Involvement of protein phosphorylation in the sensitivity of photosystem II to strong illumination

Four types of differently phosphorylated hylakoids isolated from field grown spinach ( Spinacia oleracea L.) were tested for the sensitivity of photosystem II (PSII) to photoinactivation. Phosphorylation of light-harvesting II complexes (LHCII) protected PSII electron transfer from photoinhibitory damage, while the phosphorylation of the PSII core polypeptides slightly accelerated the decline of electron transfer during high irradiance treatment. Dephosphorylation of the CP43 apoprotein and PsbH protein by an alkaline phosphatase resulted in an extreme sensitivity of the thylakoids to strong illumination. The PSII photoinactivation of thylakoids with the impaired oxygen-evolving complex was found to be independent of phosphorylation.
The thylakoids of the thermophilic cyanobacterium Synechococcus elongates were used in order to compare the plants with an organism where LHCII complexes are missing and the PSII core proteins are not phosphorylated.     

Reconstitution of iron-sulfur center B of Photosystem I damaged by mercuric chloride

Incubation of thylakoid membranes from spinach with low concentrations of mercuric chloride induces the loss of one of the iron-sulfur centers, F_B, in Photosystem I (PS I) and inhibits the electron transfer from PS I to the soluble electron carrier, ferredoxin. Reconstitution of this damaged iron-sulfur center has been carried out by incubating treated thylakoid membranes with exogenous FeCl₃ and Na₂S in the presence of-mercaptoethanol under anaerobic conditions. Low temperature EPR measurements indicate that center F_B is largely restored. Kinetic experiments show that the restored F_B can be photoreduced from P700. However, these reconstituted thylakoid membranes are still incompetent in the photoreduction of ferredoxin and NADP⁺, even though ferredoxin binding to the modified membranes was not impaired, indicating additional changes in the structure of the PS I complex must have occurred.     

Embryogenesis and plant regeneration of spinach (Spinacia oleracea L.) from hypocotyl segments

A system for somatic embryogenesis and plant regeneration of spinach from hypocotyl segments has been established. Callus was induced on solid media supplemented with 8.5–15.0 mg.l⁻¹ of indole-3-acetic acid and 3.46–34.64 mg.l⁻¹ gibberellic acid. Callus was then subcultured on different media (solid or liquid) with or without IAA, or continuously maintained on the initiating media. Somatic embryos were obtained in subcultures on IAA-containing media as well as in long-term cultures on initiating media. The best results were achieved in liquid subcultures. About 60% of plantlets survived after transplanting in pots.     

Light-induced fluorescence quenching in the light-harvesting chlorophyll a/b protein complex

Summary Irradiation of the principal photosystem II light-harvesting chlorophyll-protein antenna complex, LHC II, with high light intensities brings about a pronounced quenching of the chlorophyll fluorescence. Illumination of isolated thylakoids with high light intensities generates the formation of quenching centres within LHC II in vivo, as demonstrated by fluorescence excitation spectroscopy. In the isolated complex it is demonstrated that the light-induced fluorescence quenching: a) shows a partial, biphasic reversibility in the dark; b) is approximately proportional to the light intensity; c) is almost independent of temperature in the range 0–30°C; d) is substantially insensitive to protein modifying reagents and treatments; e) occurs in the absence of oxygen. A possible physiological importance of the phenomenon is discussed in terms of a mechanism capable of dissipating excess excitation energy within the photosystem II antenna.Abbreviations chla chlorophyll a - chlb chlorophyll b - F₀ fluorescence yield with reaction centers open - F_m fluorescence yield with reaction centres closed - F_i fluorescence at the plateau level of the fast induction phase - LHC II light-harvesting chlorophyll a/b protein complex II - PS II photosystem II - PSI photosystem I - Tricine N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]glycine     

Continuous shoot growth monitoring in hydroponics

A weighing apparatus for automatic recording of fresh weight of shoots of spinach plants ( Spinacia oleracea L., cv. Subito) growing in nutrient solution is described. The system was tested for 17 days in a controlled environment and enabled the determination of the relative growth rate (RGR) of the shoot fresh weight. Results from three consecutive growth experiments demonstrated diurnal fluctuations in the relative growth rate of the shoot fresh weight. In general, relative growth rates were between 0.32 and 0.36 day⁻¹ 16 days after sowing and decreased to between 0.11 and 0.18 day⁻¹ during the 12 following days. The variance between three replicate growth curves was compared with the variance of a growth function fitted through destructively obtained spinach shoot weight data.     

Influence of nitrate concentration at the root surface on yield and nitrate uptake of kohlrabi (Brassica oleracea gongyloides L.) and spinach (Spinacia oleracea L.)

The objective of this study was to determine if plant roots have to take up nitrate at their maximum rate for achieving maximum yield. This was investigated in a flowing-solution system which kept nutrient concentrations at constant levels. Nitrate concentrations were maintained in the range 20 to 1000 μM. Maximum uptake rate for both species was obtained at 100 μM. Concentrations below 100 μM resulted in decreases in uptake rate per cm root (inflow) for both spinach and kohlrabi by 1/3 and 2/3, respectively. However, only with kohlrabi this caused a reduction in N uptake and yield. Thus indicating that this crop has to take up nitrate at the maximum inflow. Spinach, however, compensated for lower inflows by enhancing its root absorbing surface with more and longer roots hairs. Both species increased their root length by 1/3 at low nitrate concentrations.     

