Somatic embryogenesis and plant regeneration in callus culture of tef, Eragrostis tef (Zucc.) Trotter

The study was carried out to establish in vitro culture conditions for plant regeneration of tef, Eragrostis tef (Zucc.) Trotter. Mature seeds of two Ethiopian varieties, DZ-01-354 and DZ-01-196, were used to initiate callus cultures on Murashige and Skoog (MS) medium with different auxins. Four- and 8-week-old calli induced on a medium with 2.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) were subcultured onto various media to induce somatic embryogenesis. Compact, nodulated, embryogenic callus was observed after transfer onto MS-callus proliferating (CP) medium. Embryogenic tissue appeared on soft and amorphous callus and developed into somatic embryos during a subsequent subculture to MS embryo-promoting (EP) media. Various growth regulator combinations were tested in CP and EP media to obtain a high efficiency of somatic embryo formation. The highest frequency of calli forming somatic embryos (56.1–68.3%) was observed when CP media with 2.0 or 4.0 mg/l 2,3,5-triiodobenzoic acid were employed and then cultures were transferred to EP media with 0.5 mg/l 2,4-D and 0.5 mg/l kinetin followed by 0.5 mg/l indole-3-acetic acid and 0.5 mg/l N⁶-benzyladenine. Plant development from somatic embryos was obtained on MS medium supplemented with 1.0 mg/l gibberellic acid. On average, 71.2% of calli displaying somatic embryos converted into plants. Regenerated plants were successfully transferred to soil. Neither chlorophyll-deficient plants nor morphological variants were found among regenerants. All regenerated plants were fertile. Received: 9 May 1997 / Revision received: 25 September 1997 / Accepted: 3 January 1998     

A genetic linkage map of tef [Eragrostis tef (Zucc.) Trotter] based on amplified fragment length polymorphism

A genetic linkage map of tef was constructed with amplified fragment length polymorphism (AFLP) markers using F₅ recombinant inbred lines (RILs) derived by single seed descent from the intraspecific cross of ’Kaye Murri’×’Fesho’. A total of 192 EcoRI/MseI primer combinations were screened for parental polymorphism. Around three polymorphic fragments per primer combination were detected, indicating a low polymorphism level in tef. Fifty primer combinations were selected to assay the mapping population, and 226 loci segregated among 85 F₅ RILs. Most AFLP loci behaved as dominant markers (presence or absence of a band), but about 15% of the loci were codominant. Significant deviations from the expected Mendelian segregation ratio were observed for 26 loci. The genetic linkage map comprised 211 markers assembled into 25 linkage groups and covered 2,149 cM of genome. AFLP is an efficient marker system for mapping plant species with low polymorphism such as tef. This is the first genetic linkage map constructed for tef. It will facilitate the mapping of genes controlling agronomically important traits and cultivar improvement in tef. Received: 27 April 1998 / Accepted: 4 January 1999     

Photosynthetic limitations in response to water stress and recovery in Mediterranean plants with different growth forms

* Whether photosynthesis is limited during water stress and recovery because of diffusive or biochemical factors is still open to debate, and apparent contradictions appear when various studies on species with different growth forms are compared. * Ten Mediterranean species, representing different growth forms, were subjected to different levels of water stress, the most severe followed by rewatering. A quantitative limitation analysis was applied to estimate the effects of water stress on stomatal (S(L)), mesophyll conductance (MC(L)) and biochemical limitations (B(L)). * Results confirmed a general pattern of photosynthetic response to water stress among C(3) plants when stomatal conductance (g(s)) is used as a reference parameter. As g(s) values decreased from a maximum to approx. 0.05 mol H(2)O m(-2) s(-1), the total photosynthetic limitation rose from 0 to approx. 70%, and this was caused by a progressive increase of both S(L) and MC(L) limitations, while B(L) remained negligible. When lower values of g(s) were achieved (total photosynthetic limitation increased from 70 to 100%), the contribution of S(L) declined, while MC(L) still increased and B(L) contributed significantly (20-50%) to the total limitation. * Photosynthetic recovery of severely stressed plants after rewatering showed a dominant role of MC(L), irrespective of the degree of photosynthesis recovery.     

