10个香橼半同胞家系子代苗抗寒性研究

对10个3年生香橼半同胞家系子代进行了抗寒性研究。结果表明,各家系子代苗之间的抗寒性有显著差异。春江7号和新世纪2号的抗寒性差,在田间-11℃自然低温下受冻级别均达Ⅲ级,耐寒指数为3.25～3.40,半致死温度分别为-7.91℃和-7.05℃。春江5号抗寒性较强,耐寒指数为2.17,半致死温度为-13.28℃。其余家系抗寒性中等,-11℃田间自然低温下受冻级别均为Ⅱ级,耐寒指数为2.33～2.86,半致死温度为-11.69℃～-9.91℃。在-3℃～-15℃低温胁迫下,家系子代苗叶片内SOD活性、可溶性蛋白质含量、脯氨酸含量及丙二醛(MDA)含量均有极显著差异,并随处理温度降低总体呈先上升后下降的趋势。SOD活性、可溶性蛋白质含量及脯氨酸含量与耐寒指数、半致死温度均呈负相关,MDA含量与耐寒指数、半致死温度呈正相关。一般SOD活性、可溶性蛋白质含量及脯氨酸含量高,MDA含量低的家系,耐寒指数、半致死温度低,抗寒性强;反之,SOD活性、可溶性蛋白质含量及脯氨酸含量低,MDA含量高的家系,耐寒指数、半致死温度高,抗寒性差。香橼各家系通过上述生理指标的综合变化来调节自身对逆境的适应性,增强抗寒性,减少逆境的伤害,为耐寒品种选育提供生理基础。     

Expression of a carrot 36 kD antifreeze protein gene improves cold stress tolerance in transgenic tobacco

Antifreeze proteins （AFPs） enable organisms to survive under cold conditions, and have great potential in improving cold tolerance of cold-sensitive plants, In order to determine whether expression of the carrot 36 kD antifreeze protein gene confers improved cold-resistant properties to plant tissues, we tried to obtain transgenic tobacco plants which expressed the antifreeze protein. Cold, salt, and drought induced promoter Prd29A was cloned using PCR from Arabidopsis. Two plant expression vectors based on pBI121 were constructed with CaMV35S：AFP and Prd29A：AFP. Tobacco plantlets were transformed by Agrobacterium-medicated transformation. PCR and Southern blotting demonstrated that the carrot 36 kD afp gene was successfully integrated into the genomes of transformed plantlets. The expression of the afp gene in transgenic plants led to improved tolerance to cold stress. However, the use of the strong constitutive 35S cauliflower mosaic virus （CaMV） promoter to drive expression of afp also resulted in growth retardation under normal growing conditions. In contrast, the expression of afp driven by the stress-inducible Prd29A promoter from Arabidopsis gave rise to minimal effects on plant growth while providing an increased tolerance to cold stress condition （2℃）. The results demonstrated the prospect of using Prd29A-AFP transgenic plants in cold-stressed conditions that will in turn benefit agriculture.     

Expression of a Carrot Antifreeze Protein Gene in Escherichia coli

The recombinant expression vectorpET43. lb-AFP, which contains full encoding region of a carrot 36 kD antifreeze protein (AFP) gene was constructed. The recombinant was transformed into expression host carrying T7 RNA polymerase gene (DE3 lysogen) and induced by 1 mmol. L-1 IPTG (isopropyl-β-D-thiogalactoside) to express 110 kD polypeptide of AFP fusion protein.The analysis of product solubility revealed that pET43. 1b-AFP was predominately soluble, and the expressed amount reached the maximum after the IPTG treatment for 3 h.     

Novel plasmodesmata association of dehydrin-like proteins in cold- acclimated Red-osier dogwood (Cornus sericea)

Dehydrins are proteins associated with conditions affecting the water status of plant cells, such as drought, salinity, freezing and seed maturation. Although the function of dehydrins remains unknown, it is hypothesized that they stabilize membranes and macromolecules during cellular dehydration. Red-osier dogwood (Cornus sericea L.), an extremely freeze-tolerant woody plant, accumulates dehydrin-like proteins during cold acclimation and the presence of these proteins is correlated with increased freeze tolerance (Karlson 2001, Sarnighausen et al. 2002, Karlson et al. 2003). Our objective was to determine the location of dehydrins in cold-acclimated C. sericea stems in an effort to provide insight into their potential role in the freeze tolerance of this extremely cold hardy species. Abundant labeling was observed in the nucleus and cytoplasm of cold-acclimated C. sericea stem cells. In addition, labeling was observed in association with plasmodesmata of cold-acclimated vascular cambium cells. The unique association of dehydrin-like proteins with plasmodesmata has not been reported previously.     

