干旱胁迫与不同储藏时间对杠柳种子萌发的影响

为了阐明干旱胁迫与不同储藏时间对杠柳种子萌发的影响机理,在室内采用不同浓度的聚乙二醇（PEG）模拟干旱胁迫条件,对3种不同储藏时间的杠柳种子进行了萌发期对比试验,观测了种子的总萌发率、发芽势、活力指数和胚根的生长情况,分析了抗旱特性,结果表明：不同程度的干旱胁迫与储藏时间对杠柳种子萌发均有一定的影响,当PEG-6000浓度为5%,对储藏1年杠柳种子的萌芽率和发芽势起促进作用,对储藏2年种子萌芽率轻度抑制,对储藏3年种子抑制影响较大,当PEG-6000浓度>5%,3种种子的萌芽率和发芽势均呈下降趋势,但当胁迫浓度增大到25%时,种子基本没有萌发。在不同的水分胁迫处理中,3种种子的活力指数和胚根生长速度一直呈下降趋势,尤其3年种子的下降幅度最大。不同储藏时间杠柳种子对干旱胁迫的适应性有差异,储藏1~2年的杠柳种子在PEG-6000浓度为5%~15%中萌发效果较好,且储藏1年种子好于2年种子,3年种子较差。3种种子的耐旱能力由强到弱依次为：1年种子>2年种子>3年种子。     

干旱胁迫对杠柳光合特性及抗氧化酶活性的影响

以盆栽杠柳幼苗为材料,用称重控水的方法设置4个含水量梯度,研究不同程度干旱胁迫对杠柳光合特性和抗氧化系统的影响,为黄河三角洲贝壳堤岛植被恢复过程中的物种培育和种植管理提供依据.结果表明:(1)随着干旱胁迫的加剧,杠柳幼苗叶片净光合速率(Pn)先增后降,蒸腾速率(Tr)和气孔导度(Gs)逐渐下降,胞间CO2浓度(Ci)先降后升,说明杠柳幼苗Pn下降在轻度干旱胁迫下主要是由气孔导度下降引起,而在重度胁迫下主要由非气孔因素引起.(2)随着干旱胁迫程度的加剧,杠柳幼苗瞬时水分利用效率(WUE)、表观光能利用效率(LUE)、羧化效率(CE)均呈现先增加后下降趋势;WUE在重度胁迫下才开始下降,比对照显著下降了39%,LUE(CE)则在中度胁迫下就开始下降,中度和重度胁迫分别比对照下降了48%(33%)、71%(69%).(3)随着干旱胁迫的加剧,杠柳叶片的超氧化物歧化酶(SOD)活性先升高后降低,过氧化物酶(POD)、过氧化氢酶(CAT)活性和MDA含量均逐渐增高,表明POD、CAT抑制杠柳叶片脂质过氧化效应不显著,SOD则抑制作用显著,且12.8%的贝壳沙含水量是SOD的耐受阈值,此时MDA含量比对照显著增加153%.     

Proline Accumulation in Leaves of Periploca sepium via Both Biosynthesis Up-Regulation and Transport during Recovery from Severe Drought

Drought resistance and recovery ability are two important requisites for plant adaptation to drought environments. Proline (Pro) metabolism has been a major concern in plant drought tolerance. However, roles of Pro metabolism in plant recovery ability from severe drought stress are largely unexplored. Periploca sepium Bunge has gained increasing attention for its adaptation to dry environments. Here, we investigated Pro metabolism in different tissues of P. sepium seedlings in the course of drought stress and recovery. We found that leaf Pro metabolism response during post-drought recovery was dependant on drought severity. Pro biosynthesis was down-regulated during recovery from -0.4 MPa but increased continually and notably during recovery from -1.0 MPa. Significant correlation between Pro concentration and Δ1-pyrroline-5-carboxylate synthetase activity indicates that Glutamate pathway is the predominant synthesis route during both drought and re-watering periods. Ornithine δ-aminotransferase activity was up-regulated significantly only during recovery from −1.0 MPa, suggesting positive contribution of ornithine pathway to improving plant recovery capacity from severe drought. In addition to up-regulation of biosynthesis, Pro transport from stems and roots also contributed to high Pro accumulation in leaves and new buds during recovery from −1.0 MPa, as indicated by the combined analysis of Pro concentration and its biosynthesis in stems, roots and new buds. Except its known roles as energy, carbon and nitrogen sources for plant rapid recovery, significant positive correlation between Pro concentration and total antioxidant activity indicates that Pro accumulation can also promote plant damage repair ability by up-regulating antioxidant activity during recovery from severe drought stress.     

