硅缓解植物镉毒害的生理生态机制

镉是对生物毒性最强的污染物之一。过量的镉能够抑制植物的生长和光合作用,干扰矿质代谢并诱发氧化胁迫。硅作为一种有益元素,主要以Si(OH)_4的形态通过主动或被动方式被植物体吸收并转运到地上部分。硅对植物镉毒害具有缓解作用,但其缓解机制在不同物种、品种或生态型之间存在显著差异,并表现出一定的硅/镉浓度依赖性。总体上可概括为避性机制和耐性机制。避性机制包括:(1)在器官水平,减少植物根系对镉的吸收及其向地上部的转运;(2)在细胞水平,增强细胞壁对镉的吸附能力,减少共质体中镉的含量。耐性机制包括:(1)诱导细胞产生小分子螯合剂,增强对镉的螯合作用,减少细胞中游离态镉的含量;(2)增强抗氧化机制,减轻氧化胁迫;(3)改善光合作用和无机营养,促进植物生长。从植物对硅的吸收和转运、镉对植物的毒害作用以及硅对缓解植物镉毒害的生理生态机制3个方面进行了综述,并基于目前的研究现状和薄弱之处,对今后的研究重点进行了展望。     

Dualities in plant tolerance to pollutants and their uptake and translocation to the upper plant parts

There is a duality in plant tolerance to pollutants and its response to the pollutants’ stress.On the one hand some plants, (hyper)tolerant to heavy metals, are able to hyperaccumulate these metals in shoots, which could be beneficial for phytoremediation purposes to clean-up soil and water. On the other hand tolerant food crops, exposed to heavy metals in their growth medium, may be dangerous as carriers of toxic metals in the food chain leading to food toxicity. There is an additional duality in plant tolerance to heavy metals and that is in food crops that are tolerant and/or hyperaccumulators, which could be used on one hand for phytoremediation, under controlled conditions and on the other hand for food fortification with essential metals.Similarly, plants are also exposed to a large number of xenobiotic organic pollutants. Because they generally cannot avoid these compounds, plants take up, translocate, metabolize and detoxify many of them. There is a large variability in tolerance (defence) mechanisms against organic pollutants among plant species. This includes production of reductants but also scavenger molecules like ascorbate and glutathione and expression of the P-450 defence system, and superfamilies of the enzymes glutathione- and glucosyl-transferases. Again, with view to organic pollutants, plant detoxification mechanisms might well protect the plant itself, but produce compounds with some deleterious potential for other organisms.In this review we discuss these dualities on the basis of examples of agricultural and ‘wild’ species exposed to metal contaminants (mainly Cd) and organic pollutants. Differences in uptake and translocation of various pollutants and their consequences will be considered. We will separately outline the effects of the organic and non-organic pollutants on the internal metabolism and the detoxification mechanisms and try to indicate the differences between both types of pollutants. Finally the consequences and solutions of these dualities in plant tolerance to pollutants will be discussed.     

Chloroplast targeting of phytochelatin synthase in Arabidopsis: effects on heavy metal tolerance and accumulation

The enzymatically synthesized thiol peptide phytochelatin (PC) plays a central role in heavy metal tolerance and detoxification in plants. In response to heavy metal exposure, the constitutively expressed phytochelatin synthase enzyme (PCS) is activated leading to synthesis of PCs in the cytosol. Recent attempts to increase plant metal accumulation and tolerance reported that PCS over-expression in transgenic plants paradoxically induced cadmium hypersensitivity. In the present paper, we investigate the possibility of synthesizing PCs in plastids by over-expressing a plastid targeted phytochelatin synthase (PCS). Plastids represent a relatively important cellular volume and offer the advantage of containing glutathione, the precursor of PC synthesis. Using a constitutive CaMV 35S promoter and a RbcS transit peptide, we successfully addressed AtPCS1 to chloroplasts, significant PCS activity being measured in this compartment in two independent transgenic lines. A substantial increase in the PC content and a decrease in the glutathione pool were observed in response to cadmium exposure, when compared to wild-type plants. While over-expressing AtPCS1 in the cytosol importantly decreased cadmium tolerance, both cadmium tolerance and accumulation of plants expressing plastidial AtPCS1 were not significantly affected compared to wild-type. Interestingly, targeting AtPCS1 to chloroplasts induced a marked sensitivity to arsenic while plants over-expressing AtPCS1 in the cytoplasm were more tolerant to this metalloid. These results are discussed in relation to heavy metal trafficking pathways in higher plants and to the interest of using plastid expression of PCS for biotechnological applications.     

