Some Factors Affecting the Hill Reaction Activity in Cotton Chloroplasts

A method of plant culture was developed for growing large leaves of glandless cotton on single stems. Chloroplasts isolated from these leaves actively reduced ferricyanide when assayed for the Hill reaction. Hill reaction activity increased 133% when the 0.5 m sucrose isolation medium was replaced with 10% (w/v) polyethylene glycol, both buffered at pH 7.6. The presence of 2 or 5% (w/v) bovine serum albumin in the sucrose buffer did not increase Hill activity. Ferricyanide reduction in the dark occurred in all assays, and the possibility of gossypol as the reductant is discussed. Half-life of the chloroplasts stored in 10% glycerol at -23 C was 23 days. The ammonium ion at 0.01 m enhanced Hill reaction activity up to 171%. Leaves containing chloroplasts with the highest Hill reaction activity were found near the 8th node below the apex. Leaf water potentials less than -28 bars reduced the activity about 50%. Daylight conditions during the winter months in the greenhouse reduced the activity about 30%.     

Luminescence Induced in Intact Leaves and Isolated Chloroplasts by Alcohols and Other Substances

Elodea leaves and spinach chloroplasts emit red light when treated with alcohols or certain other solvents for chlorophyll. The intensity of the light, the lag phase and the threshold concentration vary considerably between different alcohols. Light emission from a leaf starts a few seconds to a few minutes after the addition of alcohol to the medium, reaches a maximum after 2–45 minutes (sometimes more) and then continues for many hours. Despite the faintness of the glow, the total number of photons given off from a leaf after addition of alcohol may exceed the number of photons given off from the same sample as long-lived afterglow after saturating irradiation with far-red light. The maximum yield of photons per chlorophyll molecule is a little more than 10^–5. The alcohol-induced luminescence is not influenced by a decrease in the oxygen tension to one fifth of the normal. Electron micrographs of treated leaves reveal that the thylakoid lipids contract to drops at the edges of the grana. Treatment of isolated chloroplasts with ethanol results in light emission when the concentration is high enough to dissolve the chlorophyll. It is estimated that the surface free energy in the thylakoid lipid-aqueous interface, due to ordinary interfacial tension, is large enough to account for the light emission observed when the interfaces contract or disappear.     

Factors Affecting the Rate of Photophosphorylation by Isolated Chloroplasts of Euglena gracilis*

SYNOPSIS The rate of photophosphorylation by Euglena chloroplasts depends not only on the physiologic stage of cell growth but also on the stage of development of chloroplasts in these cells at the time of harvesting. Both of these processes can be markedly influenced by a number of environmental factors; they are affected neither in a parallel manner nor completely independent of each other. In addition, the rate of photophosphorylation of chloroplasts can also be greatly affected by the conditions employed for their isolation. After investigating the various environmental factors both during cell growth and chloroplast isolation, we have developed a procedure which increased the photophosphorylation rate of our chloroplast preparations more than 5-fold, giving a specific activity in the range of 100-150 μmoles ATP/mg chlorophyll/hr routinely. The procedure is simple, needs no special equipment and requires only 2 or 3 days for cell growth.     

Factors Affecting Cell Fusion Induced by Sendai Virus

Cell fusion mediated by exogenous Sendai virus appears to occur in four temperature-dependent stages. The first two, which include viral adsorption, are pH dependent and can be inhibited by viral antibody. Viral envelope constituents remain detectable on the cell surface during the third stage and disappear only when cell-to-cell fusion supervenes. The relationship of these interactions to possible mechanisms of cell fusion are discussed.     

Induction Kinetics of Millisecond-Delayed Luminescence in Intact Bryopsis Chloroplasts

The induction curve of delayed luminescence emitted from 0.5to 2.5 ms after excitation of dark-adapted intact chloroplastsof the green alga, Bryopsis maxima, showed three transient peaks,L₁, L₂ and L₃ (in order of appearance), at about 0.1, 1 and5 s after theonset of intermittent illumination. Intact chloroplastswere needed for L₂ to appear, whereas L₁ and L₃ were presentin hypotonically treated chloroplasts. L₁ and L₂ are related to the electric field generated acrossthe thylakoid membranesbecause the two peaks parallelled theappearance of the first and second peaks of electrochromic absorptionchanges at 560 nm and they were totally abolished by valinomycinand CCCP. A smaller contribution to the L₁ and L₂ of the protonactivity gradient across the membranes, or of pH changes insideor outside the membranes, was suggested by the partial suppressionof the transient by NH₄CI. L₃ is related to the proton gradient or pH changes because thetransient was inhibited by NH₄CI and CCCP but enhanced by N,N'-dicyclohexylcarbodiimide.In the presenceof valinomycin, which somewhat lowered the peakheight of L₃, the kinetics of delayed luminescence parallelledthat of fluorescence. Electrogenic reactions which occur sequentiallyduring the dark to light transition of the photosynthetic machineryin intact chloroplasts is discussed in connection with transientchanges in delayed luminescence. (Received November 8, 1982; Accepted May 21, 1983)     

