Cold pretreatment to enhance green plant regeneration from rye anther culture

Cold pretreatment of detached tillers of rye, Secale cereale, was tested under two light regimes. The ryes included two spring and two winter cultivars. Significant increases in green plant regeneration were recorded in each experiment when cold pretreatments of two to four weeks were applied. Dim light during the stress period improved green plant regeneration for two of the four cultivars tested. The highest regeneration rate, 30.6 green plants per 100 plated anthers, was reached following three weeks at +4 °C under dim light, for spring rye Jo02. Starvation stress applied to plated anthers in mannitol medium suppressed anther response. This revised version was published online in June 2006 with corrections to the Cover Date.     

Production of large number of doubled haploid plants from barley anthers pretreated with high concentrations of mannitol

Pretreatment with increasing concentrations of mannitol, from 0.3 to 0.7 M, was used to induce stress in cultured anthers of barley (Hordeum vulgare L.). Three cultivars with varying degrees of androgenetic ability were studied. A positive linear relationship was found between concentration of mannitol in the pretreatment medium and the number of regenerated green doubled haploid plants in all the cultivars. The pretreatment also resulted in an increasing proportion of embryos to dividing microspores, and in green to albino plantlets. The optimum length of the pretreatment seemed to be genotype dependent. When Ficoll was used as an alternative stress agent a differential genotype response was observed.Abbreviations BAP N⁶ benzyl-aminopurine - IAA indol acetic acid     

Drought enhances maize chilling tolerance. II. Photosynthetic traits and protective mechanisms against oxidative stress

In the present research we studied the photosynthetic traits and protective mechanisms against oxidative stress in two maize ( Zea mays L.) genotypes differing in chilling sensitivity (Z7, tolerant and Penjalinan, sensitive) subjected to 5°C for 5 days, with or without pretreatment by drought. The drought pretreatment decreased the symptoms of chilling injury in Penjalinan plants estimated as necrotic leaf area and maximum quantum yield of photosystem II. Furthermore, drought pretreatment diminished the level of lipid peroxidation caused by chilling in Penjalinan plants. After one day of recovery from chilling the Z7 and drought-pretreated Penjalinan plants showed higher net photosynthesis rates than the non-drought-pretreated Penjalinan plants, thereby decreasing the probability of generating reactive oxygen species. The greater net photosynthesis was correlated with the greater NADP-malate dehydrogenase activity. No differences in either the de-epoxidation state of the xanthophyll cycle or the antioxidant enzyme activities were found among the chilled groups of plants. However, a drastic decrease in ascorbate content was observed in chilled Penjalinan plants without drought pretreatment. As we found an increase of H₂O₂ content after drought pretreatment, we suggest its involvement as a signal in the drought-enhanced chilling tolerance of maize.     

Production of doubled haploids in durum wheat (<Emphasis Type="Italic">Triticum durum</Emphasis> Desf.) through culture of unpollinated ovaries

The objective of this work was to produce doubled haploid plants from durum wheat through gynogenesis using unpollinated ovary culture of three local Tunisian genotypes (Jenah Khotifa, Hmira, Azizi) and three improved cultivars (Karim, Khiar, Razzek). A total of 12,000 unpollinated ovaries were cultured in this study. Spikes were either pretreated at 4°C for 14 days or at 4°C in a mannitol solution (0.3 M) for 7 days. Induction was performed using two media. We showed that ovary development, callus and plantlet regeneration was influenced significantly by genotype and growth conditions. The highest regeneration frequency was obtained when the microspore population was in the late mononucleate to binucleate stage. Our results suggested that the cold pretreatment for 14 days was more efficient than the cold treatment in a mannitol solution. Furthermore, the addition of 2,4-D, vitamins and glutamine, and the use of maltose as sugar source in media improved the ovary culture. When the unpollinated ovaries were cultured under the conditions found to be optimal in the present study, a total of 84 plants were produced, all green and haploid. The best levels for regenerated plants were obtained with the cultivars Khiar (3.5%), Hmira (3.1%) and Karim (1.5%). Fertile doubled haploid plants were obtained by colchicine treatment. This result represents a modern tool for breeders to produce durum wheat homozygous lines in a few months.     

