd stress. In this study, we identified a nucleus-localized NAC-type transcription factor, ANAC004, which is induced by Cd and involved in regulating Cd resistance in Arabidopsis. First, anac004 mutants exhibited Cd sensitive phenotype and accumulated more Cd (12-23% higher than wild type in roots and shoots); plants overexpressing ANAC004 showed the opposite phenotype and with lower Cd accumulation. Second, ANAC004 enhanced Cd fixation in cell wall hemicellulose, thus reducing Cd2+ influx into root cells. Third, ANAC004 was involved in the process of vacuolar Cd compartmentalization by regulating the genes associated with Cd detoxification (PCS1/2, NAS4, ABCC1/2/3, MTP1/3, IREG2 and NRAMP3/4). Fourth, ANAC004 reduced root-to-shoot Cd translocation through down-regulated Cd translocation-related genes (HMA2 and HMA4). Finally, the expression of genes related to ABA synthesis (AAO3, MCSU, and NCED3) and the activities of antioxidant enzymes (SOD, POD and CAT) were all reduced in anac004 mutants, leading to reduced levels of endogenous ABA and increased accumulation of reactive oxygen species (O2.- and H2O2) and MDA, which ultimately weakened resistance to Cd. Our results suggest that ANAC004 decreases Cd accumulation in Arabidopsis through enhancing cell wall Cd immobilization, increasing vacuolar Cd detoxification, and inhibiting Cd translocation, thus improving Cd resistance, processes that might be mediated by ABA signaling and antioxidant defense systems.