Qualitative and Quantitative Evaluation of Gall Induced by <Emphasis Type="Italic">Pseudophacopteron alstonium</Emphasis> Yang et Li 1983 (Hemiptera: Psyllidae: Phacopteronidae) as Plant Parasite,in <Emphasis Type="Italic">Alstonia scholaris</Emphasis> Leaves

Alstonia scholaris (Dr C. Alston, 1685–1760) (Family Apocynaceae) (Chattim tree), commonly known as devil tree, is an evergreen tropical tree. The tree is native to India and also found in Sri Lanka, Southern China, throughout Malaysia to northern Australia. This plant is seriously damaged by formation of tumor like galls across the Kolkata city,West Bengal which affects its ornamental and medicinal value. Gall is formed by ovipositing adults of Pseudophacopteron alstonium Yang et Li 1983 (Hemiptera: Psyllidae: Phacopteronidae) and results in destruction of host plant. The nymphal stage undergoes moulting through first instar to third instar to reach the adult within galls. It is observed that highly infested leaves can bear 60–80 galls. The gallmaker Pseudophacopteron sp. stresses the host organ, and the host counters it with physiological activities supplemented by newly differentiated tissues. In infested leaves, chlorophyll and carbohydrate contents decreased sequentially with the age of the gall. There were no significant changes in protein and total amino acid content in gall tissue. But total lipid content was highest in mature galled leaves. Increased phenolic content after psylloid herbivory, which exerted oxidative stress on the host plants, was observed in gall infested leaves as compared to fresh ungalled leaves of Alstonia scholaris. Moisture content was highest in ungalled healthy leaves than the young galled, mature galled and perforated galled leaves.     

Changes in photosynthesis of alpine plant <Emphasis Type="Italic">Saussurea superba</Emphasis> during leaf expansion

The native alpine plant Saussurea superba is widely distributed in Qinghai–Tibetan Plateau regions. The leaves of S. superba grow in whorled rosettes, and are horizontally oriented to maximize sunlight exposure. Experiments were conducted in an alpine Kobresia humilis meadow near Haibei Alpine Meadow Ecosystem Research Station (37°29′–37°45′N, 101°12′–101°33′E; alt. 3200 m). Leaf growth, photosynthetic pigments and chlorophyll fluorescence parameters were measured in expanding leaves of S. superba. The results indicate that leaf area increased progressively from inner younger leaves to outside fully expanded ones, and then slightly decreased in nearly senescent leaves, due to early unfavorable environmental conditions, deviating from the ordinary growth pattern. The specific leaf area decreased before leaves were fully expanded, and the leaf thickness was largest in mature leaves. There were no significant changes in the content of chlorophylls (Chl) and carotenoids (Car), but the ratios of Chl a/b and Car/Chl declined after full expansion of the leaves. The variation of Chl a/b coincided well with changes in photochemical quenching (q _P) and the fraction of open PSII reaction centers (q _L). The maximum quantum efficiency of PSII photochemistry after 5 min dark relaxation (F _(v)/F _(m)) continuously increased from younger leaves to fully mature leaves, suggesting that mature leaves could recover more quickly from photoinhibition than younger leaves. The light-harvesting capacity was relatively steady during leaf expansion, as indicated by the maximum quantum efficiency of open PSII centers (\(F_{\text{v}}^{{\prime }}\)/\(F_{\text{m}}^{{\prime }}\)). UV-absorbing compounds could effectively screen harmful solar radiation, and are a main protection way on the photosynthetic apparatus. The decline of q _P and q _L during maturation, together with limitation of quantum efficiency of PSII reaction centers (L _(PFD)), shows a decrease of oxidation state of Q_A in PSII reaction centers under natural sunlight. Furthermore, light-induced (Φ _NPQ) and non-light-induced quenching (Φ _NO) were consistent with variation of L _(PFD). It is concluded that the leaves of S. superba could be classified into four functional groups: young, fully expanded, mature, and senescent. Quick recovery from photoinhibition was correlated with protection by screening pigments, and high level of light energy trapping was correlated with preservation of photosynthetic pigments. Increasing of Φ _NPQ and Φ _NO during leaves maturation indicates that both thermal dissipation of excessive excitation energy in safety and potential threat to photosynthetic apparatus were strengthened due to the declination of q _P and q _L, and enhancement of L _(PFD).     