Phytochelatins are involved in differential arsenate tolerance in Holcus lanatus

Arsenate tolerance is conferred by suppression of the high-affinity phosphate/arsenate uptake system, which greatly reduces arsenate influx in a number of higher plant species. Despite this suppressed uptake, arsenate-tolerant plants can still accumulate high levels of As over their lifetime, suggesting that constitutive detoxification mechanisms may be required. Phytochelatins are thiol-rich peptides, whose production is induced by a range of metals and metalloids including arsenate. This study provides evidence for the role of phytochelatins in the detoxification of arsenate in arsenate-tolerant Holcus lanatus. Elevated levels of phytochelatin were measured in plants with a range of tolerance to arsenate at equivalent levels of arsenate stress, measured as inhibition of root growth. The results suggest that arsenate tolerance in H. lanatus requires both adaptive suppression of the high-affinity phosphate uptake system and constitutive phytochelatin production.     

Antioxidant and photosynthetic response of a purple-leaved and a green-leaved cultivar of sweet basil (Ocimum basilicum) to boron excess

Boron (B) toxicity induces oxidative stress and alterations in the photosynthetic process. The occurrence of visible symptoms depends on plant species and even on cultivar. However, limited information is available concerning antioxidant responses to B toxicity; therefore a study was carried out to assess the role of antioxidants in hydroponically grown sweet basil submitted to B excess. Two cultivars were compared: the purple-leaved ‘Red Rubin’ that shown scarce symptoms of B-induced toxicity and the green-leaved ‘Tigullio’ in which they were evident.Sweet basil plants were grown in “floating raft system” for 20 days with 0.2 (control), 2 and 20 mg L⁻¹ of B in the nutrient solution. At the end of treatments visible symptoms of damage were evaluated and some parameters were measured: growth, leaf B accumulation, gas exchange and chlorophyll fluorescence, pigment, phenols and malondialdehyde (MDA) concentrations, total non-enzymatic antioxidant ability, antioxidant molecules and enzymes.B excess negatively affected growth and photosynthesis in both cultivars but differential mechanisms were recorded. ‘Tigullio’ exhibited a larger B accumulation in leaves as compared to ‘Red Rubin’. Moreover, in ‘Red Rubin’ plants a greater constitutive content of ascorbic acid, glutathione, anthocyanins and, consequently, a stronger antioxidant ability than ‘Tigullio’ were recorded. MDA test confirmed that the extent of oxidative stress was larger in ‘Tigullio’ than in ‘Red Rubin’. A general stimulation of antioxidant enzymes occurred by increasing B concentration in the growth medium. Notable, anthocyanins were likely involved in the B tolerance shown by ‘Red Rubin’ in consideration of their antioxidant properties and because of the role of these compounds in photoprotection. This paper represents a contribution to understanding the role of antioxidant compounds in plant tolerance to B toxicity.     

