Changes on protein expression associated with salinity tolerance in Brassica cell cultures

The synthesis of proteins from salt-tolerant Brassica oleracea L. var. botrytis L. subvar. cauliflora (Gars.) DC. (cauliflower) cell cultures is modified in relation to controls in several features. There are nine newly induced polypeptides in tolerant cultures (absent in control conditions). Some of them are only present under low salt levels (85 mM NaCl). Another group seems to be representative of moderate and high salt levels (170 and 255 mM NaCl), and a third group is present in all the salt conditions tested. On the other hand, the synthesis of most of the polypeptides present in control conditions is modified in salt-tolerant cultures by increasing, decreasing or stopping their synthesis in any of the tested conditions. The relationship between these changes in Brassica and other plant systems is discussed.     

耐盐水稻种质资源的筛选

用0.8% NaCl溶液和国际水稻所水稻耐盐性9级评价方法对前人选留的38份水稻耐盐种质资源(用0.5%NaCl盐土筛选)进行了重复筛选,以期获得极端耐盐水稻种质资源.同时研究了不同盐浓度对水稻发芽率的影响.结果表明:高浓度盐溶液显著抑制水稻种子的发芽率,但品种间有较大差异;筛选出苗期极端耐盐品种6份,其中1级1份,2级4份,3级1份;极端耐盐水稻品种苗高的增长受高浓度盐溶液抑制,但品种间差异不显著;而盐溶液对极端耐盐水稻品种出叶速率的影响因品种而异.     

Comparative proteomic analysis of salt-responsive proteins in canola roots by 2-DE and MALDI-TOF MS

Salinity stress is a major abiotic stress that affects plant growth and limits crop production. Roots are the primary site of salinity perception, and salt sensitivity in roots limits the productivity of the entire plant. To better understand salt stress responses in canola, we performed a comparative proteomic analysis of roots from the salt-tolerant genotype Safi-7 and the salt-sensitive genotype Zafar. Plants were exposed to 0, 150, and 300 mM NaCl. Our physiological and morphological observations confirmed that Safi-7 was more salt-tolerant than Zafar. The root proteins were separated by two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry was applied to identify proteins regulated in response to salt stress. We identified 36 and 25 protein spots whose abundance was significantly affected by salt stress in roots of plants from the tolerant and susceptible genotype, respectively. Functional classification analysis revealed that the differentially expressed proteins from the tolerant genotype could be assigned to 14 functional categories, while those from the susceptible genotype could be classified into 9 functional categories. The most significant differences concerned proteins involved in glycolysis (Glyceraldehyde-3-phosphate dehydrogenase, Fructose-bisphosphate aldolase, Phosphoglycerate kinase 3), stress (heat shock proteins), Redox regulation (Glutathione S-transferase DHAR1, L-ascorbate peroxidase), energy metabolism (ATP synthase subunit B), and transport (V-type proton ATPase subunit B1) which were increased only in the tolerant line under salt stress. Our results provide the basis for further elucidating the molecular mechanisms of salt-tolerance and will be helpful for breeding salt-tolerant canola cultivars.     

西藏小麦耐盐性鉴定及分析

在3种不同盐胁迫浓度下对161份来源于西藏的小麦材料进行了苗期耐盐性鉴定,其中有147份西藏半野生小麦及14份密穗小麦.以普通小麦耐盐品种茶淀红麦和盐敏感品种PI94341为对照.在147份西藏半野生小麦中有14份表现为耐盐,占9.5%.14份密穗小麦中没有发现耐盐材料.在这161份材料中还发现了3份对盐胁迫浓度梯度不敏感的品种.结果表明,我国特有的西藏半野生小麦中蕴藏有丰富的耐盐种质,它们可供小麦耐盐育种用做亲本.     