土壤水分胁迫下沙漠葳的光合生理特征

以从美国西部引进的沙生灌木——沙漠葳（Chilopsis linearis）的2年生实生苗为材料,通过盆栽试验于7-9份进行轻度、中度和重度土壤水分胁迫处理并分析其光合生理响应特征.结果显示：（1）60 d中度或重度土壤水分胁迫使沙漠葳的净光合速率（Pn）、水分利用效率（WUE）和光补偿点（LCP）显著降低,暗呼吸速率（DRR）减弱,而气孔导度（Cs）增大,气孔限制值（Ls）变小.（2）Pn日变化在7月份的轻度和中度土壤水分胁迫下表现为双峰曲线,其余月份的胁迫处理均为单峰曲线,同期内各胁迫处理Pn峰值出现的时间基本相同,而且8月份各水分胁迫的Pn值显著高于其它月份;WUE的日变化趋势和Pn日变化相似,而且土壤水分胁迫越严重,其水分利用效率越低;各水分胁迫处理的Pn和Tr光响应拟合曲线均基本呈抛物线形或倒抛物线形.（3）在轻度和中度土壤水分胁迫的初期,Pn降低主要受气孔限制因素的影响,随着胁迫期的延长和胁迫的加重,Pn降低由气孔限制为主逐步转向以非气孔限制为主.研究表明,沙漠葳对干旱胁迫具有一定的适应能力,但长期中度或重度干旱胁迫都会影响沙漠葳的正常生长发育,使其光合生产力大大降低.     

Photosynthetic responses to temperature of phosphoenolpyruvate carboxykinase type C4 species differing in cold sensitivity

Photosynthetic rates, the activities of key enzymes associated with the C₄ cycle and ribulose-1,5-bisphosphate carboxylase (RuBPCase), and the levels of metabolites involved in the C₄ cycle were compared between the two phosphoenolpyruvate carboxykinase (PCK) type C₄ species Spartina anglica, which is cold-tolerant, and Zoysia japonica, which is cold-sensitive, during exposure to low temperature. Plants of both species grown outside in summer were placed in a growth chamber at 27/20 °C day/night temperatures. After 1 week, plants were exposed to 20/17 °C for 1 week and then to 10/7 °C for 2 weeks. Photosynthetic rates in Z. japonica decreased progressively to about 25% during the chilling treatments. In contrast, S. anglica exhibited a 43% increase in photosynthetic rates after exposure to 20 °C for 1 week, which remained relatively constant thereafter. Consistent with these observations, most of the C₄ enzymes and RuBPCase in Z. japonica declined. Phosphoenolpyruvate carboxylase (PEPC) and PCK activities declined particularly drastically during the treatments. However, the activities of these enzymes in S. anglica showed either a slight increase or decrease upon a mild cold treatment, and remained relatively constant during further chilling treatments. There was a sharp decline in phosphoenolpyruvate in Z. japonica after exposure to 10 °C. On the other hand, metabolite levels in S. anglica were largely unaffected by the chilling treatments. These results suggest that the drastic declines of both PEPC and PCK activities may be important limiting factors responsible for cold sensitivity in C₄ photosynthesis of Z. japonica.     

Stomata response to changes in temperature and humidity in wheat cultivars grown under contrasting climatic conditions

Stomatal response to changes in temperature and humidity was studied in wheat (Triticum aestivum L.) cv. Iren’ cultivated under conditions of high water supply and cv. Kazakhstanskaya 10, which is relatively drought tolerant. Experiments were performed under both laboratory and field conditions. It was demonstrated that stomata of cv. Kazakhstanskaya 10 plants closed rapidly with reducing humidity (the response of the first type), whereas, in cv. Iren’, this response was less expressed and, under conditions of a high water content in soil, stomatal conductance could increase in response to reduced humidity (the response of the second type). At an increased stomatal conductance and transpiration, water content in cv. Iren’ plants was maintained due to the increase in hydraulic conductance and water inflow from the roots. A possible role of the first-type response (rapid stomata closure) for growth maintenance under drought and of the second-type response (a parallel increase in the stomatal and hydraulic conductance) for providing of rapid growth and high productivity under sufficient water supply is discussed. A possibility to use the type of stomata behavior for cultivar assessment is considered.     

