Salt tolerance of Rhizobium species in broth cultures

Salt tolerance of five rhizobia strains was examined in broth cultures. Five levels of NaCl concentration were used and the optical density was taken as a measure for the vigour of bacterial growth. Rhizobium leguminosarum and R. meliloti were tolerant to high levels of salinity and growth curves in saline broth showed a similar pattern to the control level. Rhizobium japonicum, cowpea Rhizobium, and R. trifolii were intolerant to salt and showed a strong growth retardation with increasing salt concentration. Growth was inhibited at high levels of salinity. It is suggested that rhizobia sensitivity to salts may be partly responsible to the inhibition of nitrogen fixation by legumes growing under salt stress.     

Salinity and its effects on the functional biology of legumes

Salinity has plagued soil fertility and drastically affected growth and survival of glycophytes in irrigated regions of the world since the beginning of recorded history. It is particularly common in arid and semi-arid areas where evapotranspiration exceeds annual precipitation, and where irrigation is therefore necessary to meet crop water needs. Salt buildup in the soils and groundwater has threatened its productivity and sustainability. Plant responses to salt stress include an array of changes at the molecular, biochemical and physiological levels. Salt stress involves a water deficit induced by the salt concentration in the rhizosphere, resulting in disruption of homeostasis and ion distribution in the cell and denaturation of structural and functional proteins. As a consequence, salinity stress often activates cell signaling pathways including those that lead to synthesis of osmotically active metabolites, specific proteins, and certain free radical scavenging enzymes that control ion and water flux and support scavenging of oxygen radicals or chaperones. ROS detoxification forms an important defense against salt stress. Legumes are a key component of sustainable agriculture and can offer many economic and environmental benefits if grown more widely in crop rotations because of their ability to fix nitrogen in the root nodules in a symbiotic interaction with soil rhizobia. Due to their capacity to grow on nitrogen-poor soils, they can be efficiently used for improving saline soil fertility and help to reintroduce agriculture to these lands. However, in legumes, salt stress imposes a significant limitation of productivity related to the adverse effects on the growth of the host plant, the root-nodule bacteria, symbiotic development and finally the nitrogen fixation capacity. This paper reviews responses of legumes to salinity stress with emphasis on physiological and biochemical mechanisms of salt tolerance.     

Global inputs of biological nitrogen fixation in agricultural systems

Biological dinitrogen (N₂) fixation is a natural process of significant importance in world agriculture. The demand for accurate determinations of global inputs of biologically-fixed nitrogen (N) is strong and will continue to be fuelled by the need to understand and effectively manage the global N cycle. In this paper we review and update long-standing and more recent estimates of biological N₂ fixation for the different agricultural systems, including the extensive, uncultivated tropical savannas used for grazing. Our methodology was to combine data on the areas and yields of legumes and cereals from the Food and Agriculture Organization (FAO) database on world agricultural production (FAOSTAT) with published and unpublished data on N₂ fixation. As the FAO lists grain legumes only, and not forage, fodder and green manure legumes, other literature was accessed to obtain approximate estimates in these cases. Below-ground plant N was factored into the estimations. The most important N₂-fixing agents in agricultural systems are the symbiotic associations between crop and forage/fodder legumes and rhizobia. Annual inputs of fixed N are calculated to be 2.95 Tg for the pulses and 18.5 Tg for the oilseed legumes. Soybean (Glycine max) is the dominant crop legume, representing 50% of the global crop legume area and 68% of global production. We calculate soybean to fix 16.4 Tg N annually, representing 77% of the N fixed by the crop legumes. Annual N₂ fixation by soybean in the U.S., Brazil and Argentina is calculated at 5.7, 4.6 and 3.4 Tg, respectively. Accurately estimating global N₂ fixation for the symbioses of the forage and fodder legumes is challenging because statistics on the areas and productivity of these legumes are almost impossible to obtain. The uncertainty increases as we move to the other agricultural-production systems—rice (Oryza sativa), sugar cane (Saccharum spp.), cereal and oilseed (non-legume) crop lands and extensive, grazed savannas. Nonetheless, the estimates of annual N₂ fixation inputs are 12–25 Tg (pasture and fodder legumes), 5 Tg (rice), 0.5 Tg (sugar cane), <4 Tg (non-legume crop lands) and <14 Tg (extensive savannas). Aggregating these individual estimates provides an overall estimate of 50–70 Tg N fixed biologically in agricultural systems. The uncertainty of this range would be reduced with the publication of more accurate statistics on areas and productivity of forage and fodder legumes and the publication of many more estimates of N₂ fixation, particularly in the cereal, oilseed and non-legume crop lands and extensive tropical savannas used for grazing.     