The short-term responses of glutathione and phytochelation synthetic pathways genes to additional nitrogen under cadmium stress in poplar leaves

Earlier it was noticed that the supplementary nitrogen to nutritive solution of the cadmium stressed (Cd-stressed) plants can alleviate the toxic effects of this metal on the plants and improve plant growth performance. But the underlying mechanisms of such detoxification effect of nitrogen were not studied. In this study, a ten-day responses of related nitrogen-synthesized genes including γ-glutamylcysteine synthetase (γ-GCs), glutathione synthetase (ECGs) and phytochelatin synthase (PCs) involved in glutathione (ECG) and phytochelation (PC) synthetic pathways were examined. The plant growth performance and leaf chlorophyll content were examined at the final harvest. It was shown that the supplement of additional nitrogen to poplar plants under cadmium stress could significantly up-regulate the expression levels of γ-GCs, ECGs and PCs genes in plant leaves during the first 12 hours. Furthermore, cadmium stressed plants with additional nitrogen supplement showed significant enhancement in growth performance and increase in leaf chlorophyll content compared to sole cadmium stressed plants. Our results suggest that additional nitrogen could stimulate a short-term defense system in poplar plants through ECG and PC synthetic pathways. It is contribute to the alleviation of the toxic symptoms in polar plants caused by cadmium stress. This study provides a potential method to render harmless cadmium toxicity in stressed plants with nitrogen fertilization.     

镉（Cd）胁迫下蓖麻蛋白质组学筛查及RcBSK7抗性功能研究

为揭示蓖麻(Ricinus communis)植株响应重金属镉(Cd)胁迫相关机制,筛选出蓖麻中参与Cd胁迫的抗性基因。本研究通过观察种子发芽及植株生长状态,最终确定以水处理的蓖麻植株为对照,研究其在3种剂量(300、700、1 000 mg·L^-1)Cd胁迫处理下的反应机制,以期为揭示蓖麻响应Cd胁迫的防御和解毒机制提供新思路。利用差异蛋白质组学分析蓖麻在Cd胁迫下的网络调控机制,即随着Cd胁迫浓度的增加,蓖麻植株分别通过阻隔根系对重金属Cd的吸收、提高自身抗氧化能力、抑制Cd²⁺运转以及诱导细胞程序性死亡等防御解毒过程以抵抗Cd胁迫损伤。根据组学分析结果筛选出差异显著基因RcBSK7,通过在拟南芥(Arabidopsis thaliana)中进行功能验证可知,该基因对提高蓖麻对Cd耐受性具有重要的作用。本研究增强了对蓖麻植株在3种Cd胁迫下多样性和复杂性的认识,为耐Cd基因鉴定和土壤中重金属污染修复提供了有价值的理论依据。     

Identification and analysis of cold stress-inducible genes in Korean rapeseed varieties

The rapeseed plant (Brassica napus) is one of the top five oil-producing crops in the world. However, its potential for global cultivation is limited due to its sensitivity to cold. We selected five varieties of rapeseed popular in Korea and analyzed their sensitivity to cold stress. Three — ‘Naehan’, ‘Sandongchae’, and ‘Yudal’ — proved more tolerant than the other two — ‘Youngsan’ and ‘Tammi’. Because rapeseed is in the same family (Brassicacae) as Arabidopsis, one would expect that both share similar genes, with protein coding sequences that show >86% homology. We used a cDNA microarray enriched with Arabidopsis cold stress-related genes to examine the molecular background for differences in sensitivity among rapeseed varieties. Their hierarchical clustering patterns for expression were highly correlated with their cold-sensitive phenotypes. For example, expression patterns were similar between ‘Naehan’ and ‘Sandongchae’ as well as between ‘Yudal’ and our Arabidopsis samples. Using co-expression gene sets developed in this laboratory, we further analyzed expression patterns for three cultivars. Three distinct gene sets were commonly enriched in ‘Naehan’ against ‘Youngsan’ and ‘Tammi’, indicating that they could be the core gene sets that confer a high degree of cold tolerance in ‘Naehan’. Knockout plants with down-regulated expression of GSTF2, TRXh5, LTI30, and LACS4 were highly cold-sensitive whereas overexpression of those four genes led to a reversal of this sensitivity. In fact, overexpression of GSTF2 and LTI30 enhanced tolerance, with plants that over-expressed GSTF2 having extremely high tolerance that enabled them to acclimate almost completely to freezing conditions. By comparison, only about 40% of the wild type demonstrated a similar acclimation. Therefore, the genes described here could be useful in furthering our knowledge about other stress responses in rapeseed, thereby facilitating the production of cold-tolerant plants.     