Variable agronomic practices, cultivar, strain source and initial contamination dose differentially affect survival of Escherichia coli on spinach

Aims: Greenhouse and field trials were conducted under different agronomic practices and inoculum doses of environmental Escherichia coli and attenuated E. coli O157:H7, to comparatively determine whether these factors influence their survival on leaves and within the rhizosphere. Methods and Results: Hydroponic conditions: E. coli spray‐inoculated at log 4 CFU ml^?1 was recovered from leaf surfaces at a mean population of 1·6 log CFU g^?1 at 15 days. E. coli O157:H7 sprayed at log 2 or 4 CFU ml^?1 levelled off on spinach leaf surfaces at a mean average population of 1·4 log CFU g^?1 after 14 days, regardless of initial dose. Quantitative recovery was inconsistent across leaf developmental age. Field conditions: Average populations of E. coli O157:H7 spray‐inoculated at log 1·45 or 3·4 CFU m^?2 levelled off at log 1·2 CFU g^?1 over a 14‐day period. Pathogen recovery from leaves was inconsistent when compared to regularly positive detection on basal shoot tissue. Pathogen recovery from soil was inconsistent among sampling locations. Moisture content varied up to 40% DW and was associated with 50% (P < 0·05) decrease in positive locations for E. coli O157:H7 but not for E. coli. Conclusions: Overall, similar populations of environmental E. coli and E. coli O157:H7 were recovered from plants despite differences in inoculum dose and agronomic conditions. Strain source had a significant impact on the quantitative level and duration of survival on leaves and in soil. Water availability appeared to be the determinant factor in survival of E. coli and E. coli O157:H7; however, E. coli showed greater environmental fitness. Significance and Impact of the Study: Persistence of surrogate, indicator E. coli and E. coli O157:H7, irrespective of variable growing conditions in spinach is predominantly limited by water availability, strain source and localization within the plant. These findings are anticipated to ultimately be adopted into routine and investigative pathogen testing protocols and mechanical harvest practices of spinach.     

Switching among natal and auxiliary hosts increases vulnerability of Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) to insecticides

The role of insecticidal application and host plant resistance in managing Spodoptera exigua has been well documented, but the effect of different host plants, on which the pest cycles its population in the field, has seldom been investigated. Therefore, we have studied the vulnerability of S. exigua against commonly used insecticides (cypermethrin, chlorpyrifos, lufenuron, and emamectin benzoate) with different mode of actions when it switches its generations from natal to auxiliary hosts and vice versa. Different field populations being established on different host plants including castor, cauliflower, cotton, okra, and spinach were collected and reared in the laboratory before insecticidal bioassays. The role of larval diet and host plant switching on their response to tolerate applied insecticides was studied using leaf‐dip bioassay methods. Host switching demonstrated a significant role in altering the vulnerability of S. exigua populations to tested insecticides. Spodoptera exigua sourced from castor, when switched host to okra and spinach, exhibited 50% higher mortality when treated with emamectin benzoate. This trend in mortality was consistent upon complete host switch cycle (natal—auxiliary—natal host). However, the highest increase (92%) in vulnerability was recorded when the larvae were shifted to spinach from cotton. In general, chlorpyrifos and lufenuron had highest efficacies in terms of larval mortality. The findings of present studies provide insights to a better understanding the behavior of polyphagous pests and the role of different host plants in altering the susceptibility of these pests against applied insecticides. Ultimately the results warrant that due consideration should be given to cropping patterns and time of host switching by pest population during planning and executing chemical control.     

Influence of Lime and Organic Matter on the Mobility of Cadmium in Cadmium-Contaminated Soil in Relation to Nutrition of Spinach

Cadmium and cadmium compounds are water soluble, mobile in most soils, bio-available, and tend to bio-accumulate. A pot culture experiment was conducted on contaminated soil to study the influence of lime and organic matter on the mobility of cadmium in spinach and its rhizosphere soil. Application of lime (50% and 100% lime requirement) and organic matter (0.5 and 1% by weight of soil) to soil decreased the availability of Cd to the soil and plant throughout the crop growth. The highest diethylene triamine penta-acetic acid (DTPA) extractable Cd was 10.84 mg kg^?1 in the treatment OM₀ L₀ (No application of organic matter and lime) at 20 days after sowing of spinach. Likewise, the highest Cd concentration in spinach roots and shoots were 19.80 and 17.0 mg kg^?1 in the treatment OM₀ L₀ at 20 days after sowing. The Cd concentration in spinach roots and shoots were decreased by 63.23 and 71.88%, respectively, in the treatment OM₁ L₁₀₀ (application of FYM at 1.0% by weight of soil and lime at 100% lime requirement) after 60 days of growth. The lowest concentrations of Cd in the soil and plant after the harvest of the crop were 2.88 and 4.27 mg kg^?1, respectively, in the treatment OM₁ L₁₀₀ and resulted in 65.75 and 71.55% decrease over control (OM₀ L₀). The highest total chlorophyll content of leaves was 2.19 mg kg^?1 of fresh weight in the treatment OM₁ L₁₀₀ at 40 days of crop growth.     

乐果对菠菜叶片POD、SOD、CAT活性及MDA含量的影响

研究了喷施乐果后不同时间及不同施用量对菠菜(Spinacia oleracea)叶片POD、SOD和CAT活性及MDA含量的影响;同时检测了喷药后不同时间菠菜中乐果的残留量。结果表明,喷施乐果可促使菠菜叶片中POD和CAT活性增强以及MDA含量明显增加,而SOD活性在部分乐果处理中呈下降趋势;随着喷药后时间的延长,乐果在菠菜中的残留量亦随之减少。