Functional redundancy between flavodiiron proteins and NDH‐1 in Synechocystis sp. PCC 6803

In oxygenic photosynthetic organisms, excluding angiosperms, flavodiiron proteins (FDPs) catalyze light‐dependent reduction of O₂ to H₂O. This alleviates electron pressure on the photosynthetic apparatus and protects it from photodamage. In Synechocystis sp. PCC 6803, four FDP isoforms function as hetero‐oligomers of Flv1 and Flv3 and/or Flv2 and Flv4. An alternative electron transport pathway mediated by the NAD(P)H dehydrogenase‐like complex (NDH‐1) also contributes to redox hemostasis and the photoprotection of photosynthesis. Four NDH‐1 types have been characterized in cyanobacteria: NDH‐1₁ and NDH‐1₂, which function in respiration; and NDH‐1₃ and NDH‐1₄, which function in CO₂ uptake. All four types are involved in cyclic electron transport. Along with single FDP mutants (?flv1 and Δflv3) and the double NDH‐1 mutants (?d1d2, which is deficient in NDH‐1_1,2 and ?d3d4, which is deficient in NDH‐1_3,4), we studied triple mutants lacking one of Flv1 or Flv3, and NDH‐1_1,2 or NDH‐1_3,4. We show that the presence of either Flv1/3 or NDH‐1_1,2, but not NDH‐1_3,4, is indispensable for survival during changes in growth conditions from high CO₂/moderate light to low CO₂/high light. Our results show functional redundancy between FDPs and NDH‐1_1,2 under the studied conditions. We suggest that ferredoxin probably functions as a primary electron donor to both Flv1/3 and NDH‐1_1,2, allowing their functions to be dynamically coordinated for efficient oxidation of photosystem I and for photoprotection under variable CO₂ and light availability.     

Multilevel regulation of non‐photochemical quenching and state transitions by chloroplast NADPH‐dependent thioredoxin reductase

In natural growth habitats, plants face constant, unpredictable changes in light conditions. To avoid damage to the photosynthetic apparatus on thylakoid membranes in chloroplasts, and to avoid wasteful reactions, it is crucial to maintain a redox balance both within the components of photosynthetic electron transfer chain and between the light reactions and stromal carbon metabolism under fluctuating light conditions. This requires coordinated function of the photoprotective and regulatory mechanisms, such as non‐photochemical quenching (NPQ) and reversible redistribution of excitation energy between photosystem II (PSII) and photosystem I (PSI). In this paper, we show that the NADPH‐dependent chloroplast thioredoxin system (NTRC) is involved in the control of the activation of these mechanisms. In plants with altered NTRC content, the strict correlation between lumenal pH and NPQ is partially lost. We propose that NTRC contributes to downregulation of a slow‐relaxing constituent of NPQ, whose induction is independent of lumenal acidification. Additionally, overexpression of NTRC enhances the ability to adjust the excitation balance between PSII and PSI, and improves the ability to oxidize the electron transfer chain during changes in light conditions. Thiol regulation allows coupling of the electron transfer chain to the stromal redox state during these changes.     

Identification of the spermatogenic stages in living seminiferous tubules of man.

Parts of human seminiferous tubules containing late spermatids with condensed nuclei (Stage II) absorbed more transmitted light than did parts at other stages. Spermatogenic stages I, III, IV and VI were identifiable by phase-contrast microscopy.     

Fertility variation in Pinus sylvestris: A TEST OF SEXUAL ALLOCATION THEORY

Sexual allocation theory assumes trade-offs and negative genetic correlations between male and female allocation in hermaphrodite plants. We tested this assumption by studying variation in male and female fertility in two experimental populations of Pinus sylvestris. In these populations, the genotypes have been vegetatively replicated, which allowed separation of the genetic and environmental components of variation. The genetic components of variation accounted for 36% of the total variation in pollen production in the two populations, and for 54% of cone production. As assumed by sexual allocation theory, the genetic correlation of pollen and cone production was negative in both populations (-0.59 and -0.15). However, positive environmental correlations between these traits (0.43 and 0.45) resulted in no phenotypic correlation in one population and a positive phenotypic correlation in the other.     