The Inhibition Analysis of Two Heavy Metal ATPase Genes (NtHMA3a and NtHMA3b) in Nicotiana tabacum

Previous studies suggested that plants detoxified mercury and cadmium through similar mechanisms. A heavy metal ATPase (adenosine triphosphatase) gene, HMA3, plays a key role in the plant's cadmium detoxification. To prove whether HMA3 also participates in mercury detoxification in plants, an experiment was designed to inhibit the expressions of HMA3 genes (NtHMA3a and NtHMA3b) in tobacco plants. Results showed that plants’ tolerance to mercury ions had not changed after the expressions of NtHMA3a and NtHMA3b were inhibited. When mercury content was measured from the whole seedlings, no differences had been observed among wild-type, NtHMA3a-NtHMA3b-RNAi, and the empty-vector transgenic plants. HMA3 was not the key gene responsible for plants’ mercury ion uptake from soil. Although the mercury content in the root was higher than that in the shoot for each seedling, in each treatment, neither in shoots nor in roots were statistical differences in mercury content found among NtHMA3a-NtHMA3b-RNAi, empty-vector transgenic, and wild-type plants. After the expressions of NtHMA3a and NtHMA3b were inhibited, the movement of mercury ions from root to shoot had not been affected. HMA3 was not the key gene responsible for mercury ion transportation from root to shoot. When mercury content was measured from the whole seedling, no significant difference had been found among wild-type, NtHMA3a-NtHMA3b-RNAi, and the empty-vector transgenic plants. For mercury ion translocation across tonoplast, the main pathway might not be HMA3, but ABC (ATP-binding cassette) transporters.     

The cytosolic O-acetylserine(thiol)lyase gene is regulated by heavy metals and can function in cadmium tolerance

Regulation of the expression of the cytosolic O-acetylserine(thiol)lyase gene (Atcys-3A) from Arabidopsis thaliana under heavy metal stress conditions has been investigated. Northern blot analysis of Atcys-3A expression shows a 7-fold induction after 18 h of cadmium treatment. Addition of 50 microm CdCl(2) to the irrigation medium of mature Arabidopsis plants induces a rapid accumulation of the mRNA throughout the leaf lamina, the root and stem cortex, and stem vascular tissues when compared with untreated plants, as observed by in situ hybridization. High pressure liquid chromatography analysis of GSH content shows a transient increase after 18 h of metal treatment. Our results are compatible with a high cysteine biosynthesis rate under heavy metal stress required for the synthesis of GSH and phytochelatins, which are involved in the plant detoxification mechanism. Arabidopsis-transformed plants overexpressing the Atcys-3A gene by up to 9-fold show increased tolerance to cadmium when grown in medium containing 250 microm CdCl(2), suggesting that increased cysteine availability is responsible for cadmium tolerance. In agreement with these results, exogenous addition of cystine can, to some extent, also favor the growth of wild-type plants in cadmium-containing medium. Cadmium accumulates to higher levels in leaves of tolerant transformed lines than in wild-type plants.     

高等植物重金属耐性与超积累特性及其分子机理研究

由于重金属污染日益严重, 重金属在土壤_植物系统中的行为引起了人们的高度重视。高等植物对重金 属的耐性与积累性, 已经成为污染生态学研究的热点。近年来, 由于分子生态学等学科的发展, 有关植物对重金属的解毒和耐性机理、重金属离子富集机制的研究取得了较大进展。高等植物对重金属的耐性和积累在种间和基因型之间存在很大差异。根系是重金 属等土壤污染物进入植物的门户。根系分泌物改变重金属的生物有效性和毒性, 并在植物吸收重金属的过程中发挥重要作用。土壤中的大部分重金属离子都是通过金属转运蛋白进入根细胞, 并在植物体内进一步转运至液泡贮存。在重金属胁迫条件下植物螯合肽 (PC) 的合成是植物对胁迫的一种适应性反应。耐性基因型合成较多的PC, 谷胱甘肽 (GSH) 是合成PC的前体, 重金属与PC螯合并转移至液泡中贮存, 从而达到解毒效果。金属硫蛋白 (MTs) 与PC一样, 可以与重金属离子螯合, 从而降低重金属离子的毒性。该文从分子水平上论述了根系分泌物、金属转运蛋白、MTs、PC、GSH在重金属耐性及超积累性中的作用, 评述了近 10年来这方面的研究进展, 并在此基础上提出存在的问题和今后研究的重点。     

