Diurnal Pattern of Translocation and Carbohydrate Metabolism in Source Leaves of Beta vulgaris L

Transitions in carbohydrate metabolism and translocation rate were studied for evidence of control of export by the sugar beet (Beta vulgaris L. Klein E.) source leaf. Steady-state labeling was carried out for two consecutive 14-hour light periods and various quantities related to translocation were measured throughout two 24-hour periods. Starch accumulation following illumination was delayed. Near the end of the light period, starch stopped accumulating, whereas photosynthesis rate and sucrose level remained unchanged. At the beginning of the dark period there was a 75-minute delay before starch was mobilized. The rate of import to the developing sink leaves at night was similar to that during the day, whereas export decreased considerably at night.

Starch accumulation and degradation seemed to be initiated in response to the level of illumination. Cessation of starch accumulation before the end of the light period was initiated endogenously. Exogenous control appeared to be mediated by the level of sucrose in the source leaf while endogenous control seemed to be keyed to photoperiod or photosynthetic duration.

     

Effect of Exogenously Supplied Foliar Potassium on Phloem Loading in Beta vulgaris L

The effect of foliar application of K⁺ on processes associated with phloem loading was investigated in source leaves of sugar beet (Beta vulgaris L.). KCI was supplied exogenously at concentrations of up to 100 millimolar in the solution bathing the abraded upper epidermis of source leaves. K⁺ added at concentrations below 30 millimolar generally promoted the rate of export of material derived from ¹⁴CO₂ but not from exogenously applied [¹⁴C]sucrose. Paralleling promotion of export, the level of material derived from photosynthesis, which was released into the bathing solution, also increased in response to addition of K⁺ to the free space. Net photosynthetic rate was not affected. K⁺ at 5 and 15 millimolar concentrations did not stimulate uptake of [¹⁴C]sucrose into source leaf discs.     

Influence of Translocation of Photosynthetic Efficiency of Phaseolus vulgaris L

Measurements of net photosynthesis show that in Phaseolus vulgaris L. the cultivar Michelite-62 exceeds the cultivar Red Kidney in net CO₂ uptake by 23 to 31%. Data on translocation of pulse label indicate that export of a pulse of photosynthetically assimilated ¹⁴C from the source leaf of either M-62 or Red Kidney follows an exponential pattern and shows an initial rapid phase followed by a second slower phase. The steeper slope for both phases in M-62 suggests its rate of translocation of pulse label is higher than that of Red Kidney. Furthermore, only 38% of the ¹⁴C remains in the leaf of M-62 after 8 hours, while Red Kidney retains up to 60% of the label. Leaf autoradiographs obtained after pulse labeling demonstrate a much faster rate of vein loading in M-62 and are considered evidence for the higher translocation efficiency of M-62. These results provide evidence for a positive correlation between photosynthetic efficiency and translocation efficiency in M-62 and Red Kidney and give support to our hypothesis that translocation is one of the important physiological factors controlling the varietal differences in photosynthetic efficiency in Phaseolus vulgaris.     

In Vivo Nitrite Reduction in Leaf Tissue of Phaseolus vulgaris L

Experiments were performed to establish a procedure for in vivo measurement of nitrite utilization by leaf tissue of bean (Phaseolus vulgaris L. cv. Top Crop).     

Heterogeneity of Glutamine Synthetase Polypeptides in Phaseolus vulgaris L

Glutamine synthetases from roots, nodules, and leaves of Phaseolus vulgaris L. have been purified to homogeneity and their polypeptide composition determined.

The leaf enzyme is composed of six polypeptides. The cytosolic fraction contains two 43,000 dalton polypeptides and the chloroplastic enzyme is formed by four 45,000 dalton polypeptides. Root glutamine synthetase consists only of the same two polypeptides of 43,000 dalton that are present in the leaf enzyme. The nodule enzyme is formed by two polypeptides of 43,000 dalton, one is common to the leaf and root enzyme but the other is specific for N₂-fixing nodule tissue. The two glutamine synthetase forms of the nodule contain a different proportion of the 43,000 dalton polypeptides.