The Role of Abscisic Acid in Induction of Androgenesis: A Comparative Study Between Hordeum vulgare L. Cvs. Igri and Digger

Under the same mannitol pretreatment and culture conditions, regeneration efficiency in the barley cultivar (cv.) Igri was about 10 times higher than in the cv. Digger, a difference only partially reflected by a difference in viable microspores after anther pretreatment. Therefore, a comparative study between cvs. Igri and Digger was carried out under various pretreatment conditions. For both cultivars, under water, CPW buffer and mannitol pretreatment conditions, there was a positive correlation between microspore viability and regeneration efficiency in that mannitol > CPW buffer >> water. Mannitol pretreatment of cv. Igri produced a much higher endogenous abscisic acid (ABA) level than as to Digger. Addition of ABA stimulated both percentages of viability and regeneration efficiency except in the case of mannitol pretreatment. Under CPW buffer pretreatment conditions, addition of ABA significantly stimulated regeneration efficiency and was ABA concentration dependent. However, cv. Digger was less responsive to ABA than cv. Igri. In both cultivars, under less optimal pretreatment conditions (e.g., water and CPW buffer), the effect of ABA was to stimulate increased percentages of viability and/or to reduce the number of binucleate microspores. Moreover, in cv. Igri, direct culture of anthers for 4 days without pretreatment caused an increased number of binucleate microspores compared with microspores with pretreatment for 4 days. These binucleate microspores showed DNA degradation in the nuclei. However, with mannitol pretreatment binucleate microspores and DNA fragmentation in the nuclei of microspores was rarely observed. On the basis of our observations, we suggest that the difference in regeneration efficiency in cv. Igri and cv. Digger is related to the differences in endogenous ABA production levels under mannitol pretreatment and responsiveness to ABA. One of the effects of ABA is likely due to an inhibition of cell death. Received May 21, 1999; accepted October 5, 1999     

不同胁迫预处理提高水稻幼苗抗寒性期间膜保护系统的变化比较

通过比较盐、冷和热激三种不同胁迫预处理提高水稻(Oryza sativa L.)抗寒性过程中膜保护系统的变化,探讨植物交叉适应机理。结果表明,水稻幼苗经不同胁迫预处理均可提高幼苗的抗寒性。与未预处理苗相比,不同胁迫(冷、热、盐)预处理之间在处理后、低温伤害后及恢复2d后的3个不同时期膜酶促和非酶促保护系统及同工酶酶谱变化各有异同,既有部分共性,也有其独特性。     

An Enhancing Effect of Exogenous Mannitol on the Antioxidant Enzyme Activities in Roots of Wheat Under Salt Stress

The role of mannitol as an osmoprotectant, a radical scavenger, a stabilizer of protein and membrane structure, and protector of photosynthesis under abiotic stress has already been well described. In this article we show that mannitol applied exogenously to salt-stressed wheat, which normally cannot synthesize mannitol, improved their salt tolerance by enhancing activities of antioxidant enzymes. Wheat seedlings (3 days old) grown in 100 mM mannitol (corresponding to −0.224 MPa) for 24 h were subjected to 100 mM NaCl treatment for 5 days. The effect of exogenously applied mannitol on the salt tolerance of plants in view of growth, lipid peroxidation levels, and activities of antioxidant enzymes in the roots of salt-sensitive wheat (Triticum aestivum L. cv. Kızıltan-91) plants with or without mannitol was studied. Although root growth decreased under salt stress, this effect could be alleviated by mannitol pretreatment. Peroxidase (POX) and ascorbate peroxidase (APX) activities increased, whereas superoxide dismutase (SOD), catalase (CAT), and glutathione reductase (GR) activities decreased in Kızıltan-91 under salt stress. However, activities of antioxidant enzymes such as SOD, POX, CAT, APX, and GR increased with mannitol pretreatment under salt stress. Although root tissue extracts of salt-stressed wheat plants exhibited only nine different SOD isozyme bands of which two were identified as Cu/Zn-SOD and Mn-SOD, mannitol treatment caused the appearance of 11 different SOD activity bands. On the other hand, five different POX isozyme bands were determined in all treatments. Enhanced peroxidation of lipid membranes under salt stress conditions was reduced by pretreatment with mannitol. We suggest that exogenous application of mannitol could alleviate salt-induced oxidative damage by enhancing antioxidant enzyme activities in the roots of salt-sensitive Kızıltan-91.     