Insight into citric acid-induced chromium detoxification in rice (Oryza sativa. L)

Abstract

High content of chromium in plants hampers plants’ metabolism, disrupts plant growth and development. Therefore, improving plants’ tolerance to Cr toxicity is very essential. In our present study, we investigated the role of citric acid (CA) on chromium detoxification in terms of stress tolerance in rice. Application of CA under Cr stress restore the growth parameters, total protein content and membrane stability confirming that CA plays important role in Cr detoxification in rice. However, supplementation of CA under Cr stress caused no significant change in root Cr content but increased shoot Cr concentration (97?µg/g) compare with Cr stressed plant (24?µg/gm), suggesting that CA alleviates Cr toxicity by its chelating properties. Moreover, Fe content showed no significant changes due to CA supplementation under Cr stress, implying that Fe regulation is not involved with CA-mediated mitigation of Cr toxicity in rice. Furthermore, increased CAT, POD, and GR activity along with raised metabolites (glutathione and proline) indicates active participation in ROS scavenging and palliate the Cr toxicity in rice.     

Environmental-dependent proline accumulation in plants living on gypsum soils

Biosynthesis of proline—or other compatible solutes—is a conserved response of all organisms to different abiotic stress conditions leading to cellular dehydration. However, the biological relevance of this reaction for plant stress tolerance mechanisms remains largely unknown, since there are very few available data on proline levels in stress-tolerant plants under natural conditions. The aim of this work was to establish the relationship between proline levels and different environmental stress factors in plants living on gypsum soils. During the 2-year study (2009–2010), soil parameters and climatic data were monitored, and proline contents were determined, in six successive samplings, in ten taxa present in selected experimental plots, three in a gypsum area and one in a semiarid zone, both located in the province of Valencia, in south-east Spain. Mean proline values varied significantly between species; however, seasonal variations within species were in many cases even wider, with the most extreme differences registered in Helianthemum syriacum (almost 30 μmol g^?1 of DW in summer 2009, as compared to ca. 0.5 in spring, in one of the plots of the gypsum zone). Higher proline contents in plants were generally observed under lower soil humidity conditions, especially in the 2009 summer sampling preceded by a severe drought period. Our results clearly show a positive correlation between the degree of environmental stress and the proline level in most of the taxa included in this study, supporting a functional role of proline in stress tolerance mechanisms of plants adapted to gypsum. However, the main trigger of proline biosynthesis in this type of habitat, as in arid or semiarid zones, is water deficit, while the component of ‘salt stress’ due to the presence of gypsum in the soil only plays a secondary role.     

Role of Phytohormones in Regulating Heat Stress Acclimation in Agricultural Crops

Heat stress (HS) seriously affects crop growth, causing significant crop yield losses worldwide. The regulatory mechanisms controlling HS tolerance in plants are not well understood. Phytohormones are important molecules for coordinating myriad of phenomena related to plant growth and development. They are also essential endogenous signaling molecules that actively mediate numerous physiological responses under abiotic stress by triggering stress-responsive regulatory genes involved in plant growth. This review updates the central role of various phytohormones—indole acetic acid, gibberellic acid, abscisic acid, cytokinins, ethylene, salicylic acid, brassinosteroids, strigolactone, and jasmonic acid—in regulating the HS response so that plants can adapt to increasing temperature stress. We also reveal how these stress-responsive phytohormones switch on various regulatory gene(s) and genes encoding antioxidants and heat shock proteins (HSPs) to combat HS in various plant species.

