低温锻炼后桑树幼苗光合作用和抗氧化酶对冷胁迫的响应

以桑树品种“秋雨”为试验材料,研究了桑树幼苗在低温锻炼、冷胁迫和常温恢复期间的光合作用和抗氧化酶活性的变化.结果表明: 12 ℃3 d低温锻炼明显提高了桑树幼苗的抗冷性.3 ℃3 d冷胁迫下,12 ℃3 d低温锻炼后的桑树幼苗叶片净光合速率(P_n)、气孔导度(G_s)和PSⅡ 最大光化学效率(F_v/F_m)明显高于对照(未经低温锻炼)处理的桑树幼苗,而且其在常温下的恢复也较对照桑树幼苗迅速.在12 ℃ 3 d低温锻炼和3 ℃ 3 d冷胁迫期间,桑树幼苗叶片脯氨酸和可溶性糖含量明显增加,而经低温锻炼的桑树幼苗叶片丙二醛(MDA)含量明显低于未经低温锻炼的桑树幼苗,经低温锻炼的桑树幼苗叶片抗坏血酸过氧化物酶(APX)活性则明显高于未经低温锻炼的桑树幼苗.说明渗透调节物质含量增加和APX活性提高在低温锻炼诱导桑树幼苗的抗冷性上发挥着重要的作用.     

Early developmental and stress responsive ESTs from mungbean, <Emphasis Type="Italic">Vigna radiata</Emphasis> (L.) Wilczek,seedlings

Although mungbean (Vigna radiata (L.) Wilczek) is commonly used as human food; the genomic resources of this species available in databases are limited. This study aims to develop expressed sequence tag (EST) resources for mungbean genes informative to early seedling development and chilling response. Two mungbean varieties that differ in disease resistance were found to also differ in their susceptibility to chilling temperatures. A total of 1,198 ESTs were obtained from one cDNA library and four PCR-select cDNA subtraction libraries; among these 523 were clustered into 136 contigs and 675 were singletons. The 811 non-redundant uniESTs were compared to GenBank using the Basic Local Alignment Search Tool (BLAST) and WU-BLAST algorithms, of these only 489 uniESTs had significant sequence homology, which may be involved in resuming the metabolic activity of seedlings, switching on photomorphogenesis, fuelling photosynthesis and/or initiating the unique developmental programs. Their encoded proteins may associate with regulatory proteins to trigger a direct stress response or participate in acclimation to environmental stressors. The uniEST platform reported will enrich the genomic resources of mungbean for functional genomic research on seedling development and chilling response of tropical crops and provide targets for improving the chilling tolerance of the tropical crops. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The after-effects of temperature and irradiance during early growth of winter oilseed rape (Brassica napus L. var. oleifera, cv. Górczański) seedlings on the progress of their cold acclimation

Experiments performed under controlled conditions showed that level of PPFD (photosynthetic photon flux density) during early seedlings growth (preceding cold acclimation at +2 °C) was not the key factor for the development of frost resistance. It did not modify the beneficial effects of prehardening (Rapacz 1997, in this issue) at moderately low (+12 °C) day temperature. Now I have shown that the increase of PPFD may replace to some extent prehardening in the development of frost resistance. It was particularly seen in non-prehardened plants, which had been grown under warm-day (+20 °C) conditions. Prehardening performed under controlled conditions, as well as seedlings growth under natural autumn conditions in the field, allowed to maintain a high net-photosynthesis rate at chilling temperatures. A net-photosynthesis rate during cold acclimation at +2 °C corresponded well with higher frost resistance. As a result, seedlings non subjected to prehardening and grown before cold acclimation under low PPFD acclimated better, if the cold treatment was applied only at nights (+20/2 °C day/night). Only under such conditions the photosynthetic rate was sufficiently high to allow plants to reach a higher level of frost resistance. All other plants acclimated better when they were exposed to the hardening temperature continuously during days and nights (+2/2 °C day/night).     

