Changes in Leaf Water Relations and Injuries in Maize Seedlings Induced by Different Chilling Conditions

The time course of injuries and electrolyte leakage as well as changes in leaf water content and relative turgidity during chilling at 5°C, 65 or 100 % relative humidity (RH) were determined for seedlings of two maize hybrids differing in chilling tolerance.
In the course of chilling of entire seedlings the statistically significant injuries and increased leakage of electrolytes occurred first, followed by significant water deficit in leaves which was independent of RH and genotype. However, during prolonged chilling high and significant correlation between the level of chilling-induced water stress and the extent of injuries was observed. Thus it has been assumed that the main factor responsible for primary chilling injuries in maize seedlings is the immediate influence of chilling temperatures on the cell membranes, and not the secondary water stress. The level of the water stress and the extent of the dehydration injuries under chilling conditions depended on the chilling sensitivity of the genotype. Differences between the examined genotypes consisted not only in delayed chilling injuries in a chilling tolerant hybrid, but also in significantly lesser effects of these injures on water relations.
However, exposure to low temperature of the aboveground part or of the roots of the seedlings only, caused first the chilling-induced marked water deficit in the leaves and next increased leakage of electrolytes. A direct effect of chilling temperature on the membranes in a situation when only a part of the seedling is being chilled, may be less harmful to the plant as a whole owing to normal functioning of the non-chilled part.     

Effects of Exposure to Short Periods of Suboptimal Temperature during Chill (5 °C) on Gas Exchange and Chlorophyll Fluorescence in Maize Seedlings (Zea Mays L.)

The effect of various periods of exposure to suboptimal temperature ('warm breaks'– WB: 14 °C for 4 h, 1 h and 0 h – control in 24 h cycles) during chilling (5 °C) of maize seedlings on the photosynthesis, the photochemical efficiency of photosystem II (Fv/Fm) and on the injuries of the cell membranes of leaves and water content in plants was compared. The measurements were conducted after 1, 3, 7 and 12 chill cycles. It was found that WB of either length distinctly diminished the chill-induced inhibition of net photosynthesis and the decrease of photochemical efficiency of PSII. The protective effect on WB on these parameters was observed shortly after completion of chilling of the plant as well as an after-effect. Daily warming up of the plants also reduced the leakage of electrolytes and diminished the water deficit of the chilled seedlings. The protective effect of WB on the measured parameters of the plants was greater in the chill-sensitive genotype than in the chill-tolerant one, especially when plants were warmed up for 4 h. The results obtained are an indication that short periods of warm weather during cold spring may diminish the injuries of the photosynthetic apparatus, as well as reduce the disturbance of water status of seedlings, contributing in this way to better condition of maize crops.     

Chilling Tolerance of 10 Maize Genotypes as Related to Chilling-induced Changes in ACC and MACC Contents

We studied chilling-induced changes of 1-aminocydopropane-1-carboxylic acid (ACC) and of 1-(malonylamino) cyclopropane-1-carboxylic acid (MACC) contents in seedlings of ten maize genotypes with different chilling tolerance. Seedlings at the third leaf stage were chilled at 5°C and at 65% RH. Immediately before and after two and five days of chilling the contents of ACC and of MACC in the third leaf were measured. Water content and – after recovery – the degree of necrotic injuries and the percentage of seedling survival were also determined.
After 2 days of chilling, the ACC content increased in all genotypes investigated. The increase was significantly higher in the sensitive genotypes than in the tolerant ones. There was a significant correlation between ACC content and necrotic injuries of seedlings. Chilling for 5 days increased the ACC content further and the difference between the two groups of genotypes still existed.
The MACC content increased after 5 days of chilling in all genotypes investigated. The increase was greater in the tolerant genotypes than in the sensitive ones. However, the difference in MACC accumulation between the two groups of genotypes investigated was not significant, and thus no correlation between MACC accumulation and chilling susceptibility was found.
The possible causes for the increase of ACC and MACC contents under chilling conditions and the possibility of using the ACC content as an indicator of chilling tolerance in maize breeding are discussed.     

