Use of somaclonal variation and in vitro selection for chilling tolerance improvement in rice

Embryo-derived calli of four rice varieties cultivated at high altitude in Burundi — Facagro 57, Facagro 76, Kirundo 3 and Kirundo 9 — were submitted to different temperature regimes. The percentage of regenerating calli greatly varied depending on variety, length of culture and callus temperature treatment. The reduction of regeneration percentages induced by low temperature was more pronounced in the more sensitive varieties. Regenerated plants (R0) and their progenies in R1, R2 and R3 were cold-screened together with control plants. In all varieties, significantly higher survival rates were obtained in R3 with in vitro plants than with control plants. Such chilling tolerance improvement was not obtained following a massal selection applied during 3 successive generations onto the control plants. In vitro plants regenerated from calli cultivated either at 25 °C, either at 4 °C, were cultivated at different altitudes in Burundi during two successive generations. For most observed traits, the in vitro plants were characterized by lower means, larger variation and higher maximum values than the control plants. The most chilling-tolerant somaclonal families were most usually characterized by extensive differences in fatty acid composition, chilling-induced electrolyte leakage and chlorophyll fluorescence, compared to the varieties they derived from. This revised version was published online in July 2006 with corrections to the Cover Date.     

Field evaluation and selection of rice somaclonal variants at different altitudes

A field experiment was designed to compare control with tissue culture-derived plants of rice, and to study the effects of natural selection pressure applied to the first generation (R0) of in vitro plants on the performance of their progeny. Control- and R0 in vitro plants of four rice varieties from middle altitude in Burundi, i.e. Facagro 57′, Facagro 76′, ‘Kirundo 3’ and ‘Kirundo 9’, were cultivated at four different altitudes (800, 1380, 1650 and 1900 m). Several parameters concerning tillering capacity, plant height development and seed production were measured. The means for most measured parameters were higher in control plants than in the in vitro plants, while the higher variation coefficients and most extreme values were usually found in the in vitro plants. For each variety, the 20 control plants and 20 in vitro plants having the highest production were selected at each altitude of 1380 and 1650 m, and their progenies were cultivated on the same site at 1580 m. Among the plants arising from 1650 m, most maximum values, higher variation coefficients and, in some cases, higher means were found in the in vitro plants. In contrast, among the plants arising from 1380 m, higher means most usually corresponded to control plants, while the tendency for maximum values and variation coefficients were unclear. These results indicate the positive effect of natural selection pressure applied in the first generation to plants rising from tissue culture. They also demonstrate that considerable variation may be generated in vitro. The interest for plant breeding purposes is discussed.     

Evaluation of chilling sensitivity in different rice varieties. Relationship between screening procedures applied during germination and vegetative growth

Summary Rice varieties were screened for chilling tolerance during germination and vegetative growth using different techniques. The effects of temperature during germination were investigated from 10 to 25°C. The screening was most effective at 10°C. Time of data collection has to be considered in order to discriminate slow germinating from chilling sensitive varieties. Chilling survival tests (10°C) applied at the 2-leaf stage revealed that different chilling durations brought complementary results: the longer treatments allowed to identify the most tolerant varieties, while shorter treatments allowed to distinguish intermediate from most sensitive rices. Results of chilling tests applied at the 2- and the 8-leaf stages showed a high correlation. Low-temperature electrolyte leakage and low-temperature fluorescence were also highly correlated to each other and to survival tests applied at the same development stage. By contrast, weak correlations were observed between low-temperature germination and plantlet chilling survival. It is concluded that low-temperature electrolyte leakage and low-temperature fluorescence are good indicators of chilling tolerance during the vegetative growth. The possibility to use a chilling screening procedure applied at this stage in order to predict the chilling tolerance at other development stages is discussed.     