Influence of anther pretreatment and culture medium composition on the production of barley doubled haploids from model and low responding cultivars

Different pretreatments were given to anthers of barley before culturing, and their effects assessed on the frequency of embryos and green doubled haploid plants produced. Mannitol pretreatment was better than cold pretreatment for some low responding cultivars. Optimal concentration of mannitol for pretreatment depended on cultivar. Low responding genotypes needed a higher concentration of mannitol than responsive ones. The addition of Ficoll to liquid medium increased the number of embryos and green plants. The influence of the growth regulators 2,4-D and TIBA was assayed using ten cultivars of barley grown in Spain. The anti-auxin TIBA gave good embryo production with some of the low responding cultivars. Two row-type cultivars always produced higher number of embryos and green plantlets than six row-type. The application of these modifications to 10 F₁ hybrids with potential agronomic value, allowed the production of almost 1000 doubled haploid plants from only 3500 anthers. Up to two doubled haploid plants per flower were produced from the cross Monlon × Sonja. This revised version was published online in June 2006 with corrections to the Cover Date.     

Chilling to zero degrees disrupts pollen formation but not meiotic microtubule arrays in Triticum aestivum L.

Throughout the wheat‐growing regions of Australia, chilling temperatures below 2 °C occur periodically on consecutive nights during the period of floral development in spring wheat (Triticum aestivum L.). In this study, wheat plants showed significant reductions in fertility when exposed to prolonged chilling temperatures in controlled environment experiments. Among the cultivars tested, the Australian cultivars Kite and Hartog had among the lowest levels of seed set due to chilling and their responses were investigated further. The developmental stage at exposure, the chilling temperature and length of exposure all influenced the level of sterility. The early period of booting, and specifically the +4 cm auricle distance class, was the most sensitive and corresponded to meiosis within the anthers. The response of microtubules to chilling during meiosis in Hartog was monitored, but there was little difference between chilled and control plants. Other abnormalities, such as plasmolysis and cytomixis increased in frequency, were associated with death of developing pollen cells, and could contribute to loss of fertility. The potential for an above‐zero chilling sensitivity in Australian spring wheat varieties could have implications for exploring the tolerance of wheat flower development to chilling and freezing conditions in the field.     

Separately and simultaneously induced dark chilling and drought stress effects on photosynthesis, proline accumulation and antioxidant metabolism in soybean

The effects of separately or simultaneously induced dark chilling and drought stress were evaluated in two Glycine max (L.) Merrill cultivars. For the separately induced dark chilling treatment (C), plants were incubated at 8 °C during 9 consecutive dark periods. During the days, plants were kept at normal growth temperatures. For the separately induced drought treatment (D), plants were maintained at normal growth temperatures without irrigation. For the simultaneously induced dark chilling and drought stress treatment (CD), plants were dark chilled without irrigation. All treatments caused similar decreases in pre-dawn leaf water potential, but resulted in distinct physiological and biochemical effects on photosynthesis. In Maple Arrow, where C had the smallest effect on photosynthesis, prolonged CD caused less inhibition of photosynthesis compared to D. Compared to Fiskeby V, the photosynthetic apparatus of Maple Arrow appears to possess superior dark chilling tolerance, a property which probably also conveyed enhanced protection against CD. Proline accumulation was prevented by CD at the ψ_PD where D already resulted in considerable accumulation. The superior capacity for proline accumulation in Maple Arrow would seem to be an important factor in its stress tolerance. Antioxidant activity evoked by CD and D was higher than for C alone. In Fiskeby V, the small increase in ascorbate peroxidase (EC 1.11.1.7) activity, which was in most cases not accompanied by increased gluthatione reductase (EC 1.6.4.2) activity, could impact negatively on its stress tolerance. These results demonstrate large genotypic differences in response to chilling and drought stress, even between soybean cultivars regarded as chilling tolerant.     

Chilling-induced leaf abscission of Ixora coccinea plants. III. Enhancement by high light via increased oxidative processes

The role of increased oxidation induced by successive stresses of chilling and high light in the induction of leaf abscission was studied in Ixora coccinea plants in relation to auxin metabolism and oxidative processes. Exposure of plants following dark chilling (7°C for 3 days) to high light (500–700 μmol m⁻² s⁻¹ photosynthetically active radiation) for 5 h at 20–25°C enhanced chilling-induced leaf abscission. This abscission was inhibited by pretreatment with the antioxidant butylated hydroxyanisole, α -naphthaleneacetic acid or the ethylene action inhibitor, 1-methylcyclopropene. The oxidative processes initiated during the low light period following the dark chilling period, such as indoleacetic acid (IAA) decarboxylation and lipid peroxidation, were further enhanced by subsequent exposure to high light. Photoinhibition, expressed by the reduction of the chlorophyll fluorescence parameter Fv/Fm, was evident following exposure to high light, irrespective of the temperature of the pretreatment, but this reduction persisted only in chilled plants. This suggests that oxidative processes generated during and after the chilling period might have inhibited the recovery from photoinhibition. The chilling stress under darkness induced a 60% reduction in superoxide dismutase (SOD) activity and significant increases (130–600%) in the activities of several other antioxidative enzymes. These data suggest that the chilling-induced reduction in SOD activity may well be responsible for the increase in the oxidative stress induced by the subsequent light treatment, as expressed by the increased enzymatic activities. Taken together, this study provides further support for the involvement of oxidative processes in the events occurring in tissues exposed to sequential chilling and light stresses, leading to reduction in free IAA content in the abscission zone and to leaf abscission.     

