Genotypic difference in salinity tolerance in quinoa is determined by differential control of xylem Na loading and stomatal density

Quinoa is regarded as a highly salt tolerant halophyte crop, of great potential for cultivation on saline areas around the world. Fourteen quinoa genotypes of different geographical origin, differing in salinity tolerance, were grown under greenhouse conditions. Salinity treatment started on 10 day old seedlings. Six weeks after the treatment commenced, leaf sap Na and K content and osmolality, stomatal density, chlorophyll fluorescence characteristics, and xylem sap Na and K composition were measured. Responses to salinity differed greatly among the varieties. All cultivars had substantially increased K⁺ concentrations in the leaf sap, but the most tolerant cultivars had lower xylem Na⁺ content at the time of sampling. Most tolerant cultivars had lowest leaf sap osmolality. All varieties reduced stomata density when grown under saline conditions. All varieties clustered into two groups (includers and excluders) depending on their strategy of handling Na⁺ under saline conditions. Under control (non-saline) conditions, a strong positive correlation was observed between salinity tolerance and plants ability to accumulate Na⁺ in the shoot. Increased leaf sap K⁺, controlled Na⁺ loading to the xylem, and reduced stomata density are important physiological traits contributing to genotypic differences in salinity tolerance in quinoa, a halophyte species from Chenopodium family.     

盐生植物盐爪爪的耐盐生理特性探讨

当前土壤盐渍化日益严重,是限制植物生长的一个主要环境因子,然而在盐碱自然环境中生长着许多耐盐植物,为更好地了解盐生植物的耐盐机理,该文从无机离子Na+,K+,Ca2+含量、脯氨酸水平、水势变化、丙二醛含量和盐胁迫的表型等生理参数以及半定量RT-PCR检测脯氨酸合成关键酶基因(P5CS)的表达规律等方面探讨盐胁迫下盐爪爪的耐盐特性。结果表明:(1)随着盐浓度的升高,Na+在根和肉质化的叶中显著地富集,且叶中积累的Na+比根中更多;(2)在盐胁迫条件下,随着盐浓度的增加,脯氨酸的含量和脯氨酸合成关键酶基因的表达显著地增强;(3)Na+和脯氨酸是植物有效的渗透调节剂,可使处于低水势的植物细胞仍能从细胞外高浓度的盐溶液中吸收水分;(4)在0和700 mmol·L-1Na Cl处理下,盐爪爪肉质化叶中丙二醛的含量较其它处理高,这表明植物在这两个处理下可能受到了氧化胁迫;(5)从盐胁迫3个月的生长表型来看,低盐环境中生长的盐爪爪植株的生物量更多,肉质化的叶嫩且绿。综上所述,结合对野外生境的调查和实验室长期的盐胁迫表型结果表明盐爪爪的生长是需盐的,相对低的盐浓度环境对盐爪爪的生长是顺境,而无盐或高浓度盐环境对于盐爪爪的生长来说都是逆境。该研究结果为全面深入研究盐爪爪的耐盐特性,以及更好地利用盐爪爪的生物和基因资源改良土壤和提高作物和林木的耐盐性奠定基础。     

Anatomical adaptations and ionic homeostasis in aquatic halophyte Cyperus laevigatus L. Under high salinities

Salinity is extremely hazardous to agriculture worldwide and its expanding constantly. Soil of almost 100 countries facing salinity problem including Pakistan. Cyperus laevigatus also act as salinity indicator species is a naturally adapted halophyte dispersed in subtropical regions of world. Six populations of C. laevigatus were collected from different saline habitats to evaluate adaptations regarding anatomical and physiological characteristics. C. laevigatus is perfectly adapted to harsh environmental conditions like dry barren soils, saline lakes, hyper-saline wetlands and salt marshes. Ecological success of this species is due to plasticity in physiological and anatomical characteristics to adapt variable environmental conditions. C. laevigatus is a halophyte, exhibited increased biomass production in moderately saline habitat. Higher uptake of K⁺ occurs to compensate the uptake of Na⁺ ion contents, a striking feature of salt-tolerant and halophytic species. Accumulation of osmoprotectants like proline, free amino acids, soluble sugar and protein contribute significantly to osmotic adjustment. Stem thickness enhanced as salinity level of habitat increased to store water in parenchymatous tissues under physiological drought. Intensive sclerification in root cortex provide mechanical strength to plant as well as prevent the radial leakage of water. Well-developed aerenchyma, increased vascular bundle area, broader vessels, small and dense stomata are critical to cope with environmental hazards. Population of Jahlar lake showing maximum biomass production indicate that this species grows better in moderate salinities. Therefore, this species will prove very useful for revegetation of salt affected rangeland and prairies by direct growth of such halophytic ecotypes.     