Variation in response of accessions of minor millets,Pennisetum americanum (L.) Leeke (pearl millet) and Eleusine coracana (L.) Gaertn (finger millet), and Eragrostis tef (Zucc.) Trotter (tef) to salinity in early seedling growth

The response to increasing NaCl concentration of seedlings of 25 accessions of Ethiopian land races of each of Pennisetum americanum (L.) Leeke (pearl millet) and Eleusine coracana (L.) Gaertn (finger millet), and 15 accessions of Eragrostis tef (Zucc.) Trotter (tef), was examined after two week's growth in NaCl solution culture. Although increasing NaCl concentration significantly reduced seedling root lengths, there was considerable variation within, and between accessions within each species.Analysis based upon a non-linear least square inversion method, using root length data, revealed significant differences in accessions of P. americanum and E. tef on the basis of the estimated salinity threshold, C _t , the NaCl concentrations at which root length begins to decrease. C _t did not differ significantly between E. coracana accessions. Estimates of C₅₀ and C₀, mininum concentrations causing a 50% decrease in root length, and zero root growth respectively, revealed differences between and within accessions for all three species. Overall, finger millet was more tolerant than tef, which was more tolerant than pearl millet. There is clear evidence that differences in tolerance are genetically based from broad sense heritability estimates.     

石灰岩土壤基质上构树幼苗接种丛枝菌根（AM）真菌的光合特征

就石灰岩适生植物构树(Broussonetia papyrifera)分别进行菌根真菌（简称AM真菌）摩西球囊霉(Glomus mosseae)、地表球囊霉(Glomus versiforme)、透光球囊霉(Glomus diaphanum)的单独接种、混合接种和不接种处理,幼苗生长3个月后测定其生长及光合生理指标,以期从光合水平上了解接种AM真菌对植物的生长、生理的响应。结果表明：接种AM真菌促进了构树幼苗生长,接种组地茎、苗高、总叶面积较对照差异性显著,但单株幼苗叶片数较对照无显著差异。接种处理导致了幼苗较高的菌根侵染率,而非接种处理侵染率为0。较对照而言,接种处理净光合速率显著提高,而蒸腾速率和气孔导度在单接种G.diaphanum下降低,其他三种处理均显著提高。接种处理光合耗水量较对照处理显著降低,提高了光合水分利用效率。接种后叶绿素a、b含量极显著提高。     

Flooding, gas exchange and hydraulic root conductivity of highbush blueberry

Highbush blueberry plants ( Vaccinium corymbosum L. cv. Bluecrop) growing in containers were flooded in the laboratory for various durations to determine the effect of flooding on carbon assimilation, photosynthetic response to varying CO₂ and O₂ concentrations and apparent quantum yield as measured in an open flow gas analysis system. Hydraulic conductivity of the root was also measured using a pressure chamber. Root conductivity was lower and the effect of increasing CO₂ levels on carbon assimilation less for flooded than unflooded plants after short-(i-2 days), intermediate-(10–14 days) and long-term (35–40 days) flooding. A reduction in O₂ levels surrounding the leaves from 21 to 2% for unflooded plants increased carbon assimilation by 33% and carboxylation efficiency from 0.012 to 0.021 mol CO₂ fixed (mol CO₂)⁻¹. Carboxylation efficiency of flooded plants, however, was unaffected by a decrease in percentage O₂, averaging 0.005 mol CO₂ fixed (mol CO₂)⁻¹. Apparent quantum yield decreased from 2.2 × 10⁻¹ mol of CO₂ fixed (mol light)⁻¹ for unflooded plants to 2.0 × 10⁻³ and 9.0 × 10⁻⁴ for intermediate- and long-term flooding durations, respectively. Shortterm flooding reduced carbon assimilation via a decrease in stomatal conductance, while longer flooding durations also decreased the carboxylation efficiency of the leaf.     