低温胁迫下籽用西瓜幼苗生理变化与耐冷性的研究

以三叶一心期的籽用西瓜幼苗为材料,采用10℃低温胁迫的方法,研究了黑籽瓜兰州大片(wb10)和红籽瓜巢湖红(wb7)随着处理天数的延长幼苗的耐冷性、光合特性、生理特性的变化规律,以探讨籽用西瓜耐低温的生理机制,从而为籽用西瓜的引种栽培和抗寒品种选育提供理论依据。结果表明:不同品种籽用西瓜幼苗的叶绿素含量、Pn、Tr、Gs、Ci随低温胁迫时间的延长而逐渐降低;Pro、MDA、相对电导率随低温胁迫时间的延长而升高。两品种的冷害指数存在一定差异,耐冷性表现为wb10>wb7。低温胁迫下,耐冷性较强的wb10幼苗叶片中的叶绿素含量、Pn、Tr、Gs、Ci均高于耐冷性较弱的wb7,而耐冷性较强的wb10幼苗叶片中MDA含量、脯氨酸含量、相对电导率均低于wb7。综合试验中各项指标认为,耐冷性与品种的来源、类型及在低温下植株的冷害指数、光合特性和细胞膜稳定性有关。     

Fluorimetric Determination of the Genetic Variability Existing for Chilling Tolerance in Sweet Sorghum and Sudan Grass

Chilling temperatures drastically inhibited the photochemical quenching of chlorophyll fluorescence (q_Q) measured in intact leaves photosynthesizing under steady-state conditions. This effect appeared, however, to be characteristic of chilling-susceptible plant species and was not observed in plants which are known to be chilling-tolerant, indicating that the measurement of q_Q can serve in practice to estimate rapidly the relative chilling tolerance of crop plants. A large number (28) of sweet sorghum (Sorghum bicolor) and Sudan grass (Sorghum sudanense) genotypes were screened for chilling tolerance using this rapid q_Q method. Although sweet sorghum and sudan grass obviously behaved as chilling-sensitive plants, a considerable variation for chilling susceptibility was observed among the different genotypes tested. Some sweet sorghum varieties, such as ‘Dale’ and ‘Keller’, and most of the sudan grass varieties appeared to possess a certain degree of resistance towards low temperature stress, indicating the existence of useful germplasms in Sorghum for improving stress tolerance.     

水稻品种对几种逆境的多重耐性及与ABA的关系

从一些耐旱和耐冷品种中筛选出几个具多重耐性的品种,其中大黄谷耐渗透胁迫和盐胁迫,芝麻糯耐盐和冷胁迫,桂溪和湘中籼2号耐渗透胁迫,也较耐冷胁迫.以大黄谷、芝麻糯和桂溪为材料,研究了多重耐性与ABA积累的关系.在渗透胁迫、盐胁迫和冷胁迫下,各水稻品种内源游离ABA含量迅速积累,耐性品种积累的ABA量都较敏感品种高,     

Electrolyte Leakage, ATP Content in Leaves and Intensity of Net Photosynthesis in Maize Seedlings at Permanent or Different Daily Exposure to Low Temperatures

Changes were determined in electrolyte leakage (EL), ATP content in leaves and intensity of net photosynthesis (FJ in maize seedlings as a result of continued exposure to low temperatures for several days or, after a cold night (5 °C, 9 h), alternating application of 5 °C and 20 °C temperatures during the lit phase of the day (h of chilling during day/night: 0/9, 5/9, 10/9, 15/9).
At continuous exposure to low temperatures, permanent reduction of ATP content occurred between days 7 and 10, while EL increased significantly between days 4 to 7 (depending on experimental conditions). Of three leaves differing in age, most injuries were found in the oldest leaf. An increased intensity of irradiation from 150 to 500 μE. m⁻².s⁻¹ caused an earlier decrease in the ATP content and a further increase in electrolyte leakage. ATP content showed a 24 h rhythm, it increased at the end of the night and decreased at the end of the day. The rhythm was particularly noticeable in the control plants (20 °C), as well as, however to a lesser extent, in plants exposed to daily fluctuating chilling temperatures. It has also been observed that higher temperatures occurring in day time may diminish electrolyte leakage induced by previous low temperatures
The intensity of F_n determined at 20 °C, 4 h after withdrawal of chilling, decreased with the prolongation of the chilling period during the day.
Thus, it may be assumed that the EL and F_n values as indicators of the plants' sensitivity are in better agreement with the induced chill doses than the ATP content in leaves, which to a greater extent depends on the interaction between chilling and other experimental factors, such as irradiation intensity or the sequence of light and darkness.     