     

Abscisic Aldehyde Is an Intermediate in the Enzymatic Conversion of Xanthoxin to Abscisic Acid in Phaseolus vulgaris L. Leaves

The enzymatic conversion of xanthoxin to abscisic acid by cell-free extracts of Phaseolus vulgaris L. leaves has been found to be a two-step reaction catalyzed by two different enzymes. Xanthoxin was first converted to abscisic aldehyde followed by conversion of the latter to abscisic acid. The enzyme activity catalyzing the synthesis of abscisic aldehyde from xanthoxin (xanthoxin oxidase) was present in cell-free leaf extracts from both wild type and the abscisic acid-deficient molybdopterin cofactor mutant, Az34 (nar2a) of Hordeum vulgare L. However, the enzyme activity catalyzing the synthesis of abscisic acid from abscisic aldehyde (abscisic aldehyde oxidase) was present only in extracts of the wild type and no activity could be detected in either turgid or water stressed leaf extracts of the Az34 mutant. Furthermore, the wilty tomato mutants, sitiens and flacca, which do not accumulate abscisic acid in response to water stress, have been shown to lack abscisic aldehyde oxidase activity. When this enzyme fraction was isolated from leaf extracts of P. vulgaris L. and added to extracts prepared from sitiens and flacca, xanthoxin was converted to abscisic acid. Abscisic aldehyde oxidase has been purified about 145-fold from P. vulgaris L. leaves. It exhibited optimum catalytic activity at pH 7.25 in potassium phosphate buffer.     

Isozymes of Glutamine Synthetase in Phaseolus vulgaris L. and Phaseolus lunatus L. Root Nodules

The glutamine synthetase (GS) isozymes in the plant fraction of nodule extracts from 62 cultivars of Phaseolus vulgaris L. and one cultivar of Phaseolus lunatus L. were analyzed by polyacrylamide gel electrophoresis. All P. vulgaris nodule extracts displayed two GS activity bands: a nodule-specific band (GS_n1) and a band (GS_n2) similar to the single band (GS_r) present in root extracts. In nodule extracts of P. lunatus, the GS_n1 band was detected, but the GS_n2 band was barely detectable. In contrast to P. vulgaris, the GS_n2 band and the GS_r band of P. lunatus appeared to be different. The electrophoretic mobility of the GS_n1 band in P. vulgaris was governed by both the plant cultivar and the development stage of the nodule. In nodule extracts of P. vulgaris and P. lunatus, the zone of GS_n1 activity coincided with six to nine distinct protein bands as revealed after treatment of gels, which had previously been stained for GS activity, with Coomassie blue. All these protein bands were shown to consist of polypeptides of identical molecular weight (approximately 47,000 daltons) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Our results indicate that P. vulgaris continuously generates isozymes of GS_n1 of increasing electrophoretic mobility during the course of nodule development.     

Genetic and phenotypic differentiation of Calluna vulgaris (L.) Hull. in Pritobolie and Europe

Geographic variation and differentiation of the chloroplast DNA haplotypes and morpho-anatomical leaf parameters were assessed in a number of eastern European groups of Calluna vulgaris (L.) Hull. populations and in the Pritobolien group of populations of this species in Western Siberia, which have long been isolated from the European populations. Sharply pronounced genetic and phenotypic distances and their gradients between the Pritobolien and European population groups were revealed. These distances were many times higher than those between the relatively homogeneous eastern European groups. The data obtained generally supported the hypothesis on the phenogenetic divergence of the Pritobolien marginal populations of C. vulgaris from the European, probably at the subspecies level.     

The influence of aluminium availability on phosphate uptake in Phaseolus vulgaris L. and Phaseolus lunatus L.

Aluminium toxicity is one of the major limiting factors of crop productivity on acid soils. High levels of available aluminium in soil may induce phosphorus deficiency in plants. This study investigates the influence of Aluminium (Al) on the phosphate (P_i) uptake of two Phaseolus species, Phaseolus vulgaris L. var. Red Kidney and Phaseolus lunatus L. The two bean species were treated first with solutions of Al at different concentrations (0, 25, 50 and 100 μM, pH 4.50) and second with solutions of P_i (150 μM) at pH 4.50. The higher the Al concentration the higher the Al concentration sorbed but P. vulgaris L var. Red Kidney adsorbed significantly more Al than P. lunatus L. Both species released organic acids: P. vulgaris L var. Red Kidney released fumaric acid and P. lunatus L. fumaric and oxalic acids which could have hindered further Al uptake.The two bean species showed a sigmoid P_i uptake trend but with two different mechanisms. P. vulgaris L var. Red Kidney showed a starting point of 3 h whereas P. lunatus L. adsorbed P_i immediately within the first minutes. In addition, P. vulgaris L var. Red Kidney presented significantly higher P_i uptake (higher uptake rate ‘k’ and higher maximum adsorption ‘a’ of the kinetic uptake model). The Al treatments did not significantly influence P_i uptake. Results suggest that P. lunatus L. might adopt an external Al detoxification mechanism by the release of oxalic acid. P. vulgaris L var. Red Kidney on the other hand seemed to adopt an internal detoxification mechanism even if the Al sorbed is poorly translocated into the shoots. More detailed studies will be necessary to better define Al tolerance and/or resistance of Phaseolus spp.     