Effects of salt stress and rhizobial inoculation on growth and nitrogen fixation of three peanut cultivars

Increasing soil salinity represents a major constraint for agriculture in arid and semi‐arid lands, where mineral nitrogen (N) deficiency is also a frequent characteristic of soils. Biological N fixation by legumes may constitute a sustainable alternative to chemical fertilisation in salinity‐affected areas, provided that adapted cultivars and inoculants are available. Here, the performance of three peanut cultivars nodulated with two different rhizobial strains that differ in their salt tolerance was evaluated under moderately saline water irrigation and compared with that of N‐fertilised plants. Shoot weight was used as an indicator of yield. Under non‐saline conditions, higher yields were obtained using N fertilisation rather than inoculation for all the varieties tested. However, under salt stress, the yield of inoculated plants became comparable to that of N‐fertilised plants, with minor differences depending on the peanut cultivar and rhizobial strain. Our results indicate that N fixation might represent an economical, competitive and environmentally friendly choice with respect to mineral N fertilisation for peanut cultivation under moderate saline conditions.     

Oxygation Enhances Growth, Gas Exchange and Salt Tolerance of Vegetable Soybean and Cotton in a Saline Vertisol

Impacts of salinity become severe when the soil is deficient in oxygen. Oxygation (using aerated water for subsurface drip irrigation of crop) could minimize the impact of salinity on plants under oxygen-limiting soil environments. Pot experiments were conducted to evaluate the effects of oxygation (12% air volume/volume of water) on vegetable soybean (moderately salt tolerant) and cotton (salt tolerant) in a salinized vertisol at 2, 8, 14, 20 dS/m ECe. In vegetable soybean, oxygation increased above ground biomass yield and water use efficiency (WUE) by 13% and 22%, respectively, compared with the control. Higher yield with oxygation was accompanied by greater plant height and stem diameter and reduced specific leaf area and leaf Na+ and Cl-concentrations. In cotton, oxygation increased lint yield and WUE by 18% and 16%, respectively, compared with the control, and was accompanied by greater canopy light interception, plant height and stem diameter. Oxygation also led to a greater rate of photosynthesis, higher relative water content in the leaf, reduced crop water stress index and lower leaf water potential. It did not, however, affect leaf Na+ or Cl- concentration. Oxygation invariably increased, whereas salinity reduced the K+ : Na+ ratio in the leaves of both species. Oxygation improved yield and WUE performance of salt tolerant and moderately tolerant crops under saline soil environments, and this may have a significant impact for irrigated agriculture where saline soils pose constraints to crop production.     

Growth and nitrogen fixation in Lotus japonicus and Medicago truncatula under NaCl stress: nodule carbon metabolism

