Non-enzymatic antioxidative defence in drought-stressed mulberry (<Emphasis Type="Italic">Morus indica</Emphasis> L.) genotypes

The present study investigated drought-induced responses of non-enzymatic antioxidants in four diverse mulberry genotypes (Morus indica L. S-36, M-5, MR-2 and V-1). Inside the glasshouse, potted plants were subjected to four water regimes for 75 days: (a) control: pots maintained at 100% pot water holding capacity (PC) (b) low water stress: 75% PC (c) medium water stress: 50% PC and (d) high water stress: 25% PC. Photosynthetic leaf gas exchange and non-enzymatic antioxidants including α-tocopherol, ascorbic acid (AA), glutathione, proline and total carotenoids were measured in leaves at regular intervals. Amongst all, V-1 was relatively drought tolerant and showed exceeded accumulation of α-tocopherol and AA-glutathione pool in association with higher carotenoids and proline contents. Susceptible S-36, M-5 and MR-2 could not induce any significant up-regulation in AA-glutathione pool leading to endogenous loss of α-tocopherol and more lipid peroxidation. Reactive oxygen species (ROS) like hydrogen peroxide (H₂O₂) and superoxide (O₂ ^{· ?}) showed apparent accumulation in water-stressed leaves and significantly contributed to lipid peroxidation in susceptible genotypes when compared to V-1. Our study demonstrated that proline, AA and glutathione were the major non-enzymatic antioxidants in mulberry with α-tocopherol and carotenoids as good additional indicators for drought stress tolerance. These non-enzymatic antioxidants can cumulatively render effective protection against oxidative damage and can be considered as reliable markers for screening drought-tolerant mulberry genotypes.     

Physiological foundation for the differences of long-term water use efficiency among Populus deltoides clones

With randomized complete block design, long-term water use efficiency (WUE_L), photosynthetic parameters (net photosynthetic rate Pn, transpiration rate Tr, stomatal conductance Gs, instantaneous water use efficiency WUE_i, maximum photosynthetic rate P max, light saturation pointLSP, carboxylic efficiency Ce), stomatal properties (frequency, size, superior/inferior stoma amount ratio) and root/shoot ratio of twelve Populus deltoids clones were studied under water stress in a greenhouse. Four water treatments were designed, namely, well-watered condition, slight water stress, moderate water stress, and severe water stress. Although the volumes of irrigated water for different treatments were the same, the irrigation intervals were strictly controlled. The results showed that long-term water use efficiency (WUE_L) differences among tested clones were obvious and became greater with the increase in water stress. J₂, J₆, J₇, J₈, and J₉ were excellent clones with high WUE_L. Stomatal frequency, stomatal size, Gs, Tr, Pn, and root/shoot ratio of J₂et al were moderate, whereas the superior/inferior ratio, P_max, LSP, Ce of J2et al were higher than other clones. The interrelations between WUE_L and the physiological parameters were analyzed. The results revealed that the WUE_L differences were induced by a series of physiological parameters, and clones with higher WUE_L always had strong photosynthetic capacity, higher WUE_i, and optimum root/shoot ratio.     

Leaf yield loss assessment due to Macrophomina root rot disease in mulberry gardens of south India

Abstract

Leaf yield loss in mulberry due to Macrophomina root rot disease was assessed in three different states of south India at field level. The highest leaf yield loss recorded was in V-1 variety (34.74%), whereas the lowest leaf yield loss was in K-2 variety (28.54%). However, the leaf yields losses in other varieties viz., MR-2 (32.90%), S-36 (32.06%), RFS-175 (31.75%) and S-13 (29.0%) recorded were medium. The average leaf yield loss was 31.49% due to root rot disease caused by M. phaseolina in mulberry.     

