Honglei Jia

Arabidopsis CROLIN1, a novel plant actin-binding protein, functions in cross-linking and stabilizing actin filaments.

Background: Higher order actin filament structures are involved in many cellular processes. Results: Arabidopsis CROLIN1 contains a predicted actin-cross-linking domain and shows F-actin binding, cross-linking, and stabilizing activities in vitro. Conclusion: CROLIN1 functions as an actin-binding and cross-linking protein. Significance: CROLIN1 is a previously undiscovered plant actin-cross-linking protein. Higher order actin filament structures are necessary for cytoplasmic streaming, organelle movement, and other physiological processes. However, the mechanism by which the higher order cytoskeleton is formed in plants remains unknown. In this study, we identified a novel actin-cross-linking protein family (named CROLIN) that is well conserved only in the plant kingdom. There are six isovariants of CROLIN in the Arabidopsis genome, with CROLIN1 specifically expressed in pollen. In vitro biochemical analyses showed that CROLIN1 is a novel actin-cross-linking protein with binding and stabilizing activities. Remarkably, CROLIN1 can cross-link actin bundles into actin networks. CROLIN1 loss of function induces pollen germination and pollen tube growth hypersensitive to latrunculin B. All of these results demonstrate that CROLIN1 may play an important role in stabilizing and remodeling actin filaments by binding to and cross-linking actin filaments.

Anisotropic Cell Expansion Is Affected through the Bidirectional Mobility of Cellulose Synthase Complexes and Phosphorylation at Two Critical Residues on CESA3.

Phosphorylation of a subunit of cellulose synthase differentially alters the rate at which cellulose synthase moves in both directions along microtubules, thereby impacting growth and development. Here we report that phosphorylation status of S211 and T212 of the CESA3 component of Arabidopsis (Arabidopsis thaliana) cellulose synthase impacts the regulation of anisotropic cell expansion as well as cellulose synthesis and deposition and microtubule-dependent bidirectional mobility of CESA complexes. Mutation of S211 to Ala caused a significant decrease in the length of etiolated hypocotyls and primary roots, while root hairs were not significantly affected. By contrast, the S211E mutation stunted the growth of root hairs, but primary roots were not significantly affected. Similarly, T212E caused a decrease in the length of root hairs but not root length. However, T212E stunted the growth of etiolated hypocotyls. Live-cell imaging of fluorescently labeled CESA showed that the rate of movement of CESA particles was directionally asymmetric in etiolated hypocotyls of S211A and T212E mutants, while similar bidirectional velocities were observed with the wild-type control and S211E and T212A mutant lines. Analysis of cell wall composition and the innermost layer of cell wall suggests a role for phosphorylation of CESA3 S211 and T212 in cellulose aggregation into fibrillar bundles. These results suggest that microtubule-guided bidirectional mobility of CESA complexes is fine-tuned by phosphorylation of CESA3 S211 and T212, which may, in turn, modulate cellulose synthesis and organization, resulting in or contributing to the observed defects of anisotropic cell expansion.

Glucose‐6‐phosphate dehydrogenase‐dependent hydrogen peroxide production is involved in the regulation of plasma membrane H+‐ATPase and Na+/H+ antiporter protein in salt‐stressed callus from Carex moorcroftii

Glucose-6-phosphate dehydrogenase (G6PDH) is important for the activation of plant resistance to environmental stresses, and ion homeostasis is the physiological foundation for living cells. In this study, we investigated G6PDH roles in modulating ion homeostasis under salt stress in Carex moorcroftii callus. G6PDH activity increased to its maximum in 100 mM NaCl treatment and decreased with further increased NaCl concentrations. K+/Na+ ratio in 100 mM NaCl treatment did not exhibit significant difference compared with the control; however, in 300 mM NaCl treatment, it decreased. Low-concentration NaCl (100 mM) stimulated plasma membrane (PM) H+-ATPase and NADPH oxidase activities as well as Na+/H+ antiporter protein expression, whereas high-concentration NaCl (300 mM) decreased their activity and expression. When G6PDH activity and expression were reduced by glycerol treatments, PM H+-ATPase and NADPH oxidase activities, Na+/H+ antiporter protein level and K+/Na+ ratio dramatically decreased. Simultaneously, NaCl-induced hydrogen peroxide (H₂O₂) accumulation was abolished. Exogenous application of H₂O₂ increased G6PDH, PM H+-ATPase and NADPH oxidase activities, Na+/H+ antiporter protein expression and K+/Na+ ratio in the control and glycerol treatments. Diphenylene iodonium (DPI), the NADPH oxidase inhibitor, which counteracted NaCl-induced H₂O₂ accumulation, decreased G6PDH, PM H+-ATPase and NADPH oxidase activities, Na+/H+ antiporter protein level and K+/Na+ ratio. Western blot result showed that G6PDH expression was stimulated by NaCl and H₂O₂, and blocked by DPI. Taken together, G6PDH is involved in H₂O₂ accumulation under salt stress. H₂O₂, as a signal, upregulated PM H+-ATPase activity and Na+/H+ antiporter protein level, which subsequently resulted in the enhanced K+/Na+ ratio. G6PDH played a central role in the process.

