Seasonal regulation of a 24-kDa protein from red-osier dogwood (Cornus sericea) xylem

Previously, we showed that an apparent cell wall-plasma membrane interaction in xylem ray parenchyma differed between cold acclimated and non-acclimated red-osier dogwood (Cornus sericea L.) (Ristic and Ashworth 1994). For the present study, a calcium chloride extraction method was used to identify cell-wall-associated xylem proteins that accumulated during periods of cold acclimation. A 24-kDa protein represented the predominant protein in both total protein and CaCl2 extracts during cold acclimation of field-grown plants. Two-dimensional gel electrophoresis separated the 24-kDa protein into four basic isoforms. The most abundant and basic isoform had a high glycine content. In-gel digestion of this basic 24-kDa isoform generated three partial peptide fragments, of which one exhibited homology to the dehydrin protein family. An anti-dehydrin polyclonal antibody cross-reacted with the 24-kDa protein, providing further evidence that this protein is related to dehydrins. The 24-kDa protein began to accumulate in late August, reached a maximum in midwinter, declined during the spring months and was absent in early summer.     

Ultrastructural and Extracellular Protein Changes in Cell Suspension Cultures of Populus euphratica Associated with Low Temperature-induced Cold Acclimation

Populus euphratica Olive is the only tree species that can grow in the saline land and also survive cold winters in northwest China, and it plays a very important role in stabilizing the vulnerable ecosystem there. A cell suspension culture was initiated from callus derived from plantlets of Populus euphratica. Cold acclimation was induced (LT50 of-17.5 ℃) in cell suspension at 4-5 ℃ in the dark for 30 days and the freezing tolerance increased from LT50 of-12.5 ℃ in nonacclimated cells to LT50 of-17.5 ℃ in cold-acclimated cells. Microvacuolation, cytoplasmic augmentation and accumulation of starch granules were observed in cells that were cold-acclimated by exposure to low temperatures. Several qualitative and quantitative changes in proteins were noted during cold acclimation. Antibodies to carrot extracellular (apoplastic) 36 kD antifreeze protein did not cross react on immunoelectroblots with extracellular proteins in cell suspension culture medium of Populus euphratica, indicating no common epitopes in the carrot 36 kD antifreeze protein and P euphratica extracellular proteins. The relationship of these changes to cold acclimation in Populus euphratica cell cultures was discussed.     

Cold acclimation induced accumulation of phenolic compounds and freezing tolerance in Ammopiptanthus mongolicus

Ammopiptanthus mongolicus, the only freezing tolerant evergreen broad-leaved shrub, local species of the Alashan desert, northwest sand area of China, can survive -30°C or even lower temperature in winter. In the present study, the secondary products phenolics in A. mongolicus cotyledons were determined to study the effects of phenolics on cold tolerance. Cytochemical localization of phenolics in cotyledon cells was observed by electron microscopy and the content of phenolic compounds was assayed by spec-trophotometric measurement. The results showed that the freezing tolerance of A. mongolicus seedlings increased after acclimation at 2-6°C for 14 days, which accompanied the increase of the content of phenolic compounds in cotyledons. Cytochemical observation showed that phenolic deposits were mainly localized in vacuoles and in close proximity to tonoplast, and also in the cytoplasm. The amount and the size of phenolics droplets increased obviously in cytoplasm and vacuoles after cold acclimation, predominantly ag-gregated along membranes of vacuoles and tonoplast. No phenolic deposits were found in cell walls. As hydrogen-or elec-tron-donating agents, phenolics may protect plant cells against reactive oxygen species formed during chilling or freezing stress and improve the freezing tolerance of cold-acclimated A. mongolicus seedlings.     

