Twenty-five cultivars were initially screened for germination at 10 °C, 30 °C, and 40 °C. Four cultivars were chosen for further study for physiological and biochemical characteristics—namely, `Texas Cream 40' (TC-40), which showed ability to germinate at very high (40 °C) and low (10 °C) temperatures; `Black Crowder' (BC), which had acceptably high germination at 40 °C, but reduced germination at 10 °C; and `Mississippi Purple' (MP), which exhibited lower germination at all temperatures tested. The main sugars present in cowpea seed were sucrose, raffinose, and stachyose. Sugar contents were affected by cultivar, type of tissue, and temperature. Sucrose contents were higher in embryo tissue of cultivars with a lower germination percentage, and reduced in the cultivar with a higher germination percentage, suggesting the use of sucrose for germination. Sucrose decreased greatly at 30 °C and increased again at 40 °C. Sucrose “de novo” synthesis was higher at higher temperature. An accumulation of sucrose was evident in embryo tissues of cultivars with reduced ability to germinate at low temperature. Raffinose and stachyose contents were higher in ungerminated seed. In germinated seed, raffinose and stachyose contents were found only in cotyledon tissues at 10 °C. The peroxidase activity was affected by cultivars, type of tissue, and temperature. The highest peroxidase activity was found at low temperature (10 °C) in embryo tissue of the cultivar with the highest germination. The result also suggests that high peroxidase activity was related to ability of seed to germinate at low temperature.
Shahidul Islam, Carmen Rafaela, and James Garner
Joyce C. Pennycooke, Ramarao Vepachedu, Cecil Stushnoff, and Michelle L. Jones
Previous studies of plant tolerance to low temperature have focused primarily on the cold acclimation response, the process by which plants increase their tolerance to freezing in response to low nonfreezing temperatures, while studies on the deacclimation process have been largely neglected. In some plants, cold acclimation is accompanied by an increase in raffinose family oligosaccharides (RFO). The enzyme α-galactosidase (EC 220.127.116.11) breaks down RFO during deacclimation by hydrolyzing the terminal galactose moieties. Here we describe the isolation of PhGAL, an α-galactosidase cDNA clone from Petunia (Petunia ×hybrida `Mitchell'). The putative α-galactosidase cDNA has high nucleotide sequence homology (>80%) to other known plant α-galactosidases. PhGAL expression increased in response to increased temperature and there was no evidence of developmental regulation or tissue specific expression. Increases in α-galactosidase transcript 1 hour into deacclimation corresponded with increases in α-galactosidase activity and a concomitant decrease in raffinose content, suggesting that warm temperature may regulate RFO catabolism by increasing the transcription of the α-galactosidase gene. This information has potential practical applications whereby α-galactosidase may be targeted to modify endogenous raffinose accumulation in tissues needed for freezing stress tolerance.
C.L. Haynes, O.M. Lindstrom, and M.A. Dirr
Decreasing photoperiods and decreasing temperatures induce cold acclimation and the accumulation of soluble sugars in many plants. Two cultivars of southern magnolia differing in cold hardiness and acclimation patterns, were monitored to determine photoperiod × temperature interaction on cold hardiness and soluble sugar content. Cold hardiness increased with low temperatures and short photoperiods. Total soluble sugars, sucrose, and raffinose consistently increased in the leaves and stems of both cultivars in response primarily to low temperature. `Little Gem' was less responsive to photoperiod than `Claudia Wannamaker'
Aly M. Lasheen and Carl E. Chaplin
Levels of soluble sugars in bark, leaves, leaf buds and flower buds of 2 cultivars of peach (Prunus persica (L.) Batsch) differing in cold hardiness were compared throughout the year. Thirteen sugars — galactose, glucose, fructose, xylose, stachyose, sucrose, raffinose, rhamnose, maltose, trehalose, arabinose, ribose and mannose — were present in measurable and variable concentrations. In general, oligosaccharides accumulated, particularly in the bark, during fall and winter, whereas monosaccharides accumulated during periods of active growth. These data do not show significant differences between the 2 cultivars regarding the accumulation of these sugars and cold hardiness.
D. L. Hughes and M. Yamaguchi
The concentration of fructose, glucose, sucrose, raffinose, and stachyose in the leaves, stem, and fruit of the muskmelon plant (Cucumis melo L.) was determined using paper and gas chromatography. Stachyose was the predominant sugar in the leaves but was absent in the developing fruit. Most of the 14C label in 14CO2-treated leaves was associated with stachyose, while the 14C label in the fruit was associated with fructose and glucose. The concentration of stachyose was highest in the leaves, intermediate in the stem, and lowest in the fruit.