PP333对黄瓜幼苗素质及抗冷能力的影响

研究了ＰＰ３３３对黄瓜幼苗素质和抗冷能力的影响,结果显示ＰＰ３３３处理降低了幼苗高度和子叶面积,但却促进了根系生长、根冠比值和叶绿素含量显增加,明显表现壮苗效果。ＰＰ３３３能有效降低黄瓜幼苗在低温下的相对电导率,维持较高的ＳＯＤ活性和Ｖｃ、ＧＳＨ含量,因而提高了幼苗的抗冷能力。     

茄子种质资源的耐冷性鉴定

采用低温胁迫下幼苗叶片冷害指数、种子活力指数、叶片电导百分率为指标,对34份不同类型的茄子种质资源进行耐冷性鉴定.结果表明,活力指数与冷害指数、电导百分率均呈极显著负相关,电导百分率与冷害指数呈极显著正相关.通过耐冷隶属函数值和聚类分析,初步筛选出11份耐冷性较强的材料.     

Leaf cold acclimation and freezing injury in C3 and C4 grasses of the Mongolian Plateau

The scarcity of C4 plants in cool climates is usually attributed to their lower photosynthetic efficiency than C3 species at low temperatures. However, a lower freezing resistance may also decrease the competitive advantage of C4 plants by reducing canopy duration, especially in continental steppe grasslands, where a short, hot growing season is bracketed by frost events. This paper reports an experimental test of the hypothesis that cold acclimation is negligible in C4 grasses, leading to greater frost damage than in C3 species. The experiments exposed six C3 and three C4 Mongolian steppe grasses to 20 d chilling or control pre-treatments, followed by a high-light freezing event. Leaf resistance to freezing injury was independent of photosynthetic type. Three C3 species showed constitutive freezing resistance characterized by <20% leaf mortality, associated with high photosynthetic carbon fixation and electron transport rates and low leaf osmotic potential. One freezing-sensitive C4 species showed the expected pattern of chilling-induced damage to photosynthesis and >95% leaf mortality after the freezing event. However, three C3 and two C4 species displayed a cold acclimation response, showing significant decreases in osmotic potential and photosynthesis after exposure to chilling, and a 30-72% reduction of leaf freezing injury. This result suggested that down-regulation of osmotic potential may be involved in the cold acclimation process, and demonstrated that there is no inherent barrier to the development of cold acclimation in C4 species from this ecosystem. Cold acclimation via osmoregulation represents a previously undescribed mechanism to explain the persistence of C4 plants in cool climates.     

香蕉肉桂醇脱氢酶基因的克隆及表达分析

肉桂醇脱氢酶(CAD)在木质素合成过程中起关键作用。通过RACE(rapid-amplification of cDNA ends)方法从香蕉根系cDNA均一化全长文库中获得一个肉桂醇脱氢酶基因,命名为MaCAD1(GenBank登录号为KF582533)。MaCAD1是香蕉MYB基因编码框全长cDNA,包含一个1 077bp的最大开放阅读框(ORF),编码358个氨基酸。蛋白质序列同源比对发现,其含有完整的醇脱氧酶的典型保守结构域,属于典型的CAD蛋白。系统进化树比对分析表明,MaCAD1与水稻OsCAD6(CAD39907)的亲缘关系较近。组织特异性研究表明MaCAD1基因组成型表达于香蕉各个组织。在耐病和感病品种中,MaCAD1均上调表达,但在耐病品种中MaCAD1在所有时间点相对于对照增加的倍数均高于感病品种,表明MaCAD1基因在香蕉的抗病性中起着重要作用,MaCAD1可以作为一个新的响应枯萎病侵染的标记基因。     

Are budburst dates,dormancy and cold acclimation in walnut trees (<Emphasis Type="Italic">Juglans regia</Emphasis> L.) under mainly genotypic or environmental control?