Contrasting patterns of sun-red and shade-green leaves of <Emphasis Type="Italic">Buxus microphylla</Emphasis> in response to gradients of excess light during winter acclimation

Leaf reddening in overwintering evergreens largely restricts their application in landscapes and is generally triggered in response to excess light. To explore how leaves respond to excess light and examine the potential relevance of leaf reddening in this process, a comparative field study was conducted on the sun leaves (SUL), shade leaves (SHL) and three levels of artificially shaded sun leaves (SSUL) of Buxus microphylla ‘Wintergreen’. The seasonal changes in leaf colorations, chlorophyll (Chl) and carotenoid contents, leaf absorbance and chlorophyll fluorescence characteristics were investigated. The results showed that SUL upregulated Chl a/b with increased reductions in Chl b compared with Chl a, accumulated red pigments in the upper palisade mesophyll with reduced absorption in blue and red light but increased absorption in green light, and additionally, significantly downregulated photochemical activities through the sustained enhancement of energy dissipation in PSII antenna (ΦD) from fall to midwinter. In the SSUL, as the light intensity decreased, all of the above processes were mitigated except that the SSUL maintained constant absorptions in blue light region and whose levels were similar to those of the SUL and SHL. In contrast, the SHL maintained relatively high levels of Chl a and Chl b, remained completely green and showed regulated ΦD and ΦE (energy dissipation in PSII reaction centers) to maintain relatively high photochemical activity in the winter. We conclude that the sun leaves downregulate Chl contents to reduce the light absorption and simultaneously enhance sustained ΦD to dissipate most of the light energy, whereas shade leaves maintain relatively high Chl contents and demonstrate regulated proportions of ΦD and ΦE to match the extent to which the absorbed light can be utilized through photochemical reactions. The accumulated red pigments in sun phenotypes may provide a shading effect on Chls by directing energy to non-photosynthetic reaction centers in the blue light region where the absorption is offset by the reduced Chls.     

Gas exchange and water relations of three <Emphasis Type="Italic">Vitis vinifera</Emphasis> L. cultivars growing under Mediterranean climate

Optical characteristics, contents of photosynthetic pigments, total soluble sugars, and starch, rates of gas exchange, chlorophyll (Chl) a fluorescence, and leaf water relations were analysed in three Vitis vinifera L. cultivars, Tinto Cão (TC), Touriga Nacional (TN), and Tinta Roriz (TR), grown in Mediterranean climate. Chl content was significantly lower in TC than in TN and TR leaves, while the Chl a/b ratio was higher. TR had the lowest net photosynthetic rate, stomatal conductance, and contents of soluble sugars and starch than TN and TC. In spite of low Chl content, TC showed the lowest photon absorbance and the highest photochemical efficiency of photosystem 2. TC had the lowest predawn and midday leaf water potential. The capability for osmotic adjustment was similar among cultivars and the calculated modulus of elasticity was higher in TC leaves. The typical lighter green leaves of TC seemed to be an adaptive strategy to high irradiance and air temperature associated to water stress.     

Effects of drought stress on fluorescence characteristics of photosystem II in leaves of Plectranthus scutellarioides

Drought stress has multiple effects on the photosynthetic apparatus. Herein, we aimed to study the effect of drought stress on fluorescence characteristics of PSII in leaves of Plectranthus scutellarioides and explore potentially underlying mechanisms. Plants of P. scutellarioides were grown in a greenhouse and subjected to drought (DS, drought-stressed) or daily irrigation (control group). Leaf chlorophyll (Chl) index and induction kinetics curves of Chl a fluorescence and the JIP-test were used to evaluate effects of drought lasting for 20 d. Our results showed that both the leaf and soil relative water content decreased with increasing treatment duration. The leaf Chl index was reduced to half in the DS plants compared with the control group after 20 d. The minimal fluorescence in the DS plants was higher than that in the control plants after 10 d of the treatment. Maximum photochemical efficiency and lateral reactivity decreased with increasing treatment duration in the DS plants. With the continuing treatment, values of absorption flux per reaction center (RC), trapped energy flux per RC, dissipated energy flux per RC, and electron transport flux per RC increased in the earlier stage in the DS plants, while obviously decreased at the later stage of the treatment. In conclusion, drought stress inhibited the electron transport and reduced PSII photochemical activity in leaves of P. scutellarioides.     