荞麦种子萌发期多种抗氧化酶活性的研究

通过研究荞麦种子萌发期(0～7 d)超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)和抗坏血酸过氧化物酶(ASP)四种抗氧化酶的酶活性变化,以及对自由基等的清除效果,分析荞麦种子内在抗氧化酶系统在萌发期对细胞膜结构的修复及保护作用。实验结果表明:荞麦种子在萌发期产生的活性氧、自由基等对四种抗氧化酶均有激活效应,其中超氧化物歧化酶(SOD)活性最先上升,其他三种酶随其后被激活,四种酶的氧化反应存在一定关联性和协同作用。苦荞的抗氧化酶活性高于甜荞。     

Presoaking with hemin improves salinity tolerance during wheat seed germination

The aim of this study was to investigate whether presoaking with hemin, an inducer of heme oxygenase-1 (HO-1), could alleviate salinity damage during wheat seed germination in comparison with the pretreatment of a well-known nitric oxide (NO) donor sodium nitroprusside (SNP). The results showed that, compared with the samples upon 150 mM NaCl salt stress alone, both 10 ??M hemin and 200 ??M SNP pretreatments could (1) significantly attenuate the inhibition of seed germination and thereafter seedling growth; (2) induce HO expression; (3) enhance amylase activity, thus accelerating the formation of reducing sugar and total soluble sugar; and (4) increase the potassium (K) to sodium (Na) ratio, particularly in the shoot parts. Hemin and SNP could also increase antioxidant enzyme activities, including superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (POD), and ascorbate peroxidase (APX), thus resulting in the alleviation of oxidative damage, as indicated by the decrease of thiobarbituric acid reactive substances (TBARS) content. Moreover, semi-quantitative RT-PCR and isozymatic analysis illustrated that hemin and SNP pretreatment were able to up-regulate the expression of Mn-SOD (especially) and Cu/Zn-SOD gene, and activate SOD isozymatic activities. Since the addition of the NO scavenger methylene blue (MB) differentially reversed the above effects, the protective roles of hemin might be related to the induction of endogenous NO signal. Meanwhile, hemin-driven NO production was confirmed. Together, these results indicated that hemin exerted an advantageous effect on enhancing salinity tolerance during wheat seed germination, which might interact with NO.     

Seasonal changes in cold tolerance,water relations and accumulation of cations and compatible solutes in Atriplex halimus L.

We determined the cold (freezing) tolerance for field-grown plants of Atriplex halimus L. (Chenopodiaceae) in relation to plant ploidy level, leaf water relations and accumulation of osmolytes. Plants were grown at two sites in Murcia (Spain), having average minimum temperatures in the coldest month of 0.6 and 12.1 °C, respectively. LT₅₀ values derived from laboratory freezing tests, using leaves taken from the plants in early winter and in spring, showed greater tolerance for winter-harvested leaves; the acclimation was more pronounced at the cold-winter site. Cold tolerance was related positively with leaf K and/or Na accumulation. Analysis of compatible organic solutes (soluble sugars, total amino acids and quaternary ammonium compounds) showed that cold tolerance (measured both as LT₅₀ and as winter freezing damage in situ) was related most closely with leaf concentrations of soluble sugars. The leaf percentage dry matter content was related to both in vitro and in vivo tolerance, while tolerance in vitro was correlated also with the osmotic (potential ψ_s) and the relative water content. The two diploid (2n = 2x = 18) populations, from Spain, showed greater cold tolerance than the three tetraploid (2n = 4x = 36) populations, from North Africa and Syria, which may be related to the latter's greater cell size and consequent dilution of osmolytes. In this halophytic species, cold tolerance, like salinity and drought tolerance, seems to depend on osmotic adjustment, driven by vacuolar accumulation of K and Na and cytoplasmic accumulation of compatible solutes.     

