High-density algal photobioreactors using light-emitting diodes

Lack of high-density algal photobioreactors (PBR) has been a limitation in exploiting the biotechnological potential of algae. Recent developments of highly efficient light-emitting diodes (LED using gallium aluminum arsenide chips) have made the development of a small LED-based PBR possible. We have calculated theoretical values of gas mass transfer requirements and light-intensity requirement to support high-density algal cultures for the 680 nm monochromatic red light from LED as a light source. A prototype PBR has been designed based on these calculations. A cell concentration of more than 2 x 10(9) cells/mL (more than 6.6% v%sol;v), cell doubling times as low as 12 h, and an oxygen production rate as high as 10 mmol oxygen/L culture/h were achieved using on-line ultrafiltration to periodically provide fresh medium. (c) 1994 John Wiley & Sons, Inc.     

Spread of beet necrotic yellow vein virus in infected seedlings and plants of sugar beet (Beta vulgaris)

Summary Infection of sugar beet roots by beet necrotic yellow vein virus (BNYVV) was investigated with transmission electron microscopy, immunogold labelling and enzyme linked immuno sorbent assay (ELISA). Here we show that infection of sugar beet roots is very fast, occurring during germination. Seedlings grown directly in infected soil showed higher BNYVV infection than plants transplanted into infected soil after seven days of initial growth in sterilized soil. The earlier the initial infection, the faster was its spread. The study showed that a few differentiated cells of the cortex and of the xylem parenchyma were the preferred sites of viral multiplication. The spread of viral infection was slow through differentiated tissues. Intact virions were frequently found in undifferentiated and mature vessel elements and xylem parenchyma, whereas they were rare in sieve elements. Virus particle number in the differentiating tracheary elements was high, suggesting that infection of the vessel elements preceded their differentiation. This would explain increased infection after early inoculation. Even the xylem tissue of the primary root was highly infected, the seedlings lacked virus particles in their hypocotyls and leaves.     

Identification of RFLP markers closely linked to the bolting gene B and their significance for the study of the annual habit in beets (Beta vulgaris L.)

The annual habit in beet is due to complete or partial absence of the vernalization requirement and can cause severe problems in the beet crop. The absolute vernalization requirement in beet is controlled by a major geneB (bolting), known to be linked to the geneR (red hypocotyl color), in linkage group I. Segregation for theB andR genes was studied in several beet progenies. Penetrance of the annual habit inBb genotypes was affected by both environmental and genetic factors. The precise location in linkage group I of the major geneB was found by restriction fragment length polymorphism (RFLP) analysis in a back-cross progeny exhibiting partial penetrance of the annual habit. The linkage value betweenB andR was in good accordance with previous estimations. Use of the closest RFLP marker (pKP591: 3.8 recombination units) allowed us to estimate the penetrance of the annual habit in this back-cross as 0.62. Evidence of pseudo-compatibility was found in the wild coastal beet (Beta vulgaris sspmaritima) used as the mother plant of the back-cross: the selfing rate was estimated as 7%.     

Construction and characterisation of a yeast artificial chromosome library containing five haploid sugarbeet (Beta vulgaris L.) genome equivalents

A yeast artificial chromosome (YAC) genomic library of Beta vulgaris was constructed in the pYAC4 vector. High-molecular-weight DNA was prepared from agarose-embedded leaf protoplasts from a triploid cultivar. The library was found to contain 33,500 clones in an ordered array of microtiter plates. Mean size of the inserts was estimated to be 135 kb, and the library should therefore represent the equivalent of five haploid genomes. The library was characterised for the presence of highly repetitive, chloroplast and single-copy sequences. In order to isolate single-copy sequences, 18 pools of DNA, each from 1920 individual YAC clones, were prepared for rapid screening of the library by the polymerase chain reaction. The results of these screenings showed that the number of isolated clones was at or near the frequency expected.     

Genotypic differences in cold tolerance are masked by high sucrose and cytokinin in shoot cultures of sugarbeet

Cold tolerance of field grown plants and shoot cultures of a commercial sugarbeet cultivar, Hilma, was compared with that of two cultivars bred for improved cold tolerance, Monofeb and Winter Hybrid 88619. Leaves of Monofeb and Winter Hybrid 88619 showed an increase in frost tolerance compared to Hilma, as assessed by electrolyte leakage measurements, in both July, and November. However, all varieties exhibited acclimation in the latter month. Similar qualitative differences between cultivars were detected in shoot cultures only when maintained on low (1%) sucrose medium, without added plant growth regulators. The use of high (3%) sucrose and benzyladenine, which releases apical dominance producing multiple shoots, each contributed to a substantial lowering of the temperature at which cold-induced damage occurred in leaves. Under these conditions varietal differences were masked. The implications of these findings in regard to in vitro selection for improved cold tolerance in organized cultures are discussed.Abbreviations BA benzyladenine - MS Murashige & Skoog (1962)     

CHARACTERIZATION OF TWO CHLORELLA VULGARIS (CHLOROPHYCEAE) STRAINS ISOLATED FROM WASTEWATER OXIDATION PONDS1

We report here on the characterization and isolation of two ecotypes of Chlorella vulgaris Beyerinck that coexist in wastewater reservoirs. One ecotype (C1) contains high amounts of chlorophyll b, is capable of autotrophic growth, and can utilize only a few organic solutes for growth. The second ecotype (C2) contains low amounts of chlorophyll b, requires vitamin B₁₂, and can support its growth with a broad range of organic compounds. Of the two ecotypes, the latter showed slower growth rates when light was the sole source of energy. Cells of C2-type Chlorella attained higher photosynthetic activities than C1-type cells at saturating irradiances. However, their low chlorophyll b content and lower light utilization efficiency suggest that C2-type Chlorella contains relatively low amounts of light-harvesting antennae, a disadvantage in severely light-limited ecosystems like wastewater reservoirs. We hypothesize that the two Chlorella types coexist by adopting different lifestyles: C1-type cells rely largely on their photosynthetic potential for energy conservation and growth, whereas C2-type cells may exploit their heterotrophic properties for this purpose.     