Monitoring cashew seedlings during interactions with the fungus <Emphasis Type="Italic">Lasiodiplodia theobromae</Emphasis> using chlorophyll fluorescence imaging

The chlorophyll (Chl) fluorescence imaging technique was applied to cashew seedlings inoculated with the fungus Lasiodiplodia theobromae to assess any disturbances in the photosynthetic apparatus of the plants before the onset of visual symptoms. Two-month-old cashew plants were inoculated with mycelium of L. theobromae isolate Lt19 or Lt32. Dark-adapted and light-acclimated whole plants or previously labelled, single, mature leaf from each plant were evaluated weekly for Chl fluorescence parameters. From 21 to 28 days, inoculation with both isolates resulted in the significantly lower maximal photochemical quantum yield of PSII (F_v/F_m) than those for control samples, decreasing from values of 0.78 to 0.62. In contrast, the time response of the measured fluorescence transient curve from dark-acclimated plants increased in both whole plants and single mature leaves in inoculated plants compared with controls. The F_v/F_m images clearly exhibited photosynthetic perturbations 14 days after inoculation before any visual symptoms appeared. Additionally, decays in the effective quantum yield of PSII photochemistry and photochemical quenching coefficient were also observed over time. However, nonphotochemical quenching increased during the evaluation period. We conclude that F_v/F_m images are the effective way of detecting early metabolic perturbations in the photosynthetic apparatus of cashew seedlings caused by gummosis in both whole plants and single leaves and could be potentially employed in larger-scale screening systems.     

Effects of drought stress on growth and chlorophyll fluorescence of Lycium ruthenicum Murr. seedlings

The present study aimed to determine effects of drought stress on Lycium ruthenicum Murr. seedlings. Our results showed that mild drought stress was beneficial to growth of L. ruthenicum seedlings. Their height, basal diameter, crown, leaf number, stem dry mass, leaf and root dry mass increased gradually when the soil water content declined from 34.7 to 21.2%. However, with further decrease of the soil water content, the growth of L. ruthenicum seedlings was limited. After 28 d of treatment, the seedlings were apparently vulnerable to drought stress, which resulted in significant leaf shedding and slow growth. However, growth was restored after rehydration. Drought treatments led to a decrease in contents of chlorophyll (Chl) a, b, and Chl (a+b) and increase in the Chl a/b ratio. After rewatering, the Chl content recovered to the content of the control plants. Under drought stress, minimal fluorescence and nonphotochemical quenching coefficient increased, thereby indicating that L. ruthenicum seedlings could protect PSII reaction centres from damage. Maximum fluorescence, maximum quantum yield, actual quantum yield of PSII photochemistry, and photochemical quenching decreased, which suggested that drought stress impacted the openness of PSII reaction centres. A comparison of these responses might help identify the drought tolerance mechanisms of L. ruthenicum. This could be the reference for the planting location and irrigation arrangements during the growing period of L. ruthenicum.     

Chlorophylls <Emphasis Type="Italic">d</Emphasis> and <Emphasis Type="Italic">f</Emphasis> and their role in primary photosynthetic processes of cyanobacteria