Comparison of G6PDH Activity and LT50 Between P.tomentosa and P.suaveolens During Freezing Acclimation

The differences of glucose-6P dehydrogenase(G6PDH) activity and freezing resistance induced by freezing acclima-tion between cuttings of freezing-sensitive P.tomentosa and freezing-resistant P.suaveolens were compared for exploring the role of G6PDH on the enhancement of freezing resistance induced by freezing acclimation.After 5d of freezing acclimation at -3℃ ,the LT50 of P.tomentosa has deereased from -6.2℃ incontrol cuttings to -14.3℃ in freezing acclimated ones,and the increase of LT50 of P.tomentosa has decreased from -6.2℃ in control cutting to -14.3℃ in freezing acclimated ones,and the increase of G6PDH activity was observed in freezing acclimated cuttings as compared with control ones.Whereas,when P.suaveolens was freezing acclimated at -20℃ for 5d,the LT50 has decreased from -27.1℃ in control cuttings to -43.5℃ in freezing acclimated ones,and the activity of G6PDH increased considerably.In addition,the increase of LT50 and the decrease of G6PDH activity resulting from 2d of deacclimation at 25℃ were found in two kinds of freezing acclimated cuttings.It is concluded that the increase in the activity of G6PDH may associate with the inherited freezing resistance of species and the enhancemen of freezing resistance of cuttings,and may play an important role in the antifreeze process under freezing temperature,which would provide the basis for the study on the molecular mechanism of freezing resistance in P.suaveolens and the cloning of gene associated with freezing resistance.     

低温锻炼与CaCl2处理对毛白杨幼苗可溶性总蛋白、CaM及抗冻性的影响

低温锻炼提高了毛白杨幼苗叶片和枝条中钙调素 (CaM)和可溶性总蛋白含量 ,同时也提高了幼苗的抗冻性 .CaCl2 处理对上述低温锻炼的作用有加强的效果 ,体现在明显提高了CaM和可溶性总蛋白含量以及幼苗抗冻性 ;钙离子螯合剂EGTA、钙离子通道阻断剂LaCl3 或钙调素拮抗剂CPZ处理则减弱了低温锻炼的效应 ,表现出CaM和可溶性总蛋白含量以及幼苗抗冻性明显降低 .进一步研究发现 ,CaCl2 处理的低温锻炼不仅能减轻低温胁迫引起的CaM和可溶性总蛋白含量的下降程度 ,而且有利于幼苗在恢复过程中CaM和可溶性总蛋白水平的迅速回升 .这表明Ca2 + CaM可能参与了CaM和可溶性总蛋白的合成及抗冻性的诱导     

在低温锻炼中毛白杨幼苗可溶性总蛋白、RNA、DNA、RNase及抗冻性的变化

低温锻炼不仅提高了毛白杨幼苗存活率和抗冻性以及RNA和可溶性总蛋白的含量 ,降低了RNase活性 ,而且能减轻低温胁迫引起的RNA和可溶性总蛋白含量的下降程度和RNase活性的提高 ,有利于幼苗在恢复过程中RNA和可溶性总蛋白水平的迅速回升以及RNase活性的降低 .进一步研究发现 ,DNA含量无论在低温锻炼中还是在低温胁迫下或是在随后的恢复生长期均保持相对稳定 ;低温锻炼所引起的RNA含量的增加 ,与RNase活性的降低呈明显的负相关 ,与可溶性总蛋白含量的增加以及幼苗抗冻性的提高成正相关 .这表明低温锻炼可能抑制RNase活性 ,有效地促进RNA含量的增加 ,而RNA可能参与了可溶性总蛋白的合成及抗冻性的诱导     

胡杨悬浮细胞原生质体游离及质膜单离子通道测定

该文以胡杨 (Populuseuphratica)悬浮细胞为材料 ,利用 1 0 %纤维素酶“onozuka”R 10、0 0 1%果胶酶Y 2 3、0 15 %离析酶R 10和 0 1%半纤维素酶的混合酶液消化细胞壁 ,得到原生质体 .并利用膜片钳细胞贴附技术分别测到质膜内向和外向单通道电流 .通道电流在不同水平上变化 ,表明并非只有一种类型的通道开放 .树木细胞原生质体的分离以及利用膜片钳测定细胞质膜离子通道的成功实践为深入研究木本植物抗盐的细胞学机制奠定了基础     