NaCl-induced changes in plasma membrane lipids and proteins of <Emphasis Type="Italic">Zea mays</Emphasis> L. cultivars differing in their response to salinity

Two contrasting maize (Zea mays L.) cultivars, i.e., Giza 2 (salt tolerant) and Trihybrid 321 (salt sensitive), were grown hydroponically to study NaCl effect (100 mM) on root plasma membrane (PM) lipid and protein alterations. The PM total sterols of Trihybrid 321 were decreased while that of Giza 2 was increased in response to salt. Salt imposition had no significant effect on PM total glycolipids and proteins of both cultivars. The PM total phospholipids were increased in Trihybrid 321 but it did not change significantly in Giza 2 after salinity stress. Molecular percentage of PM phospholipids and fatty acids of both cultivars was different in absence (0 mM) and presence (100 mM) of salt. The most abundant phospholipids in untreated Trihybrid 321 PM were phosphatidylglycerol (PG), phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylserine (PS), which changed into PG, PS, phosphatidylinositol (PI) and PC after salt treatment. However, the dominant phospholipids of the control PM of Giza 2 were PC, PE, PS and PG, which changed into PG, PE, PS and diphosphatidylglycerol (DPG) after salt imposition. Over 60% of the total fatty acids were saturated in control and salinized PM of both cultivars, which was increased after salt stress. The predominant fatty acid in the control and salinized PM of Trihybrid 321 was C18:1 and C17:0, respectively. However, in control and treated PM of Giza 2, the predominant fatty acid was C17:0 and C20:0, respectively. Qualitative and quantitative differences in PM protein patterns were found in both cultivars with and without salt. PM lipid changes enhanced membrane integrity, reflected in different ion accumulation (Mansour et al. 2005), and hence salt tolerance of Giza 2.     

Spermine accumulation under salt stress

Polyamines have long been recognized to be linked to stress situations, and it is generally accepted that they have protective characteristics. However, little is known about their physiological relevance in plants subjected to long-term salt stress. In order to precise their importance, two rice (Oryza sativa) cultivars differing in their salt tolerance were salinized for 7, 14 and 21 days. The activities of some of the enzymes involved in polyamine metabolism, free polyamines and proline contents were evaluated. Arginine decarboxylase and S-adenosyl-L-methionine decarboxylase activities were reduced in both cultivars as a consequence of salt treatment. However, spermidine synthase activity was reduced in the salt tolerant cultivar (var Giza) but not in the salt sensitive (var El Paso), while no polyamine oxidase activity was detected. During the salinization period, putrescine and spermidine levels decreased in both cultivars, although less dramatically in Giza. Simultaneously, spermine accumulations occur in both varieties, while proline accumulation was major in the sensitive one. However, spermine accumulation induced by treatment with spermidine synthase inhibitor cyclohexylamine, determined no reduction in leaf injury associated with salt stress in both cultivars. The data presented suggest that spermine accumulation is not a salt tolerance trait.     

甜菜耐盐性筛选及其幼苗对盐胁迫的响应

为了探讨甜菜耐盐机理、不同耐盐等级甜菜盐耐性机制差异,对40份甜菜品种（系）用300mmol·L-1NaCl胁迫处理,通过芽期相对盐害率和苗期盐害指数确定不同品种耐盐等级,将不同耐盐等级材料进行NaCl浓度梯度（150、300mmol·L-1）处理,测定植株鲜重、超氧化物歧化酶（SOD）活性、丙二醛（MDA）含量和叶片质膜透性。试验结果如下：40份甜菜材料经过芽期和苗期耐盐性筛选最终确定耐盐等级极强的品种（系）1份（‘KWS0143’）,耐盐等级强的品种（系）6份（‘ACERO’等）,耐盐等级中等的品种（系）14份（‘HI0183’等）,耐盐等级弱的品种（系）5份（‘OVITO’等）;在同一盐胁迫时间下,不同耐盐等级材料的植株鲜重随盐浓度的增加呈下降趋势,品种‘KWS0143’和‘ACERO’变化幅度较小,品种‘OVATIO’下降幅度最大;不同耐盐等级材料随盐浓度增加和胁迫时间延长超氧化物歧化酶（SOD）活性呈现先上升而后下降的趋势,耐盐性极强的品种酶活性显著高于其他耐盐等级材料;不同耐盐等级材料随盐浓度增加和胁迫时间延长丙二醛（MDA）含量和叶片质膜透性均呈上升趋势,耐盐等级弱的品种（系）明显高于其他各个材料。     

