Continuous Light Conditions Research Articles

Impact of Photoperiod on the Circadian Cellulose and Cellulase Rhythms in the Digestive System of Silkworm, Bombyx Mori

Circadian cellulose and cellulase rhythms were studied in the digestive system of Bombyx mori under 12 hr light-dark cycle (LD), continuous light (LL) and continuous dark (DD) conditions. Since, the symbiotic microbes are associated with cellulose digestion in the silkworm; both the rhythms are presumed to have microbial origin. The rhythms were analyzed in terms of phase response curves and interpreted as cellulose synthetic cycles (CS cycles) and cellulase enzyme synthetic cycles (CES cycles) in gut wall and cellulose uptake cycles (CU cycles) and cellulase enzyme release cycles (CER cycles) in gut lumen. The cellulose rhythm in the gut wall maintained 7 CS cycles each with a duration of 3.4 hr and the 24 hr free running time remained unchanged under altered photoperiodic conditions. In gut lumen, the rhythm showed 7 CU cycles, each with a duration of 3.4 hr under LD and 6 CU cycles of 4.0 hr each under LL and DD, with the result, the 24 hr free running period of the rhythm was clock shifted to 28 hr both under LL and DD. The cellulase rhythm in the gut wall maintained 7 CES cycles with a duration of 3.4 hr each under LD and DD and 9 cycles each with a duration of 2.7 hr under LL. The 24 hr free running time of the cellulase rhythm of LD and DD, was thus advanced to 19 hr under LL. The cellulase in gut lumen maintained 6 CER cycles with a duration of 4.0 hr each, under LD, LL and DD and hence the 24 hr free running time of the rhythm was not affected. Further analysis of data showed that cellulose synthesis was stimulated by light cues, while its uptake from gut lumen through enhanced microbial cellulase activity was favoured by both diet and light cues.

Spatiotemporal dynamics of circadian clock in lettuce

Abstract Plant circadian clock controls many physiological events by synchronization with environmental changes. The clock can be controlled artificially by imposing various light conditions in a closed system for plant cultivation in plant factory. The plant circadian rhythm is formed by enormous self-sustained cellular circadian oscillators, so that the synchronization of circadian oscillators is of great importance to the formation of individual circadian rhythms. Therefore, the study of synchronization phenomena is important for precise control of the circadian rhythm. In this study, we have investigated the spatiotemporal dynamics of circadian oscillators in lettuce (Lactuca sativa L.). Bioluminescence of transgenic lettuce carrying a CCA1::LUC construct as a reporter of a circadian gene expression was measured in the leaf and root. We observed phase wave propagation in the leaf with a phase delay in the region of the primary vein, with the primary vein region showing lower amplitude than the other regions. Wave propagation occurred from the edge of the leaf inward. In addition, a striped wave traveled from the base to the tip along the roots, which were grown under continuous dark or light conditions. The features observed in the lettuce plants could be explained by phase oscillator models established in the study of Arabidopsis thaliana. The results of this study show the possibility of applying circadian clock control in a model plant for plant production.

Effects of light intensities and photoperiods on growth and proteolytic activity in purple non-sulfur marine bacterium, <i>Afifella marina</i> strain ME (KC205142)

Afifella marina strain ME (KC205142), a purple non-sulfur bacterium was isolated from mangrove habitats of Sabah. The effects of light intensities and photoperiods on proteolytic activity in Afifella marina strain ME (KC205142) were investigated. Secretion of proteolytic enzymes in Afifella marina was preliminarily assessed by skim milk agarose media. Subsequently, light intensities, such as, dark, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500 and 5000 lux were used to evaluate the effects on proteolytic activity in Afifella marina strain ME under anaerobic condition. After that, the effect of photoperiods on proteolytic activity was monitored under anaerobic light condition (3000 lux) at 0 h (0L/24D), 6 h (6L/18D), 12 h (12L/12D), 18 h (18L/6D) and 24 h (24L/0D) of photoperiod. The highest proteolytic activity of 74.67 U was recorded at 3000 lux illumination light intensity. The proteolytic activity in bacterium Afifella marina strain ME was positively associated with the dry cell weight. The proteolytic activity of 72.67 U in bacterium Afifella marina strain ME at 18 h (18L/6D) photoperiod is not significantly different (p > 0.05) from proteolytic activity of 74.67 U recorded at continuous light (24L/0D) condition. Light intensity of 3000 lux, culture period of 48 h and a photoperiod of 18 h (18L/ 6D) were the optimum parameters for proteolytic activity in bacterium Afifella marina strain ME.