As observed for most stresses, tree frost resistance can be split into two main processes: avoidance and tolerance. Avoidance of freezing is achieved by introducing species only in the climatic context in which the probability of freezing events is very low for the sensitive stages of buds or stems; i.e., when good synchronism exists between the annual cycle and the critical climatic periods. Buds become able to grow only after chilling requirements have been satisfied (endodormancy released) during winter; they subsequently break after heat requirements have been completed (end of ecodormancy) in early spring. Actually, this period is often subject to more or less severe freezing events. Trees are also able to adjust their freezing tolerance by increasing their capacity of extracellular freezing and decreasing the possibility of intracellular freezing through the process of frost acclimation. Both freezing resistance processes (avoidance and tolerance) are environmentally driven (by photoperiod and temperature), but there are also genotypic effects among species or cultivars. Here, we evaluated the degree to which differences in dormancy release and frost acclimation were related to environmental and genetic influences by comparing trees growing in common garden conditions. This investigation was carried out for two winters in lowland and mountain locations on different walnut genotypes differing significantly for budburst dates. Chilling requirement for endodormancy release and heat requirement during ecodormancy were evaluated in all situations. In addition, frost acclimation was assessed by the electrolyte leakage method on stems from the same trees before leaf fall through budburst. No significant differences were observed in chilling requirements among genotypes. Moreover, frost acclimation dynamics were similar between genotypes or locations when expressed depending on chilling units accumulated since 15 September as a time basis instead of Julian day. The only exception was for maximal frost hardiness observed during winter with the timber-oriented being significantly more resistant than fruit-oriented genotypes. Heat requirement was significantly different among genotypes. Thus, growth was significantly faster in fruit-oriented than in wood-oriented genotypes. Furthermore, among wood-oriented genotypes, differences in growth rate were observed only at cold temperatures. Frost acclimation changes differed significantly between fruit- and wood- walnuts from January through budburst. In conclusion, from September through January, the acclimation dynamic was driven mainly by environmental factors whereas from January through budburst a significant genotype effect was identified in both frost tolerance and avoidance processes.     

Chill-responsive dehydrins in blueberry: Are they associated with cold hardiness or dormancy transitions?

To survive winters, woody perennials of temperate zones must enter into endodormancy. Resumption of spring growth requires sufficient exposure to low temperature (chill units, CUs) in winter (chilling requirement), which also plays a role in the development of cold hardiness (cold acclimation). Physiological studies on dormancy breaking have focused on identifying markers, such as appearance or disappearance of proteins in response to varying degrees of chill unit accumulation. However, whether these changes are associated with dormancy transitions or cold acclimation is not clear. In the present study, greenhouse-grown blueberry (Vaccinium section Cyanococcus) plants were used to address this question. Three blueberry cultivars, Bluecrop, Tifblue, and Gulfcoast having chilling requirement of approximately 1 200, 900 and 600 CUs, respectively, were first exposed to 4°C for long enough to provide chill units equivalent to one-half of their respective chilling requirement. This treatment was expected to result in cold acclimation. A fraction of plants was then subjected to a 15/12°C (light/dark) regime for 2 weeks, a treatment expected to be “dormancy-neutral” but cause deacclimation. Before and after each treatment, cold hardiness and dormancy status of floral buds were determined; proteins were extracted from the buds collected on the same sampling date, and separated by one-dimensional SDS-PAGE. Dehydrin-like proteins were identified by immunoblotting, using anti-dehydrin antiserum. Results indicate that the chilling treatment resulted in cold acclimation as indicated by increased bud hardiness in all three cultivars. Data also indicate a distinct accumulation of three dehydrin-like proteins of 65, 60, and 14 kDa during cold acclimation. The cold hardiness and levels of dehydrin proteins decreased during the exposure to 15/12°C for 2 weeks. Results also confirmed that this treatment had no negative effect on chill unit accumulation. Densitometric scans of protein gels indicated a close association between the abundance of dehydrins and degree of cold hardiness in these cultivars. In addition, levels of the dehydrin proteins and cold hardiness remained about the same between 100% and >100% satisfaction of chilling requirement. These results suggest that changes in dehydrin expression are more closely associated with cold hardiness than with dormancy transitions.     

Alleviating effects of exogenous Gamma-aminobutiric acid on tomato seedling under chilling stress

Low temperature during germination and early seedling growth is one of the most significant limiting factors in the productivity of plants. Tomato seedling germination is sensitive to chilling stress. Gamma-aminobutyric acid (GABA), as a non-protein amino acid, involved in various stress tolerances in plants. In this study, 5-day old tomato seedlings were exposed to chilling stress (2 ± 0.05 °C for 48 h) and then the effects of 0, 100, 250, 500 and 750 μmolL⁻¹ concentrations of GABA on electrolyte leakage, proline and malondialdehyde (MDA) content were investigated. The resultS showed that the antioxidant enzyme activity, electrolyte leakage, MDA and proline content were significantly reduced by GABA treatments. However under chilling stress seedlings treated with GABA exhibited significantly higher sugar and proline contents as compared to un-treated seedlings. These results suggest that GABA treatment protects tomato seedlings from chilling stress by enhancing some antioxidant enzymes activity and reducing MDA content which results in maintaining membrane integrity.     