The Effect of Short Warm Breaks during Chilling on Photosynthesis and the Activity of Antioxidant Enzymes in Plants Sensitive to Chilling

The effect of short warm breaks (from 15 min to 5 h) during chilling of three chilling-sensitive species (tomato, maize and soybean) was investigated. Injuries, intensity of net photosynthesis and antioxidant enzyme activity were measured. Throughout chilling treatment, plants were warmed by transferring them during the last few hours of the light phase from chilling temperature (5 °C for tomato and maize, 2 °C for soybean) to 20 °C. After warming, seedlings were moved back to chilling conditions. Warm breaks of 5 h almost entirely prevented the appearance of injuries, as measured by changes in leakage of electrolytes and tissue water content, during 12 days of chilling. Even a 15-min warm break ensured a significant decrease in injuries in chilled maize seedlings compared to continuously chilled seedlings. Inhibition of gas exchange and fluorescence in seedlings of two maize genotypes differing in chilling resistance was, to a small extent, prevented by 1-h warm breaks, while 4-h warm breaks reduced inhibition significantly. The length of the warm break (1 or 4 h) had no influence on changes in SOD activity compared to continuously chilled plants, but warm breaks of 4 h produced a significant increase in CAT activity. The possible influence of an alternative pathway in preventing injuries is discussed.     

Effects of Exogenous Abscisic Acid on Antioxidant System in Weedy and Cultivated Rice with Different Chilling Sensitivity under Chilling Stress

Two chilling‐tolerant genotypes, that is, weedy rice WR03‐45 and cultivated rice Lijiangxintuanheigu and two chilling‐sensitive genotypes, that is, weedy rice WR03‐26 and cultivated rice Xiuzinuo were used in this study to investigate the effects of exogenous abscisic acid (ABA) on protection against chilling damage as well as on changes in physiological features. The results showed that under chilling stress the increased levels of superoxide radical (), hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) in WR03‐45 and Lijiangxintuanheigu were lower than those in WR03‐26 and Xiuzinuo. Activities of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR)) and non‐enzymatic antioxidants (ascorbate acid (AsA) and reduced glutathione (GSH)) were enhanced in WR03‐45 and Lijiangxintuanheigu, whereas they were decreased significantly in WR03‐26 and Xiuzinuo. Application of exogenous ABA reduced the chilling damage in the four genotypes. The pre‐treatment with ABA decreased the levels of , H₂O₂ and MDA caused by chilling stress in the four genotypes through increasing the activities of SOD, CAT, APX, GR and the contents of AsA and GSH in the four genotypes under chilling stress. Moreover, pre‐treatment with Fluridone, the ABA biosynthesis inhibitor, prohibited the effects of ABA through enhancing the oxidative damages and suppressing the antioxidant defence systems under chilling stress. The results indicate the mechanism for rice with chilling tolerance is to enhance the capacity of antioxidant defence systems under chilling stress. Furthermore, ABA plays important roles in the tolerance of rice against chilling stress for it could induce the capacity of whole antioxidant defence systems including enzymatic and non‐enzymatic constitutions under chilling stress.     

The Effect of Short Warm Breaks during Chilling on Water Status, Intensity of Photosynthesis of Maize Seedlings and Final Grain Yield

The effects of short-term exposure of seedlings to suboptimal temperature (14 °C for 1 or 4 h in 24 h cycles) during chilling (5 °C for 12 days) on the water status and intensity of photosynthesis of tolerant (TG) and chilling-sensitive (SG) maize genotypes were studied. Daily warming for 1 or 4 h resulted in a decrease in the hydration of the seedlings to 31.1 % and 61.5 % (SG) and 14.8 % (TG) and 39.1 % (SG), respectively, in comparison with the continuously chilled control. During warming for 4 h, both genotypes absorbed water from soil in amounts that partly compensated for its loss through transpiration, after the plants had been moved to the lower temperature. A protective effect of shorter warming (1 h) on the hydration of the seedlings was a result of a strong, stomatal limitation of transpiration during the initial days of chilling. Warming for 1 or 4 h also increased the ability of TG stomata to close in reaction to water deficit in chilling conditions. The effect of increased temperature delayed the decrease of P_N in leaves and limited RGR inhibition of the seedling mass caused by chilling. Daily warming of plants at the seedling phase (14 and 20 °C for 1 or 4 h) reduced the unfavourable effect of chilling (5 °C for a period of 8 days) on the final yield, the filling of caryopses and their number in a cob after growth in natural conditions.     