Effects of temperature, light regime and substrates on the production and germination of Stenocarpella maydis pycnidiospores

This study aimed to evaluate the production and germination of Stenocarpella maydis pycnidiospores under in vitro conditions as affected by substrate composition (sorghum, wheat, black oat or barley), light regime (continuous dark, 12-h light dark -1 or continuous light conditions), and incubation temperature (21, 24, 27, 30 or 33oC). Each substrate (20 g of grain) was soaked in 100 mL of water for 24h and sterilized twice for 20 min at 127oC. Three plugs (5 mm diameter) of a single-spored culture of S. maydis were used as inocula for each substrate. Assessments of pycnidiospore production per gram of grain and percent germination were made 14 days after inoculation. Barley, black oat or wheat grains were the best substrates for the mass production of S. maydis pycnidiospores and the maintenance of high germination rates. The highest pycnidiospore production (67,600 pycnidiospores g -1 ) was obtained using barley grain as a substrate with incubation at 27oC under a 12-h light dark -1 cycle.

Use of Linear and Weibull Functions to Model Ascorbic Acid Degradation in Chinese Winter Jujube during Postharvest Storage in Light and Dark Conditions

Degradation of ascorbic acid (AA) in Zhanhua winter jujube stored at selected temperatures under continuous light and darkness conditions was described by first-order kinetics and Weibull distribution function, and the temperature dependence of AA loss was studied by the Arrhenius equation. The high correlation coefficients (R 2 > 0.99) indicated the acceptability of Weibull model for predicting the degradation of AA. Activation energy was determined to be 10.586 and 11.868 kcal/mol under continuous light condition and 12.636 and 13.190 kcal/ mol under continuous darkness condition according to first-order and Weibull models, respectively. This study revealed that the loss of AA in continuous darkness was more temperature-dependent than the other samples. In continuous light, AA degradation may be favored even if at low temperatures. Therefore, a low temperature and continuous darkness condition is recommended to preserve AA content in jujube fruits during postharvest storage.