Pollen competition and seed-siring success in Picea abies

The aim of the present work was to study pollen-tube competition in Picea abies. Controlled crossings were performed with pollen mixtures including pairs of pollen lots with fast and slowly elongating pollen-tubes. Paternity analysis using isozyme markers was performed on the progenies in order to study whether the in vitro pollen-germination vigour corresponds to the proportion of seeds sired by the pollen donor. Paternal success was found to be unequal, 15 out of 23 crossings producing progeny that differed significantly from the hypothetical ratio of 1:1. The paternal contribution in the majority of the crossings was as expected: the pollen parent with more-vigorous in vitro germination sired more seeds than the less-vigorous pollen. In the case of two pollen mixtures, however, the seed-siring success summed over the maternal trees was the opposite to the expected value. Despite these aberrations, the results support the hypothesis that pollen-tube competition is one of the factors contributing to male fitness in P. abies. However, when all the other factors affecting pollination and seed set under natural conditions are taken into account, it is clear that the seed-siring success of a particular paternal genotype cannot be predicted reliably by measuring only the in vitro pollen vigour. Received: 2 July 2001 / Accepted: 7 August 2001     

The Production of Transgenic Scots Pine (Pinus Sylvestris L.) via the Application of Transformed Pollen in Controlled Crossings

The study demonstrates the production of a transgenic Scots pine (Pinus sylvestris L.) seedling through the application of transformed pollen in controlled crossings. The pollen lots were transformed by particle bombardment, resulting in transient transformation frequencies varying from 15 to 49% of the germinated pollen grains, and bombarded pollen was used to pollinate megasporangiate strobili. Progeny was screened by histochemical, GUS assays, and selected seedlings were further analysed by PCR. PCR amplification revealed the presence of both the nptII and gusA genes in one seedling (23/237). Results were confirmed by Southern blot analysis. The morphology and growth of this transgenic seedling was normal. Although the transformation frequency of recovered plants was very low (1/14999), the present protocol suggests that production of transgenic Scots pine is possible without the use of any tissue culture methods or the involvement of marker genes, for selection of transformants.     

Crosstalk between chloroplast thioredoxin systems in regulation of photosynthesis

Thioredoxins (TRXs) mediate light‐dependent activation of primary photosynthetic reactions in plant chloroplasts by reducing disulphide bridges in redox‐regulated enzymes. Of the two plastid TRX systems, the ferredoxin‐TRX system consists of ferredoxin‐thioredoxin reductase (FTR) and multiple TRXs, while the NADPH‐dependent thioredoxin reductase (NTRC) contains a complete TRX system in a single polypeptide. Using Arabidopsis plants overexpressing or lacking a functional NTRC, we have investigated the redundancy and interaction between the NTRC and Fd‐TRX systems in regulation of photosynthesis in vivo. Overexpression of NTRC raised the CO₂ fixation rate and lowered non‐photochemical quenching and acceptor side limitation of PSI in low light conditions by enhancing the activation of chloroplast ATP synthase and TRX‐regulated enzymes in Calvin–Benson cycle (CBC). Overexpression of NTRC with an inactivated NTR or TRX domain partly recovered the phenotype of knockout plants, suggesting crosstalk between the plastid TRX systems. NTRC interacted in planta with fructose‐1,6‐bisphosphatase, phosphoribulokinase and CF₁γ subunit of the ATP synthase and with several chloroplast TRXs. These findings indicate that NTRC‐mediated regulation of the CBC and ATP synthesis occurs both directly and through interaction with the ferredoxin‐TRX system and is crucial when availability of light is limiting photosynthesis.     

Electrical activity and intraluminal pressure of the cauda epididymidis of the rat

Smooth muscle electrical activity was recorded simultaneously with intraluminal pressure in the cauda epididymidis and epididymal vas of the rat in vitro. Spontaneous electrical activity spread over distances of several centimetres and was associated with a pressure wave recorded through an open-ended catheter in the distal cauda and epididymal vas. Transmission of pressure wave was gradually damped by filling of the catheter with spermatozoa.     

Examination of living Sertoli cells in normal and hypospermatogenic men.

The functional state of living Sertoli cells can be assessed by using transillumination and phase-contrast microscopy. In tissue from hypospermatogenic men the Sertoli cells contained more lipid droplets than in normal cases and sometimes whole degenerative cells.     

Variation in pollen viability among Picea abies genotypes – potential for unequal paternal success

An in vitro germination method was used to study variation in pollen viability, that is pollen-tube growth rate together with germination percentage, among the Picea abies genotypes in a seed orchard. The method permits easy, rapid screening of large numbers of genotypes. Significant variation in pollen viability among the genotypes was evident, the differences among the pollen-lot means being 7–10–fold in different years. No correlation was found between the average pollen viability and the phenology, growth or growing-site characteristics of the pollen donors. However, there appeared to be pollen lots that either benefit from a higher germination temperature or else germinate faster at lower temperatures. The significant variation in pollen viability among the pollen donors indicates a potential for male gametophyte competition. This, together with the observed genotype-environment interactions in pollen performance, may contribute to the variable genetic composition of seed produced in the seed orchard. Received: 9 December 1999 / Accepted: 22 December 1999