Water-use efficiency of a mallee eucalypt growing naturally and in short-rotation coppice cultivation

This study compared mature Eucalyptus kochii subsp. plenissima trees in inner regions or edges of natural bushland to young trees belt-planted through cleared agricultural land as uncut saplings or regenerating coppice over 2.7 years at Kalannie, Western Australia (320 mm annual rainfall). We assessed the ability of the species to alter its gas exchange characteristics, leaf physical attributes, and water-use efficiency of foliar carbon assimilation (WUE _i) or of total dry matter production (WUE _DM). Stomatal conductance (g _s) varied five-fold between treatment means, with coppices exhibiting greatest values and mature bush least. Photosynthetic rates followed this trend. Leaf photosynthetic capacity estimated by chlorophyll content varied 1.3-fold parallel with variations in leaf thickness, with coppices rating lowest and mature edge trees most highly. WUE _i varied 1.5-fold between treatments and was greatest in mature inner-bush and edge trees. Leaf photosynthetic capacity and g _s were both correlated with WUE _i. Carbon isotope composition (δ¹³C values) of new shoot dry matter produced early in a seasonal flush were similar to those of root starch but when averaged over the whole season correlated well with WUE _i and gas exchange characteristics of trees of each treatment. Coppices showed poorest WUE _i and most negative shoot tip δ¹³C but their WUE _DM was high. This discrepancy was suggested to relate to carbon allocation strategies in coppices favouring fast growth of replacement shoots but not of roots. Physiology of coppice growth of E. kochii is usefully geared towards both rapid and water-use efficient production of woody biomass in water limited environments.     

Growth, gas exchange, and water-use efficiency response of two young apple cultivars to drought stress in two scion-one rootstock grafting system

The combination of two scion-one rootstock was used for two apple cultivars, ‘Pink Lady’ and ‘Qinguan’, budded on the same, one-year-old Malus hupehensis (Pamp.) Rehd. to reduce the impact of root and pot size and in order to understand the growth, water-use efficiency (WUE), and chlorophyll fluorescence characteristics. The two-scion grafted trees were planted in plastic pots under two water regimes, i.e. 70% field capacity (FC) and 55% FC. Results indicated that different scions were affected differently by drought stress. ‘Pink Lady’ had higher net photosynthetic rate (P _N), stomatal conductance (g _s), and transpiration rate (E) compared with ‘Qinguan’ under both water treatments. However, ‘Qinguan’ had lower minimal fluorescence (F₀), higher maximum fluorescence (F_m), and higher maximum photochemical efficiency of photosystem II (F_v/F_m) than ‘Pink Lady’ at 55% FC. Moreover, ‘Qinguan’ had larger shoot dry mass (ShDM) and higher intrinsic WUE_I than ‘Pink Lady’ under both water status. Gas-exchange and growth parameters, except for P _N and scion diameter, were significantly affected by the cultivar and water treatment. At 70% FC, ShDM was significantly correlated with WUE_I. Moreover, WUE_I was negatively linearly correlated with g _s at either 70 or 55% FC. These results might indicate that ‘Pink Lady’ was more sensitive to drought than ‘Qinguan’. ‘Qinguan’ apple was able to improve WUE more than ‘Pink Lady’ under both well-watered and drought conditions. The growth parameters and photosynthetic capacity of two different scions showed that the combination of double scion-one rootstock might eliminate the influences of the rootstock and pot size.     

Peanut Photosynthesis Under Drought and Re-Watering

The photosynthetic response of three Arachis hypogaea L. cultivars (57-422, 73-30, and GC 8-35) grown for two months was measured under water available conditions, severe water stress, and 24, 72, and 93 h following re-watering. At the end of the drying cycle, all the cultivars reached dehydration, relative water content (RWC) ranging between 40 and 50 %. During dehydration, leaf stomatal conductance (g _s), transpiration rate (E), and net photosynthetic rate (P _N) decreased more in cvs. 57-422 and GC 8-35 than in 73-30. Instantaneous water use efficiency (WUE_i) and photosynthetic capacity (P _max) decreased mostly in cv. GC 8-35. Except in cv. GC 8-35, the activity of photosystem 1 (PS1) was only slightly affected. PS2 and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) were the main targets of water stress. After re-watering, cvs. 73-30 and GC 8-35 rapidly regained g _s, E, and P _N activities. Twenty-four hours after re-watering, the electron transport rates and RuBPCO activity strongly increased. P _N and P _max fully recovered later. Considering the different photosynthetic responses of the studied genotype, a general characterisation of the interaction between water stress and this metabolism is presented.     