둥근마(Dioscorea opposita Thunb.) 기내 소괴경 생산을 위한 배양환경과 배지의 교체효과

둥근마 액체배양에서 소괴경의 생산을 위하여 배양온도와 일장의 전환과 배양 배지의 교체 효과를 조사하였다. 둥근마 액체배양에서 식물체의 증식을 위한 일장시간은 6주의 배양기간 동안 <TEX>$25^{\circ}C$</TEX>에서 16시간보다는 24시간이 효과적이었다. 기내소괴경의 생산을 위하여 배양기간을 12주로 연장하여 배양하였으며, 배지교체의 효과를 검토하기 위하여, 배양 중기인 배양 후 6주째에 새로운 배지로 교체하였더니, 배지를 교체하지 않은 것보다 소괴경의 형성수가 평균 5개씩 더 많았고, 소괴경무게도 70% 이상 증가하였다. 배양환경의 전환이 소괴경 형성에 미치는 효과를 알아보기 위하여, 12주의 배양기간 중에 식물체 생장이 활발한 초기 6주간은 <TEX>$25^{\circ}C$</TEX>에서 24시간의 일장환경에서 배양하고, 이후 6주간의 배양온도와 일장시간을 전환하였다. 배양온도는 <TEX>$25^{\circ}C$</TEX>가 <TEX>$15^{\circ}C$</TEX>보다 효과적이었고, <TEX>$25^{\circ}C$</TEX>에서 일장시간은 24시간이 암처리 또는 8시간 일장처리보다 효과가 있었다. 12주간의 배양기간중에 배지 교체의 적절한 시기는 배양 후 2, 4, 6, 8, 10주째 또는 배지 무교체 처리구중에서, 생체중, 마디수, 소괴경수에서는 2주 또는 6주째에 교체하는 것이 좋았고, 소괴경중은 8주째 교체 처리구에서 가장 무거웠으나 소괴경의 수와 무게를 모두 고려하였을 때, 초기 배양 후 6주째에 배지 교체하는 것이 가장 효율적이었다. 12주간의 배양기간 중 배양 후 6주째에 교체할 배지의 성분은 sucrose 30, 60 <TEX>$g{\cdot}L^{-1}$</TEX> 또는 sucrose 60 g + BA 0.1 <TEX>$mg{\cdot}L^{-1}$</TEX> 함유 배지 중에서 sucrose 60 <TEX>$g{\cdot}L^{-1}$</TEX>이 함유된 배지로 교체하였을 때, 소괴경의 총무게가 가장 무거워 소괴경의 비대에 가장 효과적이었다. 따라서, 둥근마의 액체배양시 <TEX>$25{\pm}1^{\circ}C$</TEX>, 24시간 광조건으로, 마디접종 후 6주째에 sucrose 60 <TEX>$g{\cdot}L^{-1}$</TEX>를 함유한 배지로 교체하여 6주간 추가 배양하는 방법이 기내 식물체와 소괴경의 생장과 형성에 가장 효과적이었다. To figure out optimum culturing condition for <TEX>$in$</TEX> <TEX>$vitro$</TEX> yam (<TEX>$Dioscorea$</TEX> <TEX>$opposita$</TEX> Thunb.) production, various media components and environmental controls were evaluated, effective temperature, light condition and timing of the liquid media replacement for multiplication of yam in liquid culture were determined in this study. There was no visible difference detected for <TEX>$15^{\circ}C$</TEX> and <TEX>$25^{\circ}C$</TEX> temperature conditions. At <TEX>$25^{\circ}C$</TEX>, continuous light condition was more effective compared to 16 hour light/ 8 hour dark condition. Effect of media replacement was tested, and approximately 5 more microtubers obtained and 70% of increase in weight was detected when media replacement was performed. Timing for media replacement was tested at 2, 4, 6, 8, and 10 weeks after initial culture for 12 weeks. Considering both number and weight of microtubers, replacement of media 6 weeks after initial culture was most effective. In terms of component of media, significant increase in weight of microtubers was observed in MS media containing sucrose <TEX>$60g{\cdot}L^{-1}$</TEX>. In summary, the most effective condition for <TEX>$in$</TEX> <TEX>$vitro$</TEX> propagation of chinese yam is replacing medium 6 weeks after initial inoculation with MS medium containing sucrose <TEX>$60g{\cdot}L^{-1}$</TEX> in continuous light condition.

Modulation of fungicidal potential of Anabaena strains by light and temperature

The regulation of fungicidal and hydrolytic enzyme activity was investigated in a set of cyanobacterial strains belonging to the genus Anabaena (Anabaena laxa RPAN8, Anabaena iyengarii RPAN9, Anabaena variabilis RPAN59 and Anabaena oscillarioides RPAN69), with A. variabilis RPAN16 serving as negative control. Time course studies undertaken with cultures incubated under different light and temperature conditions revealed enhancement in growth and fungicidal activity under continuous light (CL) and light dark (LD, 16:8) conditions and temperature of 30 °C and 40 °C. A significant increase of 3-18 % in chitosanase activity was recorded in all the 4-week-old cultures under CL condition and at 40 °C. Endoglucanase activity of RPAN8 and 9 was twofolds higher than the other strains under all light/dark conditions and temperature in the 4-week-old cultures, while continuous dark (CD) enhanced CMCase activity in RPAN69. This study provided useful information regarding the most suitable conditions of light and temperature for maximizing hydrolytic enzyme activity and fungicidal activity, as a prelude to their effective use as biocontrol agents.