干旱胁迫下外源ABA对观赏海棠叶片可溶性蛋白和脱水素积累的影响

以抗旱性较强的观赏海棠品种‘王族’和抗旱性较弱的品种‘红丽’幼树为试材,采用"喷施ABA+干旱处理"和"喷水+干旱处理(对照)"2个处理组,测试不同干旱条件下ABA处理的观赏海棠叶片相对含水量(RWC)、相对电导率(REC)、可溶性蛋白含量(SPC)、可溶性蛋白和脱水素表达情况。结果显示:与对照相比,ABA处理的‘王族’叶片RWC显著升高,REC和SPC显著降低,而‘红丽’的相应指标变化则相反。SDS-PAGE分析表明:ABA处理分别引起了干旱处理过程中‘王族’叶片14.8,18.0,46.2,50.9,78.1,145.2ku和‘红丽’叶片14.4,18.0,32.8,46.2,89.6,141.0ku可溶性蛋白的变化。免疫蛋白印迹法测定结果表明:干旱处理分别诱导‘王族’叶片14.8,51.0ku和‘红丽’叶片14.4,32.8ku脱水素的积累;ABA处理显著地的诱导2品种叶片干旱处理开始时脱水素的积累,并降低干旱处理7天后‘王族’叶片14.8,51.0ku和‘红丽’叶片14.4ku的积累。ABA能够诱导正常水分条件下观赏海棠叶片脱水素的表达,但当植株处于干旱胁迫时,ABA抑制其叶片中被诱导的脱水素的积累,并且此抑制作用与品种间抗旱性呈正相关性。     

Vitality characterization of stressed trees based on non-destructive and real-time monitoring of stem water content

The detection of stem water content is necessary as it is an important indicator for measuring woody plant vitality. However, the relationship between stem water content, determined by non-destructive, real-time, and long-term monitoring, and woody plant vitality remains undefined. In this study, the response of woody plant vitality to stem water content under different stress (freeze–thaw, pest, or drought) was analysed by mining the dynamic characteristics of the stem water content in different woody plants at the temporal scales of year, month, and day. Compared with unstressed trees, stressed trees had contrasting diurnal patterns. The stem water content in Populus koreana Rehd. during the freeze period was much lower than that during the thaw period, and opposite diurnal variation trends were observed during the freeze and thaw periods. The stem water content in infected Lagerstroemia indica was lower than that in uninfected L. indica, and the amplitude of the diurnal variation curve was lower in infected than in uninfected L. indica. Under drought stress, the more severe the water shortage, the lower the stem water content in Malus micromalus. When it was below a certain threshold, the diurnal variation trend was opposite to that without water shortage. In conclusion, stem water content dynamics can be used to evaluate the cold, pest, and drought response of trees, which could monitor tree health and guide forest assessment.     

Effects of "near-lethal" stress on bud dormancy and stem cold hardiness in red-osier dogwood

We studied the effects of "near-lethal" (NL, 47 degrees C for 1 h) heat stress, applied to intact shoots of red-osier dogwood (Cornus sericea L.) during early (October), deep (November) or late (December) dormancy, on bud dormancy release and development of stem tissue cold hardiness under natural conditions and at a constant temperature of 0 or 23 degrees C in the dark. The NL heat-stress treatment overcame bud dormancy when applied during the early and late stages of dormancy. During October and December, all plants in the 23 degrees C + dark post-stress environment broke bud within 35 and 12 days, respectively, whereas the corresponding values for days to bud break in the control plants were more than 150 and 110 days, respectively. Application of NL heat stress during deep dormancy caused only slightly earlier bud break compared to the control plants. In the 0 degrees C + dark post-stress environment, all NL heat-treated plants died within 9 weeks. Under natural post-stress conditions, bud break in plants receiving NL heat stress during early and deep dormancy occurred at the same time as in control plants, whereas bud break of plants receiving NL heat stress during late dormancy occurred 55 days earlier than in control plants. Under both natural and 23 degrees C + dark post-stress conditions, cold hardiness of plants receiving NL heat stress during early dormancy was similar to that of controls. Application of NL heat stress during deep dormancy hastened the rate of deacclimation under the 23 degrees C + dark post-stress conditions but had no effect on deacclimation under natural post-stress conditions. Application of NL heat stress during late dormancy enhanced deacclimation of plants in both the 23 degrees C + dark and natural post-stress environments.     