Abscisic Acid May Alter the Salicylic Acid‐Related Abiotic Stress Response in Maize

The effect of 0, 0.05 or 0.1 mm abscisic acid treatment on chilling tolerance and salicylic acid‐related responses was investigated in young maize seedlings (Zea mays L., hybrid Norma). Although the pre‐treatment of maize seedlings with abscisic acid slightly decreased the chlorophyll content, it also reduced the level of chilling injury caused by 6 days of cold treatment at 5 °C. Under normal growth conditions, increased levels of bound salicylic acid and of bound ortho‐hydroxycinnamic acid were observed in the leaves during abscisic acid treatment. In the roots, abscisic acid did not affect the free and bound salicylic acid levels, but increased the amount of free and bound ortho‐hydroxycinnamic acid. The activity of glutathione‐S‐transferase increased on the 3rd day of abscisic acid treatment, whereas it did not change when followed by cold stress, compared with the control leaves. In the roots, the activities of glutathione reductase, glutathione‐S‐transferase and ascorbate peroxidase increased during the abscisic acid treatment, and those of glutathione‐S‐transferase and ascorbate peroxidase were also stimulated when abscisic acid pre‐treatment was followed by cold stress, compared with the control roots. Our results suggest that an overlap may exist between the abscisic acid‐induced cold acclimation and the salicylic acid‐related stress response.     

Improved frost tolerance and winter hardiness in proline overaccumulating winter wheat mutants obtained by in vitro-selection is associated with increased carbohydrate,soluble protein and abscisic acid (ABA) levels

In previous studies in vitro-selection of proline overaccumulating lines of winter wheat (Triticum sativum L. cv. Jo 3063) with increased frost tolerance was reported. These traits were found to be genetically stable. In the present study the improvement of frost tolerance (winter hardiness) under field conditions is confirmed for F₇ progenies of the mutants. Moreover, the mutants accumulated higher levels of glucose and fructose, soluble protein and abscisic acid (ABA) in addition to proline than the wild type under cold hardening conditions in a growth chamber as well as under cold hardening field conditions. ABA and proline levels peaked when the temperature decreased, whereas carbohydrate levels increased more slowly at decreasing temperature. Soluble protein levels also increased during cold hardening, but in addition showed sharp declines during frost periods. Increased carbohydrate levels of the mutants were associated with lower osmotic potential values. The differences in carbohydrate, protein and ABA levels between the mutants and the wild type are probably due to pleiotropic effects of the mutation.     

脱落酸对低温胁迫下玉米幼苗生长和光合特性的影响

脱落酸（ABA）是作物生产中广泛应用的一种植物激素。探究ABA对低温胁迫下玉米幼苗光合特性的影响,为阐明低温胁迫下玉米幼苗叶片光合结构的防御保护机制提供理论依据。以玉米品种“久龙5号”为试验材料,采用盆栽试验,探讨了低温胁迫0、2、4、6、8d下不同浓度脱落酸对玉米幼苗生长及光合特性的影响。结果表明,低温胁迫显著抑制玉米幼苗的正常生长,导致SPAD值下降,叶片净光合速率（P_n）、蒸腾速率（T_r）、气孔导度（G_s）、PSⅡ光化学最大效率（F_v/F_m）和潜在光化学效率（F_v/F_o）水平下降,初始荧光（F_o）显著升高。外源喷施适当浓度的ABA能够显著缓解由低温胁迫导致的玉米叶片P_n、T_r和G_s下降幅度,提高F_v/F_m和F_v/F_o,降低F_o水平。由此可见,外源施加适当浓度的ABA能提高幼苗叶片的光合能力,提高PSⅡ反应中心活性,促进物质积累,提高玉米幼苗的耐冷性。同时,外源施加ABA存在一定的浓度效应,即“低促高抑”。本研究所设计浓度范围中15mg/L ABA浓度对光合抑制缓解效果最显著。     

Genome-wide association study for frost tolerance in rapeseed/canola (Brassica napus) under simulating freezing conditions