The finding of unique Chl d- and Chl f-containing cyanobacteria in the last decade was a discovery in the area of biology of oxygenic photosynthetic organisms. Chl b, Chl c, and Chl f are considered to be accessory pigments found in antennae systems of photosynthetic organisms. They absorb energy and transfer it to the photosynthetic reaction center (RC), but do not participate in electron transport by the photosynthetic electron transport chain. However, Chl d as well as Chl a can operate not only in the light-harvesting complex, but also in the photosynthetic RC. The long-wavelength (Q_y) Chl d and Chl f absorption band is shifted to longer wavelength (to 750 nm) compared to Chl a, which suggests the possibility for oxygenic photosynthesis in this spectral range. Such expansion of the photosynthetically active light range is important for the survival of cyanobacteria when the intensity of light not exceeding 700 nm is attenuated due to absorption by Chl a and other pigments. At the same time, energy storage efficiency in photosystem 2 for cyanobacteria containing Chl d and Chl f is not lower than that of cyanobacteria containing Chl a. Despite great interest in these unique chlorophylls, many questions related to functioning of such pigments in primary photosynthetic processes are still not elucidated. This review describes the latest advances in the field of Chl d and Chl f research and their role in primary photosynthetic processes of cyanobacteria.     

The Chlorophyll <Emphasis Type="Italic">a</Emphasis> Fluorescence Characteristic in Different Types of Leaf Senescence in <Emphasis Type="Italic">Alhagi sparsifolia</Emphasis>

Senescence is both a highly controlled and a strictly regulated process that is gene dependent. To study the PSII reaction in different types of leaf senescence processes, stem girdling was performed on Alhagi sparsifolia to investigate the leaf status in the control, natural senescence, and girdling-induced senescence leaves. The results showed that during senescence, leaf soluble sugar content, starch content, and the energy absorbed by the unit reaction center (ABS/RC) increased; whereas leaf photosynthetic rate, photosynthetic pigment content, maximum photochemical efficiency (φ _Po), and energy used by the acceptor site in electron transfer (ET_o/RC) decreased. The result of the present research implied that stem girdling significantly accelerated leaf senescence, which was due to the accumulation of carbohydrate. Natural senescence is a highly controlled process, which is an ordered process played by genes, whereas girdling-induced senescence is a disordered one. In addition, natural senescence slightly inhibits the acceptor site of PSII but did not damage the donor site of PSII. Conversely, girdling-induced senescence not only damaged the donor site of PSII (for example, oxygen-evolving complex), but also significantly inhibited the acceptor site of PSII. Moreover, both types of senescence led to an increase in the energy absorbed by the unit reaction center (ABS/RC), which subsequently resulted in an increasing excitation pressure in the reaction center (DI_o/RC), as well as additional saved carotenoid for absorbing residual light energy and quenching reactive oxygen species during senescence.     

Activity of the System for Chlorophyll Biosynthesis and Structural and Functional Organization of Chloroplasts in a Plastome <Emphasis Type="Italic">en:chlorina-5</Emphasis> Sunflower Mutant

Chlorophyll (Chl) deficiency in leaves of a plastome sunflower (Helianthus annuus L.) en:chlorina-5 mutant is due to the formation of smaller chloroplasts with a markedly reduced membrane system, as compared to the parent 3629 line. Abnormalities in the structure of the photosynthetic apparatus in the mutant can be mainly attributed to changes in the formation of photosystem I and its light-harvesting complexes. Chl deficiency in en:chlorina-5 correlated with its lower capability of synthesizing the first specific Chl precursor, 5-aminolevulinic acid (ALA) in the light. Light-independent stages of Chl biosynthesis in the mutant had the same efficiency as in leaves of the parent line. ALA formation in darkness and its conversion into protochlorophyllide did not depend on the extent of photosynthetic membrane development and photosynthetic activity.     

Use of the atLEAF+ chlorophyll meter for a nondestructive estimate of chlorophyll content