Photosynthetic and stomatal responses ofLarix kaempferi seedlings to short-term waterlogging

The effects of short-term waterlogging on net photosynthesis, stomatal conductance and shoot water status of 2 year old seedlings ofLarix kaempferi (Lamb.) Carr. were studied under controlled environmental conditions. Waterlogging for 8 days induced significant stomatal closure and reduced net photosynthesis. After 3 days of waterlogging, stomatal conductance was reduced to 35% of that of non-waterlogged plants, and net photosynthesis was reduced to 25% of the pre-waterlogged level. At the beginning of waterlogging, slight shoot dehydration was observed. However during the latter stage of the waterlogging shoot dehydration disappeared and stomatal conductance and net photosynthesis increased. No adaptive morphological changes to waterlogging were observed in the stems and roots. Recovery of stomatal conductance and net photosynthesis after drainage was not observed within 11 days.     

Seasonal patterns of photosynthetic response and acclimation to elevated carbon dioxide in field-grown strawberry

Strawberry (Fragaria × ananassa) plants were grown in field plots at the current ambient [CO₂], and at ambient + 300 and ambient + 600 μmol mol⁻¹ [CO₂]. Approximately weekly measurements were made of single leaf gas exchange of upper canopy leaves from early spring through fall of two years, in order to determine the temperature dependence of the stimulation of photosynthesis by elevated [CO₂], whether growth at elevated [CO₂] resulted in acclimation of photosynthesis, and whether any photosynthetic acclimation was reduced when fruiting created additional demand for the products of photosynthesis. Stimulation of photosynthetic CO₂ assimilation by short-term increases in [CO₂] increased strongly with measurement temperature. The stimulation exceeded that predicted from the kinetic characteristics of ribulose-1,5-bisphosphate carboxylase at all temperatures. Acclimation of photosynthesis to growth at elevated [CO₂] was evident from early spring through summer, including the fruiting period in early summer, with lower rates under standard measurement conditions in plants grown at elevated [CO₂]. The degree of acclimation increased with growth [CO₂]. However, there were no significant differences between [CO₂] treatments in total nitrogen per leaf area, and photosynthetic acclimation was reversed one day after switching the [CO₂] treatments. Tests showed that acclimation did not result from a limitation of photosynthesis by triose phosphate utilization rate at elevated [CO₂]. Photosynthetic acclimation was not evident during dry periods in midsummer, when the elevated [CO₂] treatments conserved soil water and photosynthesis declined more at ambient than at elevated [CO₂]. Acclimation was also not evident during the fall, when plants were vegetative, despite wet conditions and continued higher leaf starch content at elevated [CO₂]. Stomatal conductance responded little to short-term changes in [CO₂] except during drought, and changed in parallel with photosynthetic acclimation through the seasons in response to the long-term [CO₂] treatments. The data do not support the hypothesis that source-sink balance controls the seasonal occurrence of photosynthetic acclimation to elevated [CO₂] in this species. This revised version was published online in June 2006 with corrections to the Cover Date.     

田间小麦叶片光合效率日变化与光合“午睡”的关系

小麦灌浆初期叶片(旗叶)晴天中午光合速率下降(“午睡”)伴随了气孔导度、胞间CO_2浓度下降,而气孔限制值中午升高,进一步证实气孑L中午关闭是光合“午睡”的一个重要原因。叶片光合效率的中午下降并非都伴随着光合“午睡”现象。当两者同时发生时,胞间CO_2浓度降低,而光合速率与气孔导度、胞间CO_2浓度之间的相关性高于光合速率与光合效率之间的相关性。这些事实表明。即使光合效率中午下降是光合“午睡”的部分原因,但较之气孔中午关闭只是一个次要原因。     

Physiological response of eight Mediterranean maquis species to low air temperatures during winter