Activities of photosystem II and antioxidant enzymes in chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L<Emphasis Type="Italic">.</Emphasis>) cultivars exposed to chilling temperatures

This study was carried out to determine the effect of chilling on both cold-acclimated and non-acclimated chickpea (Cicer arietinum L.) cultivars (Gökçe and Can?tez 87). Chickpea seedlings grown in soil culture for 12 days were subjected to chilling temperatures (2 and 4°C for 12 days) after maintaining in cold-acclimation (10°C, 7 days) or non-acclimation (25°C, 7 days) periods. The lowest values of growth parameters were obtained with cold-acclimated plants, whereas non-acclimated plants exhibited the lowest water content values, especially at 2°C. There was no effect of cold-acclimation period on chlorophyll fluorescence parameters. Plants subjected to chilling temperatures after cold-acclimation were more tolerant with respect to chlorophyll fluorescence parameters, and Gökçe had better photosystem II (PSII) photochemical activity. In the chilling treatments, total chlorophyll (a + b) content reduced, especially at 2°C, while anthocyanin and flavonoid contents increased to a greater extent in Gökçe and carotenoid content of the cultivars did not change. Malondialdehyde (MDA) content was higher for Can?tez 87, mostly at 2°C, while proline accumulation was greater for Gökçe. The cold-acclimation period led to a remarkable increase in antioxidant enzyme activities of both cultivars. The superoxide dismutase (SOD) activity was much higher in Gökçe for both chilling temperatures and the ascorbate peroxidase (APX) activity increased only in the cold-acclimated 4°C treatments. Similarly, with APX activity, the glutathione reductase (GR) and peroxidase (POD) activities of cultivars were higher in cold-acclimated plants at both the chilling temperatures, mostly in Gökçe. The results of this study indicate that cold-acclimation increased the cultivars ability to withstand the chilling temperatures. The lower MDA content and higher antioxidant and photochemical activities in Gökçe indicated an enhanced chilling tolerance capacity of this cultivar to protect the plant from oxidative damage.     

Influence of photosynthetic photon flux densities before and during long-term chilling on xanthophyll cycle and chlorophyll fluorescence quenching in leaves of tomato (Lycopersicon hirsutum)

A high-altitude ecotype of tomato ( Lycopersicon hirsutum f. typicum Humb. and Bonpl.) has previously been shown to resist further loss of photosynthetic function after three to four days of chilling stress. This study examined the influence of PPFD prior to, and during chilling on the development of protective zeaxanthin and energy-dependent quenching mechanisms in this ecotype. Five-week-old tomato plants were acclimated to either low PPFD (60 μmol m⁻² s⁻¹) or high PPFD (550 μmol m⁻² S⁻¹) at 25/20°C (day/night) for three days, and then exposed to a temperature of 5/5°C and a PPFD of either 60 or 550 μmol m⁻² s⁻¹ for three days. The plants acclimated to low PPFD had lower Chl a/b ratio, and lower level of total Chl per leaf area, total xanthophyll cycle pool and β-carotene. The capacity of their photosynthetic system to resist photoinhibition and to recover photosynthetic function was also lower compared to that of the plants acclimated at high PPFD but exposed to the same chilling stress. In the plants chilled at low PPFD, energy-dependent quenching preceded the formation of zeaxanthin on the first day of chilling and there was an overall reduction in the conversion of violaxanthin to zeaxanthin as compared to the plants chilled at high PPFD. During the last day of chilling-induced photoinhibition, energy-dependent quenching in any of the treatments did not increase, but zeaxanthin levels increased continuously throughout the three days of chilling. Our results suggest that light-acclimation before chilling affects the capacity of the plants to resist chilling-induced photoinhibition. In addition, photoinhibitory quenching appears to be a major component for quenching excessive energy at the latter stage of long-term chilling.     