Preliminary antiproliferative effects of some species of Terminalia, Combretum and Pteleopsis collected in Tanzania on some human cancer cell lines

Methanolic extracts (25 microug/ml) of species belonging to the genera of Combretum, Terminalia and Pteleopsis, collected during a field expedition in Tanzania in 1999, were screened for their antiproliferative and cytotoxic effects against three human cancer cell lines (HeLa, cervical carcinoma; T 24, bladder carcinoma; and MCF 7, breast carcinoma). A leaf extract of Combretum fragrans and a fruit extract of C. zeyheri gave the strongest antiproliferative and cytotoxic effects of all the twenty-four extracts screened in this investigation. In contrast to the highly powerful leaf extract of C. fragrans, the root extract of this species gave no cytotoxic effects against the investigated cancer cell lines at a concentration of 25 microg/ml. The other investigated species of Combretum and Terminalia differed greatly in their cytotoxic potential. Root extracts of Terminalia sambesiaca and T. sericea gave the strongest cytotoxic effects of the five species of Terminalia used in this study. Eight of the twenty-four investigated plant extracts showed pronounced cytotoxic effects (<30% proliferation compared to the control) against the T 24 bladder cancer cells, seven against the HeLa cells and four against the MCF 7 cells.     

Autumn freeze testing of one‐year reciprocal families of Pinus sylvestris (L.)

Reciprocal families of Scots pine, Pinus sylvestris (L.), from a Swedish seed orchard were artificially freeze tested to ‐10°C at the end of the first growth period. The degree of freezing damage was used as an indication of the cold acclimatization achieved at the time of freeze testing. Both one‐year cold acclimatization and one‐year height were mainly additively inherited. Specific combining ability, reciprocal and maternal effects were small. On the family level, freezing damage was non‐significantly correlated with field survival and field height after ten years. One‐year height was positively correlated with 10‐year field height and negatively correlated with field survival on the family level. Within families, plants from early germinated seeds cold acclimated earlier and were higher at the end of the first growth period than plants from lately germinated seeds. The results indicate that conclusions made from first‐year cold acclimatization studies can be influenced by variation in the rate of seed germination and seedling/germinant development.     

Effects of atmospheric humidity and acclimation temperature on the temperature response of photosynthesis in young Larix decidua Mill

Larch (Larix decidua Mill.) seedlings of a low altitude (600 m) Austrian provenance were raised outdoors and acclimated in chambers for 14 to 24 days during August and September at either 8 degrees C and an atmospheric saturation vapor pressure deficit (DeltaW) of 2.5 Pa kPa(-1), or 24 degrees C and a DeltaW of 6.2 Pa kPa(-1). Subsequently, their rates of photosynthesis, dark respiration and transpiration were measured at temperatures between 5 and 30 degrees C with DeltaW either maintained below 10 Pa kPa(-1) or allowed to increase with temperature up to 38 Pa kPa(-1). Below 15 degrees C the photosynthetic rate of cold-acclimated plants was higher, but above 15 degrees C it was lower, than that of warm-acclimated plants. Temperature acclimation caused a greater shift in the temperature optimum for photosynthesis when DeltaW was kept small than when it was allowed to increase with temperature. When DeltaW was kept small, leaf conductance of cold-acclimated plants, unlike that of warm-acclimated plants, did not increase with temperature above 15 degrees C. When DeltaW increased with temperature, leaf conductance of cold-acclimated plants decreased more rapidly with temperature than that of warm-acclimated plants. Low temperature acclimation increased the rate of photosynthesis below 15 degrees C without affecting leaf conductance, which indicates that there was an adaptation in leaf internal processes. Further evidence of a metabolic adaptation to acclimation temperature is that dark respiration of cold-acclimated plants was twice that of warm-acclimated plants at all temperatures.     