We analyzed the physiological response of the Mediterranean evergreen species (Arbutus unedo L., Cistus incanus L., Erica arborea L., Erica multiflora L., Phillyrea latifolia L., Pistacia lentiscus L., Quercus ilex L., and Rosmarinus officinalis L.) to winter low air temperatures. In occasion of two cold events, in February 2005 (T _min = 1.8 °C), and January 2006 (T _min = 3.1 °C and minimum T _air = −0.40 °C during the nights preceding the measurements), R. officinalis, C. incanus, and E. multiflora had the highest net photosynthetic rate (P _N) decrease (73 %, mean value) with respect to the winter P _N maximum, followed by A. unedo (62 %), P. latifolia and P. lentiscus (54 %, mean value), E. arborea (49 %), and Q. ilex (44 %). Among the considered species, Q. ilex was able to maintain P _N near the maximum for 150 min during the day, A. unedo, P. lentiscus, E. arborea, P. latifolia, E. multiflora, and R. officinalis for 60 min, and C. incanus for 30 min. The calculated mean winter daily P _N ranged from 7.9±0.6 (Q. ilex) to 2.8±0.5 (R. officinalis) μmol(CO₂) m⁻² s⁻¹. During the study period, chlorophyll (Chl) content decreased by 36 % on an average in the two cold events, and the carotenoid (Car) to Chl ratio increased by 133 % in Q. ilex, having the highest value in January 2006. Principal component analysis underlined the highest cold resistance of Q. ilex by high P _N and high Car/Chl ratio. On the contrary, R. officinalis and C. incanus had the lowest cold resistance by the highest P _N decrease and the lowest Car/Chl (C. incanus). Thus, winter stress could be an additional limitation to Mediterranean evergreen species production, and the capacity of the species to maintain P _N near 90–100 % during winter is determinant for biomass accumulation.     

Photosynthetic performance at low temperature of Bouteloua gracilis Lag., a high-altitude C4 grass from the Rocky Mountains, USA

The mechanisms controlling the photosynthetic performance of C₄ plants at low temperature were investigated using ecotypes of Bouteloua gracilis Lag. from high (3000 m) and low (1500 m) elevation sites in the Rocky Mountains of Colorado. Plants were grown in controlled‐environment cabinets at a photon flux density of 700 μ mol m^?2 s^?1 and day/night temperatures of 26/16 °C or 14/7 °C. The thermal response of the net CO₂ assimilation rate (A) was evaluated using leaf gas‐exchange analysis and activity assays of ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco), phosphoenolpyruvate carboxylase (PEPCase) and pyruvate,orthophosphate dikinase (PPDK). In both ecotypes, a reduction in measurement temperature caused the CO₂‐saturated rate of photosynthesis to decline to a greater degree than the initial slope of A versus the intercellular CO₂ response, thereby reducing the photosynthetic CO₂ saturation point. As a consequence, A in normal air was CO₂‐saturated at sub‐optimal temperatures. Ecotypic variation was low when grown at 26/16 °C, with the major difference between the ecotypes being that the low‐elevation plants had higher A; however, the ecotypes responded differently when grown at cool temperature. At temperatures below the thermal optimum, A in high‐elevation plants grown at 14/7 °C was enhanced relative to plants grown at 26/16 °C, while A in low‐elevation plants grown at 14/7 °C was reduced compared to 26/16 °C‐grown plants. Photoinhibition at low growth temperature was minor in both ecotypes as indicated by small reductions in dark‐adapted F_v/F_m. In both ecotypes, the activity of Rubisco was equivalent to A below 17 °C but well in excess of A above 25 °C. Activities of PEPCase and PPDK responded to temperature in a similar proportion relative to Rubisco, and showed no evidence for dissociation that would cause them to become principal limitations at low temperature. Because of the similar temperature response of Rubisco and A, we propose that Rubisco is a major limitation on C₄ photosynthesis in B. gracilis below 17 °C. Based on these results and for theoretical reasons associated with how C₄ plants use Rubisco, we further suggest that Rubisco capacity may be a widespread limitation upon C₄ photosynthesis at low temperature.     