Seasonal changes in cold tolerance,water relations and accumulation of cations and compatible solutes in Atriplex halimus L.

We determined the cold (freezing) tolerance for field-grown plants of Atriplex halimus L. (Chenopodiaceae) in relation to plant ploidy level, leaf water relations and accumulation of osmolytes. Plants were grown at two sites in Murcia (Spain), having average minimum temperatures in the coldest month of 0.6 and 12.1 °C, respectively. LT₅₀ values derived from laboratory freezing tests, using leaves taken from the plants in early winter and in spring, showed greater tolerance for winter-harvested leaves; the acclimation was more pronounced at the cold-winter site. Cold tolerance was related positively with leaf K and/or Na accumulation. Analysis of compatible organic solutes (soluble sugars, total amino acids and quaternary ammonium compounds) showed that cold tolerance (measured both as LT₅₀ and as winter freezing damage in situ) was related most closely with leaf concentrations of soluble sugars. The leaf percentage dry matter content was related to both in vitro and in vivo tolerance, while tolerance in vitro was correlated also with the osmotic (potential ψ_s) and the relative water content. The two diploid (2n = 2x = 18) populations, from Spain, showed greater cold tolerance than the three tetraploid (2n = 4x = 36) populations, from North Africa and Syria, which may be related to the latter's greater cell size and consequent dilution of osmolytes. In this halophytic species, cold tolerance, like salinity and drought tolerance, seems to depend on osmotic adjustment, driven by vacuolar accumulation of K and Na and cytoplasmic accumulation of compatible solutes.     

Genetics of drought tolerance during seed germination in tomato: inheritance and QTL mapping.

A BC1 population (N = 1000) of an F1 hybrid between a stress-sensitive Lycopersicon esculentum breeding line (NC84173; maternal and recurrent parent) and a germination stress-tolerant Lycopersicon pimpinellifolium accession (LA722) was evaluated for seed germination rate under drought stress (DS) (14% w/v polyethyleneglycol-8000, water potential approximately -680 kPa), and the most rapidly germinating seeds (first 3% to germinate) were selected. The 30 selected BC1 seedlings were grown to maturity and self pollinated to produce BC1S1 progeny seeds. Twenty of the 30 selected BC1S1 progeny families were evaluated for germination rate under DS and their average performance was compared with that of a "nonselected" BC1S1 population of the same cross. Results indicated that selection for rapid germination under DS significantly improved progeny germination rate under DS (selection gain = 19.6%), suggesting a realized heritability of 0.47 for rate of germination under DS in this population. The 30 selected BC1 plants were subjected to restriction fragment length polymorphism (RFLP) analysis, and marker allele frequencies for 119 RFLP markers which spanned 1153 cM of the 12 tomato chromosomes were determined. A distributional extreme marker analysis, which measures statistical differences in marker allele frequencies between a selected and a nonselected population, detected four quantitative trait loci (QTLs) for rate of germination under DS in this population. Of these, two QTLs, located on chromosomes 1 and 9, were contributed by the L. pimpinellifolium donor parent and had larger effects than the other two QTLs, located on chromosomes 8 and 12, which were contributed by the L. esculentum recurrent parent. A few BC1S1 families were identified with all or most of the identified QTLs and with germination rates comparable with that of LA722. These families should be useful for the development of germination drought-tolerant tomato lines using marker-assisted selection (MAS). The overall results indicate that drought tolerance during seed germination in tomato is genetically controlled and potentially could be improved by directional phenotypic selection or MAS.     