Effects of Temperature Acclimation Pretreatment on the Ultrastructure of Mesophyll Cells in Young Grape Plants （Vitis vinifera L. cv. Jingxiu） Under Cross-Temperature Stresses

Leaves from annual young grape plants （Vitis vinifera L. cv. Jingxiu） were used as experimental materials. The ultrastructural characteristics of mesophyll cells in chilling-treated plants after heat acclimation （HA） and in heat-treated plants after cold acclimation （CA） were observed and compared using transmission electron microscopy. The results showed that slight injury appeared in the ultrastructure of mesophyll cells after either HA （38℃ for 10 h） or CA （8℃ for 2.5 d）, but the tolerance to subsequent extreme temperature stress was remarkably improved by HA or CA pretreatment. The increases in membrane permeability and malondialdehyde concentration under chilling （0℃） or heat （45℃） stress were markedly inhibited by HA or CA pretreatment. The mesophyll cells of plants not pretreated with HA were markedly damaged following chilling stress. The chloroplasts appeared irregular in shape, the arrangement of the stroma lamellae was disordered, and no starch granules were present. The cristae of the mitochondria were disrupted and became empty. The nucleus became irregular in shape and the nuclear membrane was digested. In contrast, the mesophyll cells of HA-pretreated plants maintained an intact ultrastructure under chilling stress. The mesophyll cells of control plants were also severely damaged under heat stress. The chloroplast became round in shape, the stroma lamellae became swollen, and the contents of vacuoles formed clumps. In the case of mitochondria of control plants subjected to heat stress, the outer envelope was digested and the cristae were disrupted and became many small vesicles. Compared with cellular organelles in control plants, those in CA plant cells always maintained an integrated state during whole heat stress, except for the chloroplasts, which became round in shape after 10 h heat stress. From these data, we suggest that the stability of mesophyll cells under chilling stress can be increased by HA pretreatment. Similarly, CA pretreatment can protect chloroplasts, mitochondria, and the nucleus against subsequent heat stress; thus, the thermoresistance of grape seedlings was improved. The results obtained in the present study are the first, to our knowledge, to offered cytological evidence of cross-adaptation to temperature stresses in grape plants.     

Genetic basis of heterosis explored by simple sequence repeat markers in a random-mated maize population

We investigated the isozyme profiles of antioxidant enzymes in cultivars and lines with different seed productivity in cool climate conditions as a step towards understanding the physiological and genetical mechanisms underlying chilling tolerance in soybean. While no difference in superoxide dismutase, or catalase isozyme profiles was observed among the cultivars and lines tested, we found polymorphism in the ascorbate peroxidase isozyme profile; there were two types, with or without a cytosolic isoform (APX1). The cultivars and lines lacking APX1 proved more tolerant to chilling temperatures, as evaluated by yielding ability. The genotype-dependent deficiency of APX1 was consistent in plants and tissues under various oxidative stress conditions including the exposure to low-temperatures. In addition, the genetic analysis of progeny derived from crossing between cultivars differing in the isozyme profile indicated that the APX1 deficiency is controlled by a single recessive gene (apx1), and is inherited independently of the genes that have previously been identified for their association with chilling tolerance. Molecular and linkage analyses suggested that the variant gene of the APX1-absent genotype coding for a cytosolic APX, which contained a single nucleotide substitution and a single nucleotide deletion in the coding region, is responsible for the genotype-dependent deficiency of APX1. The association of APX1 deficiency with chilling tolerance is discussed in detail.     

Increased chilling tolerance and altered carbon metabolism in tomato leaves following application of mechanical stress

We investigated the effects of brushing on the chilling tolerance and metabolism of nonstructural carbohydrates (soluble sugars and starch) in tomato leaves before, during and after a chilling stress. Tomato plants ( Lycopersicon esculentum Mill. cv. Caruso) were cultivated either without mechanical stress application (control plants) or with daily brushing treatments for 15 days (brushed plants), prior to a 7-day chilling treatment (8/5°C day/night). Brushing resulted in shorter plants with a 34% reduction in leaf dry weight per area and a 59% reduction of soluble sugars and starch, on a dry weight basis. The sugar to starch ratio was not affected by brushing. A greater chilling tolerance in the brushed plants was demonstrated by the maintenance of a significantly higher PSII efficiency in brushed plants (42%) compared to that of the control plants (30%) after 7 days of chilling treatment, less visible damage to the leaf tissue, and a more rapid resumption of growth during 3 days of recovery as compared to control plants. During the chilling treatment levels of soluble sugars per leaf dry weight increased 15-fold in the brushed plants and 5-fold in control plants. In the present study we have demonstrated that brushing can increase chilling tolerance in tomato plants. The observed differences in chilling tolerance and concentration of soluble sugars in the leaves may indicate an involvement of soluble sugar levels in acclimation to chilling.     