High Salt and High pH Tolerance of New Isolated Rhizobium etli Strains from Egyptian Soils

Saline and alkaline soils are major problems contributing to the low productivity of common bean (Phaseolus vulgaris) in arid and semi-arid regions such as Egypt. Therefore our study was directed toward selecting strains more tolerant to these environmental stresses. Among seven Rhizobium etli strains isolated from Egyptian soils, we found a high degree of diversity. Strains EBRI 21 and EBRI 26 are highly tolerant to a salt concentration up to 4% NaCl. A positive correlation was found between the salt tolerance and the adaptation to alkaline pH (9). Strains EBRI 2 and EBRI 26 were adapted to elevated temperatures (42°C). The minimum level of low pH for the majority of Rhizobium etli strains from Egypt was pH 4.7 while the Colombian strain Rhizobium tropici CIAT 899 survived well at pH 4. At 0.4% NaCl, the symbiotic efficiency of the salt-tolerant strain EBRI 26 was superior in cultivar Giza 6 compared with the salt-sensitive strain EBRI 2 (18.2 compared with 13.9 nM C₂H₄ h^–1 mg^–1 nodule fresh weight). In the bean cultivar Saxa, nitrogen fixation was much more affected by high salt concentration (0.4% NaCl) than in the cultivar Giza 6 with both strains (3.9 and 3.8 nM C₂H₄ h^–1 mg^–1 nodule fresh weight, respectively). In general, stress of alkalinity had a less detrimental effect on nodulation and N₂ fixation than stress of salinity.     

Influence of host cultivars and Rhizobium species on the growth and symbiotic performance of Phaseolus vulgaris under salt stress

In order to study the effect of salt stress on the Rhizobium-common bean symbiosis, we investigated the response of both partners, separately and in symbiosis. The comparison of the behaviour of five cultivars of Phaseolus vulgaris differing in seed colour, growing on nitrates and different concentrations of NaCl, showed genotypic variation with respect to salt tolerance. Coco Blanc was the most sensitive cultivar, whereas SMV 29-21 was the most tolerant one. At the Rhizobium level, two strains previously selected for their salt tolerance were used: Rhizobium tropici strain RP163 and Rhizobium giardinii strain RP161. Their relative growth was moderately decreased at 250mM NaCl, but they were able to grow at a low rate in the presence of 342 mM NaCl. Their viability at the minimal inhibitory concentration was slightly affected. The effect of salinity on Rhizobium-plant association was studied by using the tolerant variety SMV 29-21 and the sensitive one Coco Blanc inoculated separately with both strains. In the absence of salinity, the strains induced a significantly higher number of nodules on the roots of the cultivar SMV 29-21 compared to those of Coco Blanc. Concerning effectiveness, both strains were similarly effective with SMV 29-21, but not with Coco Blanc. In the presence of salinity, Coco Blanc was more severely affected when associated with RP163 than with RP161. Salinity affected the nodulation development more than it affected the infection steps. Neither of the two strains was able to nodulate SMV 29-21 under saline conditions, in spite of the fact that this was considered the most salt-tolerant variety. The unsuccessful nodulation of SMV 29-21 could be related to the inhibition by salt of one or more steps of the early events of the infection process. In conclusion, N-fixing plants were found to be more sensitive to salt stress than those depending on mineral nitrogen. Evidence presented here suggests that a best symbiotic N2 fixation under salinity conditions could be achieved if both symbiotic partners, as well as the different steps of their interaction (early events, nodule formation, activity, etc.), are all tolerant to this stress.     

Salt Tolerant and Sensitive Rice Varieties Display Differential Methylome Flexibility under Salt Stress