Lotus japonicus and Medicago truncatula model legumes, which form determined and indeterminate nodules, respectively, provide a convenient system to study plant-Rhizobium interaction and to establish differences between the two types of nodules under salt stress conditions. We examined the effects of 25 and 50mM NaCl doses on growth and nitrogen fixation parameters, as well as carbohydrate content and carbon metabolism of M. truncatula and L. japonicus nodules. The leghemoglobin (Lb) content and nitrogen fixation rate (NFR) were approximately 10.0 and 2.0 times higher, respectively, in nodules of L. japonicus when compared with M. truncatula. Plant growth parameters and nitrogenase activity decreased with NaCl treatments in both legumes. Sucrose was the predominant sugar quantified in nodules of both legumes, showing a decrease in concentration in response to salt stress. The content of trehalose was low (less than 2.5% of total soluble sugars (TSS)) to act as an osmolyte in nodules, despite its concentration being increased under saline conditions. Nodule enzyme activities of trehalose-6-phosphate synthase (TPS) and trehalase (TRE) decreased with salinity. L. japonicus nodule carbon metabolism proved to be less sensitive to salinity than in M. truncatula, as enzymatic activities responsible for the carbon supply to the bacteroids to fuel nitrogen fixation, such as sucrose synthase (SS), alkaline invertase (AI), malate dehydrogenase (MDH) and phosphoenolpyruvate carboxylase (PEPC), were less affected by salt than the corresponding activities in barrel medics. However, nitrogenase activity was only inhibited by salinity in L. japonicus nodules.     

Effects of salinity, external K+/Na+ ratio and soil moisture on growth and ion content of Sesbania rostrata

Growth of Sesbania rostrata was decreased gradually with increase in root medium salinity (mixed salts or NaCl alone). Soil moisture or anoxia did not affect plant growth significantly. Higher K⁺/Na⁺ ratios in plant tissues compared to those in the root medium were found under different salinities. This indicated a high K⁺-Na⁺ selectivity, a characteristic generally considered unique to halophytes. S. rostrata is moderately salt tolerant and may be utilized as forage crop and green manure on saline land. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of NaCl on growth and nitrogen fixation and assimilation of inoculated and KNO3 fertilized Vicia faba L. and Pisum sativum L. plants

Faba bean (Vicia faba L. var. minor cv. Alborea) and pea (Pisum sativum L. cv. Lincoln) plants, inoculated with Rhizobium leguminosarum biovar. viciae strain GRA19, were treated with salt (100 mM NaCl) and/or nitrate (8 mM KNO₃) to test whether plants grown with inorganic-nitrogen are more tolerant to salinity than plants entirely reliant upon fixed nitrogen. According to the growth inhibition recorded, pea plants dependent on dinitrogen fixation proved more tolerant to salt stress than those N-fertilized, in contrast to results obtained for faba bean plants. This study therefore confirms that plants dependent on nitrogen fixation are not always more sensitive to salinity than are N-fertilized plants. Nitrate addition did not reduce the specific nitrogenase activity in pea, but did in faba bean. However, nodulation was inhibited in both legumes. The specific nitrogenase activity was more affected by salt treatment in N-fertilized plants for both legumes. The activity of the enzymes mediating ammonium assimilation in nodules (GS, NADH-GOGAT) was inhibited by salt stress both in N-fixing and in N-fertilized pea and faba bean plants.     

The effects of salinity and sodicity upon nodulation and nitrogen fixation in chickpea (Cicer arietinum)

Production of grain legumes is severely reduced in salt-affected soils because their ability to form and maintain nitrogen-fixing nodules is impaired by both salinity and sodicity (alkalinity). Genotypes of chickpea, Cicer arietinum, with high nodulation capacity under stress were identified by field screening in a sodic soil in India and subsequently evaluated quantitatively for nitrogen fixation in a glasshouse study in a saline but neutral soil in the UK. In the field, pH 8.9 was the critical upper limit for most genotypes studied but genotypes with high nodulation outperformed all others at pH 9.0-9.2. The threshold limit of soil salinity for shoot growth was at ECe 3 dS m(-1), except for the high-nodulation selection for which it was ECe 6. Nodulation was reduced in all genotypes at salinities above 3 dS m(-1) but to a lesser extent in the high-nodulation selection, which proved inherently superior under both non-saline and stress conditions. Nitrogen fixation was also much more tolerant of salinity in this selection than in the other genotypes studied. The results show that chickpea genotypes tolerant of salt-affected soil have better nodulation and support higher rates of symbiotic nitrogen fixation than sensitive genotypes.     