Competing carboxylases: circadian and metabolic regulation of Rubisco in C3 and CAM Mesembryanthemum crystallinum L.

The temporal co-ordination of ribulose 1·5-bisphosphate carboxylase/oxygenase (Rubisco) and phosphoenolpyruvate carboxylase (PEPc) activities by Mesembryanthemum crystallinum L. in C(3) and crassulacean acid metabolism (CAM) modes was investigated under conventional light-dark (LD) and continuous light (LL) conditions. When C(3) , net CO(2) assimilation rate increased during each subjective night under LL with maximum carboxylation unrelated to Rubisco activation state. The CAM circadian rhythm of CO(2) uptake was more pronounced, with CO(2) assimilation rate maximal towards the end of each subjective night. In vivo and in vitro techniques were integrated to map carboxylase enzyme regulation to the framework provided by CAM LL gas exchange activity. Rubisco was activated in vitro throughout each subjective dark period and consistently deactivated at each subjective dawn, similar to that observed at true dawn in constitutive CAM species. Instantaneous carbon isotope discrimination showed in vivo carboxylase co-dominance during the CAM subjective night, initially by Rubisco and latterly C(4) (PEPc), despite both enzymes seemingly activated in vitro. The circadian rhythm in titratable acidity accumulation was progressively damped over successive subjective nights, but maintenance of PEPc carboxylation capacity ensures that CAM plants do not become progressively more 'C(3) -like' with time under LL.

Determining Degradation and Synthesis Rates of Arabidopsis Proteins Using the Kinetics of Progressive 15N Labeling of Two-dimensional Gel-separated Protein Spots

The growth and development of plant tissues is associated with an ordered succession of cellular processes that are reflected in the appearance and disappearance of proteins. The control of the kinetics of protein turnover is central to how plants can rapidly and specifically alter protein abundance and thus molecular function in response to environmental or developmental cues. However, the processes of turnover are largely hidden during periods of apparent steady-state protein abundance, and even when proteins accumulate it is unclear whether enhanced synthesis or decreased degradation is responsible. We have used a (15)N labeling strategy with inorganic nitrogen sources coupled to a two-dimensional fluorescence difference gel electrophoresis and mass spectrometry analysis of two-dimensional IEF/SDS-PAGE gel spots to define the rate of protein synthesis (K(S)) and degradation (K(D)) of Arabidopsis cell culture proteins. Through analysis of MALDI-TOF/TOF mass spectra from 120 protein spots, we were able to quantify K(S) and K(D) for 84 proteins across six functional groups and observe over 65-fold variation in protein degradation rates. K(S) and K(D) correlate with functional roles of the proteins in the cell and the time in the cell culture cycle. This approach is based on progressive (15)N labeling that is innocuous for the plant cells and, because it can be used to target analysis of proteins through the use of specific gel spots, it has broad applicability.

Effects of thermophotoperiod on growth parameters of Helicoverpa armigera

AbstractThe effects of photoperiod and temperature on growth parameters of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) were investigated under laboratory conditions by exposing its larvae to fluctuating and alternating temperature regimes. Our data made evident that the interplay between photoperiod and temperature has a direct effect on growth parameter responses of H. armigera. However, the type of temperature regime (fluctuating or alternating) may enhance or diminish the effects of photoperiod. With fluctuating temperature, larval developmental time was significantly reduced with increasing photophase, irrespective of the diapause status later in the pupal stage; the lowest value was observed under continuous light conditions. With alternating temperature, larval developmental time was significantly decreased with the reduction in temperature amplitude, and with the coincidence of the thermophase with the scotophase. At both temperature regimes, no significant differences in pupal weight were observed between diapausing and non‐diapausing individuals, although, in most treatments, the diapausing pupae tended to be heavier than the non‐diapausing ones. The increased pupal weight of individuals destined for diapause is probably related to their longer larval developmental time. Knowing the effects of these factors on the variation of H. armigera growth is necessary in explaining phenomena associated with immature stages, and can lead to a more profound understanding of the potential for this insect to evolve in response to environmental changes.