Soil water deficits decrease the internal conductance to CO2 transfer but atmospheric water deficits do not

The internal conductance to CO₂ supply from substomatal cavitiesto sites of carboxylation poses a large limitation to photosynthesis.It is known that internal conductance is decreased by soil waterdeficits, but it is not known if it is affected by atmosphericwater deficits (i.e. leaf to air vapour pressure deficit, VPD).The aim of this paper was to examine the responses of internalconductance to atmospheric and soil water deficits in seedlingsof the evergreen perennial Eucalyptus regnans F. Muell and theherbaceous plants Solanum lycopersicum (formerly Lycopersiconesculentum) Mill. and Phaseolus vulgaris L. Internal conductancewas estimated with the variable J method from concurrent measurementsof gas exchange and fluorescence. In all three species steady-statestomatal conductance decreased by 30% as VPD increased from1 kPa to 2 kPa. In no species was internal conductance affectedby VPD despite large effects on stomatal conductance. In contrast,soil water deficits decreased stomatal conductance and internalconductance of all three species. Decreases in stomatal andinternal conductance under water deficit were proportional,but this proportionality differed among species, and thus therelationship between stomatal and internal conductance differedamong species. These findings indicate that soil water deficitsaffect internal conductance while atmospheric water deficitsdo not. The reasons for this distinction are unknown but areconsistent with soil and atmospheric water deficits having differingeffects on leaf physiology and/or root–shoot communication. Key words: Carbon dioxide, drought, internal conductance, mesophyll conductance, photosynthesis, stomatal conductance, transfer conductance, vapour pressure deficit, water deficit Received 11 October 2007; Revised 9 November 2007 Accepted 15 November 2007     

Stomatal response of hybrid poplar to incident light, sudden darkening and leaf excision

Responses of abaxial and adaxial stomata of Populus trichocarpa Torr. & Gray. × P. deltoides Bartr. (ex Marsh.) cv. Unal to incident light, sudden darkening and leaf excision in the light and in the dark were studied on 5-year-old trees in the field using diffusion porometry. Stomatal closure in the dark was found to be incomplete in most cases studies. Stomata closed after leaf excision in the dark within 90 min. Stomatal closure after darkening of an entire tree or an entire branch (white the rest of the tree was in the light) was slower, and complete stomatal closure was noticed only for adaxial stomata after 3 h. Adaxial stomata were more reactive and sensitive than abaxial stomata to sudden darkening and leaf excision in the light and the dark. In all treatments, stomatal response was more responsive in mature leaves than in young, still expanding leaves.     

Photosynthetic Responses of Four Hosta Cultivars to Shade Treatments

The effects of shade on the gas exchange of four Hosta cultivars were determined under differing irradiances (5, 30, 50, and 100 % of full irradiance) within pots. Irradiance saturation ranged between 400–800 mol m^–2 s^–1 among the four cultivars, of which H. sieboldiana cv. Elegans and H. plantagenea cv. Aphrodite exerted the lowest saturation and compensation irradiances. The maximal photosynthetic rate (P _max) was significantly higher in shade than in full irradiance in Elegans and Aphrodite, and was at maximum in seedlings grown in 30 % of full irradiance. The best shade treatment for cvs. Antioch and Golden Edger was 50 % of full irradiance. The diurnal gas exchange patterns in four cultivars were greatly influenced by the irradiance. Single-peak patterns of net photosynthetic rate (P _N) and stomatal conductance (g _s) were observed under 5 and 30 % full irradiance for all the cultivars while Elegans and Aphrodite suffered from midday depression in 50 % of full irradiance. Under open sky, all four cultivars showed two-peak patters in their diurnal gas exchange, but the midday depression was less in Antioch and Golden Edger than in Elegans and Aphrodite. According to their photosynthetic responses to shade, the shade tolerance of the four cultivars was in the order: Elegans>Aphrodite>Antioch>Golden Edger.