江门市园林植物寒害浅析

2008年1月下旬江门市受到了连续低温天气的影响,园林植物发生了较严重的寒害。作者对植物受害情况进行了系统地研究,结合近年来江门市园林植物引种的情况,分析发生寒害的原因,探讨防止园林植物发生寒害的对策,并有针对性地提出了灾后复产方案,对于搞好城市园林绿化建设和管理具有一定的指导意义。     

Excitation energy partitioning and quenching during cold acclimation in Scots pine

We studied the influence of two irradiances on cold acclimation and recovery of photosynthesis in Scots pine (Pinus sylvestris L.) seedlings to assess mechanisms for quenching the excess energy captured by the photosynthetic apparatus. A shift in temperature from 20 to 5 degrees C caused a greater decrease in photosynthetic activity, measured by chlorophyll fluorescence and oxygen evolution, in plants exposed to moderate light (350 micromol m(-2) s(-1)) than in shaded plants (50 micromol m(-2) s(-1)). In response to the temperature shift, maximal photochemical efficiency of photosystem II (PSII), measured as the ratio of variable to maximal chlorophyll fluorescence (Fv/Fm) of dark-adapted samples, decreased to 70% in exposed seedlings, whereas shaded seedlings maintained Fv/Fm close to initial values. After a further temperature decrease to -5 degrees C, only 8% of initial Fv/Fm remained in exposed plants, whereas shaded plants retained 40% of initial Fv/Fm. Seven days after transfer from -5 to 20 degrees C, recovery of photochemical efficiency was more complete in the shaded plants than in the exposed plants (87 and 65% of the initial Fv/Fm value, respectively). In response to cold stress, the estimated functional absorption cross section per remaining PSII reaction center increased at both irradiances, but the increase was more pronounced in exposed seedlings. Estimates of energy partitioning in the needles showed a much higher dissipative component in the exposed seedlings at low temperatures, pointing to stronger development of non-photochemical quenching at moderate irradiances. The de-epoxidation state of the xanthophyll cycle pigments increased in exposed seedlings at 5 degrees C, contributing to the quenching capacity, whereas significant de-epoxidation in the shaded plants was observed only when temperatures decreased to -5 degrees C. Thermoluminescence (TL) measurements of PSII revealed that charge recombinations between the second oxidation state of Mn-cluster S2 and the semireduced secondary electron acceptor quinone Q(B)- (S2Q(B)-) were shifted to lower temperatures in cold-acclimated seedlings compared with control seedlings and this effect depended on irradiance. Concomitant with this, cold-acclimated seedlings demonstrated a significant shift in the S2 recombination with primary acceptor Q(A)- (S2Q(A)-) characteristic TL emission peak to higher temperatures, thus narrowing the redox potential gap between S2Q(B)- and S2Q(A)-, which might result in increased probability for non-radiative radical pair recombination between the PSII reaction center chlorophyll a (P680+) and Q(A)- (P680+)Q(A)-) (reaction center quenching) in cold-acclimated seedlings. In Scots pine seedlings, mechanisms of quenching excess light energy in winter therefore involve light-dependent regulation of reaction center content and both reaction center-based and antenna-based quenching of excess light energy, enabling them to withstand high excitation pressure under northern winter conditions.     

垂榆冻害调查及其影响因素的分析

对长春市4个不同地点(路段)栽植垂榆的受冻害状况进行了调查与分析,结果表明:4个调查地点垂榆的冻害率分别为67.0%、31.0%、72.0%和92.0%,平均值为65.5%;冻害指数分别为31.0、11.4、49.2和64.9,平均值为39.1。2009~2010年冬春季节持续低温的累积作用是导致发生冻害的主要因素,老龄植株生长后期的嫁接不亲和作用与砧木个体抗寒能力的遗传差异亦是影响因素之一。     

杉木苗期抗寒性的遗传变异和家系选择

近20年来,我国杉木遗传改良工作进展很快,但杉木育种目标多局限于速生丰产,过于单一。目前杉木的栽培区域不断从中心产区向边缘产区扩展延伸,出现了对不良气候、土壤条件的适应性问题。例如,1987年冬浙江各地苗圃遭受早霜冻害十分严重,使上百万株杉苗     