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.     

低温对大白菜幼苗生长及生理指标的影响

比较不同基因型大白菜对低温冷害的响应,为黑龙江大白菜种植及耐低温品种选育提供依据。白菜幼苗生长至3叶1心时,转入人工气候箱中进行低温处理3天,对6个白菜品种进行生物产量及生理性状测定与分析。结果表明：低温胁迫条件下,6 个供试品种生物产量及叶绿素含量均呈下降趋势。除‘新锉四号’外,其余5 个大白菜品种的可溶性蛋白含量在低温处理下均高于常温对照。除‘抗病91-12’外,其余5 个品种的过氧化物酶活性在低温处理下均高于常温对照。6 个大白菜品种耐冷性能力依次为：‘新北京三号’＞‘农学昌五小根’＞‘牡丹江二号’＞‘义和九号’＞‘抗病91-12’＞‘新锉四号’。因此,‘新北京三号’可作为耐冷性品种在黑龙江地区进行种植。     

田间高温发生对不同葡萄叶片组织结构的影响

为了分析高温发生前后葡萄叶片生长发育特点，本研究通过对田间高温发生前后不同葡萄的叶片组织结构变化进行分析。结果发现：高温发生后‘申丰’、‘沪培1号’、‘醉金香’、‘巨峰’、‘巨玫瑰’的叶片、栅栏组织和海绵组织的厚度有不同程度的破坏，表现出CTR值减少，SR值增大；而高温后、‘申华’、‘申玉’和‘夏黑’叶片、栅栏组织和海绵组织的厚度变化不明显，CRT值和SR值在高温发生前后变化不明显，表现出较强的抗热能力。综合可知，较其他5个品种而言，‘申华’、‘申玉’和‘夏黑’对高温具有较强的适应性。     

Evaluation of seedling cold tolerance in rice cultivars: a comparison of visual ratings and quantitative indicators of physiological changes

Rice (Oryza sativa L.) is sensitive to prolonged exposure to low temperature, which at the seedling stage can result in significant chilling injury and mortality. The objective of this study was to quantify physiological and biochemical changes in rice seedlings undergoing chilling stress and compare those changes with visual evaluation of tolerance. Seedlings from the cultivars M-202 (tolerant) and IR50 (sensitive) were subjected to 9°C for 14 days in a controlled environment chamber. Leaf tissues were harvested at various time points for determination of electrolyte leakage, proline, malondialdehyde, ascorbic acid and reduced glutathione. Significant differences between M-202 and IR50 were detected in electrolyte leakage, proline, and ascorbic acid starting at 7 days with IR50 exhibiting higher levels of these indicators. Most IR50 seedlings were dead at 14 days. A set of fifty rice accessions including M-202 and IR50 was evaluated at 10 days of cold treatment to examine the correlation of visual ratings with the physiological indices. Visual ratings were most highly correlated with electrolyte leakage and least correlated with proline content. Based on visual ratings and the physiological indices, we identified several cultivars that outperformed M-202 in cold tolerance while IR50 had the lowest tolerance of the cultivars tested.     

