Photosynthetic pigments and photosynthetic products of endophyte-infected and endophyte-free Lolium perenne L. under drought stress conditions

Endophyte-infected (EI) seeds of Lolium perenne L. were used to attain endophyte-free (EF) population by heating the seeds at 43°C for 15 min and then 57°C for 25 min. Relative water content (RWC), chlorophyll, soluble sugar and starch content of EI and EF populations under normal and drought stress conditions were compared to investigate the effect of endophyte infection on the host plant. Under severe stress, RWC of EI leaf was significantly higher than that of EF leaf, i.e. EI plants took more advantages over EF plants in water-holding ability. Under mild stress, endophyte could enhance soluble sugars in host plants to improve their osmotic ability. With stress intensification, the improvement of endophyte no longer existed, and more photosynthetic products (such as starch) accumulated in EI plants to survive through the undesirable conditions. In the next spring, EI populations will recover more rapidly than EF populations. The biomass of a population is closely related to its photosynthesis. Under severe stress, EI population significantly accumulated more biomass than EF population. As far as photosynthetic pigments were concerned, contents of Chla, Chlb and Car of EI plants were close to those of EF plants, which suggested that endophyte infection didn’t alleviate photosynthetic pigments from being destroyed by drought stress, and endophyte might improve photosynthesis ability of its host plant in other ways.     

Photosynthetic pigments and photosynthetic products of endophyte-infected and endophyte-free Lolium perenne L.under drought strees conditions

Endophyte-infected fED seeds of Lolium perenne L.were used to attain endophyte-free fED population by heating the seeds at 43℃ for 15 min and then 57℃ for 25 min.Relative water content(RWC),chlorophyll,soluble sugar and starch content of EI and EF populations under normal and drought stress conditions were compared to investigate the effect of endophyte infection on the host plant.Under severe stress.RWC of EI leaf was significantly higher than that Of EF leaf,i.e.EI plants took more advantages over EF plants in water-holding ability.Under mild stress,endophytc could enhance soluble sugars in host plants to improve their osmotic ability.With stress intensification,the improvement of endophyte no longer existed,and more photosynthetic products(such as starch)accumulated in EI plants to survive through the undesirable condidons.In the next spring,EI populations will recover more rapidly than EF populations.The biomass of a population is closely related to its photosynthesis.Under severe stress,EI population significantly accumulated more biomass than EF population.As far as photosynthetic pigments were concerned,contents of Chla,Chlb and Car of EI plants were close to those of EF plants,which suggested that endophyte infection didn't alleviate photosynthetic pigments from being destroyed by drought stress,and endophyte might improve photosynthesis ability of its host plant in other ways.     

Effects of endophytic fungi on some drought tolerance mechanisms of tall fescue in a hydroponics culture

Neotyphodium, a seed-transmissible nonpathogenic fungal endophyte (symbiont) is considered beneficial because endophyte-infected grasses are more drought-tolerant, produce more dry matter, utilize soil nitrogen more efficiently, and deter insects. In this study, the effects of endophytes on physiological mechanisms of drought tolerance in tall fescue (Festuca arundinacea Schreb.) were studied in a greenhouse. Two clonally propagated genotypes of tall fescue (F. arundinacea Schreb.), naturally containing endophyte (EI), and their endophyte-free ramets (EF) were tested at three water stress treatments exerted by PEG 6000 in a hydroponics system. Relative water content (RWC), cell membrane stability (CMS), proline and chlorophyll contents in plant leaves were measured during water stress treatments. After harvest, K⁺, Ca²⁺, and Mg²⁺ contents were measured in plant roots and shoots. After 20 days under stress conditions, plants were transferred to basal hydroponics medium, and their survival after stress relief was evaluated. The results showed that endophyte considerably contributes to host grass water stress tolerance. Both genotypes of EI and EF plants did not differ in RWC, but, regardless of the infection status, genotype 75 had the higher RWC than genotype 83. EI clones of both genotypes maintained slightly higher chlorophyll content and membrane stability than EF clones, although these differences were not significant. The EI plants of genotype 83 concentrated significantly more proline than EF plants, but in the genotype 75, differences between EI and EF clones were not significant. Plant mineral absorption was also influenced by the endophyte presence. EI clones had the higher concentrations of K⁺ in the shoots of both genotypes. The Mg²⁺ and Ca²⁺ contents in EF plants of both genotypes were higher than EI plants in the roots, but in the shoots there were no differences between EI and EF clones. EI clones survived longer after stress removal. These results strongly suggest that Neotyphodium endophytes exert their effects on tall fescue drought tolerance through alteration of various physiological mechanisms involved. Published in Russian in Fiziologiya Rastenii, 2009, Vol. 56, No. 4, pp. 563–570. This test was submitted by the authors in English.     