Exogenous H<Subscript>2</Subscript>O<Subscript>2</Subscript> improves the chilling tolerance of manilagrass and mascarenegrass by activating the antioxidative system

The effect of foliar pretreatment by hydrogen peroxide (H₂O₂) at low concentrations of 0, 5, 10, and 15 mM on the chilling tolerance of two Zoysia cultivars, manilagrass (Zoysia matrella) and mascarenegrass (Zoysia tenuifolia), was studied. The optimal concentration for H₂O₂ pretreatment was 10 mM, as demonstrated by the lowest malondialdehyde (MDA) content and electrolyte leakage (EL) levels and higher protein content under chilling stress (7°C/2°C, day/night). Prior to initiation of chilling, exogenous 10 mM H₂O₂ significantly increased catalase (CAT), ascorbate peroxidase (APX), glutathione-dependent peroxidases (GPX), and glutathione-S-transferase (GST) activities in manilagrass, and guaiacol peroxidase (POD), APX, and glutathione reductase (GR) activities in mascarenegrass, suggesting that H₂O₂ may act as a signaling molecule, inducing protective metabolic responses against further oxidative damage due to chilling. Under further stress, optimal pretreatments alleviated the increase of H₂O₂ level and the decrease of turfgrass quality, and improved CAT, POD, APX, GR, and GPX activities, with especially significant enhancement of APX and GPX activities from the initiation to end of chilling. These antioxidative enzymes were likely the important factors for acquisition of tolerance to chilling stress in the two Zoysia cultivars. Our results showed that pretreatment with H₂O₂ at appropriate concentration may improve the tolerance of warm-season Zoysia grasses to chilling stress, and that manilagrass had better tolerance to chilling, as evaluated by lower MDA and EL, and better turfgrass quality, regardless of the pretreatment applied.     

Degradation of leaf polar lipids during chilling and post-chilling rewarming of Zea mays genotypes reflects differences in their response to chilling stress. The role of galactolipase

Degradation of leaf polar lipids [monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), sulfoquinovosyldiacylglycerol (SQDG) and phosphatidylglycerol (PG)] and chlorophyll (Chl) were studied in four Zea mays genotypes differing in chilling susceptibility following dark chilling and post-chilling rewarming at original growth conditions. Assessment of visual chilling injury symptoms during post-chilling rewarming differentiated maize inbred lines into chiling-sensitive (CS) CM7 and Co151 lines and chillingtolerant (CT) S215 and EP1 lines. Severity of chilling injury in CS and CT inbreeds were correlated with the extent of Chl and polar lipids degradation. Chilling for either 4 or 6 days followed by 4 days of rewarming caused more extensive degradation of total polar lipids content in CS than in CT lines. MGDG decreased mostly during chilling whereas DGDG dropped during rewarming only. Chl content was not affected during chilling but its large decrease, greater in CS than in CT lines, was observed upon rewarming. Extent of polar lipids breakdown in CS and CT inbreeds during chilling and post-chilling rewarming is correlated with galactolipase activity in chloroplasts (Kaniuga et al., 1998) and visual assessment of chilling injury. In view of the data it is likely that contribution of galactolipase activity induced during low-temperature stress of CS plants is an important factor responsible for thylakoid lipid degradation and development of chilling injury as postulated previously (Kaniuga 1997). It is suggested that genetically engineered reduction of galactolipase activity or elimination of the factors(s) involved in induction/stimulation of its activity during chilling might increase tolerance of CS species to chilling stress.     

Cold-induced sudden reversible lowering of in vivo chlorophyll fluorescence after saturating light pulses : a sensitive marker for chilling susceptibility

In chilling-sensitive plants (Glycine max, Saintpaulia ionantha, Saccharum officinarum) a sudden reversible drop in chlorophyll fluorescence occurs during photosynthetic induction immediately following saturating light pulses at low temperatures in the range 4 to 8°C. A comparison of two soybean cultivars of different chilling sensitivities revealed that this phenomenon, termed lowwave, indicates specific thresholds of low temperature stress. Its occurrence under controlled chilling can be regarded as a quantitative marker for screening chilling susceptibility in angiosperms.