Improving crop salt tolerance using transgenic approaches: An update and physiological analysis

Salinization of land is likely to increase due to climate change with impact on agricultural production. Since most species used as crops are sensitive to salinity, improvement of salt tolerance is needed to maintain global food production. This review summarises successes and failures of transgenic approaches in improving salt tolerance in crop species. A conceptual model of coordinated physiological mechanisms in roots and shoots required for salt tolerance is presented. Transgenic plants overexpressing genes of key proteins contributing to Na⁺ ‘exclusion’ (PM-ATPases with SOS1 antiporter, and HKT1 transporter) and Na⁺ compartmentation in vacuoles (V-H⁺ATPase and V-H⁺PPase with NHX antiporter), as well as two proteins potentially involved in alleviating water deficit during salt stress (aquaporins and dehydrins), were evaluated. Of the 51 transformations, with gene(s) involved in Na⁺ ‘exclusion’ or Na⁺ vacuolar compartmentation that contained quantitative data on growth and include a non-saline control, 48 showed improvements in salt tolerance (less impact on plant mass) of transgenic plants, but with only two tested in field conditions. Of these 51 transformations, 26 involved crop species. Tissue ion concentrations were altered, but not always in the same way. Although glasshouse data are promising, field studies are required to assess crop salinity tolerance.     

Soil drenching of paclobutrazol: An efficient way to improve quinoa performance under salinity

Salinity extent and severity is rising because of poor management practices on agricultural lands, possibility lies to grow salt‐tolerant crops with better management techniques. Therefore, a highly nutritive salt‐tolerant crop quinoa with immense potential to contribute for future food security was selected for this investigation. Soil drenching of paclobutrazol (PBZ; 20 mg l^?1) was used to understand the ionic relations, gaseous exchange characteristics, oxidative defense system and yield under saline conditions (400 mM NaCl) including normal (0 mM NaCl) and no PBZ (0 mg l^?1) as controls. The results revealed that salinity stress reduced the growth and yield of quinoa through perturbing ionic homeostasis with the consequences of overproduction of reactive oxygen species (ROS), oxidative damages and reduced photosynthesis. PBZ improved the quinoa performance through regulation of ionic homeostasis by decreasing Na⁺, Cl^?, while improving K⁺, Mg²⁺ and Ca²⁺ concentration. It also enhanced the antioxidative system including ascorbic acid, phenylalanine ammonia‐lyase, polyphenol oxidase and glutathione peroxidase, which scavenged the ROS (H₂O₂ and O₂^?‐) and lowered the oxidative damages (malondialdehyde level) under salinity in roots and more specifically in leaf tissues. The photosynthetic rate and stomatal conductance consequently improved (16 and 21%, respectively) in salt‐stressed quinoa PBZ‐treated compared to the non‐treated ones and contributed to the improvement of panicle length (33%), 100‐grain weight (8%) and grain yield (38%). Therefore, PBZ can be opted as a shotgun approach to improve quinoa performance and other crops under high saline conditions.     