Genetic diversity analysis for narrow-leafed lupin (Lupinus angustifolius L.) by SSR markers

Molecular Biology Reports - Narrow-leafed lupin (Lupinus angustifolius L.) is used as grain legumes, fodder for livestock and green manure in the world and has a great potential to be developed as...     

Response of Contrasting Rice Genotypes to Zinc Sources under Saline Conditions

Abiotic stresses are among the major limiting factors for plant growth and crop productivity. Among these, salinity is one of the major risk factors for plant growth and development in arid to semi-arid regions. Cultivation of salt tolerant crop genotypes is one of the imperative approaches to meet the food demand for increasing population. The current experiment was carried out to access the performance of different rice genotypes under salinity stress and Zinc (Zn) sources. Four rice genotypes were grown in a pot experiment and were exposed to salinity stress (7 dS m⁻¹), and Zn (15 mg kg⁻¹ soil) was applied from two sources, ZnSO₄ and Zn-EDTA. A control of both salinity and Zn was kept for comparison. Results showed that based on the biomass accumulation and K⁺/Na⁺ ratio, KSK-133 and BAS-198 emerged as salt tolerant and salt sensitive, respectively. Similarly, based on the Zn concentration, BAS-2000 was reported as Zn-in-efficient while IR-6 was a Zn-efficient genotype. Our results also revealed that plant growth, relative water content (RWC), physiological attributes including chlorophyll contents, ionic concentrations in straw and grains of all rice genotypes were decreased under salinity stress. However, salt tolerant and Zn-in-efficient rice genotypes showed significantly higher shoot K⁺ and Zn concentrations under saline conditions. Zinc application significantly alleviates the harmful effects of salinity by improving morpho-physiological attributes and enhancing antioxidant enzyme activities, and the uptake of K and Zn. The beneficial effect of Zn was more pronounced in salt-tolerant and Zn in-efficient rice genotypes as compared with salt-sensitive and Zn-efficient genotypes. In sum, our results confirmed that Zn application increased overall plant’s performance under saline conditions, particularly in Zn in-efficient and tolerant genotypes as compared with salt-sensitive and Zn efficient rice genotypes.     

Biological nitrogen fixation in resource-poor agriculture in South Africa

Farm lands of resource-poor communities in South Africa are depleted of nutrients due to continuous mono-cropping, limited use of fertilisers, and sometimes leaching caused by high rainfall. Despite the well-known advantages of biological nitrogen fixation (BNF) in cropping systems, less than 10% of the grain crops planted annually in these areas are legumes. Using a participatory research and development approach, resource-poor farmers were introduced to conservation agriculture (CA) practices, including BNF, that promoted zero (or reduced) tillage, increased retention of soil cover, as well as crop diversification. Because crop rotation and intercropping of legumes with cereals are known to contribute to soil fertility while enhancing food security, resource-poor farmers from various Provinces in South Africa were trained on the benefits of legume culture for eight years. As a result, these resource-poor farmers did not only get training in inoculation techniques, but were also supplied with inoculants for use on their farms. Data collected from Farmers Demonstration Trials at Belvedere, Dumbarton and Lusikisiki, showed that the grain and fodder yield of maize planted after legumes, and maize intercropped with legumes, were comparable to those of maize receiving high N fertilizer dose (i.e. 54 kg N at planting and 54 kg N as top-dressing). The same data further showed thatRhizobium inoculation, when combined with application of low levels of P and K, significantly increased crop yields within farmers’ trial plots. BNF therefore offers a great opportunity for resource-poor farmers in South Africa to increase their crop yields and thus improve the quality of their livelihoods through the adoption of affordable and sustainable biological technologies that enhance soil fertility.     