DNA methylation has been referred as an important player in plant genomic responses to environmental stresses but correlations between the methylome plasticity and specific traits of interest are still far from being understood. In this study, we inspected global DNA methylation levels in salt tolerant and sensitive rice varieties upon salt stress imposition. Global DNA methylation was quantified using the 5-methylcytosine (5mC) antibody and an ELISA-based technique, which is an affordable and quite pioneer assay in plants, and in situ imaging of methylation sites in interphase nuclei of tissue sections. Variations of global DNA methylation levels in response to salt stress were tissue- and genotype-dependent. We show a connection between a higher ability of DNA methylation adjustment levels and salt stress tolerance. The salt-tolerant rice variety Pokkali was remarkable in its ability to quickly relax DNA methylation in response to salt stress. In spite of the same tendency for reduction of global methylation under salinity, in the salt-sensitive rice variety IR29 such reduction was not statistically supported. In ‘Pokkali’, the salt stress-induced demethylation may be linked to active demethylation due to increased expression of DNA demethylases under salt stress. In ‘IR29’, the induction of both DNA demethylases and methyltransferases may explain the lower plasticity of DNA methylation. We further show that mutations for epigenetic regulators affected specific phenotypic parameters related to salinity tolerance, such as the root length and biomass. This work emphasizes the role of differential methylome flexibility between salt tolerant and salt sensitive rice varieties as an important player in salt stress tolerance, reinforcing the need to better understand the connection between epigenetic networks and plant responses to environmental stresses.     

水稻资源芽期和苗期耐盐碱性综合评价及耐盐基因分析

培育耐盐碱水稻品种是应对全球人口日益增长的重要途径之一。文中以21份耐盐碱性不同的水稻品种(系)为材料,在芽期和苗期设置6个不同盐碱浓度处理,测定了发芽势、发芽率、芽长、根长、根数、芽鲜重和苗鲜总重等指标,以各指标盐害率的平均值作为耐盐碱性的综合评价标准。结果表明随着盐碱浓度的提升,对种子萌发和生长的抑制越明显。在1%NaCl加0.25%NaHCO3溶液处理下,发芽率盐害率变异最大,为0%–89.80%。在所有浓度处理下,各性状指标的盐害率都具有相似的变化趋势。筛选到4份综合耐盐碱能力强(大酒谷、日本晴、魔王谷和02428)和7份弱的种质资源。比较了4份耐盐碱强和3份耐盐碱弱的资源耐盐基因序列:OsHAL3和OsRR22基因在7份材料中没有差异,SKC1和DST基因在耐盐碱强和耐盐碱弱的品种之间有明显的变异。研究结果为进一步挖掘水稻耐盐碱基因和培育耐盐碱水稻新品种提供了种质资源和理论基础。     

不同耐盐性花生品种对NaCl胁迫的光合和抗逆生理响应特征

以较耐盐花生品种‘花育25’、‘鲁花12’和盐敏感品种‘海花1’、‘花育20’为材料,采用盆栽试验,设置0、1.0、2.0、3.0 g/kg土壤NaCl胁迫浓度梯度,测定其净光合速率、表观量子效率、气孔导度等光合特性,以及抗氧化酶活性和渗透调节物质含量等指标,明确NaCl胁迫条件下不同耐盐性花生品种光合和生理生化特性的适应特征。结果表明：(1)NaCl胁迫明显抑制各品种花生叶片光合作用,净光合速率随盐胁迫浓度的升高呈明显降低的趋势。(2)各品种花生叶片净光合速率均先随光照强度的增强而升高,当光强达到一定数值时趋于平稳;光补偿点和光饱点因品种和盐胁迫浓度差异较大,较高的盐胁迫浓度使叶片光补偿点升高,盐敏感品种的光饱和点降低。(3)盐胁迫条件下,各品种叶片表观量子效率和最大净光合速率均随盐胁迫强度的增加呈显著降低趋势,盐敏感品种利用弱光的能力在低盐胁迫下强于耐盐品种,其最大净光合速率在较高盐胁迫浓度(3.0 g/kg)下明显低于耐盐品种,但两类品种的叶片表观量子效率降幅相近(78.65%~88.00%)。(4)在NaCl胁迫下,耐盐品种叶片自由水含量显著高于盐敏感品种;在2.0～3.0 g/kg NaCl胁迫下,耐盐品种叶片SOD、CAT、POD活性和MDA含量的升降幅度均低于盐敏感品种;耐盐品种在NaCl浓度低于2.0 g/kg时的抗氧化能力明显高于盐敏感品种。研究发现,盐胁迫下花生品种抗盐耐逆的主要生理响应特征是提高光补偿点和最大净光合速率,增强叶片持水能力和物质代谢能力,以及提升抗氧化和渗透调节能力。     