Comparative Water Relations of Phaseolus vulgaris L. and Phaseolus acutifolius Gray

Leaf area expansion, dry weight, and water relations of Phaseolus vulgaris L. and P. acutifolius Gray were compared during a drying cycle in the greenhouse to understand the characteristics which contribute to the superior drought tolerance of P. acutifolius. Stomates of P. acutifolius closed at a much higher water potential than those of P. vulgaris, delaying dehydration of leaf tissue. P. acutifolius had a more deeply penetrating root system, which also contributes to its drought tolerance. Root-shoot ratios did not differ between the two species either under well watered or water stressed conditions. Leaf osmotic potential was also similar in the two species, with no apparent osmotic adjustment during water stress. These results indicate that P. acutifolius postpones dehydration and suggest that sensitive stomates and a deeply penetrating root system are characteristics which, if incorporated into cultivated beans, might increase their drought tolerance.     

Interspecific hybridization of Phaseolus vulgaris L. and Phaseolus angustissimus A. Gray using in vitro embryo culture

The introgression of new desirable characters is very necessary in common bean (Phaseolus vulgaris L.)Interspecific hybridization of P. vulgaris L. with P. angustissimus A. Gray, a wild species with narrow leaves, could be successfully achieved for the first time using embryo culture. Sixteen to twenty-three-day old embryos could grow to plantlets when they were cultivated on modified Monnier's medium.The leaf shape of the hybrid plants obtained showed intermediary traits between the two parents.A chromosome stock doubling could be achieved for one embryo.     

Zinc Nutrition and Starch Metabolism in Phaseolus vulgaris L

The effect of Zn nutrition on leaf starch metabolism was studied in two navy bean (Phaseolus vulgaris L.) varieties, Sanilac and Saginaw. Sanilac is much more susceptible to Zn deficiency than is Saginaw. The variables examined in these two strains were starch content, the activity of soluble starch synthetase (ADP-glucose: starch α-4-glucosyltransferase, EC 2.4.1.b), and the size and number of starch grains. All of these variables decreased during Zn deficiency. The reductions were much greater in Sanilac than in Saginaw. Thus, positive correlations exist between the relative changes in these variables in Sanilac and Saginaw under low Zn and their genotypic difference in growth response to low Zn. These results are taken to purport that the above observations most likely represent characteristic responses to Zn deficiency. We therefore suggest that, as a possible role in plant metabolism, Zn is involved in starch formation.     

8-[C]Benzylaminopurine Translocation in Phaseolus vulgaris

[8-¹⁴C]Benzylaminopurine (BA) translocation was studied in whole plants of Phaseolus vulgaris L. under three different light regimes (continuous light, 8-hour light + 16-hour dark, dark). Applications were made to the apex, to a cotyledonary leaf, or to the root system. Results showed that no BA basipetal translocation occurred, however BA is easily absorbed by the root system and is translocated acropetally.     

Photosynthesis, Carbohydrate Metabolism, and Export in Beta vulgaris L. and Phaseolus vulgaris L. during Square and Sinusoidal Light Regimes

Rates of photosynthesis, sucrose synthesis, starch accumulation and degradation were measured in sugar beet (Beta vulgaris L.) and bean (Phaseolus vulgaris L.) plants under a square-wave light regime and under a sinusoidal regime that simulated the natural daylight period. Photosynthesis rate increased in a measured manner in direct proportion to the increasing light level. In contrast to this close correspondence between photosynthesis and light, a lag in photosynthesis rate was seen during the initial hour under square-wave illumination. The leaf appeared to be responding to limits set by carbon metabolism rather than by gas exchange or light reactions. Under the sinusoidal regime starch degradation occurred during the first and last 2 hours of the photoperiod, likely in response to photosynthesis rate rather than directly to light level. Starch broke down when photosynthesis was below a threshold rate and accumulated above this rate. Under square-wave illumination, accumulation of starch did not begin until irradiance was at full level for an hour or more and photosynthesis was at or near its maximum. Under a sinusoidal light regime, sucrose synthesis rate comprised carbon that was newly fixed throughout the day plus that from starch degradation at the beginning and end of the day. Synthesis of sucrose from recently fixed carbon increased with increasing net carbon fixation rate while its formation from degradation of starch decreased correspondingly. The complementary sources of carbon maintained a relatively steady rate of sucrose synthesis under the changing daytime irradiance.     

