Functional screening for salinity tolerant genes from <Emphasis Type="Italic">Acanthus ebracteatus</Emphasis> Vahl using <Emphasis Type="Italic">Escherichia coli</Emphasis> as a host

Salinity reduces plant growth and crop production globally. The discovery of genes in salinity tolerant plants will provide the basis for effective genetic engineering strategies, leading to greater stress tolerance in economically important crops. In this study, we have identified and isolated 107 salinity tolerant candidate genes from a mangrove plant, Acanthus ebracteatus Vahl by using bacterial functional assay. Sequence analysis of these putative salinity tolerant cDNA candidates revealed that 65% of them have not been reported to be stress related and may have great potential for the elucidation of unique salinity tolerant mechanisms in mangrove. Among the genes identified were also genes that had previously been linked to stress response including salinity tolerance, verifying the reliability of this method in isolating salinity tolerant genes by using E. coli as a host.     

Salt stress tolerance; what do we learn from halophytes?

Abiotic environmental stresses can give rise to morphological, biochemical and molecular changes that negatively affect plant growth and productivity. Among these stresses, soil salinity is the major threat. To deal and control effects of high salinity on plants, it is important to understand their responses to salt stress that disturbs the homeostatic equilibrium at cellular and molecular levels. In this regard halophytes (salt tolerant plants) can provide superior models for the study of salt stress defense parameters compared to salt sensitive species (glycophytes). Halophytes use highly developed, complex systems to tolerate salinity by maintaining a low cytosolic Na⁺/K⁺ ratio, sequestration of Na⁺ into vacuoles that then provides the osmotic potential sustaining water influx. Under low intensity stress conditions that moderately and/or transiently affect ion imbalance, the set of responses all plants initiate will be mostly to engage measures that assure ion balance. High salinity, especially over a prolonged time period, will challenge plant survival, which then requires different strategies that employ a variety of mechanisms. Plasticity and connectivity of these diverse mechanisms is engrained in species- and family-specific evolutionary history and their genetic complexity. Highlighting differences in the genetic and biochemical makeup between glycophytes and halophytes allows for comparisons between their approaches towards high salinity. This review provides a brief overview about different strategies and mechanism used by plants to avoid or confine adverse effects of high salinity.     

Bioregulators: unlocking their potential role in regulation of the plant oxidative defense system

Key message

Plant bioregulators play an important role in managing oxidative stress tolerance in plants. Utilizing their ability in stress sensitive crops through genetic engineering will be a meaningful approach to manage food production under the threat of climate change.

Abstract

Exploitation of the plant defense system against oxidative stress to engineer tolerant plants in the climate change scenario is a sustainable and meaningful strategy. Plant bioregulators (PBRs), which are important biotic factors, are known to play a vital role not only in the development of plants, but also in inducing tolerance in plants against various environmental extremes. These bioregulators include auxins, gibberellins, cytokinins, abscisic acid, brassinosteroids, polyamines, strigolactones, and ascorbic acid and provide protection against the oxidative stress-associated reactive oxygen species through modulation or activation of a plant’s antioxidant system. Therefore, exploitation of their functioning and accumulation is of considerable significance for the development of plants more tolerant of harsh environmental conditions in order to tackle the issue of food security under the threat of climate change. Therefore, this review summarizes a new line of evidence that how PBRs act as inducers of oxidative stress resistance in plants and how they could be modulated in transgenic crops via introgression of genes. Reactive oxygen species production during oxidative stress events and their neutralization through an efficient antioxidants system is comprehensively detailed. Further, the use of exogenously applied PBRs in the induction of oxidative stress resistance is discussed. Recent advances in engineering transgenic plants with modified PBR gene expression to exploit the plant defense system against oxidative stress are discussed from an agricultural perspective.

     

Plant functional diversity in tropical Andean páramos

ABSTRACT

Background: Tropical high mountains present extreme daily temperature variations, frequent high air evaporative demands and seasonal differences in soil water availability. Plants have adapted to these conditions through different avoidance-tolerance mechanisms. This review focuses on plant-growth forms and their adaptive strategies.

Aims: This integrated review of páramo plant traits aims at contributing to understanding the functioning of plant-growth forms and their significance on ecosystem properties under environmental climate and land-use changes.

Methods: Plant responses are presented along avoidance-tolerance gradients considering three main aspects: freezing resistance, water relations and gas exchange characteristics. Results from 45 herbaceous and 42 woody species along elevational gradients in the Venezuelan high Andes were analysed.

Results: Leaf supercooling is the common avoidance response of woody plants to night-time freezing temperatures, while herbaceous plants tolerate frost. Trees and caulescent rosettes maintain more positive leaf water potentials under water deficit conditions compared to more tolerant herbaceous species. All plant growth-forms showed strong stomatal control under dry-season conditions.