     

Barley plant response to cadmium action as dependent on plant age

Age-dependent differences in the responses of 3-day-old (the phase of seed germination) and 7-day-old (the phase of seedlings) of barley (Hordeum vulgare L., cv. Zazerskii 85) plants to treatment with cadmium (100 μM). Four-day-long exposure to the cadmium solution retarded root growth and suppression of the photosynthetic apparatus in germinating seeds, whereas in seedlings, the roots continued to grow actively and parameters characterizing functioning of the photosynthetic apparatus were close to control values. In the presence of cadmium, in the roots of plants the level of HvPCS gene encoding phytochelatin synthase and the content of phytochelatins (PC) increased independently of the plant age. Distinctly, changes in the content of reduced glutathione (GSH) depended on the plant age: in 3-day-old seedlings the content of GSH reduced after 40-day-long exposure to cadmium; in contrast, in 7-day-old plants, it increased. We concluded that differences in the tolerance to cadmium of plants differing in age are related to their anatomic, morphological, physiological, and biochemical properties corresponding to particular developmental stages and also to differences in functioning of different mechanisms of metal detoxification, in particular, to activation of GSH synthesis and a great increase in the content of PC in the older plants.     

Cadmium and copper uptake and accumulation by Sesbania rostrata seedling, a N-fixing annual plant: implications for the mechanism of heavy metal tolerance

Sesbania rostrata,an annual tropical legume,has been found to be tolerant to heavy metals,with an unknown mechanism.It is a promising candidate species for revegetation at mine tailings.In this study,sequential extractions with five buffers and strong acids were used to extract various chemical forms of cadmium and copper in S.rostrata,with or without Cd or Cu treatments,so that the mechanisms of tolerance and detoxification could be inferred.Both metals had low transition rates from roots to the aboveground of S.rostrata.The transition ratio of Cd (4.00%) was higher than that of Cu (1.46%).The proportion of NaCl extracted Cd (mostly in proteinbinding forms) increased drastically in Cd treated plants from being undetectable in untreated plants.This suggests that Cd induced biochemical processes producing proteinlike phytochelatins that served as a major mechanism for the high Cd tolerance of S.rostrata.The case for Cu was quite different,indicating that the mechanism for metal tolerance in S.rostrata is metal-specific.The proportion of water-insoluble Cu (e.g.oxalate and phosphate) in roots increased significantly with Cu treatment,which partially explains the tolerance of S.rostrata to Cu.However,how S.rostrata copes with the high biotic activity of inorganic salts of Cu,which increased in all parts of the plant under Cu stress,is a question for future studies.Sesbania rostrata is among the very few N-fixing plants tolerant to heavy metals.This study provides evidence for the detoxification mechanism of metals in Sesbania rostrata.     

Plant molybdoenzymes and their response to stress

Molybdenum-containing enzymes catalyse basic reactions in the nitrogen, sulphur and carbon metabolism. Mo-enzymes contain at their catalytic sites an organometallic structure termed the molybdenum cofactor or Moco. In higher plants, Moco is incorporated into the apoproteins of four enzymes: nitrate reductase (EC 1.6.6.1-3; NR), xanthine dehydrogenase (EC 1.1.1.204; XDH), aldehyde oxidase (EC 1.2.3.1; AO) and sulphite oxidase (EC1.8.3.1; SO). Molybdoenzymes in plants are key enzymes in nitrate assimilation, purine metabolism, hormone biosynthesis, and most probably in sulphite detoxification. They are considered to be involved in stress acclimation processes and, therefore, elucidation of the mechanisms of their response to environmental stress conditions is of agricultural importance for the improvement of plant stress tolerance. Here we would like to give a brief functional and biochemical characteristic of the four plant molybdoenzymes and to focus mainly on their sensitivity to environmental stress factors.