Factors Affecting the Enhancement of Oxidative Stress Tolerance in Transgenic Tobacco Overexpressing Manganese Superoxide Dismutase in the Chloroplasts

Two varieties of tobacco (Nicotiana tabacum var PBD6 and var SR1) were used to generate transgenic lines overexpressing Mn-superoxide dismutase (MnSOD) in the chloroplasts. The overexpressed MnSOD suppresses the activity of those SODs (endogenous MnSOD and chloroplastic and cytosolic Cu/ZnSOD) that are prominent in young leaves but disappear largely or completely during aging of the leaves. The transgenic and control plants were grown at different light intensities and were then assayed for oxygen radical stress tolerance in leaf disc assays and for abundance of antioxidant enzymes and substrates in leaves. Transgenic plants had an enhanced resistance to methylviologen (MV), compared with control plants, only after growth at high light intensities. In both varieties the activities of FeSOD, ascorbate peroxidase, dehydroascorbate reductase, and monodehydroascorbate reductase and the concentrations of glutathione and ascorbate (all expressed on a chlorophyll basis) increased with increasing light intensity during growth. Most of these components were correlated with MV tolerance. It is argued that SOD overexpression leads to enhancement of the tolerance to MV-dependent oxidative stress only if one or more of these components is also present at high levels. Furthermore, the results suggest that in var SR1 the overexpressed MnSOD enhances primarily the stromal antioxidant system.     

Environmental Factors Affecting the Compressive Strength of Microbiologically Induced Calcite Precipitation-Treated Soil

Some microorganisms such as Sporoscarcina pasteurii precipitate calcium carbonate and are suitable for biocementation. This study aimed to investigate the effects of several factors including concentration of bacteria, chemical reactants, temperature, and pH on precipitation of calcium carbonate. The results showed that after 7 and 14 days of curing, the compressive strength of silty clay soil samples increased steadily as pH increased from 5 to 9. It was observed that pH plays an important role in biocementation. The highest compressive strength (i.e. 92 kPa) was observed when the soil was treated with 50 ml of bacterial solution after 14 days of curing. In addition, it was observed that the highest compressive strength of samples was achieved when the temperature was 40°C.     

Iron Deficiency Induced Changes on Electron Transport Rate in Pisum Sativum Chloroplasts

Iron deficiency induced decrease in the rate of whole electron transport chain in chloroplasts of pea (Pisum sativum L.). Such reduction was mainly due to the loss of photosystem (PS) 2 activity. The same result was obtained when the ratio of variable to maximum chlorophyll fluorescence (F_v/F_m) was evaluated. The loss in PS 2 activity was primarily due to a loss of 33, 23 and 17 kDa polypeptides. In contrast, iron deficiency induced the synthesis of 28 and 29 kDa polypeptides.     

Factors Affecting Grain Enlargement in Wheat

When most of the grains were removed from wheat ears the remaininggrains developed increased numbers of aleurone and endospermcells, and the increased grain volume was accompanied by anenlarged endosperm cavity. The giant grains usually synthesizedstarch more rapidly than normal grains. Flag leaf photosynthesiswas unaffected and additional sugar was retained in the culmand glumes. Total nitrogen and free amino acid content increased.Gibberellin and auxin concentrations were greater in the glumesof the partly degrained ears, and auxin in the grains was alsogreater than in equivalent grains from intact ears.     

Factors Affecting Transfection in Bacillus stearothermophilus

The conditions for the infection of Bacillus stearothermophilus 4S with TP-1C phage deoxyribonucleic acid (DNA) are described. Cells from log-phase cultures are the most susceptible to phage DNA infection (transfection). A cellular component (competence factor) which enhances transfection is released into the culture medium during the transition period between the log and stationary phase of growth. Transfection is stimulated in the order of decreasing effectiveness, by Fe(3+), Mn(2+), and Mg(2+). The efficiency of transfection is the highest in cells growing at 60.5 C and does not occur in cells growing at 67 C although the cells are growing normally. A cellular component (competence factor) of this organism, which is released into the culture medium, advances by 40 min some step in the uptake of phage DNA.     

Factors Affecting Egg-ratio in Planktonic Rotifers

Edmondson’s egg ratio (number of amictic eggs per female) is an important life history variable, which has been in wide use to understand and predict patterns of population growth in planktonic rotifers under field conditions. It is also useful as an indicator of the health of rotifers under culture conditions. Generally, an inverse relationship exists between the egg ratio and the density of females in a population. A number of abiotic and biotic factors influence the egg ratio. For example, temperature can cause marked changes in the egg ratio by influencing the frequency of egg production and the hatching times of parthenogenetic eggs. Also, preferential feeding on ovigerous females of rotifers by invertebrate predators such as Asplanchna will lower the egg ratios of the population. The easy detachment of eggs, as may be the case in some members of the Brachionidae especially during enhanced reproduction when food levels are high, may also cause an underestimation of the egg ratio. In this review, we discuss the egg ratio of selected rotifer species in relation to the role of diel changes in egg production, the frequency or the intensity of feeding, the problems of distinguishing between different egg types and the negative effect of stressors such as toxicants and diet quality.     