Unravelling the genetic complexity of sorghum seedling development under low‐temperature conditions

Sorghum is a promising alternative to maize for bioenergy production in Europe; however, its use is currently limited by poor adaptation to low temperatures during and after germination. We collected multi‐trait phenotype data under optimal and suboptimal temperatures in a genetically diverse recombinant inbred line (RIL) mapping population showing contrasting segregation patterns for pre‐ and post‐emergence chilling tolerance. Germination, emergence, seedling development, root architecture and seedling survival were assessed in two different seedlots. Emergence and root establishment were found to be the key determinants of development and survival under chilling stress. Highly interactive epistatic quantitative trait loci (QTL) hotspots, including a previously unknown QTL on Sb06 with a significant effect on prolonged chilling survival, were found to regulate different physiological mechanisms contributing to maintenance of growth and development despite the chilling temperatures. The major QTL regions harbour promising candidate genes with known roles in abiotic stress tolerance. Identification of loci in the QTL hotspot regions conferring maintenance of cell division and growth under early chilling stress represents a promising step towards breeding for successful establishment of sorghum in temperate climates.     

Environmental effects on the early stages of tassel morphogenesis in maize (Zea mays L.)

The effects of various environmental conditions on the initiation of tassel branches (NTB) and spikelet‐pairs (NSP) were examined in the stress‐sensitive maize inbred F53. Chilling induced the most important effect, with a dramatic decrease in both NTB and NSP, provided it was applied at the end of the vegetative phase and start of the floral transition phase. The primary cause of chilling‐induced abortion of the tassel branches could be oxidative stress in the leaves, since lowering light irradiance during chilling greatly reduced the effect of cold. The comparison of inbreds F53 and F2 revealed that both genotypes exhibited a similar period of cold sensitivity at the floral transition phase, although F2 was considered from field observations as a stress‐insensitive genotype (at least for tassel development). However, our results also showed a chilling acclimation response in inbred F2 but not in inbred F53. The similarities with the work by Lejeune & Bernier (1996 Plant, Cell and Environment 19, 217–224.) concerning the effect of chilling on ear initiation in the sensitive inbred, B22, are emphasized.     

Relationships between circadian rhythm of chilling resistance and acclimation to chilling in cotton seedlings

Cotton (Gossypium hirsutum L. cv. Deltapine 50) seedlings grown under light-dark cycles of 12:12h at 35°C showed rhythmic daily changes in chilling resistance. Chilling treatment (5°C, 48h) started at the beginning or middle of the daily light period resulted in a substantial growth inhibition of the seedlings upon return to 35°C whereas when chilling was started at the beginning or middle of the dark period the subsequent growth of the seedlings was much less inhibited. This rhythm in chilling resistance persisted under continuous light for three 24-h periods, indicating that it is of an endogenous nature. Seedlings grown under continuous light from germination showed no daily changes in resistance, but a rhythm was initiated by introduction of a dark period of 6h or longer. In 24-h cycles with different light and dark periods, maximal resistance was reached just before the start of dark period. Seedlings grown at 35°C could be acclimated to chilling by exposure to low, non-damaging temperatures (25–15°C). A short-term (6h) exposure to 25°C started at the resistant phase resulted in a large increase in resistance during the following otherwise sensitive phase. The resistance induced by the low temperature matched or slightly exceeded the maximal resistance reached during the resistant phase of the daily rhythm of chilling. The low-temperature-induced resistance and the daily rhythmic increase in resistance were not additive, indicating a common mechanism for the two kinds of resistances. An adaptive advantage of a combination of a rapid temperature-induced acclimation and the daily rhythmic increase in resistance is suggested.     