Rapeseed/canola seedlings can be easily damaged by spring frost, which can rupture the cells and kill the plant. Genetic variations for frost tolerance have known to exist within rapeseed/canola gene pool. A genome-wide association study (GWAS) was conducted using 231 diverged rapeseed/canola germplasm to find the significant markers of the freezing tolerance traits. The genotypes were obtained from 21 countries and comprised of spring, winter and semi-winter growth types. The genotypes were evaluated in plant growth chamber under simulated freezing conditions. Highly significant genotypic variation was observed for the freezing tolerance. The best three freezing tolerant germplasms (Rubin, KSU-10, and AR91004) were winter type, while the four most freezing susceptible germplasms (Polo Canada, Prota, Drakkar, and BO-63) were all spring type. No geographical or growth habit type clusters were identified by structure analysis in this mixed population. One QTL was identified that was located on chromosome A02. Six freezing/abiotic stress tolerance genes have been identified in this study.     

Changes in Hormonal Balance: A Possible Mechanism of Pre‐Sowing Chilling‐Induced Salt Tolerance in Spring Wheat

There is a lack of knowledge about factors contributing to the chilling‐induced alleviatory effects on growth of plants under salt stress. Thus, the primary objective of the study was to determine whether chilling‐induced changes in endogenous hormones, ionic partitioning within shoots and roots and/or gaseous exchange characteristics is involved in salt tolerance of two genetically diverses of wheat crops. For this purpose, the seeds of two spring wheat (Triticum aestivum) cultivars, MH‐97 (salt intolerant) and Inqlab‐91 (salt tolerant) were chilled at 3°C for 2 weeks. The chilled, hydroprimed and non‐primed (control) seeds of the two wheat cultivars were sown in both Petri dishes in a growth room and in the field after treatment with 15 dS m^?1 NaCl salinity. Chilling was very effective in increasing germination rate and subsequent growth when compared with hydropriming and control under salt stress. Results from field experiments clearly indicated the efficacy of chilling over hydropriming in improving shoot dry biomass and grain yield in either cultivar, particularly under salt stress. This increase in growth and yield was related to increased net photosynthetic rate, greater potential to uptake and accumulate the beneficial mineral elements (K⁺ and Ca²⁺) in the roots and reduced uptake and accumulation of toxic mineral element (Na⁺) in the shoots of both wheat cultivars when grown under salt stress. Salt‐stressed plants of both wheat cultivars raised from chilled seed had greater concentrations of indoleacetic acid, abscisic acid, salicylic acid and spermine when compared with hydropriming and control. Therefore, induction of salt tolerance by pre‐sowing chilling treatment in wheat could be attributed to its beneficial effects on ionic homeostasis and hormonal balance. The results presented are also helpful to understand the chilling‐induced cross adaptation of plants in natural environments. Moreover, efficacy of pre‐sowing chilling treatment over hydropriming suggested its commercial utilization as a low risk priming treatment for better wheat crop production under stressful environments.     

玉米自交系苗期耐冷性鉴定

采用低温下发芽鉴定,田间分期播种和人工低温处理完整植株等方法,研究了8个玉米自交系苗期耐冷性反应.结果表明:玉米自交系的耐冷性可以分为强、中、弱等类型.玉米自交系的主要耐冷特征有耐冷性总指数、发芽临界温度、早播幼苗地上部分干重、早播叶片束缚水含量、叶片膜透性相对电导值达50%的时间、叶片超弱发光强度达1/2Max的时间和植株干枯率达50%的时间等指标.常温下游离脯氨酸含量与耐冷性呈正相关,低温下脯氨酸积累量与耐冷性呈负相关.     

Activation of Antioxidant System by KCl Improves the Chilling Tolerance in Hybrid Maize