Biochemical responses to drought stress in mulberry (<Emphasis Type="Italic">Morus alba</Emphasis> L.): evaluation of proline,glycine betaine and abscisic acid accumulation in five cultivars

Five popularly grown mulberry cultivars (K-2, MR-2, TR-10, BC2-59 and S-13) were subjected to drought stress by withholding irrigation, to obtain leaf water potentials (Ψ_w) ranging from −0.75, −1.50 and −2.25 MPa. Accumulation of proline, glycine betaine and abscisic acid (ABA) were quantified in control and water stressed mulberry leaves. The activities of enzymes involved in proline accumulation including glutamate dehydrogenase (EC1.4.1.2-4), pyrroline-5-carboxylate synthetase (EC 1.2.1.41), pyrroline-5-carboxylate reductase (EC1.5.1.2), ornithine transaminase (EC 2.6.1.13) were significantly enhanced in the leaves of all the cultivars with decreasing leaf water potentials, while the activities of proline dehydrogenase (EC 1.5.1.2) were reduced with progressive increase in water stress. Accumulation of proline, glycine betaine and abscisic acid was relatively higher in S-13 and BC2-59 compared to K-2, MR-2 and TR-10 under water deficit conditions. Our results demonstrate that S-13 and BC2-59 have superior osmoprotectant mechanisms under water-limited growth regimes.     

Tradeoffs between nitrogen- and water-use efficiency in dominant species of the semiarid steppe of Inner Mongolia

In water-limited environments, photosynthetic carbon gain and loss of water by transpiration are in a permanent tradeoff as both are contrarily regulated by stomata conductance. In semiarid steppe grasslands water limitation may covary with nitrogen limitation. Steppe grassland species are capable of optimizing their use of limiting resources, water and nitrogen, but regulation is still poorly understood. In a two-year experiment with addition of water (irrigation simulating a wet year) and nitrogen (0, 25, and 50 kg urea-N?ha^?1) we assessed intrinsic water use efficiency (WUE_i), nitrogen use efficiency (NUE), and related plant functional traits (PFTs) of four dominant C₃ species (Leymus chinensis, Agropyron cristatum, Stipa grandis, and Artemisia frigida). Water and N fertilizer supplementation significantly increased plant primary production, and N effect was more pronounced under irrigated conditions. Parallel with the responses of plant production, a strong tradeoff between WUE_i and NUE was detected: water supply increased NUE but decreased WUE_i, whereas N addition slightly increased WUE_i at the expense of NUE. This tradeoff occurred at the leaf level, and involved the responses of leaf N concentration and specific leaf area. WUE_i of species changed among treatments in a predictable manner by the parameter of leaf N content per area. Dominant plant species commonly achieved a higher utilization efficiency of the more limiting resource via altering PFTs, which was an important mechanism of adaptation to variable resource limitation in semiarid grasslands.     

Seasonal water-use efficiency and chlorophyll fluorescence response in alpha grass (<Emphasis Type="Italic">Stipa tenacissima</Emphasis> L.) is affected by tussock size

Abstact  Twelve randomly chosen Stipa tenacissima L. individuals were grouped into three tussock size classes, small (ST), medium (MT), and large (LT) with 5.6±0.8, 34.1±4.2, and 631.9±85.8 g of dry green foliar matter, respectively, in three plots with different S. tenacissima cover. Instantaneous (WUE_i) and long-term (WUE_l) water-use efficiencies were measured in two seasons of contrasting volumetric soil water content (early winter 21.0±0.8 % and summer 5.8±0.3 %). Maximum photochemical efficiency of photosystem 2 and stomatal conductance in summer assessed the extent of water and irradiance stress in tussocks of different size. WUE_i was lower in MT and ST “water spender” strategies than in LT during the high water-availability season. In summer net photosynthetic rate and WUE_i were higher and photoinhibition was lower in LT than in MT and ST. Significant spatial variability was found in WUE_i. Water uptake was competitive in stands with denser alpha grass and more water availability in summer, reducing their WUE_i. However, WUE_l showed a rising tendency when water became scarce. Thus it is important to explicitly account for plant size in ecophysiological studies, which must be combined with demographic information when estimating functional processes at stand level in sequential scaling procedures.     