Oil content and lipid composition of safflower (Carthamus tinctorius) irrigated with saline water under greenhouse and field conditions

An intensive process of land deterioration of some regions in Uzbekistan including the Aral Sea basin has led to a significant increase in soil salinity levels and consequently to a considerable reduction of total fertile soil area, as these lands are of little use for plant growth. The area is estimated to be more than 1.4 million ha of seabed. As a result, there was an immediate need to cultivate new crops capable of stopping the movement of sands and the enlargement of saline soils. Safflower (Carthamus tinctorius) is considered to be a moderately salt‐tolerant crop and, as such, one of a few crops well suited to the cropping systems of salt‐affected soils. It is used in Uzbekistan as a reserve crop when the culture of the main crop fails. In spite of the great economic importance of this oil, there is almost no available information in the literature on the effect of salinity on oil quality and its chemical ingredients. The purpose of the present study was to determine, in greenhouse and field experiments, how irrigation with saline water would influence content of oil, lipids and other lipophylic components in safflower. We found that irrigation of safflower with moderate concentrations of saline water seems feasible, as far as oil and lipid composition is concerned. Consequently, safflower might be a potential crop for lands of little use for plant growth in Uzbekistan or other similar sites in the world.     

Mutation breeding of grain legumes

Summary Grain legumes are an important group of crop plants. They provide an essential source of protein food for many developing countries, but their production has gone down in favour of more profitable crops like cereals. Therefore, genetic improvement of grain legumes is urgently needed. The primary aim of grain legume breeding must be the increase of production through adaptation to more advanced cropping schemes and reduction of crop losses. Symbiotic nitrogen fixation as developed by natural evolution does not always seem to be compatible with the needed substantial increase in yield: It is not supplying sufficient nitrogen and supplementation by fertilizer is rather uneconomic. By genetic manipulation of the plant's regulatory system nitrogen fixation may become more effective and tolerant to high soil nitrogen levels. Through a number of mutation breeding projects in different countries involving all important grain legume species it has been proven that mutation induction is a good tool for supplementing the genetic variation available from natural evolution and from selection by man. High-yielding cultivars have been developed from induced mutants, which eventually also possess a more efficient nitrogen fixation capacity.     

Azolla: Botany,physiology, and use as a green manure

This is a comprehensive review of literature pertaining to the aquatic fern Azolla and its nitrogen-fixing algal symbiont, Anabaena azollae. The preceding decade has witnessed an explosive growth in research on Azolla, and hopefully this paper will facilitate those efforts. The paper is broken into three major categories: botany, physiology and biochemistry, and agriculture. The botany section includes a world distribution map and reference tables for the 6 Azolla species, and includes the first review of literature on Anabaena azollae. The physiology and biochemistry section covers the range of topics from environmental factors to life processes and nitrogen fixation. Tables on the effect of growth regulators and on the rate of nitrogen fixation measured by acetylene reduction are presented. The agriculture section draws extensively from literature published in the People’s Republic of China and in the Democratic Republic of Vietnam. The major focus of this section is on the history and management practices for Azolla cultivation as a green manure for rice. The effect of weed suppression, use as a fish food and animal fodder, and the insects and diseases of Azolla are also discussed.     

Green manure technology: Potential,usage, and limitations. A case study for lowland rice