Hydrogen sulfide - cysteine cycle system enhances cadmium tolerance through alleviating cadmium-induced oxidative stress and ion toxicity in Arabidopsis roots

Cadmium (Cd2+) is a common toxic heavy metal ion. We investigated the roles of hydrogen sulfide (H2S) and cysteine (Cys) in plant responses to Cd2+ stress. The expression of H2S synthetic genes LCD and DES1 were induced by Cd2+ within 3 h, and endogenous H2S was then rapidly released. H2S promoted the expression of Cys synthesis-related genes SAT1 and OASA1, which led to endogenous Cys accumulation. The H2S and Cys cycle system was stimulated by Cd2+ stress, and it maintained high levels in plant cells. H2S inhibited the ROS burst by inducing alternative respiration capacity (AP) and antioxidase activity. H2S weakened Cd2+ toxicity by inducing the metallothionein (MTs) genes expression. Cys promoted GSH accumulation and inhibited the ROS burst, and GSH induced the expression of phytochelatin (PCs) genes, counteracting Cd2+ toxicity. In summary, the H2S and Cys cycle system played a key role in plant responses to Cd2+ stress. The Cd2+ tolerance was weakened when the cycle system was blocked in lcddes1-1 and oasa1 mutants. This paper is the first to describe the role of the H2S and Cys cycle system in Cd2+ stress and to explore the relevant and specificity mechanisms of H2S and Cys in mediating Cd2+ stress.

cGMP and ethylene are involved in maintaining ion homeostasis under salt stress in Arabidopsis roots

AbstractKey messagecGMP promotes ethylene production and enhances the perception of ethylene. Endogenous ethylene or cGMP accumulation maintains ion homeostasis to enhancing salt resistance.etr1-3is insensitive to cGMP under salt stress.Abstract In the present study, we presented a signaling network involving ethylene and cGMP in salt resistance pathway of Arabidopsis roots. Results showed that the ethylene-insensitive mutant etr1-3 was more sensitive to salt stress than the wild type (WT). etr1-3 displayed a greater electrolyte leakage, thiobarbituric acid reactive substances and Na+/K+ ratio, but a lower plasma membrane (PM) H+-ATPase activity compared to WT under the different NaCl contents. Application of 1-aminocyclopropane-1-carboxylic acid (ACC, an ethylene precursor) or 8-Br-cGMP (the cGMP analog) alleviated NaCl-induced injury by maintaining a lower Na+/K+ ratio and increasing PM H+-ATPase activity in WT, but not in etr1-3. Roots treated with 8-Br-cGMP could promote ethylene production and enhance the expression of ACC synthase gene in WT. In addition, the 8-Br-cGMP action in NaCl stress was inhibited by aminooxyacetic acid (an inhibitor of ethylene biosynthesis), but 6-Anilino-5,8-quinolinedione (Ly83583, a guanylate cyclase inhibitor) could not affect ACC action in WT. These results suggest that ethylene functions as a downstream signal of cGMP that stimulates the PM H+-ATPase activity, which finally results in regulating ion homeostasis in Arabidopsis tolerance to salt. Moreover, cGMP enhanced the perception of ethylene in Arabidopsis under salt stress, which reversed the salt-induced increase of ETR1 and increased ERF1 at the transcript levels in WT. In a word, cGMP modulates salt resistance pathway of ethylene through regulating biosynthesis and perception of ethylene in Arabidopsis roots.

Effects of salicylic acid, Fe(II) and plant growth-promoting bacteria on Cd accumulation and toxicity alleviation of Cd tolerant and sensitive tomato genotypes

In this study, we investigated the ameliorative effects of salicylic acid (SA), metal ion (Fe(II)), and plant growth-promoting bacteria Burkholderia sp. D54 (B) on two tomato genotypes with different Cd tolerances under Cd stress, viz. Liger (Cd tolerant) and Tabd (Cd sensitive). The plant biomass, Cd accumulation, antioxidative response, pigment content and photosynthetic performance were determined. According to the results, exogenous application of SA, Fe(II) and Burkholderia sp. D54 or their complex effectively reduced Cd accumulation and increased biomass of root, stem and leaves in both Cd sensitive and Cd tolerant genotypes. Among all treatments, SA+Fe+B exerted the best performance. Burkholderia sp. D54 effectively alleviated Cd-induced oxidative toxicity in both tomato genotypes, while SA ameliorated oxidative stress in Cd sensitive genotype. Photosynthetic pigment content and photosynthetic rate of Cd tolerant genotype was increased by all treatments, but only SA and Burkholderia sp. D54 treatment increased pigment contents and photosynthetic performance in Cd sensitive genotypes. All treatments significantly decreased Cd accumulation in both tomato genotypes. The effect of Cd reduction was Fe+SA+B>SA>Fe>B. Taken together, our results indicated that exogenous application of SA, Fe(II) and Burkholderia sp. D54 could alleviate the Cd toxicity in both Cd sensitive and Cd tolerant genotypes, although the extent varies.