内生真菌感染对干旱胁迫下黑麦草生长的影响

 内生真菌是生活在健康植物的茎叶内,形成不明显感染的一类真菌。以黑麦草（Lolium perenne L.）为实验材料,研究在不同强度的干旱胁迫下内生真菌（Neotyphodium lolii）侵染对其叶片延伸生长、分蘖数和生物量的影响。结果表明,与非感染种群相比,内生真菌感染对黑麦草叶片延伸速率无明显促进作用;内生真菌感染种群具有明显较多的分蘖数;在重度胁迫并经过恢复期后,内生真菌感染种群具有较高的根冠比。因而内生真菌可能通过提高植物的分蘖能力和促进有机物向根系的分配来促进宿主植物的营养生长并提高其抗旱性     

不同氮素营养条件下内生真菌感染对黑麦草光合和蒸腾速率及生物量的影响

以黑麦草(Lolium perenne L.)为实验材料,研究在不同氮素营养条件下内生真菌(Neotyphodium lolii)感染对其光合特性及生物量的影响。结果表明：1)整个实验中,内生真菌感染对黑麦草净光合速率、蒸腾速率以及水分利用效率均无显著影响,但在中氮和高氮水平时,感染种群的净同化累积值有超过非感染种群的趋势;2)在本实验的第一处理期和第二处理期,内生真菌感染对黑麦草绿色部分的干物质产量无显著影响,但在第三处理期感染种群的绿色部分的干物质产量显著高于非感染种群。     

干旱胁迫下内生真菌感染对黑麦草实验种群光合、蒸腾和水分利用的影响

 以黑麦草(Lolium perenne L．)为实验材料,研究在不同强度的干旱胁迫下内生真菌(Neotyphodium lolii (原 Acremonium lolii))感染对其净同化速率、蒸腾速率和水分利用效率的影响。结果显示：1)在干旱胁迫前期,内生真菌感染(EI)种群和非感染(EF)种群之间的群体净同化速率无显著差异;到胁迫后期,在重度胁迫下EI种群的净同化速率高于EF种群;复水后,各个胁迫强度EI和EF种群的净同化速率均迅速恢复,差异消失;2)在群体蒸腾速率上,干旱胁迫对其影响大于内生真菌的影响;3)在群体水分利用效率上,只是在重度胁迫后期,EI种群才高于EF种群。     

Endophytic benefit for a competitive host is neutralized by increasing ratios of infected plants

