Endogenous Cytokinins in Solanum tuberosum as Influenced by Photoperiod and Temperature

Using the cucumber cotyledon bioassay, in combination with paper and column chromatographic techniques, 4 compounds exhibiting cytokinin activity were detected in extracts of Katahdin potato plant tissues. Two of these compounds were significantly higher in plants grown under inducing conditions (26°C day and 12°C night with an 8-h photoperiod) than those grown under non-inducing conditions (28°C day and 25°C night with a 16-h photoperiod). One compound increased 10-fold in above-ground tissue after only 2 Inductive cycles. Both compounds reached their maximum values in below-ground tissue after 6 days under inducing conditions, with tuber initiation observed after 8 to 10 days.     

Plant Hormones Isolated from “Katahdin” Potato Plant Tissues and the Influence of Photoperiod and Temperature on Their Levels in Relation to Tuber Induction

Qualitative and quantitative analyses were carried out on vegetative tissues of potato (Solanum tuberosum cv. “Katahdin”) in search of natural products thought to play a role in tuber induction. Tissues were obtained from plants initially grown in a growth chamber under noninducing conditions (30°C day and 28°C night with an 18-h photoperiod), and then half of the plants were moved to inducing chambers (28°C day and 13°C night with a 10-h photoperiod) for 10 days prior to tissue harvest. Plants from each chamber were then harvested at 2-day intervals for 10 days, separated into above- and belowground portions, and the lyophilized tissues were extracted and subjected to rigorous purification and separation using high-performance liquid chromatography. This was followed by identification and quantification using combined gas chromatography-mass spectrometry. Compounds isolated and identified included gibberellic acid; cytokinins cis-zeatin riboside, trans-zeatin, trans-zeatin riboside, and isopentenyladenine; and jasmonates jasmonic acid, tuberonic acid and its methyl ester, methyl 7-isocucurbate, and 9,10-dihydromethyljasmonate. Methyl 7-isocucurbate and 9,10-dihydromethyljasmonate were detected for the first time in potato tissue as endogenous compounds. Cytokinin and jasmonate levels generally increased under inducing conditions, whereas gibberellic acid levels declined progressively during the 10-day sampling period. Only gibberellic acid, jasmonic acid, and cis-zeatin riboside levels were significantly influenced by induction.     

Interaction between day length and phytohormones in the control of potato tuberization in the in vitro culture

We studied the interaction of the day length, cytokinins, and gibberellins in the control of tuberization in potato (Solanum tuberosum L, cv. Desire) plants and derived transgenic plants with the inserted PHYB gene from Arabidopsis encoding the synthesis of phytochrome B apoprotein and put under the control of the 35S CaMV promoter. Plantlets were cultured in vitro on hormone-free MS medium containing 5% sucrose and kinetin (1 mg/l) or/and GA (0.5 and 1.0 mg/l), at long day (LD, a 16-h photoperiod), short day (SD, a 10-h photoperiod), or continuous darkness conditions. The content of cytokinins (Ck, zeatin, and zeatin riboside) in various plant organs was determined by the immunoenzyme method, and GA activity was measured in bioassay with dwarf pea. Potato plant transformation with the PHYB gene enhanced substantially tuber initiation inhibition by LD. Kinetin addition to culture medium enhanced tuberization and reduced Ck content in aboveground shoots and Ck redistribution in the favor of underground organs. GA addition to the culture medium suppressed tuberization and induced Ck accumulation in aboveground organs. We concluded that Ck role in tuberization depends on their predominant localization in above- or underground potato organs. The involvement of Ck and GA in the competitive relations between growing tubers and shoots is considered.     

Photosynthetic rates of citronella and lemongrass

Ten selections of citronella (Cymbopogon nardus [L.] Rendle) were grown at 32/27, 27/21, or 15/10 C day/night temperatures, and plants from three populations of lemongrass (Cymbopogon citratus [D.C.] Stapf from Japan or Sri Lanka and Cymbopogon flexuosus [D.C.] Stapf from India) were grown at 8- or 15-hour photoperiods. Net photosynthetic rates of mature leaves were measured in a controlled environment at 25 C and 260 microeinsteins per meter² per second. Rates declined with increasing leaf age, and from the tip to the base of the leaf blade. Rates for citronella leaves grown at 15/10 C were extremely low for all selections. Highest rates of net photosynthesis were recorded for four selections grown at 27/21 C and for two selections grown at 32/27 C. Lemongrass grown at 8-hour photoperiod had higher photosynthetic rates than that grown at 15-hour photoperiod.     