Molecular Cloning and Functional Analysis of a Na<Superscript>+</Superscript>/H<Superscript>+</Superscript> Antiporter Gene <Emphasis Type="Italic">ThNHX1</Emphasis> from a Halophytic Plant <Emphasis Type="Italic">Thellungiella halophila</Emphasis>

Na⁺/H⁺ exchanger catalyzes the countertransport of Na⁺ and H⁺ across membranes. Using the rapid amplification of cDNA ends method, a Na⁺/H⁺ antiporter gene (ThNHX1) was isolated from a halophytic plant, salt cress (Thellungiella halophila). The deduced amino acid sequence contained 545 amino acid residues with a conserved amiloride-binding domain (⁸⁷LFFIYLLPPI⁹⁶) and shared more than 94% identity with that of AtNHX1 from Arabidopsis thaliana. The ThNHX1 mRNA level was upregulated by salt and other stresses (abscisic acid, polyethylene glycol, and high temperature). This gene partially complemented the Na⁺/Li⁺-sensitive phenotype of a yeast mutant that was deficient in the endosomal–vacuolar Na⁺/H⁺ antiporter ScNHX1. Overexpression of ThNHX1 in Arabidopsis increased salt tolerance of transgenic plants compared with the wild-type plants. In addition, the silencing of ThNHX1 gene in T. halophila caused the transgenic plants to be more salt and osmotic sensitive than wild-type plant. Together, these results suggest that ThNHX1 may function as a tonoplast Na⁺/H⁺ antiporter and play an important role in salt tolerance of T. halophila. Chunxia Wu, Xiuhua Gao, and Xiangqiang Kong contributed equally to this work.     

The role of cotyledon metabolism in the establishment of quinoa (Chenopodium quinoa) seedlings growing under salinity

A growth chamber experiment was conducted to assess the effect of salinity on emergence, growth, water status, photosynthetic pigments, osmolyte accumulation, and ionic content of quinoa seedlings (Chenopodium quinoa). The aim was to test the hypothesis that quinoa seedlings are well adapted to grow under salinity due to their ability to adjust the metabolic functionality of their cotyledons. Seedlings were grown for 21 days at 250 mM NaCl from the start of the germination. Germination percentage and cotyledon area were not affected by salt whereas seedling height decreased 15%. FW increased in both control and salt-treated cotyledons, but the increase was higher under salinity. DW only increased in salt-treated cotyledons. The DW/FW ratio did not show significant differences between treatments. Relative water content, chlorophyll, carotenoids, lipids, and proteins were significantly lower under salinity. Total soluble sugars, sucrose and glucose concentrations were higher in salt-treated than in control cotyledons. Ion concentration showed a different distribution pattern. Na⁺ and Cl^? concentrations were higher under salinity, while an inverse result was observed for K⁺ concentration. Proline and glycinebetaine concentrations increased under salinity, but the increase was higher in the former than the latter. The osmoprotective role of proline, glycinebetaine, and soluble sugars is discussed.     

淋洗与植物作用耦合对盐渍化土壤的改良效应

以甘肃秦王川引大灌区盐渍化土壤为背景,以当地5种耐盐植物为材料,采用根袋法盆栽试验动态研究了淋洗结合植物种植对盐渍化土壤改良的效应。结果表明:与种前相比,单纯的淋洗作用对土壤pH值影响不大,而淋洗结合植物种植明显降低了土壤pH值,且根际土壤pH值小于非根际土壤的,5种耐盐植物中霸王根际土壤pH值降低幅度最大,达0.6个单位。K+、Ca2+、Na+、Mg2+、Cl-和SO2-4在5种植物根际土壤中均有不同程度的富集,富集程度因物种的不同而不同,随培养时间的延长而呈波动状态。5种供试植物和对照组土壤中的6种主要的可溶性盐分离子随淋洗次数和培养时间的延长呈下降趋势。在培养120d后,单纯淋洗的土壤中K+、Ca2+、Na+、Mg2+、Cl-和SO2-4的含量相比种前平均分别降低了33.3%、26.1%、35.6%、32.5%、35.5%和36.3%,植物吸收带走的上述各离子的含量平均分别占种前的46.2%、8.1%、30.2%、7.2%和21.6%,其中霸王吸收带走的盐分离子最多,而淋洗结合种植植物的土壤中上述各离子的含量与种前相比平均分别降低了67.25%、63.73%、83.8%、67.5%、81.55%和78.46%,由此可见,淋洗结合植物种植的脱盐效果优于单纯淋洗,且土壤中主要的盐分离子Na+、Cl-和SO2-4的含量降低幅度最大,通过计算得出,在Cl-、SO2-4和Na+减少的总量中还有37.73%的Na+、38.22%的Cl-和35.14%的SO2-4的减少量是由植物根系的物理化学作用机制引起的。     