As maize is a chilling-sensitive crop, low temperatures during the early stages of development can be injurious to crop growth and development. Prime mechanism behind chilling-induced damage is oxidative stress. This study was undertaken to improve the chilling tolerance in hybrid maize by seed priming with KCl. For priming, seeds of the maize hybrid Hycorn 8288 were soaked in 50, 100 and 150 mg l⁻¹ aerated solution of KCl for 24 h and then re-dried close to original weight. Primed and untreated seeds were sown at 27 °C (optimal temperature) and at 15 °C (chilling stress) under controlled conditions. Seed priming improved the performance of maize under both normal and stress conditions. It was found that the chilling tolerance in maize is well associated with the enhanced capacity of the anti-oxidative system. Priming with KCl significantly improved the chilling tolerance mainly by the activation of antioxidants including catalase, superoxide dismutase and ascorbate peroxidase enzymes. KCl treatments also improved the germination rate and time, root and shoot length, and fresh and dry weights of seedlings compared with control. Soluble sugars and α-amylase activity determined as general metabolic indicators of stress were also improved by seed priming with KCl. Other possible bases of chilling tolerance in maize included maintenance of high tissue water contents, reduced electrolyte leakage and carbohydrate metabolism. Seed treatment with 100 mg l⁻¹ KCl was the best treatment to improve the performance of hybrid maize both under normal and chilling stress conditions.     

Response of maize inbred lines to a defoliation treatment inducing tolerance to cold at germination

Defoliation during maize (Zea mays L.) kernel development has been observed to induce tolerance to cold of germinating seeds in responsive genotypes. The objectives of this study were to evaluate the response to defoliation of immature embryo and mature seed germinability at cold and to verify if the response was influenced by the developmental stage at which the treatment was applied. In three environments, six inbred lines (B73, IABO78, Lo1016, Lo964, Mo17, Os420) were defoliated (D) approximately 20 days after pollination (DAP) or not defoliated (ND). Immature embryos were excised three days after defoliation and germinated in vitro at 9 or 25 ^∘C. At maturation, kernel germination was tested at the same temperatures. Defoliation improved cold tolerance and mean time to germination (MTG) at 9 ^∘C of both embryos and kernels of Lo1016. To study the effect of kernel developmental stage on response to defoliation, plants of B73, Lo1016 and Lo964 were defoliated at 15, 18, 21, 24, 27, 30, 33, 36, and 39 DAP, or not defoliated. At the same DAP, immature grains were analyzed for dry weight, water and abscisic acid (ABA) content. In Lo1016, low amounts of kernel ABA were detected at all stages, while in Lo964 and B73 ABA increased during development. Lo1016 mature kernels showed an improvement of cold tolerance due to defoliation at all times, while the other genotypes did not. In conclusion inbred lines showed variability for mature seed and immature embryo tolerance to cold at germination and for the ability to acquire tolerance after defoliation.     

玉米耐寒性鉴定研究进展

玉米是对低温逆境较为敏感的重要的禾谷类作物。早春低温冷害是影响玉米产量水平提高的重要因素之一。选育和创造耐寒玉米种质是减少此种危害的有效途径。耐寒性鉴定则是耐寒育种研究的基础。综述了国内外玉米耐寒性的鉴定方法和评价指标,分析了目前玉米耐寒性鉴定存在的问题,探讨了未来研究的方向,为深入开展玉米耐寒性研究提供参考。     

Glycinebetaine Improves Chilling Tolerance in Hybrid Maize

Plant growth and development is hampered by various environmental stresses including chilling. We investigated the possibility of improving chilling tolerance in hybrid maize by glycinebetaine (GB) seed treatments. Maize hybrid (Hycorn 8288) seeds were soaked in 50, 100 and 150 mg l^?1 (p.p.m.) aerated solution of GB for 24 h and were dried back. Treated and untreated seeds were sown at 27 °C (optimal temperature) and at 15 °C (chilling stress) under controlled conditions. Germination and seedling growth was significantly hindered under chilling stress. Moreover, chilling stress significantly reduced the starch metabolism and relative water contents (RWC), and increased the membrane electrolyte leakage. However, activities of antioxidants (catalase, superoxide dismutase and ascorbate peroxidase) were increased under stress conditions. Seed treatments with GB improved the germination rate, root and shoot length, seedling fresh and dry weights, leaf and root scores, RWC, soluble sugars, α‐amylase activity and antioxidants significantly compared with untreated seeds under optimal and stress conditions. Induction of chilling tolerance was attributed to maintenance of high tissue water contents, reduced membrane electrolyte leakage, and higher antioxidant activities and carbohydrate metabolism. Seed treatment with 100 mg l^?1 GB was the best treatment for improving the performance of hybrid maize under normal and stress conditions compared with control and other levels used.     