Effects of sucrose concentration and exogenous hormones on growth and periplocin accumulation in adventitious roots of Periploca sepium Bunge

Periploca sepium adventitious roots were cultured on 0.5 Murashige and Skoog solid media supplemented with exogenous hormones of different types and various concentrations, and with sucrose of different concentrations. Auxins (indole butyric acid (IBA) and naphthalene acetic acid (NAA)) and cytokinins (6-benzylaminopurine (BA) and kinetin (KT)) were selected as exogenous hormones for adventitious root proliferation. Compared with other hormones, IBA was the suitable auxin for adventitious root proliferation. Under this circumstance, every root explant generates 10?C15 adventitious roots (1- to 2-cm long) after 30?days. However, nothing but callus was induced on the root explants when NAA was added into the medium and the same result was achieved when auxins (IBA or NAA) were added into the media together with cytokinins (BA or KT). The suitable concentration of IBA for adventitious root proliferation was 1?C2?mg/l, when every root explant generated 10?C20 adventitious roots (1- to 2-cm long). The optimum concentration of IBA for periplocin accumulation was 1?mg/l, when the periplocin content reached 95.46???g/g. With regard to the investigation of sucrose concentration, 2?C3% (w/v) sucrose was favorable for adventitious root proliferation as every root explant in this concentration generated 10?C20 adventitious roots (1- to 2-cm long). The highest periplocin content (101.56???g/g) was achieved at 5% (w/v) sucrose, whereas the periplocin content at 5% (w/v) sucrose did not show significant difference from the periplocin content (95.38 and 98.47???g/g, respectively) at 3% (w/v) or 4% (w/v) sucrose.     

Effects of dynamic changes of nutrients on adventitious roots growth and periplocin accumulation in culture of Periploca sepium Bunge

In this study, the dynamics of biomass production, accumulation of periplocin, medium conditions and consumption of carbon, nitrogen and phosphate were investigated in adventitious roots culture of Periploca sepium in shake flasks over a period of 4 weeks. The biomass reached the maximum peak on day 24 (2.46 and 0.213 g of fresh and dry weight, respectively). Similarly, periplocin production got to a peak of 0.083 mg g^?1 on day 24, simultaneously. PH in medium had a decrease tendency at the beginning and then remained stable around 4.0, however, EC declined during the whole culture with the nutrients consumed. Sucrose was almost used up on the first 12 days which led to the increase in glucose and fructose. In case of nitrogen, consumption of ammonium is faster than nitrate at the beginning. Phosphate was almost consumed during the first 8 days. Based on the nutrients consumption, the adding time of nutrients (1/2 MS medium and 30 g L^?1 sucrose) was investigated and it obtained highest content of periplocin (0.106 mg g^?1) and yield (0.513 mg L^?1) on day 12.     

Involvement of compatible solutes in chill hardening-induced chilling tolerance in Jatropha curcas seedlings

Low temperature is a major environmental factor that affects metabolism, growth, development, distribution and production of chilling-sensitive plant, and J. curcas L. is a sustainable energy plant with great potential for biodiesel production due to the fact that its seed contains high oil content, which has attracted much attention worldwide. Our previous work found that the chill hardening improved the chilling tolerance of J. curcas seedlings (Ao et al. in Acta Physiologiae Plantarum 35:153–160, 2013), but its mechanism still remains elusive. In present work, the mechanism of chill hardening-induced chilling tolerance was further investigated in J. curcas seedlings. The results showed that chill hardening at 12 °C for 2 days markedly lowered osmotic and water potentials, which, in turn, maintained relative higher pressure potential in leaves of J. curcas seedlings compared with the control seedlings without chill hardening. In addition, chill hardening gradually increased compatible solutes proline, betaine and total soluble sugar contents compared with the control. When the control and hardened seedlings were subjected to chilling stress at 1 °C for 1–7 days, the chill-hardened seedlings significantly accumulated higher proline, betaine and total soluble sugar contents, which decreased osmotic and water potentials, and maintained higher pressure potential. To further understand the pathways of accumulation of compatible solutes, measurement of activities of ?¹-pyrroline-5-carboxylate synthetase (P5CS), glutamate dehydrogenase (GDH), ornithine aminotransferase (OAT), arginase, proline dehydrogenase (ProDH) and betaine dehydrogenase (BADH) showed that the chill hardening at 12 °C for 2 days obviously increased the activities of P5CS, GDH, OAT, arginase and BADH, as well as lowered ProDH activity both in leaves and stems of J. curcas seedlings to some extent as compared with the control. When the control and hardened seedlings were exposed to chilling stress at 1 °C for 1–7 days, the chill-hardened seedlings generally maintained significantly higher activities of P5CS, GDH, OAT, arginase and BADH. All above-mentioned results illustrated that the chill hardening could induce an accumulation of compatible solutes in leaves of J. curcas seedlings and compatible solutes play important roles in chill hardening-induced chilling tolerance.     