A genetic link between leaf carbon isotope composition and whole‐plant water use efficiency in the C4 grass Setaria

Genetic selection for whole‐plant water use efficiency (yield per transpiration; WUE_plant) in any crop‐breeding programme requires high‐throughput phenotyping of component traits of WUE_plant such as intrinsic water use efficiency (WUE_i; CO₂ assimilation rate per stomatal conductance). Measuring WUE_i by gas exchange measurements is laborious and time consuming and may not reflect an integrated WUE_i over the life of the leaf. Alternatively, leaf carbon stable isotope composition (δ¹³C_leaf) has been suggested as a potential time‐integrated proxy for WUE_i that may provide a tool to screen for WUE_plant. However, a genetic link between δ¹³C_leaf and WUE_plant in a C₄ species has not been well established. Therefore, to determine if there is a genetic relationship in a C₄ plant between δ¹³C_leaf and WUE_plant under well watered and water‐limited growth conditions, a high‐throughput phenotyping facility was used to measure WUE_plant in a recombinant inbred line (RIL) population created between the C₄ grasses Setaria viridis and S. italica. Three quantitative trait loci (QTL) for δ¹³C_leaf were found and co‐localized with transpiration, biomass accumulation, and WUE_plant. Additionally, WUE_plant for each of the δ¹³C_leaf QTL allele classes was negatively correlated with δ¹³C_leaf, as would be predicted when WUE_i influences WUE_plant. These results demonstrate that δ¹³C_leaf is genetically linked to WUE_plant, likely to be through their relationship with WUE_i, and can be used as a high‐throughput proxy to screen for WUE_plant in these C₄ species.     

黑杨无性系间长期水分利用效率差异的生理基础

试验于2003年6月10日至8月20日在温室中进行,采用完全随机区组设计,4个水分处理（充分供水、轻度胁迫、中度胁迫、重度胁迫）,6次重复.各水分处理严格控制浇水间隔：充分供水,每2d浇水1次;轻度胁迫,每3d浇水1次;中度胁迫,每4d浇水1次;重度胁迫,每5d浇水1次.每次浇水的量相同,500ml.试验材料为美洲黑杨杂交无性系,父本为哈佛杨（Populus deltoides Bartr.CI.‘Harvard＇）,母本为山海关杨（Populus deltoides Bartr.CV.‘shanhaiguanensis＇）,共10个无性系,编号为J1～J10.以当前优良品系108杨（Populus euramericana CV.‘114/69＇）和中林46杨（Populus deltoides Bartr.CV.‘Zhonglin 46＇）为对照,编号为CK1、CK2.测定了12个无性系间长期水分利用效率（WUEL）、光合参数（净光合速率Pn、蒸腾速率Tr、气孔导度Gs、瞬时水分利用效率WUEi、最大净光合速率Pmax、光饱和点LSP、羧化效率Ce）及气孔特征参数（气孔频度、气孔大小、上/下表皮气孔数目比）、根冠比的差异,并深入分析了光合参数与WUEL、气孔特征参数与WUEL、根冠比与WUEL及光合参数之间的相互关系.结果表明：无性系间WUEL存在差异,这种差异随水分胁迫的加剧而更加显著.无性系间WUEL的差异在4个水分处理下表现出一致性,即WUEL最优的都是J2,其次为J6、J7、J8、J9.高WUEL的J2等无性系其气孔频度、气孔大小、Gs、Tr、Pn和根冠比在所有无性系中都处于适中的位置.它们的上/下表皮气孔数目比大,Pmax、LSP、Ce明显优于对照.Pmax高、LSP高、Ce大,表明高WUEL的J2等无性系光合能力强.Gs、Tr、Pn适中,表明J2等无性系在保证具有高WUEi的同时能保持较强的光合能力.根冠比适中,表明J2等无性系的光合产物在根、冠之间分配合理,能有充足的根系分布保证水分的供应,同时又有较多的光合产物积累在地上部分.气孔特征参数的差异是导致无性系间光合能力和WUEi差异,并最终导致WUEL差异的一个主要因子.气孔频度和气孔大小适中的无性系,其Gs、Tr、Pn适中,WUEi较高;上/下表皮气孔数目比大的无性系则具有较强的光合能力.无性系间WUEL的差异是一系列具有差异的生理参数共同作用的结果,高WUEL的无性系通常光合能力强、WUEi高、根冠比适中.今后工作中选育高WUE的无性系时,除需关注单个生理参数的突出作用外,更应关注生理参数的协同作用.     