Conclusions: Páramo plant growth-forms may be separated according to an avoidance-tolerance gradient in response to water deficit and low temperature resistance. Woody growth-forms tend to avoid both freezing and water stress, while herbaceous forms tolerate frost and resist an unfavourable water status. Grasses and cushion plants are at the tolerant extreme of the gradient and coincide in that both reach the highest elevations in the páramo. Andean giant rosettes are freezing avoidant, particularly susceptible to water deficit and the most vulnerable, of all growth-forms, to changing environmental conditions.     

Yeast functional screen to identify genetic determinants capable of conferring abiotic stress tolerance in <Emphasis Type="Italic">Jatropha curcas</Emphasis>

Background  

Environmentally inflicted stresses such as salinity and drought limit the plant productivity both in natural and agricultural system. Increasing emphasis has been directed to molecular breeding strategies to enhance the intrinsic ability of plant to survive stress conditions. Functional screens in microorganisms with heterologous genes are a rapid, effective and powerful tool to identify stress tolerant genes in plants. Jatropha curcas (Physic nut) has been identified as a potential source of biodiesel plant. In order to improve its productivity under stress conditions to benefit commercial plantations, we initiated prospecting of novel genes expressed during stress in J. curcas that can be utilized to enhance stress tolerance ability of plant.     

Correction to: The biotechnological importance of the plant-specific NAC transcription factor family in crop improvement

Climate change, malnutrition, and food insecurity are the inevitable challenges being faced by the agriculture sector today. Plants are susceptible to extreme temperatures during the crucial phases of flowering and seed development, and elevated carbon levels also lead to yield losses. Productivity is also affected by floods and droughts. Therefore, increasing plant yield and stress tolerance are the priorities to be met through novel biotechnological interventions. The contributions of NAC genes towards enhancing plant survivability under stress is well known. Here we focus on the potential of NAC genes in the regulation of abiotic stress tolerance, secondary cell wall synthesis, lateral root development, yield potential, seed size and biomass, ROS signaling, leaf senescence, and programmed cell death. Once naturally tolerant candidate NAC genes have been identified, and the nature of their association with growth and fitness against multi-environmental stresses has been determined, they can be exploited for building inherent tolerance in future crops via transgenic technologies. An update on the latest developments is provided in this review, which summarizes the current understanding of the roles of NAC in the establishment of various stress-adaptive mechanisms in model and food crop plants.

     

Antiporter NHX2 differentially induced in Mesembryanthemum crystallinum natural genetic variant under salt stress

Plants exhibit several mechanisms to survive under high salinity conditions. The uptake and compartmentalization of Na⁺ ion by the NHX antiporter is a crucial mechanism in homeostasis maintenance. Therefore, we evaluated McNHX2 gene expression and several physiological responses induced in three natural genetic variants of ice plants under salt stress. Based on morphology and growth behavior of wild type populations from an arid region of northwestern Mexico, we identified three ice plant natural genetic variants and called P0, P9, and P11. Several physiological parameters, such as water potential, relative water content, chlorophyll, and Na⁺ and K⁺ ion contents from all natural genetic variants exhibited a differential response under high salinity conditions. Specifically, the P0 variant showed lower water potential changes and least perturbation of Na⁺/K⁺ ratio than those of the P9 and P11 variants under saline conditions, suggesting that the P0 variant is the most salt tolerant. Unexpectedly, McNHX2 expression was repressed in the P11 variant while it was upregulated in the P0 and P9 variants under saline treatments. The McNHX2 gene was sequenced showing 15 introns and 16 exons; polymorphisms were found among the cDNAs sequences from the three natural genetic variants. All these data suggest that differential responses to high salinity involve different mechanisms operating in each variant for counteracting saline stress effects.

     

Expression of cold and drought regulatory protein (CcCDR) of pigeonpea imparts enhanced tolerance to major abiotic stresses in transgenic rice plants

Main conclusion

Transgenic rice expressing pigeonpea Cc CDR conferred high-level tolerance to different abiotic stresses. The multiple stress tolerance observed in CcCDR -transgenic lines is attributed to the modulation of ABA-dependent and-independent signalling-pathway genes.

Stable transgenic plants expressing Cajanus cajan cold and drought regulatory protein encoding gene (CcCDR), under the control of CaMV35S and rd29A promoters, have been generated in indica rice. Different transgenic lines of CcCDR, when subjected to drought, salt, and cold stresses, exhibited higher seed germination, seedling survival rates, shoot length, root length, and enhanced plant biomass when compared with the untransformed control plants. Furthermore, transgenic plants disclosed higher leaf chlorophyll content, proline, reducing sugars, SOD, and catalase activities, besides lower levels of MDA. Localization studies revealed that the CcCDR-GFP fusion protein was mainly present in the nucleus of transformed cells of rice. The CcCDR transgenics were found hypersensitive to abscisic acid (ABA) and showed reduced seed germination rates as compared to that of control plants. When the transgenic plants were exposed to drought and salt stresses at vegetative and reproductive stages, they revealed larger panicles and higher number of filled grains compared to the untransformed control plants. Under similar stress conditions, the expression levels of P5CS, bZIP, DREB, OsLEA3, and CIPK genes, involved in ABA-dependent and-independent signal transduction pathways, were found higher in the transgenic plants than the control plants. The overall results amply demonstrate that the transgenic rice expressing CcCDR bestows high-level tolerance to drought, salt, and cold stress conditions. Accordingly, the CcCDR might be deployed as a promising candidate gene for improving the multiple stress tolerance of diverse crop plants.