Leaf endophytes such as Epichloë can affect the competitive ability of host grasses, but the reported responses are inconsistent. We hypothesized that this inconsistency is caused, at least in part, by the following two aspects. One is that a competitive advantage might occur as a result of an increase in storage compounds for both growth and defense. Another is that the effect of the endophyte might be related to both water availability and host density. In a greenhouse experiment, we compared the competitive abilities of endophyte-infected (EI) and endophyte-free (EF) Leymus chinensis, a dominant grass native to the Inner Mongolia Steppe of China, subjected to ten treatments comprised of a factorial combination of two levels of water availability (well-watered and drought) and five proportions of EI to EF plants (12:0, 4:8, 6:6, 8:4, 0:12). The results showed that the competitive ability of EI plants was higher than that of EF under drought. Here, greater belowground biomass and water use efficiency might contribute to better competitiveness of EI plants. When competing under well-watered conditions, endophyte infection did not provide a benefit to the host plant in biomass accumulation, but more carbon was allocated to defense (total phenolics) in EI plants. This scenario could help EI plants suffer less damage than EF when exposed to herbivores in natural habitats. The competitive ability of EI plants was regulated by EI:EF ratios. Competitive ability of EI plants was higher than that of EF plants in mixtures with lower numbers of EI plants, but the beneficial effect of endophyte infection was neutralized in mixtures with higher numbers of EI plants. Overall, endophyte infection improved the competitive ability of the host under either drought or well-watered conditions but in the presence of herbivore, only this benefit was neutralized by increasing ratios of EI plants. We suspect that both the conditional beneficial effects and stabilizing effects of density differences are likely to keep the endophyte infection rate of L. chinensis at an intermediate level.     

Endophyte-Mediated Effects on the Growth and Physiology of Achnatherum sibiricum Are Conditional on Both N and P Availability

The interaction of endophyte–grass associations are conditional on nitrogen (N) availability, but the reported responses of these associations to N are inconsistent. We hypothesized that this inconsistency is caused, at least in part, by phosphorus (P) availability. In this experiment, we compared the performance of endophyte-infected (EI) and endophyte-free (EF) Achnatherum sibiricum subjected to four treatments comprising a factorial combination of two levels of N (N+ vs. N−, i.e. N supply vs. N deficiency) and two levels of P (P+ vs. P−, i.e. P supply vs. P deficiency) availability. The results showed that A. sibiricum–Neotyphodium associations were conditional on both N and P availability, but more conditional on N than P. Under N+P− conditions, endophyte infection significantly improved acid phosphatase activity of EI plants, such that the biomass of EI plants was not affected by P deficiency (i.e. similar growth to N+P+ conditions), and resulted in more biomass in EI than EF plants. Under N−P+ conditions, biomass of both EI and EF decreased compared with N+P+; however, EI biomass decreased slowly by decreasing leaf N concentration more rapidly but allocating higher fractions of N to photosynthetic machinery compared with EF plants. This change of N allocation not only improved photosynthetic ability of EI plants but also significantly increased their biomass. Under N−P− conditions, EI plants allocated higher fractions of N to photosynthesis and had greater P concentrations in roots, but there was no significant difference in biomass between EI and EF plants. Our results support the hypothesis that endophyte–grass interactions are dependent on both N and P availability. However, we did not find a clear cost of endophyte infection in A. sibiricum.     

不同浓度和形态磷处理下内生真菌感染对高羊茅的影响

以感染内生真菌(endophyte-infected,EI)和不感染内生真菌(endophyte-free,EF)的高羊茅(Festuca arundinacea Schreb.)为材料,在温室沙培条件下研究内生真菌对高羊茅适应缺磷及利用不同形态磷肥的影响。结果表明,1)缺磷条件下,高羊茅EI和EF植株生长差异不显著;正常供磷条件下,高羊茅EI植株拥有更多分蘖数和绿叶数。说明正常供磷条件下内生真菌改善了宿主高羊茅的生长。2)与水溶性磷相比,高羊茅根有机酸和酸性磷酸酶(acid phosphatase,APase)活性在难溶性磷条件下显著增加,而根总酚含量无显著变化。在水溶性磷条件下,高羊茅EI植株根总酚含量显著高于EF植株,此时EI植株比EF植株拥有更多分蘖数和绿叶数,说明在水溶性磷条件下内生真菌对宿主地上部生长具有一定贡献。在难溶性磷条件下,虽然高羊茅EI植株根总酚含量仍然高于EF植株,但同时EI植株根有机酸含量显著低于EF植株,因此内生真菌感染只是增大了宿主植物的根冠比,而对分蘖数和绿叶数等无显著影响,说明内生真菌对宿主利用难溶性磷贡献不大。可见,内生真菌对宿主植物的生长在水溶性磷条件下更有利。     