Photoperiod-modulated instar-specific clock-shifting in the circadian protein and amino acid rhythms in the larval segmental muscle of Bombyx mori

The photoperiod-modulated clock-shifting in the circadian protein rhythm was studied in the segmental muscle of Bombyx mori. The analysis of phase response curves of the fourth instar rhythm revealed that the muscle tissue completes six protein synthetic cycles (PS cycles) under normal 12 hr light and 12 hr dark cycle (LD), 8 cycles each under continuous light (LL) and continuous dark (DD) conditions. The fifth instar protein rhythm showed seven PS cycles each under LD and DD conditions, but only six under LL. The protein rhythm gets clock-shifted in instarspecific and photoperiod-specific fashions. In the fourth instar, both LL and DD conditions advanced the 24 hr free running time of the rhythm by six hours and set it at 18 hr, but in the fifth instar it is delayed by 4 hr and set at ~28 hr under LL, but remained unchanged under DD. Comparative analysis of protein and amino acid profiles shows that the photoperiod modulates the protein rhythm by altering the rate of amino acid mobilization.

Leaf variegation in the rice zebra2 mutant is caused by photoperiodic accumulation of tetra-cis-lycopene and singlet oxygen.

In field conditions, the zebra2 (z2) mutant in rice (Oryza sativa) produces leaves with transverse pale-green/yellow stripes. It was recently reported that ZEBRA2 encodes carotenoid isomerase (CRTISO) and that low levels of lutein, an essential carotenoid for non-photochemical quenching, cause leaf variegation in z2 mutants. However, we found that the z2 mutant phenotype was completely suppressed by growth under continuous light (CL; permissive) conditions, with concentrations of chlorophyll, carotenoids and chloroplast proteins at normal levels in z2 mutants under CL. In addition, three types of reactive oxygen species (ROS; superoxide [O₂⁻], hydrogen peroxide [H₂O₂], and singlet oxygen [¹O₂]) accumulated to high levels in z2 mutants grown under short-day conditions (SD; alternate 10-h light/14-h dark; restrictive), but do not accumulate under CL conditions. However, the levels of lutein and zeaxanthin in z2 leaves were much lower than normal in both permissive CL and restrictive SD growth conditions, indicating that deficiency of these two carotenoids is not responsible for the leaf variegation phenotype. We found that the CRTISO substrate tetra-Cis-lycopene accumulated during the dark periods under SD, but not under CL conditions. Its accumulation was also positively correlated with ¹O₂ levels generated during the light period, which consequently altered the expression of ¹O₂-responsive and cell death-related genes in the variegated z2 leaves. Taking these results together, we propose that the z2 leaf variegation can be largely attributed to photoperiodic accumulation of tetra-cis-lycopene and generation of excessive ¹O₂ under natural day-night conditions.