氯化钙对低温胁迫下番茄幼苗的影响

为探究氯化钙如何影响番茄冷胁迫过程,得出氯化钙提高番茄幼苗耐冷性机制,进行了冷害指数调查、生理指标测定。使用光照培养箱低温处理耐冷型‘O-33-1’和冷敏感型‘NY2000’,依照氯化钙处理浓度的不同记为CK、T₁、T₂、T₃,对不同处理调查冷害指数,测定叶绿素含量,测定丙二醛(MDA)、过氧化物酶(POD)、超氧化物歧化酶(SOD)活性。研究表明,冷胁迫外源喷施氯化钙时,‘O-33-1’冷害指数、MDA含量、POD含量先下降后升高,叶绿素a含量、叶绿素b含量、SOD活性先升高后下降;‘NY2000’冷害指数先下降后上升,SOD活性先升高后下降,MDA含量不断升高,叶绿素a含量、叶绿素b含量基本不变,POD则表现得没有规律。不同品种响应氯化钙提高耐冷性表现出差异;低温胁迫下,叶绿素的降解受到抑制从而提高了番茄幼苗耐冷性;耐冷性的提高与外源氯化钙导致抗氧化酶活性提高有关。     

复合盐碱胁迫对鲜食葡萄生理生化指标的影响

为筛选出较耐盐碱的新引鲜食葡萄品种,以10个葡萄品种为材料,研究不同浓度复合盐碱（盆土干重的0.29%、0.58%、0.87%）胁迫30天后葡萄生理生化指标的变化特征,对所有指标进行相关性分析,并采用主成分分析法综合评价其耐盐碱能力。结果表明,随着盐碱浓度的增加,根系活力和叶片相对含水量呈逐渐下降趋势,相对电导率、丙二醛和脯氨酸含量呈逐渐上升趋势,超氧化物歧化酶活性呈先上升后下降或持续上升2种趋势。相关性分析发现,叶片丙二醛含量、根系丙二醛含量、叶片脯氨酸含量三者之间两两极显著正相关,根系超氧化物歧化酶活性与叶片丙二醛含量、叶片超氧化物歧化酶活性显著负相关。主成分分析法提取出特征值大于1的3个主成分,累计贡献率77.99%,最终得出10个葡萄品种耐盐碱性顺序由强到弱依次为‘紫甜无核’‘、黑脆无核’‘、户太8号’‘、早夏无核’‘、丝路红玫瑰’‘、阳光玫瑰’‘、深红玫瑰’‘、夏黑’‘、甜蜜蓝宝石’‘、浪漫红颜’。     

不同梨品种需冷量与南京地区促成栽培扣棚时期的比较研究

为了探明适宜在南京地区设施促成栽培中使用的梨品种及其扣棚升温时期,2011—2012年用犹他(Utah)模型、0~7.2℃模型、≤7.2℃模型对栽培在南京地区37个梨品种的需冷量进行了测定。结果表明：‘爱宕’最低,以犹他模型计算为207 C.U。‘砀山酥’、‘鸭梨’、‘沙01’、‘六月酥’、‘早美酥’等需冷量高。不同梨品种需冷量分布广泛,但主要分布在300~700 C.U之间。相比而言,南京地区梨需冷量的研究采用犹他模型为妥。成熟期及其原产地与需冷量无必然的内在联系。南京冬季气候温和,日均温度波动较小,用于设施促成栽培需冷量小于400 C.U的早熟及中熟品种可在1月初覆膜升温;需冷量在400~700 C.U之间的早熟品种可在2月初覆膜升温。     

吉林省主栽水稻品种孕穗期耐冷性比较

为了对吉林省水稻品种进行合理布局,降低低温冷害风险,对目前吉林省主要水稻栽培品种孕穗期的耐冷性进行研究。选用目前吉林省的7个主栽水稻品种（‘长白9’,‘长白16’,‘吉粳803’,‘吉粳105’,‘吉粳88’,‘吉粳83’,‘吉粳81’）进行孕穗期16℃低温处理,通过线性内插法确定了低温处理6~14天连续9天的水稻空壳率,进一步利用统计回归方法建立了16℃低温处理下不同水稻品种处理天数与空壳率的相互关系。结果表明;所有参试品种在处理10~13天时水稻空壳率达到最高,随后无明显变化。其中耐冷性弱的‘长白19’的空壳率最早达到最大值,并且冷水反应比较敏感。耐冷性最强的‘吉粳81’,冷水反应比较迟钝。其余的品种介于二者之间。因此,品种间耐低温能力存在显著差异,选育耐低温品种是提高水稻耐冷性的一种有效途径。     