Photosynthesis, water use efficiency and stable carbon isotope composition are associated with anatomical properties of leaf and xylem in six poplar species

Although fast‐growing Populus species consume a large amount of water for biomass production, there are considerable variations in water use efficiency (WUE) across different poplar species. To compare differences in growth, WUE and anatomical properties of leaf and xylem and to examine the relationship between photosynthesis/WUE and anatomical properties of leaf and xylem, cuttings of six poplar species were grown in a botanical garden. The growth performance, photosynthesis, intrinsic WUE (WUE_i), stable carbon isotope composition (δ¹³C) and anatomical properties of leaf and xylem were analysed in these poplar plants. Significant differences were found in growth, photosynthesis, WUE_i and anatomical properties among the examined species. Populus cathayana was the clone with the fastest growth and the lowest WUE_i/δ¹³C, whereas P. × euramericana had a considerable growth increment and the highest WUE_i/δ¹³C. Among the analysed poplar species, the highest total stomatal density in P. cathayana was correlated with its highest stomatal conductance (g_s) and lowest WUE_i/δ¹³C. Moreover, significant correlations were observed between WUE_i and abaxial stomatal density and stem vessel lumen area. These data suggest that photosynthesis, WUE_i and δ¹³C are associated with leaf and xylem anatomy and there are tradeoffs between growth and WUE_i. It is anticipated that some poplar species, e.g. P. × euramericana, are better candidates for water‐limited regions and others, e.g. P. cathayana, may be better for water‐abundant areas.     

Evaluation of Certain Cowpea Genotypes for Partial Resistance to Powdery Mildew (Erysiphe polygoni)

Partial resistance to powdery mildew (Erysiphe polygoni) in twenty one cowpea cultivars was assessed over two to three seasons under field conditions. Mildew incidence was recorded as number of leaflets affected/plant and severity as AUDPC or infection rate using Gompertz transformation. Irrespective of the parameter used to measure mildew resistance or the season, mean response of cvs V-105, V-269, V-276, V-282 and V-385 showed clear slow-mildewing, while RC-48, S-488, TVX-944-02E, V-27, V-36 and V-118 were rated as susceptible. Mildew resistance in some cultivars was negatively correlated with that for the leaf rust (Uromyces vignae). AUDPC was a more robust disease parameter than infection rate and showed a good correlation (r²= 0.79) with incidence.     

Inoculation with arbuscular mycorrhizal fungi alleviates harmful effects of drought stress on damask rose

This study was conducted to examine the role of arbuscular mycorrhiza fungi (AMF) in alleviating the adverse effects of drought stress on damask rose (Rosa damascena Mill.) plants. Four levels of drought stress (100, 75, 50, and 25% FC) were examined on mycorrhizal and non-mycorrhizal plants in pots filled with sterilized soil. Our results showed that increasing drought stress level decreased all growth parameters, nutrient contents, gas exchange parameters, and water relations indicators. Under different levels of drought stress, mycorrhizal colonization significantly increased all studied parameters. P_n, g_s, and E of the mycorrhizal plants was higher than those of non-mycorrhizal plants under different levels of drought stress. The increase in those rates was proportional the level of the mycorrhizal colonization in the roots of these plants. Majority of growth, nutrition, water status and photosynthetic parameters had a great dependency on the mycorrhizal colonization under all levels of drought stress. The results obtained in this study provide a clear evidence that AMF colonization can enhance growth, flower quality and adaptation of rose plants under different drought stress levels, particularly at high level of drought stress via improving their water relations and photosynthetic status. It could be concluded that colonization with AMF could help plants to tolerate the harmful effects caused by drought stress in arid and semi-arid regions.     

Variation in Photosynthetic Rates and Biomass Productivity among Four Mulberry Cultivars

Among four mulberry (Morus alba L.) cultivars (K-2, MR-2, BC2-59, and S-13), highest net photosynthetic rate (P _N) was observed in BC2-59 while the lowest rates were recorded with K-2. Significant differences among the four cultivars were found in leaf area, biomass production, activities of ribulose-1,5-bisphosphate carboxylase and sucrose phosphate synthase, and glucose and sucrose contents. The P _N and the activities of photosynthetic enzymes in the four cultivars were significantly correlated with the growth and biomass production measured as leaf yield, total shoot mass, and aerial plant biomass.     