Dimerization and blue light regulation of PIF1 interacting bHLH proteins in Arabidopsis

Phytochrome Interacting Factor 1 (PIF1), a basic helix-loop-helix (bHLH) protein, functions as a negative regulator of various facets of photomorphogenesis. To indentify PIF1-interacting proteins, we performed yeast two-hybrid screening using PIF1 as a bait and identified a group of proteins including PIF1 itself, PIF3 and long hypocotyl in far-red 1 (HFR1), an atypical HLH protein. Directed yeast two-hybrid interaction assays showed that PIF1 can form heterodimers with all other PIFs as well as with HFR1. PIF1 and PIF3 interacted with each other in both in vitro and in vivo co-immunoprecipitation assays. PIF1-PIF3 heterodimer also bound to a G-box DNA sequence element in vitro. To understand the biological significance of these interactions, a pif1pif3 double mutant was obtained and characterized. Analyses of the single and double mutants showed that PIF3 plays a prominent role in repressing photomorphogenesis under continuous blue light conditions. pif1 and pif3 showed additive phenotypes more prominently under discontinuous blue light conditions. Similar to PIF1, PIF3 was also rapidly phosphorylated, poly-ubiquitylated and degraded in response to blue light. PIF3 also interacted with phytochromes in response to blue light. A PIF3 mutant defective in interaction with both phyA and phyB displayed reduced degradation under blue light, suggesting that phy-interaction was necessary for the blue light-induced degradation of PIF3. Taken together, these data suggest a combinatorial control of photomorphogenesis by bHLH proteins in response to light in Arabidopsis.

Circadian transcriptional regulation by the posttranslational oscillator without de novo clock gene expression in Synechococcus

Circadian rhythms are a fundamental property of most organisms, from cyanobacteria to humans. In the unicellular obligately photoautotrophic cyanobacterium Synechococcus elongatus PCC 7942, essentially all promoter activities are controlled by the KaiABC-based clock under continuous light conditions. When Synechococcus cells are transferred from the light to continuous dark (DD) conditions, the expression of most genes, including the clock genes kaiA and kaiBC, is rapidly down-regulated, whereas the KaiC phosphorylation cycle persists. Therefore, we speculated that the posttranslational oscillator might not drive the transcriptional circadian output without de novo expression of the kai genes. Here we show that the cyanobacterial clock regulates the transcriptional output even in the dark. The expression of a subset of genes in the genomes of cells grown in the dark was dramatically affected by kaiABC nullification, and the magnitude of dark induction was dependent on the time at which the cells were transferred from the light to the dark. Moreover, under DD conditions, the expression of some dark-induced gene transcripts exhibited temperature-compensated damped oscillations, which were nullified in kaiABC-null strains and were affected by a kaiC period mutation. These results indicate that the Kai protein-based posttranslational oscillator can drive the circadian transcriptional output even without the de novo expression of the clock genes.

Phytochrome interacting factors (PIFs) are essential regulators for sucrose-induced hypocotyl elongation in Arabidopsis

Phytochrome interacting factors (PIFs) are members of a subfamily of basic helix-loop-helix transcript factors and have been proposed to act as positive regulators of hypocotyl elongation under normal condition. Here, we show that PIF1, 3, 4, 5 together play a central role in sucrose-induced hypocotyl elongation. When seedlings grown in light were transferred to darkness, exogenously applied sucrose significantly induced hypocotyl elongation in wild type Col-0, but this effect was impaired in all tested pif mutants, especially in the quadruple mutant pif1pif3pif4pif5 (pifq). Subsequent experiments showed that under various light/dark (L/D) cycle conditions sucrose still markedly induced hypocotyl elongation in Col-0, but exhibited little effects in pifq. Phytohormone gibberellins (GAs) have been proven to be required for sucrose-induced hypocotyl elongation, but application of GA(3) (an active form of GAs) was not able to rescue the impairment observed in pifq, suggesting that impairment of sucrose-induced hypocotyl elongation in pifq is not due to the reduced endogenous GAs. Interestingly, through RT-PCR assay, we found that sucrose up-regulated the transcript level of PIF1, 3, 4, 5 in darkness. Furthermore, this effect was dependent on the presence of GAs. Additionally, under continuous light condition, sucrose markedly inhibited the hypocotyl elongation in Col-0 but not in pifq, whereas exogenous GA(3) could recover the repression in Col-0 but only showed slight effect in pifq. These results collectively indicate that PIFs together with GAs control the effect of sucrose on hypocotyl elongation in Arabidopsis seedlings.