The role of root apoplastic transport barriers in salt tolerance of rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Increasing soil salinity reduces crop yields worldwide, with rice being particularly affected. We have examined the correlation between apoplastic barrier formation in roots, Na⁺ uptake into shoots and plant survival for three rice (Oryza sativa L.) cultivars of varying salt sensitivity: the salt-tolerant Pokkali, moderately tolerant Jaya and sensitive IR20. Rice plants grown hydroponically or in soil for 1 month were subjected to both severe and moderate salinity stress. Apoplastic barriers in roots were visualized using fluorescence microscopy and their chemical composition determined by gas chromatography and mass spectrometry. Na⁺ content was estimated by flame photometry. Suberization of apoplastic barriers in roots of Pokkali was the most extensive of the three cultivars, while Na⁺ accumulation in the shoots was the least. Saline stress induced the strengthening of these barriers in both sensitive and tolerant cultivars, with increase in mRNAs encoding suberin biosynthetic enzymes being detectable within 30 min of stress. Enhanced barriers were detected after several days of moderate stress. Overall, more extensive apoplastic barriers in roots correlated with reduced Na⁺ uptake and enhanced survival when challenged with high salinity. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

大豆出苗期和苗期对盐胁迫的响应及耐盐指标评价

比较了4个大豆品种出苗期和苗期的耐盐性,测定150 mmol/L NaCl胁迫下的株高、下胚轴长、侧根数、地上干/鲜重、根干/鲜重、MDA含量、SOD活性、游离Pro含量,并将幼苗移栽到田间生长至成熟。结果表明:出苗期和苗期盐胁迫下4个品种的株高都显著降低、地上干/鲜重和根干/鲜重降低;出苗期胁迫侧根数减少,下胚轴长降低;而苗期胁迫侧根数增加,下胚轴长升高。未胁迫条件下,出苗期和苗期耐盐性强的品种22021-1的MDA含量和SOD活性高于耐盐性弱的品种22293-1。胁迫后,22021-1的MDA含量降低、SOD活性升高,其MDA含量分别比对照低51.03%和21.45%,SOD活性比对照高5.85%和45.77%;22293-1的MDA含量出苗期比对照高58.97%,苗期基本无变化,SOD活性出苗期和苗期升高都不显著。MDA和SOD可以作为大豆耐盐性筛选指标。早期的短时胁迫对不同耐盐性大豆品种的经济产量影响不同。     

Assessment of Genetic Diversity Among Rice Genotypes with Differential Adaptations to Salinity Using Physio-morphological and Molecular Markers

Breeding for salt tolerance using traditional screening and selection methods have been limited by the complex and polygenic nature of salt tolerance trait. This study was designed to evaluate some of the premium Basmati rice varieties for salt tolerance and to characterize genetic diversity among the rice varieties with different adaptations to saline soils using microsatellite (SSR) and ISSR markers. Plants of nine rice varieties including salt tolerant, salt sensitive and traditional Basmati, were grown in hydroponics using Yoshida solution containing 0 (control, pH 5.0) and 30 mM NaCl (Electrical conductivity 4.8 d/S, pH 5.0) and assessed for salinity tolerance on 1–9 scale as per IRRI standard evaluation system using seedling growth parameters, visual salt injuries and Na-K ratio. Physio-morphological studies showed that traditional Basmati rice varieties (Basmati 370 and HBC19) were more sensitive than the salt sensitive control variety, MI-48. SSR as well as ISSR marker systems generated higher levels of polymorphism and could distinguish between all the 9 rice cultivars. A total of 299 (225 polymorphic) and 437 (430 polymorphic) bands were detected using 28 UBC ISSR primers and 100 welldistributed mapped SSR markers, respectively. ISSR and SSR marker data-sets showed moderate levels of positive correlation (Mantel test, r = 0.43). The ISSR and SSR marker data analyzed using clustering algorithms showed two distinct clusters separating the Basmati (Basmati 370, HBC19 and CSR-30) from other non-aromatic indica (IR36, Pokkali, CSR10 and MI-48) rice varieties indicating greater divergence between Basmati and non-aromatic indica rice genotypes. Marker analysis showed a close relationship among the two traditional (Basmati 370 and HBC19) and cross-bred (CSR30) Basmati rice varieties and greater diversity between the two salt-tolerant genotypes, Pokkali and BR4-10.     