Comparison of three cold hardiness tests for conifer seedlings

Greenhouse-cultured, container-grown ponderosa pine (Pinus ponderosa var. scopulorum Engelm.), interior Douglas-fir (Pseudotsuga menziesii var. glauca (Beissn.) Franco), and Engelmann spruce (Picea engelmannii (Parry) Engelm.) were cold acclimated and deacclimated in growth chambers over 19 weeks. Cold hardiness was measured weekly by a whole-plant freeze test and by two quick tissue tests: freeze-induced electrolyte leakage of needles, and differential thermal analysis of buds. The whole-plant freeze test provided results in 7 days, and indicated differences in cold hardiness among stems, buds, and needles. Although the whole-plant freeze test could accurately measure cold hardiness, it was not precise, and it required destructive sampling. Results from freeze-induced electrolyte leakage and differential thermal analysis were available in 2 days and 1 hour, respectively. The freeze-induced electrolyte leakage test was a precise, sensitive and objective predictor of changes or differences in tissue cold hardiness. To determine actual cold hardiness, results could be calibrated to the response of the same tissue in the whole-plant freeze test. The speed and objectivity of differential thermal analysis made this test useful for rapid, general assessment of cold hardiness status, but calibration was difficult, and precision varied.     

低温胁迫下梁山慈竹体细胞突变体植株叶绿素荧光响应

以冷驯化的两种梁山慈竹(Dendrocalamus farinosus)体细胞突变体植株及梁山慈竹实生植株为材料,研究了梯度降温处理(依次为4、0、?5、?10和?15℃)对其叶绿素荧光参数的影响。结果表明,实生植株在?5℃胁迫下死亡,而突变体植株No.101-1b和No.101-1c直到?15℃胁迫下才出现死亡。低温胁迫后,突变体植株和实生植株的最大光化学效率(Fv/Fm)、光系统Ⅱ有效光量子产量(YⅡ)和光化学淬灭系数(qP)均有降低,下降幅度排序为实生植株>No.101-1b>No.101-1c。非光化学猝灭系数(NPQ)显著高于胁迫前,以No.101-1c表现最为突出。品系和温度2个因素及两者交互作用对叶绿素荧光的影响均达到显著水平。总结得出,突变体植株No.101-1c具有较高的耐寒性,本研究可为耐寒梁山慈竹体细胞突变体植株的筛选提供科学依据。     

Adaptive significance of C partitioning and regulation of specific leaf area in Betula pendula

Carbon allocation and regulation of specific leaf area (sigma) define key processes underlying the adaptation of plants to varying habitats. In this study, the general principles governing adaptation and a dynamic optimality model of plant adaptation are reviewed. The central new elements of this model are: (i) differential root carbon costs for maintaining a defined nutrient status; (ii) a simple formula for optimal sigma at steady-state as a function of nitrogen (N) status and irradiance; and (iii) generic rules for the time propagation of adapting traits. The model was applied to a large data set compiled by Ingestad et al. (1995) and McDonald et al. (1986a, 1986b) for birch seedlings (Betula pendula Roth) during stationary logarithmic growth and during transient changes in response to a range of irradiances and nutrient supply rates. In the stationary case, large variations in the fraction of leaf dry mass to total dry mass (f(L)), sigma and N concentration were simulated with high accuracy. The independently calibrated model described the temporal response of seedlings following a sharp decrease in N supply, which includes phenomena such as the temporary C accumulation in leaves and damped oscillations in N concentration. Dynamics in sigma were more sensitive to variation in light than in N supply. Nevertheless, adaptive adjustments in f(L), sigma and N concentration were strongly coupled, underlining the relevance of a whole-plant perspective when modeling plant growth and regulation. The high coincidence between model calculations and measured values supports the notion that plant acclimation can be both understood and predicted as a growth-optimizing mechanism.     