First Report of Sclerotinia Rot on Blueberry Caused by Sclerotinia sclerotiorum in Argentina.

In October 2007, blighted shoots were observed on highbush blueberry (Vaccinium corymbosum L. cv. O'Neal) plants in La Plata, Buenos Aires Province, Argentina. Isolations from surface-disinfested shoots onto carrot agar and Spezieller Nahrstoffarmer Agar (SNA) consistently yielded white colonies that produced black sclerotia, mainly near the edge of the culture plates, after 7 days. Sclerotia were transferred to SNA tubes and kept at 5°C for several months. The germination of sclerotia produced numerous 6 mm long initials, stipitate pale brown cup-shaped apothecia (10 × 6 mm) with eight-spored asci (137 × 7 μm) at 18°C and continuous light conditions. Asci with uniseriate ascospores were cylindrical and narrow at the base. Ascospores (11 to 12 × 4 μm) were hyaline, unicellular, smooth, and ellipsoid. This isolated fungus was morphologically identified as Sclerotinia sclerotiorum (Lib.) de Bary (2,3). The isolate was deposited in the IMYZA Microbial Collection as INTA-IMC 87. Mycelium was cultured in 100 ml of Czapek's-Dox medium, supplemented with sucrose, peptone, yeast extract, sodium nitrate, and vitamins (1), for 3 days and fungal DNA was obtained using a DNA extraction kit. ITS1 and ITS2 of ribosomal genes were amplified by PCR using universal primers (4) and the PCR product was sequenced. A BLAST algorithm search revealed 100% identity of the sequence with 12 GenBank entries for S. sclerotiorum. The nucleotide sequence was deposited in the GenBank with Accession No. JF277567. Pathogenicity testing was achieved by placing detached leaves of cvs. Emerald, Misty, and Start on water agar (WA) plates, inoculating with 9-mm2 mycelial blocks, and incubating at 20°C with 12 h of light. Young shoots of highbush blueberry, Misty and O'Neal, were inoculated by the cut shoot method with micropipette tips filled with mycelium and kept under greenhouse conditions at 24°C and 14 h of light. On control plants, WA blocks or WA-filled micropipette tips were used. Leaf blight was observed after 5 to 6 days and sclerotia appeared after 7 days on inoculated tissues. Shoot blight was recorded on inoculated plants after 5 days. The fungus was reisolated from inoculated tissues, with no symptoms showing on controls. To our knowledge, this is the first report of Sclerotinia rot caused by S. sclerotiorum in blueberry in Argentina.

Tissue specificity and diurnal change in gene expression of the sucrose phosphate synthase gene family in rice

The rice genome contains 5 isogenes for sucrose phosphate synthase (SPS), the key enzyme in sucrose synthesis; however, little is known about their transcriptional regulation. In order to determine the expression patterns of the SPS gene family in rice plants, we conducted an expression analysis in various tissues and developmental stages by real-time quantitative RT-PCR. At the transcript level, the rice SPS genes, particularly SPS1, were preferentially expressed in source tissues, whereas SPS2, SPS6, and SPS8 were expressed equally in source and sink tissues. We also investigated diurnal changes in SPS gene expression, SPS activity, and soluble sugar content in leaf blades. Interestingly, the expression of all the SPS genes, particularly that of SPS1 and SPS11, tended to be higher at night when the activation state of the SPS proteins was low, and the mRNA levels of SPS1 and SPS6 were negatively correlated with sucrose content. Furthermore, the temporal patterns of SPS gene expression and sugar content under continuous light conditions suggested the involvement of endogenous rhythm and/or sucrose sensing in the transcriptional regulation of SPS genes. Our data revealed differential expression patterns in the rice SPS gene family and part of the complex mechanisms of their transcriptional control.