The growing concern about the sustainability of tropical agricultural systems stands in striking contrast to a world-wide decline in the use of soil-improving legumes. It is timely to assess the future role that soil-improving legumes may play in agricultural systems. This paper reviews recent progress, potential, and limitations of green manure technology, using lowland rice cropping systems as the example.Only a few legume species are currently used as green manures in lowland rice. Sesbania cannabina is the most widely used pre-rice green manure for rice in the humid tropics of Africa and Asia. Astragalus sinicus is the prototype post-rice green manure species for the cool tropics. Stem-nodulating S. rostrata has been most prominent in recent research. Many green manure legumes show a high N accumulation (80–100 kg N ha^-1 in 45–60 days of growth) of which the major portion (about 80%) is derived from biological N₂ fixation. The average amounts of N accumulated by green manures can entirely substitute for mineral fertilizer N at current average application rates. With similar N use efficiencies, green manure N is less prone to loss mechanisms than mineral N fertilizers and may therefore contribute to long-term residual effects on soil productivity.Despite a high N₂-fixing potential and positive effects on soil physical and chemical parameters, the use of green manure legumes for lowland rice production has declined dramatically world-wide over the last 30 years. Land scarcity due to increasing demographic pressure and a relatively low price of urea N are probably the main determining factors for the long-term reduction in pre-rice green manure use. Post-rice green manures were largely substituted for by high-yielding early-maturing grain legumes. Unreliability of green manure performance, non-availability of seeds, and labor intensive operations are the major agronomic constraints. The recognition and extrapolation of niches where green manures have a comparative advantage may improve an often unfavorable economic comparison of green manure with cash crop or fertilizer N. Socio-economic factors like the cost of land, labor, and mineral N fertilizer are seen to determine the cost-effectiveness and thereby farmers' adoption of sustainable pre-rice green manure technology. Hydrology and soil texture determine the agronomic competitiveness of a green manure with N fertilizers and with alternative cash crops. In general, the niches for pre-rice green manure are characterized by a relatively short time span available for green manure growth and a soil moisture regime that is unfavorable for cash crops (flood-prone rainfed lowlands with coarse-textured soils).Given the numerous agronomic and socio-economic constraints, green manure use is not seen to become a relevant feature of favourable rice-growing environments in the foreseeable future. However, in environments where soil properties and hydrology are marginal for food crop production, but which farmers may be compelled to cultivate in order to meet their subsistence food requirements, green manures may have a realistic and applicable potential.     

Sustainable wheat (Triticum aestivum L.) production in saline fields: a review

Abstract

A large part of global agricultural fields, including the wheat (Triticum aestivum L.) ones, are subjected to various stresses including salinity. Given the increasing world population, finding methods and strategies that can alleviate salinity stress on crop yield production is of outmost importance. The presented review has consulted more than 400 articles related to the clean and sustainable production of wheat in saline fields affected by biological, environmental, economical, and social parameters including the important issue of climate change (global warming). The negative effects of salt stress on plant growth and the techniques, which have been so far detected to alleviate salinity stress on wheat growth have been analyzed and presented. The naturally tolerant species of wheat can use a range of mechanisms to alleviate salinity stress including sodium exclusion, potassium retention, and osmoregulation. However, the following can be considered as the most important techniques to enhance wheat tolerance under stress: (1) the biotechnological (crop breeding), biological (soil microbes), and biochemical (seed priming) methods, (2) the use of naturally tolerant genotypes, and (3) their combined use. The proper handling of irrigation water is also an important subject, which must be considered when planting wheat in saline fields. In conclusion, the sustainable and cleaner production of wheat under salt stress is determined by a combination of different parameters including the biotechnological techniques, which if handled properly, can enhance wheat production in saline fields.     

Physiological adaptive mechanisms of plants grown in saline soil and implications for sustainable saline agriculture in coastal zone

There is large area of saline abandoned and low-yielding land distributed in coastal zone in the world. Soil salinity which inhibits plant growth and decreases crop yield is a serious and chronic problem for agricultural production. Improving plant salt tolerance is a feasible way to solve this problem. Plant physiological and biochemical responses under salinity stress become a hot issue at present, because it can provide insights into how plants may be modified to become more tolerant. It is generally known that the negative effects of soil salinity on plants are ascribed to ion toxicity, oxidative stress and osmotic stress, and great progress has been made in the study on molecular and physiological mechanisms of plant salinity tolerance in recent years. However, the present knowledge is not easily applied in the agronomy research under field environment. In this review, we simplified the physiological adaptive mechanisms in plants grown in saline soil and put forward a practical procedure for discerning physiological status and responses. In our opinion, this procedure consists of two steps. First, negative effects of salt stress are evaluated by the changes in biomass, crop yield and photosynthesis. Second, the underlying reasons are analyzed from osmotic regulation, antioxidant response and ion homeostasis. Photosynthesis is a good indicator of the harmful effects of saline soil on plants because of its close relation with crop yield and high sensitivity to environmental stress. Particularly, chlorophyll a fluorescence transient has been accepted as a reliable, sensitive and convenient tool in photosynthesis research in recent years, and it can facilitate and enrich photosynthetic research under field environment.