At present, chlorophyll meters are widely used for a quick and nondestructive estimate of chlorophyll (Chl) contents in plant leaves. Chl meters allow to estimate the Chl content in relative units - the Chl index (CI). However, using such meters, one can face a problem of converting CI into absolute values of the pigment content and comparing data acquired with different devices and for different plant species. Many Chl meters (SPAD-502, CL-01, CCM-200) demonstrated a high degree of correlation between the CI and the absolute pigment content. A number of formulas have been deduced for different plant species to convert the CI into the absolute value of the photosynthetic pigment content. However, such data have not been yet acquired for the atLEAF+ Chl meter. The purpose of the present study was to assess the applicability of the atLEAF+ Chl meter for estimating the Chl content. A significant species-specific exponential relationships between the atLEAF value (corresponding to CI) and extractable Chl a, Chl b, Chl (a+b) for Calamus dioicus and Cleistanthus sp. were shown. The correlations between the atLEAF values and the content of Chl a, Chl b, and Chl (a+b) per unit of leaf area was stronger than that per unit of dry leaf mass. The atLEAF value- Chl b correlation was weaker than that of atLEAF value-Chl a and atLEAF value-Chl (a+b) correlations. The influence of light conditions (Chl a/b ratio) on the atLEAF value has been also shown. The obtained results indicated that the atLEAF+ Chl meter is a cheap and convenient tool for a quick nondestructive estimate of the Chl content, if properly calibrated, and can be used for this purpose along with other Chl meters.     

Solute patterns and diurnal variation of photosynthesis and chlorophyll fluorescence in Korean coastal sand dune plants

Four plant species, Elymus mollis Trin., Carex kobomugi Ohwi, Glehnia littoralis F. Schmidt ex Miq., and Vitex rotundifolia L.f., are dominant perennial species in coastal sand dunes of Korea. We examined a physiological adaptation of these species by measurements of diurnal variation in photosynthesis and chlorophyll (Chl) fluorescence and solute patterns in leaves during one season (June), which is favorable for plant growth of all four species. All four species adopted different strategies in order to utilize radiation and to maintain water status under a fluctuating microclimate. Although the lowest water contents among four plant species was found, E. mollis with a high Chl and K⁺ content showed better photosynthetic performance, with high stomatal conductance (g _s), net photosynthetic rate (P _N), instantaneous carboxylation efficiency (CE), and water-use efficiency. Midday depression of P _N in E. mollis and G. littoralis, without a reduction of gs, was associated with a reduction in CE and maximum photochemical efficiency of PSII, indicating nonstomatal limitation. Photosynthesis depression in both C. kobomugi and V. rotundifolia, with relatively low g _s values, could be attributed to both stomatal and nonstomatal limitations. The high storage capacity for inorganic ions in E. molli, C. kobomugi, and G. littoralis may play an efficient role in regulating photosynthesis and maintaining leaf water status through stomatal control, and can also play an important role in osmotic adjustment.     

Effect of day/night temperature difference on chlorophyll content,photosynthesis and fluorescence parameters of tomato at fruit stage

In order to investigate the effect of day/night temperature difference (DIF) on photosynthetic characteristics of tomato plants (Solanum lycopersicum, cv. Jinguan 5) at fruit stage, an experiment was carried out in climate chambers. Five day/night temperature regimes (16/34, 19/31, 25/25, 31/19, and 34/16°C) with respective DIFs of -18, -12, 0, +12, and +18 were used and measured at mean daily temperature of 25°C. The results showed that chlorophyll (Chl) a, Chl b, net photosynthetic rate (P_N), stomatal conductance (g_s), maximum quantum yield of PSII photochemistry (F_v/F_m), effective quantum yield of PSII photochemistry (?_PSII), and photochemical quenching (qp) significantly increased under positive DIF, while they decreased with negative DIF. In contrast, the Chl a/b ratio and nonphotochemical quenching (NPQ) decreased under positive DIF, while increased with negative DIF. Chl a, Chl b, P_N, g_s, F_v/F_m, ?_PSII, and qp were larger under +12 DIF than those at +18 DIF, while Chl a/b and NPQ showed an opposite trend.     

Effect of high light intensity on the photosynthetic apparatus of two hybrid lines of <Emphasis Type="Italic">Paulownia</Emphasis> grown on soils with different salinity

The objective of this investigation was to evaluate the simultaneous action of light stress and salinity. Pulse amplitude modulated chlorophyll fluorescence, P₇₀₀ redox state, and pigment analysis were used to assess the impact of high light intensity on Paulownia tomentosa × fortunei and Paulownia elongata × elongata grown on soils with different salinity. It was found that light stress reduced the amount of pigments and the efficiency of photochemical energy conversion, inhibited the maximum and the effective quantum yields of PSII photochemistry, decreased photochemical quenching and photosynthetic rate. Data also showed influence on the primary quinone acceptor (Q_A) reoxidation, which led to the restriction of the electron flow from Q_A to plastoquinone and stimulation of the cyclic electron flow. The possible reasons for the increased effects of the light stress under conditions of high salt concentration in soil for Paulownia tomentosa × fortunei are discussed.     