Constitutive expression of the GIGANTEA Ortholog Affects Circadian Rhythms and Suppresses One-shot Induction of Flowering in Pharbitis nil, a Typical Short-day Plant

GIGANTEA (GI) is a key regulator of flowering time, which is closely related to the circadian clock function in Arabidopsis. Mutations in the GI gene cause photoperiod-insensitive flowering and altered circadian rhythms. We isolated the GI ortholog PnGI from Pharbitis (Ipomoea) nil, an absolute short-day (SD) plant. PnGI mRNA expression showed diurnal rhythms that peaked at dusk under SD and long-day (LD) conditions, and also showed robust circadian rhythms under continuous dark (DD) and continuous light (LL) conditions. Short irradiation with red light during the flower-inductive dark period did not change PnGI expression levels, suggesting that such a night break does not abolish flowering by affecting the expression of PnGI. In Pharbitis, although a single dusk signal is sufficient to induce expression of the ortholog of FLOWERING LOCUS T (PnFT1), PnGI mRNA expression was not reset by single lights-off signals. Constitutive expression of PnGI (PnGI-OX) in transgenic plants altered period length in leaf-movement rhythms under LL and affected circadian rhythms of PnFT mRNA expression under DD. PnGI-OX plants formed fewer flower buds than the wild type when one-shot darkness was given. In PnGI-OX plants, expression of PnFT1 was down-regulated, suggesting that PnGI functions as a suppressor of flowering, possibly in part through down-regulation of PnFT1.

Cultural Characteristics of Ophiocordyceps heteropoda Collected from Korea

Isolates of Ophiocordyceps heteropoda (Kobayasi) collected from Mt. Halla on Jeju-do, Korea were tested for mycelial growth on different agar media and in the presence of different carbon and nitrogen sources. Similarly, isolates were also incubated at different temperatures as well as under continuous light and dark conditions. Growth was better on Hamada agar, basal medium, and malt-yeast agar, but poor on Czapek-Dox agar. Different carbon sources such as dextrin, saccharose, starch, lactose, maltose, fructose, and dextrose resulted in better growth. Complex organic nitrogen sources such as yeast extract and peptone revealed the most effective growth. Mycelial growth was best at 25°C. The growth rate was faster in the dark than the light, but mycelial density was less compact in the dark.

Feeding Time Synchronizes Clock Gene Rhythmic Expression in Brain and Liver of Goldfish (Carassius auratus)

Little is known about the feeding time dependence of clock gene expression in fish. The aim of the present study was to investigate whether a scheduled feeding time can entrain the rhythmic expression of several clock genes (period and cryptocrome) in the brain and liver of a teleost, the goldfish. Fish maintained under continuous light (LL) conditions were divided into 3 groups. Two groups were fed daily at 1000 h and 2200 h, respectively, and the third group was subjected to a random schedule regime. After 30 days, the fishes under 24-h food deprivation were sacrificed through a 24-h cycle, and clock gene expression in the optic tectum, hypothalamus, and liver was quantified by real-time PCR. The findings pointed to differences between the central and peripheral tissues studied. In the absence of a light-dark cycle (constant light), a scheduled feeding regime was necessary and sufficient to maintain both the rhythmic expression of several clock genes in the optic tectum and hypothalamus, as well as daily rhythms in locomotor activity. In contrast, neither locomotor activity nor clock gene expression in brain tissues was synchronized in randomly fed fish. However, in the liver, most of the clock genes studied presented significant daily rhythms in phase (related to the time of the last meal) in all 3 experimental groups, suggesting that the daily rhythm of clock genes in this organ only depends on the last meal time. The data suggest that, as in mammals, the smooth running of the food entrainable oscillator (FEO) in fish involves the rhythmic expression of several clock genes (Per1 and Cry3) in the central and peripheral structures. The results also indicate that the food anticipatory activity (FAA) in goldfish is not only the result of rhythmic clock gene expression in the liver because rhythmic clock gene expression was observed in randomly fed fishes, while FAA was not observed.