Characteristics of Crassulacean Acid Metabolism in the Succulent C(4) Dicot, Portulaca oleracea L

Crassulacean acid metabolism (CAM) was investigated in leaves and stems of the succulent C₄ dicot Portulaca oleracea L. Diurnal acid fluctuations, CO₂ gas exchange, and leaf resistance were monitored under various photoperiod and watering regimes. No CAM activity was seen in well watered plants grown under 16-hour days. Under 8-hour days, however, well watered plants showed a CAM-like pattern of acid fluctuation with amplitudes of 102 and 90 microequivalents per gram fresh weight for leaves and stems, respectively. Similar patterns were also observed in detached leaves and defoliated stems. Leaf resistance values indicated that stomata were open during part of the dark period, but night acidification most likely resulted from refixation of respiratory CO₂. In water-stressed plants maximum acid accumulations were reduced under both long and short photoperiods. At night, these plants showed short periods of net CO₂ uptake and stomatal opening which continued all night long during preliminary studies under natural environmental conditions. Greatest acid fluctuations, in P. oleracea, with amplitudes of 128 microequivalents per gram fresh weight, were observed in water-stressed plants which had been rewatered, especially when grown under short days. No net CO₂ uptake took place, but stomata remained open throughout the night under these conditions. These results indicate that under certain conditions, such as water stress or short photoperiods, P. oleracea is capable of developing an acid metabolism with many similarities to CAM.     

Photoperiodic and Hormonal Control of Tuberization in Potato Plants Transformed with the <Emphasis Type="Italic">PHYB</Emphasis> Gene from <Emphasis Type="Italic">Arabidopsis</Emphasis>

We studied photoperiodic and hormonal regulation of tuberization in wild-type potato (Solanum tuberosum L., cv, Desiree) plants and derivative transgenic plants harboring the PHYB gene from Arabidopsis, which encodes the phytochrome B apoprotein, under the control of the cauliflower mosaic virus 35S promoter. Plants were cultured on hormone-free Murashige and Skoog nutrient medium containing 5% sucrose or on the same medium supplemented with 1 mg/l kinetin under conditions of long day (LD, 16 h), short day (SD, 10 h), or SD with interrupted long night. We estimated cytokinins (zeatin and zeatin riboside) in underground and aboveground plant organs by the ELISA technique and GA activity in a bioassay with dwarf pea seedlings. Under LD conditions, transgenic plants produced substantially less tubers than wild-type plants. Kinetin addition to the culturing medium resulted in stimulation of tuberization under LD conditions, especially pronounced in the PHYB plants. The content of cytokinins and the activity of GA were much higher under LD conditions, especially in leaves. The total level of both phytohormones was higher in transformed as compared to wild-type plants. A relation of phytochrome-dependent tuberization to the hormonal status of underground and above-ground plant organs and possible reasons for kinetin stimulatory effect on this process are discussed.     

The effect of phosphorus and nitrogen deficiency on growth of seedlings of spring barley in dependence on irradiance: Content of chlorophyll,nitrogen and phosphorus

Seedlings of spring barley,Hordeum vulgare L. cv. Mirena, were grown in complete mineral solution (Richter’s solution: R) or in solution lacking either phosphorus (R-P) or nitrogen (R-N) at low (LI: 28 W m^?2) or high (HI: 122 W m^?2) irradiance. Plants were kept in controlled environment chamber with 16 h photoperiod and 20 °C/15 °C day/night temperature. The experiment was terminated after 15 days when plants grown under R -N nad HI conditions died. The content of chlorophyll was estimated during the plant growth and content of nitrogen and phosphorus was determined at the end of the experiment. Deficiency of N and P induced higher chlorophyll formation at low irradiance. The efficiency of nitrogen and phosphorus utilization,i.e. ratio of plant dry mass/weight of N and P, respectively, per plant, was higher at HI in all experimental variants. Extremely high value of P utilization was found in plants grown under P-deficiency (850) as compared to the control (80). Understanding of interactions between the irradiance and deficiency of mineral nutrients is necessary for optimization of fertilization and understanding the mechanisms of action of mineral substances on plant structures and functions.     