Neotyphodium Coenophialum-Infected Tall Fescue and Its Potential Application in the Phytoremediation of Saline Soils

The growth response of endophyte-infected (EI) and endophyte-free (EF) tall fescue to salt stress was investigated under two growing systems (hydroponic and soil in pots). The hydroponic experiment showed that endophyte infection significantly increased tiller and leaf number, which led to an increase in the total biomass of the host grass. Endophyte infection enhanced Na accumulation in the host grass and improved Na transport from the roots to the shoots. With a 15 g l^?1 NaCl treatment, the phytoextraction efficiency of EI tall fescue was 2.34-fold higher than EF plants. When the plants were grown in saline soils, endophyte infection also significantly increased tiller number, shoot height and the total biomass of the host grass. Although EI tall fescue cannot accumulate Na to a level high enough for it to be termed a halophyte, the increased biomass production and stress tolerance suggested that endophyte / plant associations had the potential to be a model for endophyte-assisted phytoextraction in saline soils.     

Gas Exchange and Chlorophyll Fluorescence in Symbiotic and Non-Symbiotic Ryegrass Under Water Stress

The symbiotic association of endophyte fungus, Neotyphodium lolii, and ryegrass improves the ryegrass resistance to drought. This is shown by a 30 % increase in the number of suckers in infected plants (E+), compared to plants lacking endophyte (E−), and by a higher water potential in the E+ than E− plants. The E+ plants have higher stomatal conductance (g _s), transpiration rate, net photosynthetic rate (P _N), and photorespiratory electron transport rate than the E− plants. The maximal photochemical efficiency (F_v/F_m) and the actual photochemical efficiency (Φ_PS2) are not affected by the endophyte fungus. The increase in P _N of the E+ plants subjected to water stress was independent from internal CO₂ concentration. An increased P _N was observed in E+ plants also in optimal water supply. Hence the drought resistance of E+ plants results in increased g _s, P _N, and photorespiratory electron transport rate. This revised version was published online in June 2006 with corrections to the Cover Date.     

Response of barley grains to the interactive e.ect of salinity and salicylic acid

Effect of grain soaking presowing in 1 mM salicylic acid (SA) and NaCl (0, 50, 100, 150 and 200 mM) on barley (Hordeum vulgare cv Gerbel) was studied. Increasing of NaCl level reduced the germination percentage, the growth parameters (fresh and dry weight), potassium, calcium, phosphorus and insoluble sugars content in both shoots and roots of 15-day old seedlings. Leaf relative water content (RWC) and the photosynthetic pigments (Chl a, b and carotenoids) contents also decreased with increasing NaCl concentration. On the other hand, Na, soluble sugars, soluble proteins, free amino acids including proline content and lipid peroxidation level and peroxidase activity were increased in the two plant organs with increasing of NaCl level. Electrolyte leakage from plant leaves was found to increase with salinity level. SA-pretreatment increased the RWC, fresh and dry weights, water, photosynthetic pigments, insolube saccharides, phosphorus content and peroxidase activity in the stressed seedlings. On the contrary, Na⁺, soluble proteins content, lipid peroxidation level, electrolyte leakage were markedly reduced under salt stress with SA than without. Under stress conditions, SA-pretreated plants exhibited less Ca²⁺ and more accumulation of K⁺, and soluble sugars in roots at the expense of these contents in the plant shoots. Exogenous application (Grain soaking presowing) of SA appeared to induce preadaptive response to salt stress leading to promoting protective reactions to the photosynthetic pigments and maintain the membranes integrity in barley plants, which reflected in improving the plant growth.     

不同氮素营养条件下内生真菌感染对黑麦草光合和蒸腾速率及生物量的影响

以黑麦草(Lolium perenne L)为实验材料,研究在不同氮素营养条件下内生真菌(Neotyphodium lolii)感染对其光合特性及生物量的影响.结果表明:1)整个实验中,内生真菌感染对黑麦草净光合速率、蒸腾速率以及水分利用效率均无显著影响,但在中氮和高氮水平时,感染种群的净同化累积值有超过非感染种群的趋势;2)在本实验的第一处理期和第二处理期,内生真菌感染对黑麦草绿色部分的干物质产量无显著影响,但在第三处理期感染种群的绿色部分的干物质产量显著高于非感染种群.     