The SIDECAR POLLEN gene encodes a microspore-specific LOB/AS2 domain protein required for the correct timing and orientation of asymmetric cell division

Control of ion loading into the xylem has been repeatedly named as a crucial factor determining plant salt tolerance. In this study we further investigate this issue by applying a range of biophysical [the microelectrode ion flux measurement (MIFE) technique for non‐invasive ion flux measurements, the patch clamp technique, membrane potential measurements] and physiological (xylem sap and tissue nutrient analysis, photosynthetic characteristics, stomatal conductance) techniques to barley varieties contrasting in their salt tolerance. We report that restricting Na⁺ loading into the xylem is not essential for conferring salinity tolerance in barley, with tolerant varieties showing xylem Na⁺ concentrations at least as high as those of sensitive ones. At the same time, tolerant genotypes are capable of maintaining higher xylem K⁺/Na⁺ ratios and efficiently sequester the accumulated Na⁺ in leaves. The former is achieved by more efficient loading of K⁺ into the xylem. We argue that the observed increases in xylem K⁺ and Na⁺ concentrations in tolerant genotypes are required for efficient osmotic adjustment, needed to support leaf expansion growth. We also provide evidence that K⁺‐permeable voltage‐sensitive channels are involved in xylem loading and operate in a feedback manner to maintain a constant K⁺/Na⁺ ratio in the xylem sap.     

Molecular characterization and functional analysis of a vacuolar Na<Superscript>+</Superscript>/H<Superscript>+</Superscript> antiporter gene (<Emphasis Type="Italic">HcNHX1</Emphasis>) from <Emphasis Type="Italic">Halostachys caspica</Emphasis>

According to sequences of several vacuolar Na⁺/H⁺ antiporter genes from Xinjiang halophytic plants, a new vacuolar Na⁺/H⁺ antiporter gene (HcNHX1) from the halophyte Halostachys caspica was obtained by RACE and RT-PCR using primers corresponding to conserved regions of the coding sequences. The obtained HcNHX1 cDNA was 1,983 bp and contained a 1,656 bp open reading frame encoding a deduced protein of 551 amino acid residues. The deduced amino acid sequence showed high identity with other NHX1 we have cloned previously from halophyte in Xinjiang desert area. The phylogenetic analysis showed that HcNHX1 formed a clade with NHX homologs of Chenopodiaceae. Expression profiles under salt treatment and ABA induction were investigated, and the results revealed that expression of HcNHX1 was induced by NaCl and ABA. To compare the degree of salt tolerance, we over-expressed HcNHX1 in Arabidopsis. Two transgenic lines grew more vigorously than the wild type (WT) under salt stress. The analysis of ion contents indicated that under salt stress, the transgenic plants compartmentalized more Na⁺ in the leaves compared with wild-type plants. Together, these results suggest that the products of the novel gene HcNHX1 from halophyte Halostachys caspica is a functional tonoplast Na⁺/H⁺ antiporter.     