Ethylene promotes pollen tube growth by affecting actin filament organization via the cGMP-dependent pathway in Arabidopsis thaliana

Ethylene and cGMP are key regulators of plant developmental processes. In this study, we demonstrate that ethylene or cGMP promote pollen tube growth in a dose-dependent manner. The etr1–1 mutant was found to be insensitive to ethylene with regard to pollen tube growth, while the growth-promoting effect of ethylene in etr2–2, ein4–4, or ein4–7 did not change, suggesting that ethylene signaling was mainly perceived by ETR1. However, the function of cGMP was not inhibited in etr1–1 and pollen tubes became insensitive to ethylene when the endogenous cGMP level was artificially decreased. This shows that cGMP is necessary for the control of pollen tube growth and that it might be a downstream component of ETR1 in the ethylene signaling pathway. Our study also found that ethylene or cGMP increase the actin bundles and elevated the percentage of relative amount of F-actin, while removal of cGMP decreased actin bundles abundance and altered the ratio of F-actin in the tip and base regions of pollen tubes. In conclusion, our data suggests that ethylene functions as the upstream signal of cGMP, and that both signals promote pollen germination and tube growth by regulating F-actin, which is essential for vesicular transport and cytoplasmic streaming.

Ethylene-induced hydrogen sulfide negatively regulates ethylene biosynthesis by persulfidation of ACO in tomato under osmotic stress

A number of recent studies identified hydrogen sulfide (H2S) as an important signal in plant development and adaptation to environmental stress. H2S has been proven to participate in ethylene-induced stomatal closure, but how the signaling pathways of H2S and ethylene interact is still unclear. Here, we reveal how H2S controls the feedback-regulation of ethylene biosynthesis in tomato (Solanum lycopersicum) under osmotic stress. We found that ethylene induced the production of H2S in guard cells. The supply of hypotaurine (HT; a H2S scavenger) or DL-pro-pargylglycine (PAG; a synthetic inhibitor of H2S) removed the effect of ethylene or osmotic stress on stomatal closure. This suggests that ethylene-induced H2S is a downstream component of osmotic stress signaling, which is required for ethylene-induced stomatal closure under osmotic stress. We further found that H2S inhibited ethylene synthesis through inhibiting the activity of 1-aminocyclopropane-1-carboxylic acid (ACC) oxidases (ACOs) by persulfidation. A modified biotin-switch method (MBST) showed that H2S can induce persulfidation of LeACO1 and LeACO2 in a dose-dependent manner, and that persulfidation inhibits the activity of LeACO1 and LeACO2. We also found that LeACO1 is persulfidated at cysteine 60. These data suggested that ethylene-induced H2S negatively regulates ethylene biosynthesis by persulfidation of LeACOs. In addition, H2S was also found to inhibit the expression of LeACO genes. The results provide insight on the general mode of action of H2S and contribute to a better understanding of a plant’s response to osmotic stress.

Effects of salicylic acid, Epi-brassinolide and calcium on stress alleviation and Cd accumulation in tomato plants

Salicylic acid (SA), Epi-brassinolide (EBL) and calcium (Ca) play crucial roles in plant development and mediate plant response to biotic and abiotic stress. This study was aimed to investigate the possible mediatory role of SA, EBL, Ca or their combination in protecting tomato plants from cadmium (Cd) toxicity. According to the results, Cd stress resulted in a significant reduction of plant dry mass, photosynthetic pigment content as well as photosynthetic rate. Exogenous application of SA decreased the malondialdehyde (MDA) level by 39.27% and increased catalase (CAT) activity by 81.17%. SA and EBL treatment significantly increased chlorophyll a (Chl a), chlorophyll b (Chl b) content, photosynthetic rate (Pn) as well as water use efficiency (WUE). SA+EBL (1:1)/Ca+SA+EBL (1:1:1) treatment obviously alleviated Cd-induced growth inhibition, the dry mass of different tomato organs were significantly increased (p < 0.05). Especially in Ca+SA+EBL treated plants, the dry mass of roots, stems and leaves increased by 141.18%, 128.57% and 118.52%, respectively. Besides, SA+EBL and Ca+SA+EBL treatments reduced the MDA level, but increased photosynthetic pigment concentration and photosynthetic efficiency. CAT activity was increased by 62.92% in Ca+SA+EBL treated plants, the WUE was increased by 557.76% in SA+EBL pretreated plants. Moreover, exogenous application of SA, SA+EBL and Ca+SA+EBL significantly decreased Cd accumulation in tomato organs (p < 0.05) compared with Cd-stressed plants. Taken together, our results indicated that exogenous application of SA, EBL and Ca individually or in combination could alleviate Cd toxicity in tomato plants, although the extent varies.