淮北地区小麦主栽品种对低温胁迫的响应及抗寒性评价

为给安徽淮北地区小麦安全生产提供指导,以当地主栽品种为材料,研究不同播期条件下小麦植株对低温胁迫的响应,结合自然低温下田间冻害情况及产量表现等,对主栽品种的抗寒性进行初步评价。结果表明,返青期-12℃以下时品种的抗寒性急剧下降;播期间小麦植株的抗寒性表现为：晚播>适播>早播;电解质渗出率均随温度的降低逐渐增大;地域间抗寒性差异明显;不同区域育成品种抗寒性强弱表现为山东省＞河南省＞安徽省;筛选出一批抗寒高产品种,如‘良星66’、‘山农20’、‘济麦22’、‘烟农19’等,可作为安徽淮北地区早播条件下种植的理想品种。     

低温胁迫对大豆花期叶片蔗糖代谢及产量的影响

为了研究低温胁迫对大豆花期产量形成的影响,以‘垦丰16’（耐冷型品种）和‘合丰50’（冷敏感型品种）为试验材料,采用盆栽试验,测定了叶片蔗糖含量以及代谢相关酶活性,成熟期测产。结果表明,大豆花期低温胁迫显著降低了大豆的株高和产量,底荚高度增加,随低温胁迫时间的延长,减产损失逐渐增加,与对照相比,‘合丰50’减产幅度为10.29%~30.88%,‘垦丰16’减产幅度为9.58%~24.22%,敏感品种的减产幅度大于耐冷品种。叶片内的淀粉和蔗糖含量随胁迫时间延长逐渐降低,果糖含量随胁迫时间延长逐渐增加,酸性转化酶、中性转化酶和蔗糖合成酶活性提高,蔗糖磷酸合成酶活性被抑制。低温恢复期间（低温处理4天后自然环境下恢复1~4天）上述指标呈现相反的变化趋势,不同品种间呈现相同的变化趋势。大豆花期遭遇低温胁迫致使蔗糖分解代谢加快,合成途径受到抑制,叶片内的蔗糖含量急剧减少,并且其危害不能在4天内恢复,这不利于大豆产量的形成,推断这是低温胁迫造成大豆减产的重要原因之一。     

山西省9个甜樱桃主栽品种需冷量研究

为了研究山西省部分甜樱桃主栽品种的需冷量,为山西省设施甜樱桃扣棚和升温时间的确定提供参考,试验以9 个山西省甜樱桃主栽品种‘红玛瑙’、‘红灯’、‘龙冠’、‘龙旭’、‘大紫’、‘美早’、‘雷尼尔’、‘早大果’、‘拉宾斯’为材料,在甜樱桃开始大量落叶时采集枝条进行低温处理,将低温积累到一定时间的甜樱桃枝条置于适宜条件下培养,3周后统计萌芽率,采用“0 ～ 7.2℃ 模型”对其需冷量进行分析。结果表明：不同甜樱桃品种需冷量有差异,供试品种的需冷量值集中分布在500～750ｈ;部分甜樱桃品种存在芽类间需冷量不同的现象;同一品种其需冷量与物候期没有相关性。     

4个桃品种抗寒性研究初报

为探讨衡量桃不同品种抗寒性的适宜指标,试验以豫农二号、红雪桃、豫花、秋蜜红4个桃品种为试材,对其在不同温度梯度下低温处理过的花朵雌蕊变褐情况进行观察以及对枝条、叶片进行相对电导率的测定。结果表明：观察低温处理的桃雌蕊的褐变发生情况,与测定叶片的相对电导率得到的结果一致,即豫农二号抗寒性强于红雪桃和秋蜜红,豫花抗寒性最弱;对休眠枝条进行相对电导率分析得出的结论则是秋蜜红与红雪桃无显著差异,秋蜜红要强于豫农二号,豫花仍为最弱。因此以不同器官相对电导率判断的品种抗寒性并不完全一致。     

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.