     

The effect of salt stress on polyamine biosynthesis and content in mung bean plants and in halophytes

The activity of L-arginine decarboxylase (EC 4.1.1.19) and L-ornithine decarboxylase (EC 4.1.1.17), polyamine content, and incorporation of arginine and ornithine into polyamines, were determined in mung bean [Vigna radiata (L.) Wilczek] plants subjected to salt (hypertonic) stress (NaCl at 0.51–2.27 MPa). Changes in enzyme activity in response to hypotonic stress were determined as well in several halophytes [Pulicaria undulata (L.), Kostei, Salsola rosmarinus (Ehr.) Solms-Laub, Mesembryanthemum forskahlei Hochst, and Atriplex halimus L.]. NaCl stress, possibly combined with other types of stress that accompanied the experimental conditions, resulted in organ-specific changes in polyamine biosynthesis and content in mung bean plants. The activity of both enzymes was inhibited in salt-stressed leaves. In roots, however, NaCl induced a 2 to 8-fold increase in ornithine decarboxylase activity. Promotion of ornithine decarboxylase in roots could be detected already 2 h after exposure of excised roots to NaCl, and iso-osmotic concentrations of NaCl and KCl resulted in similar changes in the activity of both enzymes. Putrescine level in shoots of salt-stressed mung bean plants increased considerably, but its level in roots decreased. The effect of NaCl stress on spermidine content was similar, but generally more moderate, resulting in an increased putrescine/spermidine ratio in salt-stressed plants. Exposure of plants to NaCl resulted also in organ-specific changes in the incorporation of both arginine and ornithine into putrescine: incorporation was inhibited in leaf discs but promoted in excised roots of salt-stressed mung bean plants. In contrast to mung bean (and several other glycophytes), ornithine and arginine decarboxylase activity in roots of halophytes increased when plants were exposed to tap water or grown in a pre-washed soil—i.e. a hypotonic stress with respect to their natural habitat. NaCl, when present in the enzymatic assay mixture, inhibited arginine and ornithine decarboxylase in curde extracts of mung bean roots, but did not affect the activity of enzymes extracted from roots of the halophyte Pulicaria. Although no distinct separation between NaCl stress and osmotic stress could be made in the present study, the data suggest that changes in polyamines in response to NaCl stress in mung bean plants are coordinated at the organ level: activation of biosynthetic enzymes concomitant with increased putrescine biosynthesis from its precursors in the root system, and accumulation of putrescine in leaves of salt-stressed plants. In addition, hypertonic stress applied to glycophytes and hypotonic stress applied to halophytes both resulted in an increase in the activity of polyamine biosynthetic enzymes in roots.     

The role of two superoxide dismutase mRNAs in rye aluminium tolerance

Aluminium (Al) is the main factor that limits crop production in acidic soils. There is evidence that antioxidant enzymes such as superoxide dismutase (SOD) play a key role against Al‐induced oxidative stress in several plant species. Rye is one of the most Al‐tolerant cereals and exudes both citrate and malate from the roots in response to Al. The role of SOD against Al‐induced oxidative stress has not been studied in rye. Al accumulation, lipid peroxidation, H₂O₂ production and cell death were significantly higher in sensitive than in tolerant rye cultivars. Also, we characterised two genes for rye SOD: ScCu/ZnSOD and ScMnSOD. These genes were located on the chromosome arms of 2RS and 3RL, respectively, and their corresponding hypothetical proteins were putatively classified as cytosolic and mitochondrial, respectively. The phylogenetic relationships indicate that the two rye genes are orthologous to the corresponding genes of other Poaceae species. In addition, we studied Al‐induced changes in the expression profiles of mRNAs from ScCu/ZnSOD and ScMnSOD in the roots and leaves of tolerant Petkus and sensitive Riodeva rye. These genes are mainly expressed in roots in both ryes, their repression being induced by Al. The tolerant cultivar has more of both mRNAs than the sensitive line, indicating that they are probably involved in Al tolerance.     

Heritable variability induced by the in vitro culture and genetic improvement of cultivated plants

Abstract

Genetic variability is found among plants derived from in vitro cultures of somatic cells. A number of different factors, such as the pre-existing genetic variation developed in vivo during tissue differentiation, the variation induced during the in vitro culture and also the selection for specific genotypes during plant regeneration, are considered as possible causes of the phenomenon.

The nature of the genetic changes induced in somaclones (variation in chromosome number, gross and cryptic chromosomal rearrangements, transposition of genetic elements, gene amplification and somatic gene rearrangements) is also discussed.