RETRACTED ARTICLE: Analysis of the physiological mechanism of salt-tolerant transgenic rice carrying a vacuolar Na+/H+ antiporter gene from Suaeda salsa

Salt stress is one of the most serious factors limiting the productivity of agricultural crops. Increasing evidence has demonstrated that vacuolar Na⁺/H⁺ antiporters play a crucial role in plant salt tolerance. In the present study, we expressed the Suaeda salsa vacuolar Na⁺/H⁺ antiporter SsNHX1 in transgenic rice to investigate whether this can increase the salt tolerance of rice, and to study how overexpression of this gene affected other salt-tolerant mechanisms. It was found that transgenic rice plants showed markedly enhanced tolerance to salt stress and to water deprivation compared with non-transgenic controls upon salt stress imposition under outdoor conditions. Measurements of ion levels indicated that K⁺, Ca²⁺ and Mg²⁺ contents were all higher in transgenic plants than in non-transformed controls. Furthermore, shoot V-ATPase hydrolytic activity was dramatically increased in transgenics compared to that of non-transformed controls under salt stress conditions. Physiological analysis also showed that the photosynthetic activity of the transformed plants was higher whereas the same plants had reduced reactive oxygen species generation. In addition, the soluble sugar content increased in the transgenics compared with that in non-transgenics. These results imply that up-regulation of a vacuolar Na⁺/H⁺ antiporter gene in transgenic rice might cause pleiotropic up-regulation of other salt-resistance-related mechanisms to improve salt tolerance.Fengyun Zhao and Zenglan Wang contributed equally to this work.     

Salinity Tolerance in Brassica Oilseeds

Brassica oilseed species now hold the third position among oilseed crops and are an important source of vegetable oil. The most common Brassica oil-seed crops grown for commercial purposes are rape seeds, (Brassica campestris L. and B. napus L.) and mustards (B. juncea (L.) Czern. & Coss. and B. carinata A.Br.). The other Brassica species such as B. nigra (L.) Koch and B. tournefortii Gouan are grown on a very small scale. Brassica napus, B. juncea, and B. carinata are amphidiploids, whereas B. campestris and B. nigra are diploid. Most of the Brassica species have been categorized as moderately salt tolerant, with the amphidiploid species being the relatively salt tolerant in comparison with the diploid species. Due to the higher salt tolerance of the amphidiploids, it has been suggested that their salt tolerance has been acquired from the A (B. campestris) and C (B. oleracea L.) genomes. However, significant inter- and intraspecific variation for salt tolerance exists within brassicas, which can be exploited through selection and breeding for enhancing salt tolerance of the crops. There are contrasting reports regarding the response of these species to salinity at different plant developmental stages, but in most of them it is evident that they maintain their degree of salt tolerance consistently throughout the plant ontogeny. The pattern of uptake and accumulation of toxic ions (Na⁺ and Cl^?), in tissues of plants subjected to saline conditions appears to be mostly due to mechanism of partial ion exclusion (exclusion of Na⁺ and/or Cl^?) in most of the species, although ion inclusion in some cases at intraspecific levels has also been observed. Maintenance of high tissue K⁺/Na⁺ and Ca^{2 +}/Na⁺ ratios has been suggested as an important selection criterion for salt-tolerance in brassicas. Osmotic adjustment has also been reported in Brassica plants subjected to saline conditions, but particularly to a large extent in salt-tolerant species or cultivars. The roles of important organic osmotica such as total soluble sugars, free amino acids, and free proline, which are central to osmotic adjustment, have been discussed. In canola, B. napus, no positive relationship has been observed between salt tolerance and erucic acid content of seed oil in different cultivars. Furthermore, glucosinolate content of the seed meal in canola generally increases with an increase in salt level of the growth medium. This review highlights the responses of potential Brassica crops to soil salinity from the whole plant to the molecular level. It also describes the efforts made during the past millennium in uncovering the mechanism(s) of salinity tolerance of these crops both at the whole plant and cellular levels. The important selection criteria, which are used by researchers to enhance the degree of salinity tolerance in brassicas, are summarized. In addition, the vital role of genetic engineering and molecular biology approaches to the improvement of salt tolerance in brassicas is emphasized.     