AM1 is a potential ABA substitute for drought tolerance as revealed by physiological and ultra-structural responses of oilseed rape

Abscisic acid (ABA) is an important signaling molecule for plants under drought tolerance. However, ABA itself has many limitations to be used in agriculture practically. Recently, AM1 (ABA-mimicking ligand) has been found to replace ABA. In this study, we have investigated AM1’s potential role for drought tolerance by growing two contrasting rapeseed (Brassica napus L.) genotypes: Qinyou 8 (drought sensitive) and Q2 (drought resistant) with exogenous ABA or AM1 application under well-watered and drought-stressed conditions. Results demonstrate that drought stress has hampered plant growth (relative height growth rate, plant biomass, leaf area), plant water status (leaf relative water content, root moisture content, leaf water potential), photosynthetic gas exchange attributes like net photosynthesis rate (Pn), stomatal conductance (Gs), intercellular CO₂ concentration (Ci), transpiration rate (E); chlorophyll fluorescence parameters like photosynthetic efficiency (Fv/Fm), effective quantum yield of PSII (Φ _PSII ), photochemical quenching coefficient (qL), electron transport rate (ETR) and chlorophyll content, especially for Qinyou 8 significantly compared to well-watered plants. Whereas increased root/shoot ratio (R/S), water use efficiency (WUE) and non-photochemical quenching (NPQ) was recorded in both genotypes under drought stress. On the other hand, exogenous ABA or AM1 treatment has regulated all the above parameters in a rational way to avoid drought stress. Chloroplast transmission electron microscope images, especially for Qinyou8, have revealed that oxidative stress induced by drought has blurred the grana thylakoids, increased the size or number of plastoglobules due to lipid peroxidation, and the presence of starch granules depict weak capacity to convert them into simple sugars for osmotic adjustment. However, intact grana thylakoid, few plastoglobules with no or very few starch granules were observed in the chloroplast from ABA- or AM1-treated plants under drought. More importantly, AM1-treated plants under drought stress have responded in an extremely similar way like ABA-treated ones. Finally, it is suggested that AM1 is a potential ABA substitute for plant drought tolerance.     

Responses of antioxidant defense system of <Emphasis Type="Italic">Helianthus annuus</Emphasis> to abscisic acid treatment under drought and waterlogging

The effect of abscisic acid (ABA) treatment on growth pigments and antioxidant defense system were investigated in seedlings of Helianthus annuus (cvs. Nantio F1 and Özdemirbey) subjected to drought and waterlogging stress. In addition, seedlings were sprayed with 10 M ABA three times every other day. Relative growth rate (RGR) was significantly reduced in both genotypes under drought stress, however, this growth inhibition was less in ABA-treated plants. Total chlorophyll content increased by drought stress in both genotypes. Ascorbate was not influenced by drought, while α-tocopherol increased in cv. Nantio F1. Ascorbate and α-tocopherol increased with drought stress in cv. Özdemirbey. ABA treatment decreased ascorbate and β-carotene contents while it increased α-tocopherol and xanthophylls contents under drought stress. The activity of superoxide dismutase (SOD) in both genotypes increased under drought stress-ABA combinations. Catalase (CAT) activity decreased under drought stress and drought-ABA combinations while it increased under waterlogging stress. Glutathione reductase (GR) activity decreased under drought stress but recovered with ABA treatment. The results suggested that ABA treatments have different effects on the components of antioxidant defense system in H. annuus genotypes and ABA may contribute drought-induced oxidative stress tolerance but not effects under waterlogging stress.     

Dendroecological and genetic insights for future management of an old-planted forest of the endangered Mediterranean fir Abies pinsapo