Sources of sucrose translocated from illuminated sugar beet source leaves

A search for source leaf sucrose pools that differed in their relation to export was carried out in photosynthesizing leaves of Beta vulgaris L. The time course of depletion of [¹⁴C]sucrose in a leaf in unlabeled CO₂ following steady state labeling provided evidence for two distinct sucrose pools. After the start of the light period, leaf blade sucrose remained constant although it exchanged between the two pools. Newly synthesized sucrose destined for export passed through one pool more rapidly than through the other. All of the leaf blade sucrose appeared to exchange with export sucrose. Modeling and regression analysis of [¹⁴C]sucrose data provided a means for estimating the size of the two pools. From 20 to 40% of the sucrose was calculated to be present in the pool that provided the less direct path to export; this was likely vacuolar sucrose. The remainder of the sucrose in the blade was probably in the cytoplasm and veins. Added amounts of leaf blade sucrose, produced in response to elevated CO₂, appeared to be stored mainly in the vacuolar compartment.     

Translocation and Metabolic Conversion of C-Labeled Assimilates in Detached and Attached Leaves of Phaseolus vulgaris L. in Different Phases of Leaf Expansion

Leaf excision greatly affected the actual levels of ¹⁴C-assimilates in laminas and petioles of primary bean leaves (Phaseolus vulgaris L.) following a transport period. However, it did not affect the percentage of starch in the insoluble residue; starch decreased from 20% of the insoluble residue after exposure to ¹⁴CO₂ to 3% after 5 hr in both attached and detached leaves. The transition from import to export of attached and detached leaves was at the same stage, i.e., when the cotyledons were 63 to 85% depleted. The composition of the ¹⁴C-assimilates in importing leaves was different from that in exporting leaves. In the former, only 5% of the soluble label was free sugar, while 74% was free sugar in the latter. The failure of importing leaves to export was not due to the labeled substances being nontransferable. Extracts from importing leaves applied to exporting leaves were exported; these extracts were high in amino acids and organic acids but low in free sugar. However, exporting leaves exposed to ¹⁴CO₂ appeared to export sugars more readily than amino acids. Cotyledon excision did not delay transition of leaves from import to export. Actually, excision seemed to enhance slightly the transition of the primary leaves from import to export.     

Isolation and Identification of the Precursor of Ethane in Phaseolus vulgaris L

Ethane production by homogenates of Phaseolus vulgaris L. cv. Harvester was studied. The precursor of ethane was identified as linolenic acid. The liberation of ethane was optimum at pH 4.2 and was highest from homogenates of leaves and apical buds. When roots were homogenized in linolenic acid solution, ethane and ethylene production were stimulated. In corn root homogenates, ethylene biosynthesis was stimulated nearly 8-fold by linolenic acid. The enzyme responsible for ethane production from oat root homogenates was soluble and had a high molecular weight.     

Structural and Physiological Changes in Sugar Beet Leaves during Sink to Source Conversion

The onset of export during leaf development was correlated with changes in metabolism and ultrastructure and with patterns of solute distribution in the developing seventh leaf of sugar beet (Beta vulgaris L.) in order to study the cause of initiation of translocation. Infrared gas analysis of carbon dioxide uptake showed a broad peak for net photosynthesis dm⁻² at 35 to 40% final laminar length. Pulse labeling with ¹⁴CO₂ demonstrated that maximum import of translocate occurred at 25% final laminar length; export was first observed at 35% final laminar length. Between 40 and 50% final laminar length a rapid increase in amount of export occurred, primarily as a result of the increase in the area of leaf which was exporting. Whole leaf autoradiography revealed that onset of phloem loading spread basipetally from the leaf tip; loading was initiated at about 22% final laminar length and was essentially complete by 50% final laminar length. Those areas which clearly exhibited loading no longer imported from other parts of the plant while the area in transition still appeared to import label from source regions.     

Atypical composition of seed proteins in cultivars ofPhaseolus vulgaris L.

In only one cultivar out of 1200 investigated cultivars ofPhaseolus vulgaris L. could we find an extreme change in the pattern of reserve proteins on the cathodic side: one of the proteins, called protein I, is completely absent in the cultivar ‘Krupnaya sakharnaya’ and is replaced on the same site by another protein,i.e. a protein completely different in its immunochemical specificity. The case is of interest from both the phylogenetic and systematic viewpoints and deserves further attention.