Seasonal and Experimentally Induced Changes in the Ultrastructure of Chloroplasts of Pinus silvestris

The ultrastructure of chloroplasts in mesophyll cells of Pinus silvesris was examined under the electron microscope. Secondary needles were regularly sampled from a tree in a natural stand for one year. Primary needles from one-year-old seedlings exposed to frost hardening and dehardening conditions in a controlled environment chamber were also studied. These seedlings were exposed to 8 or 55 W m^-2. All needles were put in fixative at the different sampling dates and stored in a refrigerator until they were prepared for electron microscopy at the end of the experimental period. During the summer the choroplasts were symmetrically shaped and heavily loaded with starch. The membrane systems were well developed and consisted of both grana and stroma thylakoids. In autumn and during early artificial frost hardening the starch content was reduced, the chloroplasts appeared amoeboid and membrane-free stroma regions were seen. Later the chloroplasts became swollen and aggregated in one part of the cell. Starch was lost and the chloroplasts aggregated earlier at 8 W m^-2 than at 55 W m^-2. During winter the stroma thylakoids were first reduced in number and later even the grana thylakoids were damaged, resulting in mostly disorganized single membranes. Also the chloroplast envelope disappeared. In spring and early summer the chloroplasts migrated to the proximity of the cell walls. The membrane systems were reorganized and starch accumulated. During the first days of artificial dehardening the photosynthetic membranes were severely damaged, especially at 55 W m^-2, but soon new membranes were formed. Starch accumulated earlier at 55 than at 8 W m^-2. The reported ultrastructural variations are discussed in relation to functional and biochemical fluctuations caused by the season or by artificial variations in the climate as demonstrated earlier.     

The P680 Difference Spectrum Induced by Continuous Light in Malaxis monophyllos Chloroplasts

In chloroplasts from M. monophyllos (L.) Sw., the difference spectrum of light-induced P680 which was measured under physiological temperature and neutral pH and in the presence of potassium ferricyanide was obtained by steady-state method. In contrast under the same conditions, the P680 signal was not observed in spinach chloroplasts. Either preilumination with white light or long duration treatment with potassium ferricyanide caused marked similar changes in the difference spectrum of M. monophyllos chloroplasts, i.e. besides the bleaching bands at wavelength near 680 mm and 440 nm were the same as the control, the remarkable bleaching bands at about 660 nm and 422 nm appeared. These results were discussed briefly.     

Updated Progress on Group II Intron Splicing Factors in Plant Chloroplasts

Group II introns are large catalytic RNAs (ribozymes) in the bacteria and organelle genomes of several lower eukaryotes. Many critical photosynthesis-related genes in the plant chloroplast genome also contain group II introns, and their splicing is critical for chloroplast biogenesis and photosynthesis processes. The structure of chloroplast group II introns was altered during evolution, resulting in the loss of intron self-splicing. Therefore, the assistance of protein factors was required for their splicing processes. As an increasing number of studies focus on the mechanism of chloroplast intron splicing; many new nuclear-encoded splicing factors that are involved in the chloroplast intron splicing process have been reported. This report reviewed the research progress of the updated splicing factors found to be involved in the splicing of chloroplast group II introns. We discuss the main problems that remain in this research field and suggest future research directions.     

Factors Affecting Chloroplast Replication in Spinach

Chloroplast replication has been studied in discs cut from thebase of young spinach leaves and cultured on sterile nutrientagar. In discs grown in a growth cabinet chloroplast numbersper cell increased logarithmically with time over a 7-day cultureperiod. Chloroplast replication proceeds in a similar way incultured discs and in intact leaves. Cytokinins do not affect chloroplast replication in this systembut they stimulate the fresh-weight growth of discs. Chloroplastreplication is temperature dependent, having an optimum at 25°C. By contrast chloroplast size is at a maximum in discscultured at 12 °C. Light stimulates chloroplast replication, a linear relationshipoccurring between chloroplast number per cell and the dailyquantity of light given to discs up to a saturating value of250 J d^–1. Daylength does not affect chloroplast formationin spinach. In a number of experiments a general relationship was establishedbetween chloroplast number per cell and cell size but no evidenceis available to suggest that this correlation is causal. Theresults of experiments in which discs were transferred fromdark to light suggest that some of the events which precedechloroplast replication may occur at similar rates in both lightand dark.     

环境因素对L-异亮氨酸发酵的影响

通过 5L自控发酵罐发酵实验 ,结合发酵过程中菌生长形态的变化 ,对L -异亮氨酸补料分批发酵进行研究 ,研究了环境因素对黄色短杆菌 (Brevibacteriumflavum)TJCN - 1的影响 ,优化出发酵最佳控制条件 ,提出分阶段发酵控制模式 ,对L -异亮氨酸生产有指导意义。