干旱胁迫下发菜光合作用相关基因差异表达分析

该研究以不同失水处理的发菜为研究材料,以充分吸水状态的发菜为对照,利用高通量测序技术和qRT PCR技术检测了干旱胁迫下发菜光合作用相关基因差异表达规律,并对光合色素和酶活在干旱胁迫下的变化进行了研究。结果表明：(1)发菜在不同程度干旱胁迫下有113个光合作用相关基因差异表达,其中失水30%、75%和100%的发菜分别有44个、74个和91个光合作用相关基因差异表达。(2)随着干旱胁迫程度的加深藻胆素、叶绿素a和类胡萝卜素含量逐渐降低,Rubisco活性随着干旱胁迫程度的增强先上升后下降,GAPDH活性随着干旱胁迫的增强呈现下降的趋势。研究表明,发菜通过光合作用相关基因的差异表达调控光合作用以适应干旱胁迫。该研究结果对进一步研究发菜干旱胁迫响应机制及其耐旱光合机理奠定了基础。     

4种茶菊对干旱胁迫的形态和生理响应

以4种茶菊幼苗为材料,采用盆栽控水法,研究了不同强度干旱胁迫下茶菊形态、生理生化及光合生长的响应特性,并对其进行耐旱性评价。结果显示,随着干旱胁迫程度的增强,各茶菊幼苗叶片数增量、叶面积、生物量、叶片相对含水量、净光合速率(Pn)、蒸腾速率(Tr)和气孔导度(Gs)降低;根冠比、丙二醛(MDA)含量、脯氨酸(Pro)含量、可溶性蛋白(SPC)含量和叶绿素(Chl)含量增加;胞间CO2(Ci)浓度先降后升;乳荷、黄滁龙株高增量持续降低,而玉人面、繁白露株高增量先升后降;玉人面叶超氧化物歧化酶活性(SOD)呈先升后降趋势,其它3个品种则持续升高。运用隶属函数法对抗旱能力进行综合评定,不同茶菊品种耐旱性由高到低为乳荷黄滁龙繁白露玉人面。     

Does habitat of <Emphasis Type="Italic">Atriplex halimus</Emphasis> L. affect plant strategy for osmotic adjustment?

The impact of water stress was analysed in the xero-halophyte Mediterranean shrub Atriplex halimus using two Tunisian populations originating from a sub-humid coastal site (Monastir) or from a semi-arid area (Kairouan). Seedlings were exposed for 10 days to nutrient solution containing either 0 or 15% polyethylene glycol. Water potential (Ψ_w), osmotic potential (Ψ_s), osmotic potential at full turgor [Ψ_s(100)], relative water content (RWC), shoot dry weight (DW) and changes in solute concentrations were quantified every 2 days throughout the stress period and inorganic solutes contents were determined at the end of the treatment. The water deficit induced a decrease in Ψ_w, Ψ_s and RWC in both populations, recorded changes being higher in plants of Monastir than those of Kairouan while the shoot dry weight was reduced in a similar extent in stressed plants from both populations. Water deficit induced an increase in proline, glycinebetaine and sugar concentrations. Proline accumulated as early as after the 24-h stress treatment while, glycinebetaine required more than 6 days of stress to accumulate. At the end of the stress period, the plants of Kairouan population accumulated higher amounts of proline than those of Monastir, while an opposite trend was reported for glycinebetaine. Both populations specifically accumulated Na⁺ in response to drought stress, suggesting that this element could play a physiological role in the stress response of this xero-halophyte species. Presented results suggest that the non-recyclable osmotic solute glycinebetaine does not necessarily preferentially accumulates in population facing permanent water stress and that other strategy than osmotic adjustment might be involved in drought tolerance of A. halimus.     