Developmental history affects the susceptibility of spinach leaves to in vivo low temperature photoinhibition

Room temperature chlorophyll a fluorescence was used to determine the effects of developmental history, developmental stage, and leaf age on susceptibility of spinach to in vivo low temperature (5°C) induced photoinhibition. Spinach (Spinacia oleracea cv Savoy) leaves expanded at cold hardening temperatures (5°C day/night), an irradiance of 250 micromoles per square meter per second of photosynthetic proton flux density, and a photoperiod of 16 hours were less sensitive than leaves expanded at nonhardening temperatures (16 or 25°C day/night) and the same irradiance and photoperiod. This differential sensitivity to low-temperature photoinhibition was observed at high (1200) but not lower (500 or 800 micromoles per square meter per second) irradiance treatment. In spite of a differential sensitivity to photoinhibition, both cold-hardened and nonhardened spinach exhibited similar recovery kinetics at either 20 or 5°C. Shifting plants grown at 16°C (day/night) to 5°C (day/night) for 12 days after full leaf expansion did not alter the sensitivity to photoinhibition at 5°C. Conversely, shifting plants grown at 5°C (day/night) to 16°C (day/night) for 12 days produced a sensitivity to photoinhibition at 5°C similar to control plants grown at 16°C. Thus, any resistance to low-temperature photoinhibition acquired during growth at 5°C was lost in 12 days at 16°C. We conclude that leaf developmental history, developmental stage, and leaf age contribute significantly to the in vivo photoinhibitory response of spinach. Thus, these characteristics must be defined clearly in studies of plant susceptibility to photoinhibition.     

Role of phytochrome B in the development of cold tolerance in cucumber plants under light and in darkness

Cold tolerance of cucumber (Cucumis sativus L.) seedlings was investigated using wild-type plants and the phytochrome B-deficient mutant (lh-mutant). Plants were subjected for 6 days to intermittent short-term cooling (12°C for 2 h per day) and to continuous chilling under conditions of 16-h photoperiod (day/night = 16/8 h) and permanent illumination. “Dehardening” process was initiated by the transfer of plants to either light or dark conditions at 23°C. It was concluded that phytochrome B participates in the development of cold tolerance in cucumber plants under stress conditions, i.e., under short-term intermittent chilling at nights and during dehardening in continuous darkness.     

Jasmonate effects on in vitro tuberization and tuber bulking in two potato cultivars (Solanum tuberosum L.) under different media and photoperiod conditions

Summary Jasmonic acid (JA) effects on in vitro tuberization of potato nodal explants cvs. Sangre and Russet Burbank were tested under liquid and solid media conditions and 0,8, and 16h photoperiod. Explants taken from stock plants grown on 2.5μM JA-supplemented medium tuberized first, particularly in darkness. The most pronounced benefits of the JA pretreatment were recorded under 16h photoperiod, which is known to inhibit tuberization. Cultivar Sangre benefited from the JA preconditioning of stock plants more than Russet Burbank. Russet Burbank required the JA supplement in tuberization media to reach the same degree of stimulation. Overall, microtubers produced either from JA preconditioned stock plants or on the JA-containing tuberization media were more uniform and larger than from other treatments. Eight hours photoperiod was by far the best treatment for the production of high-quality uniform microtubers. JA conditioning of stock plants prior to taking explants for tuberization is being proposed as a treatment enhancing the quality of microtubers.     

Starch Accumulation in Leaves of Potato (Solanum tuberosum L.) during the First 18 Days of Photoperiod Treatment

Potato (Solanum tuberosum L.) plants were grown under long days(LD) of 18 h before a subset of the plants was transferred to10-h photosynthetic periods with either a dark night (SD) oran 8-h dim photoperiod extension with incandescent lamps (DE).Temperature was constant at 21 °C. Leaves were sampled atthe beginning and end of the high density light period for starchanalyses. Potato leaves accumulated starch more rapidly underSD than under LD; and this difference continued after a secondmajor sink, the tuber, began to develop. Starch accumulationover 10 h in SD leaves was three times higher than in LD leaves,even after 17 d of treatment. By this time SD gave higher wholeplant relative growth rates than LD, and the tuber mass of SDplants exceeded 30% of their total plant biomass. The DE treatmentresulted in starch accumulation intermediate to the LD and SDtreatments. Genotypes likewise differed: the earlier genotype,more strongly induced to tuberize, had higher leaf starch accumulationthan the later genotype. The effects of photoperiod and genotypewere also present when potatoes were grown at 27 °C, a temperatureunfavourable for tuberization under LD. Thus the formation ofa strong tuber sink was consistently associated with more rapidleaf starch accumulation. Potato, Solanum tuberosum L., cv. Norchip, photoperiod, temperature, genotype, starch accumulation, partitionin     