Variation of leaf traits and pigment content in three species of steppe plants depending on the climate aridity

Mesophyll structure and content of photosynthetic pigments in the leaves of three species of steppe plants, Centaurea scabiosa L., Euphorbia virgata Waldst. et Kit., Helichrysum arenarium (L.) Moench, were investigated in four geographical sites of the Volga region and the Urals located in the forest-steppe and steppe zones. Variations of the studied parameters between geographical points depended both on the species and on the structural organization of the leaf. The highest level of variation was observed for leaf area and pigment content per unit leaf area, the size and the number of chloroplasts in the cell changed to a lesser extent. The leaf thickness, leaf area and mesophyll cell sizes mostly depended on the plant species. C. scabiosa had large leaves (40–50 cm²) with large thickness (280–290 μm) and large mesophyll cells (up to 15000 μm³). The leaves of H. arenarium and E. virgata were ten times smaller and characterized by 1.5 times smaller thickness and 2?3 times smaller cell size. Geographical location and climate of the region affected leaf density, proportion of partial tissue volume, and the ratio of the photosynthetic pigments. In the southern point of Volga region with the highest climate aridity, all studied species were characterized by maximum values of volumetric leaf density (LD), due to the high proportion of sclerenchyma and vascular bundles, and specificity of the mesophyll structure. With the decline in latitude, chlorophyll (Chl) and carotenoid (Car) contents in leaf area were reduced, the ratio Chl/Car was increased, and the ratio Chl a/b was declined. The reduction of the pigment content in the leaf in all species was associated with a reduction in the amount of Chl per chloroplast, and for C. scabiosa and H. arenarium it was associated also with the reduction of chloroplast amount in the leaf area. In turn, chloroplast number per leaf area and the total cell area (A_mes/A) depended on the ratio of the number and size of mesophyll cells inherent to this plant species. At the same time, we found a similar mechanism of spatial organization of leaf restructuring for all studied species—decrease in A_mes/A was accompanied by increasing in the proportion of intercellular air spaces in the leaf. It is concluded that variations in structural and functional parameters of the photosynthetic apparatus of steppe plants were associated with plant adaptation to climate features. General direction of the changes of leaf parameters of the studied species with aridity was the increase of LD and the decrease of pigment content per leaf area however the cellular mechanisms of changes in the pigment content and integral parameters of mesophyll were determined by the plant species properties.     

Variation of photosynthetic characteristics and yield in wild and cultivated species of yams (<Emphasis Type="Italic">Dioscorea spp.</Emphasis>) from Koraput,India

Variations in leaf gas-exchange characteristics, PSII activity, leaf pigments, and tuber yield were investigated in seven wild and one cultivated species of Dioscorea from Koraput, India, in order to find out their overall adaptability to the environment. The leaf photosynthetic rate, transpiration, stomatal conductance, water-use efficiency, carboxylation efficiency, and photosynthetic pigments were significantly higher in some wild species compared to the cultivated species. In addition, some wild species showed better photochemical efficiency of PSII, photochemical quenching, and electron transport rate in comparison to cultivated one. Furthermore, leaf dry matter accumulation and tuber yield was also higher in some wild species compared to the cultivated species. Taken together, the wild species, such as D. oppositifolia, D. hamiltonii, and D. pubera, showed the superior photosynthetic efficiency compared to the cultivated D. alata and they could be used for future crop improvement programs.     