干旱-复水处理对杠柳幼苗光合作用及活性氧代谢的影响

采用人工控制土壤水分试验,以80%田间持水量处理为对照,研究了3次干旱-复水处理对2年生杠柳幼苗叶片光合作用,根、茎、叶膜脂过氧化和抗氧化系统的影响.结果表明: 在干旱条件下, 杠柳叶片相对含水量和光合速率(P_n)显著降低,光合色素含量升高. 干旱复水后,叶片相对含水量完全恢复,反映了杠柳较强的旱后修复能力;叶绿素含量和P_n均明显高于对照,表现出补偿效应,适度干旱诱导了杠柳的抗旱适应性.干旱胁迫使P_n下降,以气孔限制为主,但在中午发生了非气孔限制;幼叶、新茎和细根中的超氧阴离子产生速率升高,丙二醛含量则降低,说明这些幼嫩组织未受到氧化伤害.不同器官中SOD、CAT和POD活性及变化趋势不同,以细根的3种保护酶的反应最为灵敏,说明细根是杠柳适应干旱环境的重要器官.器官间的相互合作与协调使杠柳能有效地适应干湿交替的干旱环境.     

缺磷对黑麦草-内生真菌共生体生长和根中酚含量的影响

培养液中缺磷或内生真菌感染对黑麦草地上部生长的影响不显著,但二者的交互作用却有着显著影响。随着磷浓度的下降,感染内生真菌的黑麦草显示出较强的生长优势,其根长和根中酚类物质的含量显著增加。     

Adaptive photosynthetic and physiological responses to drought and rewatering in triploid <Emphasis Type="Italic">Populus</Emphasis> populations

Cuttings of Populus cathayana Rehd, originating from three triploid and one diploid populations with the same parents but different gamete origins, were used to examine physiological responses to drought stress and rewatering by exposure to three progressive water regimes. Progressive drought stress significantly decreased the leaf relative water content (RWC), photosynthesis, and chlorophyll fluorescence parameters, and increased the relative electrolyte leakage, malondialdehyde (MDA), free proline (Pro), and antioxidant enzymes, such as superoxide dismutase, peroxidase, and catalase, in the four populations evaluated. However, compared to the diploid population, triploid populations showed lower relative electrolyte leakage and MDA, higher RWC and Pro content, and more efficient photosynthesis and antioxidant systems under the same water regime. Our data indicated that triploid populations possessed more efficient protective mechanisms than that of diploid population with gradually increasing drought stress. Moreover, some triploid genotypes were less tolerant to water stress than that of diploids due to large intrapopulation overlap.     

Rapid upregulation ofDehyrin3 andDehydrin4 in response to dehydration is a characteristic of drought-tolerant genotypes in barley

The identification of molecular markers and marker-aided selection are essential to the efficient breeding of drought-tolerant plants. However, because that characteristic is controlled by many quantitative trait loci, such markers that can screen and trace desirable barley genotypes in a segregating population or germplasm have not yet been determined. Relative water content has been used to estimate drought tolerance in plants because it is highly correlated with the drought index of yield. To develop reliable gene-specific markers for identifying tolerant versus susceptible genotypes, we performed suppression subtractive hybridization to identify candidate genes. We used two domestic barley cultivars, one having the highest RWC (drought-tolerant ‘Chalbori’) and the other having the lowest (drought-susceptible ‘Daebaekbori’). In response to dehydration at the early seedling stage, rapid upregulation ofDehydrin3 (Dhn3) andDhn4 occurred in the drought-tolerant genotypes, but not in the susceptible ones. Similar results were obtained with mature plants growing under frequent drought stress in the greenhouse. In addition,Dhn3 andDhn4 conferred higher drought tolerance when they were over-expressed in transgenicArabidopsis. Thus, in addition to using assessments of RWC, we propose thatDhn3 andDhn4 expressions can serve as drought-induced gene-specific markers to determine drought-tolerant barley genotypes at the seedling stage.