Salinity-induced enhancement of L-myo-inositol 1-phosphate synthase in rice (Oryza sativa L.)

The salt-tolerant varieties of rice (Oryza sativa L.) exhibit enhanced activity of the chloroplast form of L-myo-inositol 1-phosphate synthase (EC 5.5.4.1) under NaCl treatment either during the seedling stage or in fully grown plants during field growth. The salt-induced enhancement was noticeable only in chloroplasts from light-grown plants. The effects of these treatments on the cytosolic inositol synthase activity were less pronounced. While the effect of salt on the activity of the two forms was marginal in the salt-sensitive varieties during seedling growth, salinity affected the chloroplast inositol synthase activity adversely in these varieties during growth of the plants under field conditions. The salt-enhanced activities of inositol synthase(s) in the highly salt-tolerant varieties studied were found to be comparable to that observed in Porteresia coarctata, a halophytic wild rice species. The implications of these findings, which suggest a role of the inositol pathway in osmoregulation, are discussed.     

Genetic divergence among Egyptian cotton genotypes under water deficit conditions

Backgroundwater shortage is the greatest significant reason controlling crop yield that harmfully affects fruit production, boll shedding, lint yield and fiber properties quality in cotton plants. Objectives: this study was carried out at Sakha Agricultural Research Station, Kafr El-Sheikh, Agric. Res. Center, Egypt, during 2014 and 2015 growing seasons. A study was undertaken in cotton (G. barbadense L.) to detect the magnitude of genetic divergence among parental cotton genotypes and their F₁s cross combinations and to select most suitable combinations for tolerant water deficient. Results and conclusion: the data showed that the first eight main principal components with eigen value more than one are extracted from the complicated components, the total cumulative variance of these eight factors amount for about 85.28% of the total multivariate variance. Suggesting that these PCA scores might be used to summarize the original variables in any further analysis of the data. The female parent 10,229 formed wide cluster having wide divergent distance from the other groups. On the same trend, the extra-long varieties Giza 96 and Giza 45 formed unique groups, cluster number 8 and cluster 1 nearly related. Cluster number 7 and cluster 4 which consisted of two genotypes for each characterized by susstable to water deficit conditions. The cross combinations Giza 77 x Dandra, Giza 77 x Pima S6, Giza 96 x Australy and Minufy x Dandra grouped at the same cluster, however the cross combinations Giza 86 x Dandra and Giza 68 × 10,229 classified into two different clusters. The data revealed that the inter cluster distance was higher than the intra cluster, indicating wide genetic divergent among the studied genotypes. Principal components analysis is useful in identifying and most influential characters affecting genetic variation of population.     

Phenolic content and antioxidant activity in two contrasting Medicago ciliaris lines cultivated under salt stress

The objective of this study was to determine more indepth physiological and antioxidant responses in two Medicago ciliaris lines (a salt-tolerant line TNC 1.8 and a salt-sensitive line TNC 11.9) with contrasting responses to 100 mM NaCl. Under salt stress, both lines showed a decrease in total biomass and in the growth rate for roots, but TNC 1.8 was less affected by salt than TNC 11.9 in that it maintained leaf growth even in the presence of added salt. In both lines, salt stress mainly affected micronutrient status (Fe, Mn, Cu and Zn) rather than K nutrition, but the tolerant line TNC 1.8 accumulated more Na in leaves and less in roots compared with TNC 11.9. Salt stress decreased total soluble sugars (TSS) in all organs of the sensitive line TNC 11.9, whereas TSS was only reduced in roots of the tolerant line. The salt-induced drop in growth was linked to an increase in lipid peroxidation in roots of both lines and in leaves of the sensitive line. The salt-tolerant line TNC 1.8 was more efficient at managing salt-induced oxidative damage in leaves and to a lesser extent in roots than the salt-sensitive line TNC 11.9, by preserving higher phenolic compound and superoxide dismutase levels in both organs.