Pinsapo fir (Abies pinsapo Boiss.) is an endangered Mediterranean conifer that has raised strong conservation interest as a paradigmatic example of species characterized by small and fragmented populations. We studied an old reforestation stand composed of A. pinsapo, Pinus nigra and Pinus sylvestris established in the 1910s in central-eastern Spain (about 500 km north of the species native distribution range), with the aim of evaluating the stand’s suitability as an ex situ conservation unit for the fir. To this end, we investigated the long-term performance of the stand and assessed genetic diversity of A. pinsapo. Tree-ring width (TRW) and carbon isotope discrimination (Δ¹³C) were used to characterise growth dynamics and intrinsic water-use efficiency (WUE_i), respectively. Furthermore, 42 pinsapo firs were genotyped at five microsatellite loci to compare their genetic variation with published data on natural populations. A. pinsapo showed ca. two-fold higher radial growth than pines in the last 80 years; however, a growth decrease was observed for all species from the 1990s onwards. Indexed TRW was positively associated with Δ¹³C at the species level, denoting inter-annual growth dependence on water availability. Overall, Δ¹³C was higher for A. pinsapo compared to pines, indicating lower WUE_i, but Δ¹³C significantly decreased over the last 50 years for all species, likely as the result of tighter stomatal regulation of water loss, resulting in WUE_i increases of about 25 %. Recently, however, A. pinsapo showed reduced WUE_i increase in concord with growth slowdown, suggesting a threshold response for stomatal regulation. Although genetic diversity of A. pinsapo was about half of natural populations, the old-planted stand could be important for the conservation of this endemic species considering its good long-term growth and physiology. The latest decrease in performance of A. pinsapo, however, asks for urgent management measures aimed at reducing the competition for water and promoting growth and natural regeneration. This study illustrates the potential of combining tree-ring-based long-term physiological information with genetic data to ascertain the prospects of artificial stands for conservation purposes.     

Water use efficiency, transpiration and net CO2 exchange of four alfalfa genotypes submitted to progressive drought and subsequent recovery

The predicted worldwide increase in arid areas and water stress episodes will strongly affect crop production. Plants have developed a wide diversity of physiological mechanisms for drought tolerance. A decline in photosynthesis and thus yield production is a common response to drought, as are increases in the water use efficiency of photosynthesis (WUE_ph) and productivity (WUE_p). The aim of our study was to determine the physiological effects (especially WUE_ph and WUE_p) of progressive drought and subsequent recovery in three cultivars adapted to a Mediterranean climate [Tafilalet (TA), Tierra de Campos (TC), and Moapa (MO)], and another representative from an oceanic climate (Europe (EU)). The accuracy of the relationships between WUE_ph or WUE_p and carbon isotope discrimination (Δ ¹³C) in shoots was also investigated as a function of water stress intensity. Mild drought (7 days of water withholding) decreased the net CO₂ exchange (A), leaf conductance to water (g) and transpiration in EU leading to an increased WUE_ph. Δ ¹³C was correlated with WUE_p but not with WUE_ph, probably due to a late decrease in g. After moderate drought (14 days), A and g decreased in all cultivars, increasing WUE_ph. In this period WUE_p also increased. Both WUE parameters were correlated with Δ ¹³C, which may indicate that the g value at the end of moderate water stress was representative of the growing period. After 21 days, TA was the most productive cultivar but under severe drought conditions there was no difference in DM accumulation among cultivars. After the recovery period, leaf area was increased but not total DM, showing that leaves were the most responsive organs to rewatering. Severe water stress did not decrease WUE_ph or WUE_p, and Δ ¹³C did not increase after recovery. This absence of a response to severe drought may indicate significant effects on the photosynthetic apparatus after 21 days of withholding water. As for mild drought, WUE_p but not WUE_ph was correlated with Δ ¹³C, supporting the view that WUE_p is a more integrative parameter than WUE_ph.     

Anthocyanin production in the hyperaccumulator plant Noccaea caerulescens in response to herbivory and zinc stress

Stomatal behavior in response to drought has been the focus of intensive research, but less attention has been paid to stomatal density. In this study, 5-week-old maize seedlings were exposed to different soil water contents. Stomatal density and size as well as leaf gas exchange were investigated after 2-, 4- and 6-week of treatment, which corresponded to the jointing, trumpeting, and filling stages of maize development. Results showed that new stomata were generated continually during leaf growth. Reduced soil water content significantly stimulated stomatal generation, resulting in a significant increase in stomatal density but a decrease in stomatal size and aperture. Independent of soil water conditions, stomatal density and length in the trumpeting and filling stages were greater than in the jointing stage. Irrespective of growth stage, severe water deficit significantly reduced stomatal conductance (G _s), decreasing the leaf transpiration rate (T _r) and net photosynthetic rate (P _n). Stomatal density was significantly negatively correlated with both P _n and T _r but more strongly with T _r, so the leaf instantaneous water use efficiency (WUE _i ) correlated positively with stomatal density. In conclusion, drought led to a significant increase in stomatal density and a reduction in stomatal size and aperture, resulting in decreased P _n and T _r. Because the negative correlation of stomatal density to T _r was stronger than that to P _n, leaf WUE _i tended to increase.