Frequently asked questions about chlorophyll fluorescence,the sequel

Using chlorophyll (Chl) a fluorescence many aspects of the photosynthetic apparatus can be studied, both in vitro and, noninvasively, in vivo. Complementary techniques can help to interpret changes in the Chl a fluorescence kinetics. Kalaji et al. (Photosynth Res 122:121–158, 2014a) addressed several questions about instruments, methods and applications based on Chl a fluorescence. Here, additional Chl a fluorescence-related topics are discussed again in a question and answer format. Examples are the effect of connectivity on photochemical quenching, the correction of F _V /F _M values for PSI fluorescence, the energy partitioning concept, the interpretation of the complementary area, probing the donor side of PSII, the assignment of bands of 77 K fluorescence emission spectra to fluorescence emitters, the relationship between prompt and delayed fluorescence, potential problems when sampling tree canopies, the use of fluorescence parameters in QTL studies, the use of Chl a fluorescence in biosensor applications and the application of neural network approaches for the analysis of fluorescence measurements. The answers draw on knowledge from different Chl a fluorescence analysis domains, yielding in several cases new insights.     

The effect of light quality on the pro-/antioxidant balance,activity of photosystem II,and expression of light-dependent genes in <Emphasis Type="Italic">Eutrema salsugineum</Emphasis> callus cells

The antioxidant balance, photochemical activity of photosystem II (PSII), and photosynthetic pigment content, as well as the expression of genes involved in the light signalling of callus lines of Eutrema salsugineum plants (earlier Thellungiella salsuginea) under different spectral light compositions were studied. Growth of callus in red light (RL, maximum 660 nm), in contrast to blue light (BL, maximum 450 nm), resulted in a lower H₂O₂ content and thiobarbituric acid reactive substances (TBARS). The BL increased the activities of key antioxidant enzymes in comparison with the white light (WL) and RL and demonstrated the minimum level of PSII photochemical activity. The activities of catalase (CAT) and peroxidase (POD) had the highest values in BL, which, along with the increased H₂O₂ and TBARS content, indicate a higher level of oxidative stress in the cells. The expression levels of the main chloroplast protein genes of PSII (PSBA and PSBD), the NADPH-dependent oxidase gene of the plasma membrane (RbohD), the protochlorophyllide oxidoreductase genes (POR B, C) involved in the biosynthesis of chlorophyll, and the key photoreceptor signalling genes (CIB1, CRY2, PhyB, PhyA, and PIF3) were determined. Possible mechanisms of light quality effects on the physiological parameters of callus cells are discussed.     

Cloning and functional analysis of two <Emphasis Type="Italic">GmDeg</Emphasis> genes in soybean [<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merr.]

Although light is the ultimate substrate in photosynthesis, strong light can also be harmful and lead to photoinhibition. The DEG proteases play important roles in the degradation of misfolded and damaged proteins. In this study, two photoinhibition-related genes from soybean [Glycine max (L.) Merr.], GmDeg1 and GmDeg2, were cloned. Bioinformatics analysis indicated that these two proteases both contain a PDZ domain and are serine proteases. The expression levels of GmDeg1 and GmDeg2 increased significantly after 12 h of photooxidation treatment, indicating that GmDeg1 and GmDeg2 might play protective roles under strong light conditions. In in vitro proteolytic degradation assays, recombinant GmDeg1 and GmDeg2 demonstrated biological activities at temperatures ranging from 20°C to 60°C and at pH 5.0 to 8.0. By contrast, the proteases showed no proteolytic effect in the presence of a serine protease inhibitor. Taken together, these results provided strong evidence that GmDeg1 and GmDeg2 are serine proteases that could degrade the model substrate in vitro, indicating that they might degrade damaged D1 protein and other mis-folded proteins in vivo. Furthermore, GmDeg1 and GmDeg2 were transformed into Arabidopsis thaliana to obtain transgenic plants. Leaves from the transgenic and wild-type plants were subjected to strong light conditions in vitro, and the PSII photochemical efficiency (Fv/Fm) was measured. The Fv/Fm of the transgenic plants was significantly higher than that of the wild-type plants at most time points. These results imply that GmDeg1 and GmDeg2 would have similar functions to Arabidopsis AtDeg1, thus accelerating the recovery of PSII photochemical efficiency.