Environmental control of growth and development of Scrophularia marilandica

Summary Seedlings of Scrophularia marilandica were grown at different combinations of day/night temperature and photoperiod under controlled conditions. The species flowered in long days. The stems of plants grown at low temperature and short photoperiod failed to elongate. Treatment with gibberellic acid (GA₃) simulated the effect of increasing temperature and photoperiod and caused stem elongation in plants which would otherwise not have elongated. Application of GA₃ to plants grown at high temperature and long photoperiod resulted in increased stem elongation and flowering. The growth retardant (2-chloroethyl)trimethylammonium chloride (CCC) had little effect on rosette plants grown at low temperature and short photoperiod. Application of CCC to +GA₃ plants grown at a higher temperature and long photoperiod gave a significant increase in stem height. The interaction between temperature and applied GA is described in an experiment using plants grown at high and low temperatures for varying periods of time.This work was supported by National Science Foundation Grant GB 17483.     

Growth responses of Typha orientalis presl to controlled temperatures and photoperiods

Experiments are described in which seedlings of Typha orientalis Presls were grown for up to 6 months under precise conditions of temperature and photoperiod; photosynthesis was by natural daylight and did not vary between treatments. Variable treatments were imposed either from the seedling stage or on large plants raised under constant conditions.In general, total dry matter production increased as photoperiod increased from 8 to 16 h and also as day or night temperature increased, maximum production occurring when there was a warm day (30 or 27°C) and a small temperature drop (to 22°C) at night. The distribution of dry matter was also markedly affected by the imposed variables, leaf growth being favoured by high temperatures (to 30°C) and long photoperiods, and production of roots and rhizomes by low temperatures (to 10°C) and short photoperiods. None of the treatments resulted in floral initiation. The results are considered in relation to growth in the natural habitat.     

Effect of temperature on nitrogenase functioning in cowpea nodules

Nitrogenase (EC 1.7.99.2) activity of a cowpea (Vigna unguiculata (L.) Walp cv Caloona) symbiosis formed with a Rhizobium strain (176A27) lacking uptake hydrogenase and maintained under conditions of a 12-hour day at an air temperature of 30°C (800-1000 microeinsteins per square meter per second) and a 12-hour night at an air temperature of 20°C showed a marked diurnal variation in ratio of nitrogen fixed to hydrogen evolved. As little as 0.3 micromole nitrogen was fixed per micromole hydrogen evolved in the photoperiod versus up to 0.6 in the dark period. In plants maintained under the same diurnal illumination regime but at constant (day and night) air temperature (30°C), this difference was abolished and a relatively constant ratio of nitrogen fixed to hydrogen evolved (around 0.3 micromole per micromole) was observed day and night. Exposure of nodulated roots to a range of temperatures maintained for 2 hours in a single photoperiod indicated that, whereas hydrogen evolution increased with increasing temperature from 15°C to a maximum around 35°C, nitrogen fixation was largely unaffected over this temperature range. Both functions of the enzyme declined sharply at temperatures above 38°C. A similar general response of nitrogen fixation to root temperature was observed in glasshouse-grown, sand-cultured plants maintained under a range of temperatures (from 15 to 35°C) for a 14-day period in mid vegetative growth. The effect of temperature on the proportion of electrons allocated to proton reduction compared with nitrogen reduction showed a linearly increasing relationship (correlation coefficient = 0.96) between 15°C and 47°C.     

Phytochrome B mediates the photoperiodic control of tuber formation in potato

To determine whether phytochrome B is involved in the response of potato plants to photoperiod, a potato PHYB cDNA fragment was inserted in the antisense orientation behind the 35S CaMV promoter in Bin19 and this construct was transformed into Solanum tuberosum ssp. andigena plants which normally require short days for tuberization. Two independent transformants were obtained that had much lower levels of PHYB mRNA and protein, and which exhibited phenotypes characteristic of phyB mutants, for example, elongated stems and decreased chlorophyll content. The level of phyA, and of several phytochrome A-controlled responses, was unaffected in these plants. The photoperiodic control of tuberization in these antisense PHYB plants was abolished, the plants tuberizing in short day, long day, or short day plus night break conditions. This result shows that phytochrome B is required for the photoperiodic control of tuberization in potato ( Solanum tuberosum ssp. andigena ) and that it regulates this developmental process by preventing tuber formation in non-inductive photoperiods rather than by promoting tuberization in inductive photoperiods.     