Fate of compatible solutes during dilution stress in Ectothiorhodospira halochloris

Abstract Ectothiorhodospira halochloris reacts upon enhancement of the water activity in the environment by excreting its major compatible solute, glycine betaine, thus decreasing the osmotic pressure inside the cell. A suddenly induced dilution stress leads to an overshoot of this reaction, so that more glycine betaine than necessary to compensate the external osmotic change is released. Subsequently the cells take up glycine betaine until they reach osmotic balance with the medium. E. halochloris possesses an active transport system that allows an uptake of glycine betaine against a concentration gradient. Glycine betaine is not metabolized in E. halochloris . Ectoine, a minor compatible solute of E. halochloris , is excreted in a similar manner to that of glycine betaine during dilution stress, whereas no excretion of the third compatible solute, trehalose, was detected.     

Fate of compatible solutes during dilution stress in Ectothiorhodospira marismortui

Abstract The proliferation of murine spleen cells stimulated by a T-cell mitogen such as phytohemagglutinin (PHA) or concanavalin A (ConA) was significantly suppressed when the mice were immunized with either the viable cells or the sonicate of Salmonella typhimurium but not of Escherichia coli . The suppression of T-cell proliferation caused by the sonicate of S. typhimurium was completely restored by addition of phorbol 12-myristate-13-acetate (PMA), an activator of protein kinase C (PKC). Western blots using anti-phosphotyrosine antibodies showed that the mitogen-induced tyrosine phosphorylation of 120-, 106-,94-,76-,68- and 57-kDa proteins in murine splenic T-cells was inhibited in the mice immunized with the viable cells but not the sonicate of S. typhimurium . These results suggest that the inhibition caused by the sonicate involves suppression of PKC activity, whilst that produced by viable cells involves down-regulation of tyrosine phosphorylation, and both inhibitions correlate with the induction of cell-mediated immunity in mice, as evidenced by the induction of delayed-type hypersensitivity reactions.     

Chickpea seed proteins: Modification during germination

Changes in the proteins of chickpea during a 12-day germination period are reported using techniques of gel filtration, DEAE-cellulose chromatography, polyacrylamide gel (PAG) electrophoresis and ultracentrifugation. In the ultracentrifuge, the total proteins of dormant seeds resolve into 3 components which have the sedimentation coefficients of 2.2 S, 6.9 S and 10.3 S respectively. On germination, the presence of fractions of lower sedimentation coefficient indicates possible degradation of these components; in the early stages, the degradation rate of the 7 S fraction is higher, while the 10 S fraction is broken down faster in the later stages. Gel filtration experiments indicate the possibility of degradation of high polymer into intermediary products. Increase in the relative mobility of protein components on PAG and elution constant on DEAE-cellulose chromatographs indicates an increase in the net negative charge of the protein fractions. The accumulation of subunits of the proteins is negligible during the germination period.     

Enhancement of tolerance of abiotic stress by metabolic engineering of betaines and other compatible solutes

The accumulation of compatible solutes, such as betaines, proline and sugar alcohols, is a widespread response that may protect plants against environmental stress. It is not yet fully understood how these compounds are involved in the stress tolerance of whole plants. Some plants have been genetically engineered to express enzymes that catalyze the synthesis of various compatible solutes. Some interventions have increased the tolerance of some crop plants to abiotic stress. Furthermore, analysis of such transgenic plants has begun to clarify the roles of compatible solutes in stress tolerance.     