Canes of three field-grown cultivars of red raspberry (Rubus idaeus L. `Maurin Makea', `Ottawa', and `Muskoka') were sampled from October to April. Carbohydrate contents of canes and flower buds were analyzed, and cold hardiness (LT50) was determined by controlled freezing. Starch, sucrose, glucose, fructose, and minor amounts of raffinose and stachyose were present in both cane and bud tissues. Glucose and fructose were the predominant sugars in buds. In canes, the proportion of sucrose of all sugars was greater than in buds. Seasonal changes in carbohydrates were related to changes in cold hardiness and mean air temperature during a 5-day period preceding sampling. Starch decreased during fall and was barely detectable in midwinter. Soluble carbohydrates accumulated to 73 to 89 mg·g-1 dry weight in canes and 113 to 131 mg·g-1 dry weight in buds in midwinter. The most striking increase occurred in the concentration of sucrose, but glucose, fructose, raffinose, and stachyose also accumulated. There was a positive correlation between LT50 and the amount of starch, but a negative correlation between LT50 and the amounts of total soluble carbohydrates, sucrose, glucose, and fructose. High levels of sucrose, total soluble carbohydrates, and a high ratio of sucrose to glucose plus fructose were characteristic of a hardy cultivar. Results are evidence of the importance of carbohydrate reserves, especially sucrose, on winter survival of red raspberry.
Manfredo J. Seufferheld and Cecil Stushnoff
Strawberry plantlets, regenerated from leaf disks, were used as a model system to study the effect of high concentrations of sugars and dehydration on survival during cryopreservation. After cold acclimation, plantlets imbibed for 3 days (one day each) in 0.5, 0.7 and 1.2 M sucrose and (1.0M sucrose + 0.2M raffinose) and desiccated to 25 % moisture (fwb) in alginate capsules consistently survived cryopreservation. Differential scanning calorimetry revealed only a very small exotherm between -20C and -28C during freezing; a glass transition at -50C and a small melting event at -10C during warming. Conversely, samples with the lowest survival rate, had a large nucleation exotherm at -30C and a devitrification exotherm between -70 and -40C. We conclude that imbibition with sugars, coupled with desiccation treatments, may be used to manipulate freeze tender tissues of strawberry to permit successful cryopreservation.
Cecil Stushnoff, Richard L. Remmele Jr., and V. Esensee
Aqueous fractions in dormant buds of Amelanchier alnifolia Nutt. `Smoky', may exist either as liquid, ice or glass phases depending on the temperature history and the water content of the tissue. Phase diagrams for these states were constructed from differential scanning calorimetry (DSC) freezing and warming scans. The diagrams show that glass transition temperatures shift to warmer temperatures as cold hardening increases and as the water content is lowered by controlled desiccation. Glass transitions were detected from -60 to -20° C, during slow freezing scans in the DSC, suggesting that survival of this extremely cold hardy tissue is based upon a potential to undergo glass transitions in the dormant state. Endogenous raffinose family oligosaccharides (RFO) increase during cold hardening, and decrease as hardiness diminishes with the onset of growth.
Donald E. Irving, Paul L. Hurst, and Jonathan S. Ragg
During this study, we divided the developmental growth pattern of buttercup squash into three phases: 1) early growth, from flowering up to 30 days after flowering; 2) maturation, from 30 days until 60 days after flowering (or harvest); and 3) ripening, from 60 days (or harvest) until ≈100 days after flowering. Harvest occurred at 48 days after flowering. Fruit growth (expansion), starch, and dry matter accumulation were largely completed during early growth, and there was a progressive decline in the respiration rate. Extractable activities of acid and alkaline invertases, sucrose synthase, alkaline α-galactosidase, and sucrose phosphate synthase (assayed with saturating substrates) were high initially but declined markedly during this phase. Glucose, fructose, and low concentrations of raffinose saccharides were present, but no sucrose was detected. During maturation, starch and dry matter remained nearly constant and sucrose began to accumulate. During ripening, starch was degraded, sucrose synthase activity was significant but relatively constant, sucrose phosphate synthase activity increased, and sucrose continued to accumulate.
R. D. Wright and L. H. Aung
Determinations of carbohydrates in the plant organs of Rhododendron spp. cv. ‘Sweetheart Supreme’ and ‘Hexe’, were made by chemical analysis, thin-layer chromatography (TLC) and gas-liquid chromatography (GLC). Reducing sugar content was 1.5 times higher in buds than in leaves for ‘Hexe’ with no significant differences for ‘Sweetheart Supreme’. Reducing sugars were also higher in the roots than stems with both cultivars. Sucrose content was 1.4 times greater in leaves than in buds of ‘Hexe’ and 1.6 times greater in ‘Sweetheart Supreme’. Starch was significantly higher in leaves and buds than in stems and roots. The predominant soluble sugars identified by TLC and GLC were sucrose, glucose, and fructose. Small but detectable amounts of raffinose and maltose and an unidentified compound were also found in the plant organs.