Role of gibberellins in stem elongation and flowering in radish

The relationship among gibberellins, CCC, vernalization, and photoperiod in the flowering response of radish, Raphanus sativus L., cv. Miyashige-sofuto, was studied. The optimal condition for flowering was vernalization and a 16-hour photoperiod; GA₃ had no additional effect. Gibberellin A₃ (60 μg total) was not able to induce flowering in nonvernalized plants grown on 8-hour days, but it did increase the percentage of nonvernalized plants that flowered under long days from 60 to 100.

Gibberellin content of vernalized seedlings increased within the first 24 hours after seedlings were transferred to the greenhouse. Content reached a peak in the first 4 days after transfer and thereafter remained constant. Essentially no gibberellin was found in 2 day-old non-vernalized (control) seedlings of comparable size to the vernalized ones. Gibberellin content in the controls reached a peak on the fourth day of growth in the greenhouse; thereafter, it decreased steadily.

Bolting was inhibited slightly by CCC when applied during vernalization; it was almost completely inhibited when CCC was applied after seed vernalization. Extraction experiments revealed that CCC actually reduced the gibberellin content when applied during or after vernalization. The dwarfing agent, however, had essentially no effect on flowering. We concluded that gibberellins likely play a direct role in bolting of `Miyashige-sofuto' radish, but probably are not directly functional in initiating flowering.

     

Causes of Flowering of Long-Day Potato Species under Short-Day and Cold-Night Conditions

Flowering and tuber formation in high-mountain potato species Solanum sparsipilum Bitt., S. acaule Bitt., S. punae Juz., S. demissum Lindl., and a tuber crop Ullucus tuberosus Caldas. were investigated. All these species are characterized by absolute requirement of long day-length for flowering and short day-length for tuberization. Plants were grown under the following conditions: natural day-length with a photoperiod of 17 h or longer (treatment 1), short days with a photoperiod of 12 h and warm nights (15–20°C) (treatment 2), and short days with cold nights (5–6°C) (treatment 3). In the first treatment, plants produced flowers but no tubers. In the second treatment, plants produced tubers but no flowers. In the third treatment, plants produced both flowers and tubers. In leaves of S. acaule and U. tuberosus, the levels of gibberellins and ABA were determined. A high activity of gibberellins in the third treatment was similar to that in the first treatment, whereas high ABA activity in the third treatment was similar to that in the second treatment. It is supposed that cold nights retard the destruction of GA in plants during the dark period of diurnal cycle and ensure a permanently high level of gibberellins, which facilitates flowering of long-day species under short-day conditions. The high level of ABA is considered a plant response to short-day conditions, which is favorable for tuberization.     

C(4) Acid Metabolism and Dark CO(2) Fixation in a Submersed Aquatic Macrophyte (Hydrilla verticillata)

The CO₂ compensation point of the submersed aquatic macrophyte Hydrilla verticillata varied from high (above 50 microliters per liter) to low (10 to 25 microliters per liter) values, depending on the growth conditions. Plants from the lake in winter or after incubation in an 11 C/9-hour photoperiod had high values, whereas summer plants or those incubated in a 27 C/14-hour photoperiod had low values. The plants with low CO₂ compensation points exhibited dark ¹⁴CO₂ fixation rates that were up to 30% of the light fixation rates. This fixation reduced respiratory CO₂ loss, but did not result in a net uptake of CO₂ at night. The low compensation point plants also showed diurnal fluctuations in titratable acid, such as occur in Crassulacean acid metabolism plants. However, dark fixation and diurnal acid fluctuations were negligible in Hydrilla plants with high CO₂ compensation points.     

Carbon dioxide compensation values in citronella and lemongrass

Carbon dioxide compensation values of mature leaves from 10 selections of citronella (Cymbopogon nardus [L.] Rendle) grown at 32/27 or 27/21 C day/night temperatures and three strains of lemongrass (Cymbopogon citratus [D.C.] Stapf. and Cymbopogon flexuosus [D.C.] Stapf.) grown at 8- or 15-hour photoperiods were measured in a controlled environment at 25 C. All leaves had low compensation values but citronella varied from 1.3 to 9.7 μl/liter and lemongrass from 0.7 to 3.5 μl/liter. Lower growing temperature generally resulted in lower compensation values for citronella but there was no consistent photoperiod effect on lemongrass.