Frost resistance and water status of winter rape leaves as affected by differential shoot/root temperature

Twenty day-old winter rape ( Brassica napus L. var. oleifera L. cv. Jantar) seedlings, grown in nutrient solution, were exposed to different shoot/root temperature (i. e. 20/20, 20/3, 3/20 and 3/3°C) for 2 or 4 weeks. Chilling treatments modified markedly the pattern of plant growth as indicated by changes in dry matter accumulation in individual plant parts (leaves, hypocotyls, roots) and decreased leaf specific area. Growth of roots was less sensitive to low temperature than that of shoots. This was reflected by a decrease in shoot/root biomass ratio. Chilling treatments increased freezing resistance, decreased water content and water potential and modified reducing sugar, soluble protein and phospholipid contents in the leaves. A biphasic character of tissue responses to chilling temperature was observed, the most remarkable changes being registered during the first 7 or 14 days of the treatment. Effects of root or shoot exposure to chilling temperature on ice nucleation temperature, LT₅₀, water potential, accumulation of sugars and phospholipids in leaves were additive. All the observations point to the important role of the root system in plant acclimation to cold. Its impact on water status of leaves is emphasized and some mechanisms of root involvement in acclimation processes are proposed.     

Electrolyte leakage and lipid degradation account for cold sensitivity in leaves of Coffea sp. plants

Five Coffea genotypes differing in their sensitivity to low positive temperatures were compared with regard to the effects of chilling on membrane integrity, as well as their ability to recover from cold-induced injury upon re-warming. Membrane damage was evaluated through electrolyte leakage, changes in membrane lipid composition and malondialdehyde (MDA) production in control conditions (25/20 degrees C, day/night), after a gradual temperature decrease period to 15/10 degrees C, after chilling treatment (3 nights at 4 degrees C) and upon re-warming to 25/20 degrees C during 6 days (recovery). C. dewevrei showed the highest electrolyte leakage at 15/10 degrees C and after chilling. This was due mainly to lipid degradation observed at 15/10 degrees C, reflecting strong membrane damage. Furthermore, MDA production after chilling conditions indicated the occurrence of lipid peroxidation. A higher susceptibility of C. dewevrei to cold also was inferred from the complete absence of recovery as regards permeability, contrary to what was observed in the remaining plants. Apoat? and Piat? presented significant leakage values after chilling. However, such effects were reversible under recovery conditions. Exposure to cold (15/10 degrees C and 3 x 15/4 degrees C) did not significantly affect membrane permeability in Catuaí and Icatú. Furthermore, no significant MDA production was observed even after chilling treatments in Apoat?, Piat?, Catuaí and Icatú, suggesting that the four genotypes had the ability to maintain membrane integrity and/or repair membrane damage caused by low temperatures. Apoat?, Piat? and, to a lower extent, Catuaí, were able to cope with gradual temperature decrease through an enhanced lipid biosynthesis. After acclimation, Piat? and Catuaí showed a lowering of digalactosyldiacylglycerol to monogalactosyldiacylglycerol ratio (MGDG/DGDG) as a result of enhanced DGDG synthesis, which represents an increase in membrane stability. The same was observed in Apoat? after chilling, in spite of phospholipids decrease. The studied parameters clearly indicated that chilling induced irreversible membrane damage in C. dewevrei. We also concluded that increased lipid synthesis, lower MGDG/DGDG ratio, and changes in membrane unsaturation occurring during acclimation to low temperatures may be critical factors in maintenance of cellular integrity under chilling.     

Oxygen-mediated cold-acclimation in cucumber (Cucumis sativus) seedlings

Cold acclimation of etiolated cucumber seedlings, consisting of cooling at 12°C for 48 h followed by a warming period at 25°C, led to tolerance to subsequent chilling at 2°C. Tolerance, as evidenced by freedom from chilling injury and continued growth, developed during the warming period in a time-course manner for 12 h but decreased with prolonged warming. A similar increase and subsequent decrease was also observed in the content of palmitic, linoleic and linolenic acids in total lipid fraction from cucumber hypocotyl tissue. During the warming period supra-ambient oxygen stimulated, whereas subambient oxygen inhibited, the increase in fatty acid content as well as development of chilling tolerance. A strong correlation between oxygen-mediated changes in fatty acid content and associated development of cold tolerance suggests that both these processes are interrelated. Cold acclimation, but not cold stress, led to an increase followed by a decrease in CO₂ evolution suggesting that a respiratory upsurge is yet another feature of cold acclimation in cucumbers.