Induction of tolerance to desiccation and cryopreservation in silver maple (Acer saccharinum) embryonic axes

Twenty percent of of the world's flowering plants produce recalcitrant seeds (i.e., seeds that cannot withstand drying or freezing). We investigated whether the embryonic axis from the normally recalcitrant seeds of silver maple (Acer saccharinum L.) can be made tolerant to desiccation (10% water content) and low temperature (-196 degrees C, cryopreservation) by pretreatment with ABA or the compound tetcyclacis, which enhances endogenous ABA concentrations. Pretreatment of axes with both ABA and tetcyclacis increased germination after desiccation and freezing to 55% from a control value of zero. Pretreatment of axes with ABA and tetcyclacis increased the ABA content of the axes, as measured by enzyme-linked immunoassay, and stimulated the synthesis of storage and dehydrin-like proteins, believed to have a role in the desiccation tolerance of orthodox seeds.     

Fall fertilization of Pinus resinosa seedlings: nutrient uptake, cold hardiness, and morphological development

? Fall fertilization may increase plant nutrient reserves, yet associated impacts on seedling cold hardiness are relatively unexplored.

? Bareroot red pine (Pinus resinosa Ait.) seedlings in north-central Minnesota, USA were fall fertilized at the end of the first growing season with ammonium nitrate (NH₄NO₃) at 0, 11, 22, 44, or 89 kg N ha^?1. Seedling morphology and cold hardiness [assessed by freeze induced electrolyte leakage (FIEL)] were evaluated six weeks after fertilization and following the second growing season.

? Seedling height and number of needle primordia increased with fertilizer rate for both sampling years. Seedlings fertilized with 44 and 89 kg N ha^?1 attained target height (15 cm) after the second growing season. Shoot and root N concentration increased after the first growing season in fall fertilized seedlings compared to controls. Fall fertilized seedlings had lower FIEL (i.e., increased cold hardiness) compared to controls when tested at ?40 °C after the first growing season, but no significant differences in FIEL of control and fertilized seedlings were observed after the second growing season.

? Results suggest that fall fertilization of red pine seedlings can help render desired target height in the nursery, while maintaining or increasing cold hardiness levels.

     

Allocation of 14C assimilated in late spring to tissue and biochemical stem components of cork oak (Quercus suber L.) over the seasons

Carbon distribution in the stem of 2-year-old cork oak plants was studied by (14)CO(2) pulse labeling in late spring in order to trace the allocation of photoassimilates to tissue and biochemical stem components of cork oak. The fate of (14)C photoassimilated carbon was followed during two periods: the first 72 h (short-term study) and the first 52 weeks (long-term study) after the (14)CO(2) photosynthetic assimilation. The results showed that (14)C allocation to stem tissues was dependent on the time passed since photoassimilation and on the season of the year. In the first 3 h all (14)C was found in the polar extractives. After 3 h, it started to be allocated to other stem fractions. In 1 day, (14)C was allocated mostly to vascular cambium and, to a lesser extent, to primary phloem; no presence of (14)C was recorded for the periderm. However, translocation of (14)C to phellem was observed from 1 week after (14)CO(2) pulse labeling. The phellogen was not completely active in its entire circumference at labeling, unlike the vascular cambium; this was the tissue that accumulated most photoassimilated (14)C at the earliest sampling. The fraction of leaf-assimilated (14)C that was used by the stem peaked at 57% 1 week after (14)CO(2) plant exposure. The time lag between C photoassimilation and suberin accumulation was ～8 h, but the most active period for suberin accumulation was between 3 and 7 days. Suberin, which represented only 1.77% of the stem weight, acted as a highly effective sink for the carbon photoassimilated in late spring since suberin specific radioactivity was much higher than for any other stem component as early as only 1 week after (14)C plant labeling. This trend was maintained throughout the whole experiment. The examination of microautoradiographs taken over 1 year provided a new method for quantifying xylem growth. Using this approach it was found that there was more secondary xylem growth in late spring than in other times of the year, because the calculated average cell division time was much shorter.