Enhancement of extractable ethylene at light/dark transition in primary leaves of paclobutrazol-treated Phaseolus vulgaris seedlings

Paclobutrazol [(2RS,3RS)-1-(4-chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-yl)pentan-3-ol], a triazole growth retardant, increased the 1-aminocyclopropane-1-carboxylic acid (ACC) level and resulted in reduced ethylene production, estimated as ethylene release in a closed system or by vacuum-extraction, in the primary leaves of Phaseolus vulgaris L. cv. Juliska seedlings exposed to light. At the light/dark transition, a definite enhancement of the endogenous ethylene level was observed by vacuum-extraction of primary leaves of treated plants and the ethylene deficiency of retardant-treated leaves ceased. The concentration of ACC after the light/dark transition followed the pattern for ethylene, and the increase in ACC content was paralleled by a decrease in malonyl-ACC.
It is concluded that the internal level of ethylene is not necessarily lower in the primary leaves of paclobutrazol-treated bean plants, but under special environmental conditions in vivo it may reach that of the control.     

Turgor-dependent unloading of assimilates from coats of developing legume seed. Assessment of the significance of the phenomenon in the whole plant

The in vivo significance of turgor-dependent unloading was evaluated by examining assimilate transport to and within intact developing seeds of Phaseolus vulgaris (cv. Redland Pioneer) and Vicia faba (cv. Coles Prolific). The osmotic potentials of the seed apoplast were low. As a result, the osmotic gradients to the seed coat symplast were relatively small (i.e. 0.1 to 0.3 MPa). Sap concentrations of sucrose and potassium in the seed apoplast and coat symplast accounted for some 45 to 60% of the osmotic potentials of these compartments. Estimated turnover times of potassium and sucrose in the seed apoplast of < 1 h were some 5 to 13 times faster than the respective turnover times in the coat symplast pools. The small osmotic gradient between the seed apoplast and coat symplast combined with the relatively rapid turnover of solutes in the apoplast pool, confers the potential for a small change in assimilate uptake by the cotyledons to be rapidly translated into an amplified shift in the cell turgor of the seed coat. Observed adjustments in the osmotic potentials of solutions infused between the coat and cotyledons of intact seed were consistent with the in vivo operation of turgor-dependent unloading of solutes from the coat. Homeostatic regulation of turgor-dependent unloading was indicated by the maintenance of apoplast osmotic potentials of intact seeds when assimilate balance was manipulated by partial defoliation or elevating pod temperature. In contrast, osmotic potentials of the coat symplast adjusted upward to new steady values over a 2 to 4 h period. The resultant downward shift in coat cell turgor could serve to integrate phloem import into the seed coat with the new rates of efflux to the seed apoplast. Circumstantial evidence for this linkage was suggested by the approximate coincidence of the turgor changes with those in stem levels of ³²P used to monitor phloem transport. The results obtained provide qualified support for the in vivo operation of a turgor homeostat mechanism. It is proposed that the homeostat functions to integrate assimilate demand by the cotyledons with efflux from and phloem import into the coats of developing legume seed.     

Turgor-dependent unloading of photosynthates from coats of developing seed of Phaseolus vulgaris and Vicia faba. Turgor homeostasis and set points

Key physiological characteristics of turgor-dependent efflux of photosynthates were examined using excised coats and cotyledons of developing Phaseolus vulgaris (cv. Redland Poineer) and Vicia faba (cv. Coles Prolific) seed during the linear phase of seed fill. Exposure to solutions of high osmotic potential inhibited net uptake of [¹⁴C]sucrose by cotyledons at developmental stages less than 60% of their final dry weight. The effect could not be fully reversed by transferring cotyledons to solutions set at lower osmotic potentials. The inhibition became apparent at osmotic potentials that were higher than those that caused stimulation of efflux from seed coats. Net [¹⁴C]sucrose uptake by cotyledons at more advanced stages of development was unaffected by external osmotic potential. Specified tissue layers were removed from seed coats by pretreatment with pectinase. Efflux studies with the pectinase-modified coats of Phaseolus and Vicia seed demonstrated that the cellular site of turgordependent efflux was the ground parenchyma and thin-wall parenchyma transfer cells, respectively. Coats subjected to long-term (hours) incubations, under hypo-osmotic conditions, exhibited the capacity for turgor regulation. This was mediated by turgor-dependent efflux of solutes. The solutes exchanged were of nutritional significance to the developing embryo. The relationship between efflux and coat turgor was characterised by a turgor-independent phase at low turgors. Once turgor exceeded a minimal value (set point), efflux increased in proportion to the magnitude of the turgor deviation (error signal) from the set point. For coats of sink-limited seed of Vicia and Phaseolus, efflux exhibited apparent saturation at turgors above 0.25 and 0.5 MPa respectively. The putative turgor set point and slope of the turgor-dependent component of efflux varied with seed development, the prevailing source/sink ratio and genetic differences in seed growth rate. The nature of these kinetic variations was compatible with the competitive ability of the seed. A turgor homeostat model is proposed that incorporates the observed functional attributes of turgor-dependent efflux. Operationally, the model provides a mechanistic basis for the integration of assimilate demand by the cotyledons with assimilate import into and unloading from the seed coat.