Regulation of cation transporter genes by the arbuscular mycorrhizal symbiosis in rice plants subjected to salinity suggests improved salt tolerance due to reduced Na<Superscript>+</Superscript> root-to-shoot distribution

Rice is a salt-sensitive crop whose productivity is strongly reduced by salinity around the world. Plants growing in saline soils are subjected to the toxicity of specific ions such as sodium, which damage cell organelles and disrupt metabolism. Plants have evolved biochemical and molecular mechanisms to cope with the negative effects of salinity. These include the regulation of genes with a role in the uptake, transport or compartmentation of Na⁺ and/or K⁺. Studies have shown that the arbuscular mycorrhizal (AM) symbiosis alleviates salt stress in several host plant species. However, despite the abundant literature showing mitigation of ionic imbalance by the AM symbiosis, the molecular mechanisms involved are barely explored. The objective of this study was to elucidate the effects of the AM symbiosis on the expression of several well-known rice transporters involved in Na⁺/K⁺ homeostasis and measure Na⁺ and K⁺ contents and their ratios in different plant tissues. Results showed that OsNHX3, OsSOS1, OsHKT2;1 and OsHKT1;5 genes were considerably upregulated in AM plants under saline conditions as compared to non-AM plants. Results suggest that the AM symbiosis favours Na⁺ extrusion from the cytoplasm, its sequestration into the vacuole, the unloading of Na⁺ from the xylem and its recirculation from photosynthetic organs to roots. As a result, there is a decrease of Na⁺ root-to-shoot distribution and an increase of Na⁺ accumulation in rice roots which seems to enhance the plant tolerance to salinity and allows AM rice plants to maintain their growing processes under salt conditions.     

Salinity tolerance of Aegilops cylindrica genotypes collected from hyper-saline shores of Uremia Salt Lake using physiological traits and SSR markers

Uremia Salt Lake, in North West Iran, has a hyper-saline water. A rare highly salinity-tolerant grass species, Aegilops cylindrica grows along its shores. Salinity tolerance of 44 genotypes of Ae. cylindrica, mainly collected from the Lake, was evaluated under control and 400 mM NaCl conditions using the physiological traits of plant height, dry weight, proline content, Na⁺ and K⁺ concentrations as well as K⁺/Na⁺ ratio. To evaluate the association between microsatellite (EST-SSR and SSR) markers and salinity tolerance, 35 primer pairs were used. Results showed a significant variation in the 44 genotypes studied in terms of their traits except for proline content. Ten most salinity-tolerant genotypes were identified based on their ability to survive, to produce the highest dry weight, and to sustain the least leaf Na⁺ concentration under salinity stress. The very high negative correlation found between Na⁺ concentration and salinity tolerance revealed the importance of individual or a combination of Na⁺ exclusion and excretion mechanisms contributing to the hyper-salinity tolerance of these genotypes. Clustering analysis based on marker data divided the 44 studied genotypes into two groups that were consistent with their saline and non-saline geographical areas. Results of molecular markers showed that four microsatellite markers (Xgwm312, Xwmc170, Xgwm291 and Xgwm410) generated a distinguished banding pattern in ten most salinity-tolerant genotypes. These results supported previous reports on their linkage with Na⁺ exclusion genes (HKT1;5 and HKT1;4) in wheat, which provided further evidence of usefulness of both genes and the linked markers to the salinity tolerance of the halophytic grass family species.     

Mechanisms of sodium uptake by roots of higher plants

The negative impact of soil salinity on agricultural yields is significant. For agricultural plants, sensitivity to salinity is commonly (but not exclusively) due to the abundance of Na⁺ in the soil as excess Na⁺ is toxic to plants. We consider reducing Na⁺ uptake to be the key, as well as the most efficient approach, to control Na⁺ accumulation in crop plants and hence to improve their salt resistance. Understanding the mechanism of Na⁺ uptake by the roots of higher plants is crucial for manipulating salt resistance. Hence, the aim of this review is to highlight and discuss recent advances in our understanding of the mechanisms of Na⁺ uptake by plant roots at both physiological and molecular levels. We conclude that continued efforts to investigate the mechanisms of root Na⁺ uptake in higher plants are necessary, especially that of low-affinity Na⁺ uptake, as it is the means by which sodium enters into plants growing in saline soils.