Salt and drought tolerance of sugarcane under iso-osmotic salt and water stress: growth,osmolytes accumulation,and antioxidant defense

In order to discriminate between the ionic and osmotic components of salt stress, sugarcane (Saccharum officinarum L. cv. Co 86032) plants were treated with salt-NaCl or polyethylene glycol-PEG 8000 solutions (?0.7 MPa) for 15 days. Both the salt and PEG treatments significantly reduced leaf width, number of green leaves, and chlorophyll stability index. Osmotic adjustment (OA) indicated that both the stresses led to significant accumulation of osmolytes and sugars. Salt stressed plants appeared to use salt as an osmoticum while the PEG stressed plants showed an accumulation of sugars. Oxidative damage to membranes was not severe in plants subjected to salt or PEG stress. The salt stressed plants showed an increase in the activities of superoxide dismutase (SOD) and ascorbate peroxidase (APX), while PEG stress led to an increase in SOD but not APX activity as compared to the control. Thus, results indicate that the iso-osmotic salt or PEG stress led to differential responses in plants especially with respect to growth, OA, and antioxidant enzyme activities.     

Osmoadaptation mechanisms in prokaryotes: distribution of compatible solutes

Microorganisms respond to osmotic stress mostly by accumulating compatible solutes, either by uptake from the medium or by de novo synthesis. These osmotically active molecules preserve the positive turgor pressure required for cell division. The diversity of compatible solutes is large but falls into a few major chemical categories; they are usually small organic molecules such as amino acids or their derivatives, and carbohydrates or their derivatives. Some are widely distributed in nature while others seem to be exclusively present in specific groups of organisms. This review discusses the diversity and distribution of known classes of compatible solutes found in prokaryotes as well as the increasing knowledge of the genes and pathways involved in their synthesis. The alternative roles of some archetypal compatible solutes not subject to osmoregulatory constraints are also discussed.     

24-Epibrassinolide alleviates salt-induced inhibition of productivity by increasing nutrients and compatible solutes accumulation and enhancing antioxidant system in wheat (Triticum aestivum L.)

Two wheat (Triticum aestivum L.) cultivars, Sids 1 and Giza 168, were grown under non-saline or saline conditions (4.7 and 9.4 dS m^?1) and were sprayed with 0.00, 0.05 and 0.10 mg l^?1 24-epibrassinolide (EBL). Salt stress markedly decreased plant productivity and N, P, and K uptake, particularly in Giza 168. A follow-up treatment with 0.1 mg l^?1 EBL detoxified the stress generated by salinity and considerably improved the above parameters, especially in Sids 1. Organic solutes (soluble sugars, free amino acids, proline and glycinebetaine), antioxidative enzymes (superoxide dismutase, peroxidase, catalase and glutathione reductase), antioxidant molecules (glutathione and ascorbate) and Ca and Mg levels were increased under saline condition, and these increases proved to be more significant in salt-stressed plants sprayed with EBL, particularly at 0.1 mg l^?1 EBL with Sids 1. Sodium concentration, lipid peroxidation, hydrogen peroxide content and electrolyte leakage were increased under salt stress and significantly decreased when 0.1 mg l^?1 EBL was sprayed. Clearly, EBL alleviates salt-induced inhibition of productivity by altering the physiological and biochemical properties of the plant.     

Induction of chilling tolerance in wheat during germination by pre-soaking seed with nitric oxide and gibberellin

Chilling depresses seed germination and seedling establishment, and is one major constraint to grain yield formation in late sown winter wheat. Seeds of winter wheat (Triticum aestivum L.) were separately pre-soaked with sodium nitroprusside (SNP, as nitric oxide donor) and Gibberellic acid (GA₃) before germination and then germinated under low temperature. SNP and GA₃ pre-treatment increased seed germination rate, germination index, weights and lengths of coleoptile and radicle, while they decreased mean germination time and weight of seeds germinating under low temperature. Exogenous NO and GA₃ increased seed respiration rate and promoted starch degradation along with increased amylase activities. In addition, efficient antioxidant systems were activated by NO, and which effectively reduced concentrations of malondialdehyde and hydrogen peroxide (H₂O₂). Seedling growth was also enhanced by exogenous NO and GA₃ as a result of improved seed germination and maintenance of better reactive oxygen species homeostasis in seedling growing under chilling temperatures. It is indicated that exogenous NO was more effective than GA₃ in alleviating chilling stress during seed germination and seedling establishment in wheat.