Cold stress tolerance of soybeans during flowering: QTL mapping and efficient selection strategies under controlled conditions

Breeding soybeans for higher latitudes requires cultivars with an increased chilling stress tolerance, especially when flowering occurs. Phenotyping in climate chambers to select for this trait is labour‐intensive and requires an optimal allocation of resources due to limited space. We screened a diversity panel of 35 early maturity cultivars and a biparental population of 103 RILs for their cold stress tolerance at flowering stage. Pod number under control and stress conditions is highly heritable and showed only a weak correlation between the two treatments. Based on different testing scenarios, we could show that testing more genotypes with less replicates yields much higher responses to selection and hence should be pursued in such climate‐controlled experiments. We identified quantitative trait loci (QTL) for pod number under both conditions (chromosomes 7 and 13) and a cold tolerance‐specific QTL (chromosome 11). Furthermore, we performed genomic predictions using different test set scenarios and prediction models, showing that genomic prediction is a promising tool to select for cold stress tolerance, particularly if known QTL can be used as fixed effects in the model.     

Physiological and Biochemical Responses of Hexaploid and Tetraploid Wheat to Drought Stress

An experiment was conducted to investigate the physiological and biochemical responses of two hexaploids viz., C 306 (water stress tolerant) and Hira (water stress susceptible), and two tetraploids, HW 24 (Triticum dicoccum) and A 9‐30‐1 (Triticum durum) wheat genotypes to water stress under pot culture condition. Water stress was imposed for a uniform period of 10 days at 50, 60 and 70 days after sowing (DAS) and observations were recorded at 60, 70 and 80 DAS. Total dry matter and plant height were recorded at harvest. Water stress caused a decline in relative water content (RWC), chlorophyll and carotenoid content, membrane stability and nitrate reductase activity and increased accumulation of proline at all stages and abscisic acid (ABA) at 80 DAS in all the genotypes. Both the tetraploids showed a lower reduction in RWC and highest ABA accumulation under water stress. Among the hexaploids Hira showed the most decline in RWC and the lowest ABA accumulation. The tetraploids also showed comparatively higher carotenoid content and membrane stability, closely followed by C 306, while Hira showed the minimum response under water stress. Nitrate reductase activity and chlorophyll content under irrigated conditions were highest in Hira but under water stress the lowest per cent decline was observed in C 306, followed by HW 24, A 9‐30‐1, and Hira. Proline accumulation under water stress conditions was highest in hexaploids C 306 and Hira and lowest in tetraploids HW 24 and A 9‐30‐1. Tetraploids HW 24, followed by A 9‐30‐1 maintained higher plant height and total dry matter (TDM) under water stress and also showed a lower per cent decline under stress than hexaploids C 306 and Hira. From the results it is clear that proline accumulation did not contribute to better drought tolerance of tetraploids than hexaploids. It is also apparent that water stress tolerance is the result of the cumulative action of various physiological processes, and all the parameters/processes may not be positively associated with the drought tolerance of a particular tolerant genotype.     

Contents of Total Phenolics and Ferulic Acid, and PAL Activity during Water Potential Changes in Leaves of Maize Single-Cross Hybrids of Different Drought Tolerance

The aim of this study was to determine whether the resistance and/or sensitivity to drought stress, can be attributed to the level of phenolic compounds in the leaves of maize genotypes. The experiments were carried out on seedlings of three maize genotypes characterized by different levels of drought resistance. Experiments with three periods of drought were conducted (8, 11 and 14 days), to obtain plants with different levels of water potential in leaves, which induced changes in the total phenolic content and ferulic acid, and l ‐phenylalanine ammonia‐lyase (PAL) activity. Only for the drought‐resistant genotype Tina, was the low water potential found to be correlated with the high level of the total phenolic content and ferulic acid, which is the main source of blue fluorescence emissions. Moreover, only for Tina were the highest intensities of blue fluorescence emission correlated with the low water potential in leaves. The phenolic compounds present in leaf tissues can protect the deeper situated mesophyll, by absorbing light reaching the leaf and transforming it into a blue fluorescence. Phenolic compounds can, in this way, function as photoprotectors limiting the excitation of chlorophyll during conditions of water deficit in leaves.