香橼优良单株半同胞家系子代抗寒性研究

在南京六合地区对3年生24个香橼优良单株半同胞家系子代进行抗寒性测定研究,结果表明家系子代间抗寒性存在着显著的差异。田间观测-11℃自然低温下春江4#和香橼4#的抗寒性最强,受冻株率分别为86.67%和100%,平均受冻级别均为Ⅰ级,耐寒指数均为2.00;新世纪8#、春江7#和新世纪2#的抗寒性最差,受冻株率均为100%,平均受冻级别均达Ⅲ级,耐寒指数均大于3.00。室内抗寒力测定供试家系子代的半致死温度在-14.84℃～-7.05℃之间。春江4#和香橼4#的抗寒力最强,半致死温度分别为-14.84℃和-13.94℃;春江7#和新世纪2#的抗寒力最差,半致死温度分别为-7.91℃和-7.05℃。抗寒性评估结果一致。根据半致死温度,结合田间抗寒性观测,供试香橼家系子代抗寒性评估排序为春江4#>香橼4#>春江6#>春江5#>新世纪9#>新世纪10#>新世纪3#>香橼1#>新世纪7#>香橼7#>新世纪8#>香橼6#>香橼5#>春江3#>新世纪4#>新世纪6#>香橼2#>春江8#>新世纪5#>春江1#>新世纪1#>春江2#>春江7#>新世纪2#。     

CaCl2在毛白杨幼苗抗冻性低温诱导中的作用

不同浓度的CaCl2 处理对毛白杨幼苗抗冻性的提高效应是不同的 ,其中 10mmol·L-1CaCl2 处理更有利于幼苗抗冻性的提高 ,而且处理最适宜时间为 5天 .低温锻炼不仅提高了毛白杨幼苗叶片和枝条中钙调素含量 ,同时也提高了幼苗抗冻性 .CaCl2 处理对低温锻炼的作用有加强效果 ,而钙离子螯合剂EGTA、钙离子通道阻断剂LaCl3 或钙调素拮抗剂CPZ处理则减弱低温锻炼效应 ,表现出CaM含量和幼苗抗冻性明显降低 .这表明Ca2 + CaM可能参与了幼苗抗冻性的诱导     

额济纳绿洲不同树龄胡杨狭叶和阔叶耐旱特征

应用压力室技术对胡杨幼苗、幼树和成熟胡杨的狭叶和阔叶测定,绘制PV曲线并利用相关公式计算不同树龄胡杨狭叶和阔叶的水分参数,同时结合角质层蒸腾和气孔蒸腾测定结果研究胡杨狭叶和阔叶耐旱特征。研究结果表明：胡杨阔叶的耐早性强于狭叶,二者的耐旱方式存在差异,阔叶渗透物质含量高,忍耐低渗透势的能力强;狭叶主要通过高弹性的细胞壁维持膨压,其忍耐低渗透势的能力不及阔叶。成熟胡杨无论狭叶还是阔叶主要的水分散失形式是气孔蒸腾,体内的水分平衡主要通过气孔进行调节。     

黑麦草抗冻蛋白基因的cDNA克隆及序列分析

对多年生黑麦草进行冷胁迫处理后,提取叶片总RNA,采用RT-PCR法获得了LpAFP成熟蛋白的cDNA克隆。序列分析结果表明,该成熟蛋白的开放读码框(ORF)长度为357 bp,编码一条由118个氨基酸组成的多肽。生物信息学分析结果表明,黑麦草成熟抗冻蛋白与其他植物抗冻蛋白的同源性较低,与鱼类抗冻蛋白和昆虫抗冻蛋白则没有同源性。     

不同水分条件下胡杨和新疆杨苗期叶绿素变化

在不同水分条件下,应用SPDA-502叶绿素测定仪测定了胡杨和新疆杨苗木旺盛生长期的叶绿素含量.结果表明:(1)在7月和8月,不同水分处理下胡杨和新疆的叶绿素含量变化存在差异,7月胡杨叶绿素含量随着水分降低呈现增长趋势,而新疆杨叶绿素含量表现出先升后降再升高的趋势.8月叶绿素的含量变化是随着水分的减少2个树种都呈现先升...     

应用一元材积表法测定胡杨林分蓄积量

通过标准地每木调查,结合一元材积表测定计算胡杨林的林分蓄积量,结果显示:胡杨的一元材积方程为V=0.000484469d2.011854,将各径阶相应单株平均材积按径阶大小顺序排列成表即为一元材积表;4块样地林分平均直径分别为:15.1、15.0、13.7、15.9cm,在株数累积分布曲线上所对应的株数累积百分数的位置...     

干旱胁迫对胡杨叶片色素及光谱特征影响的研究进展

胡杨作为塔里木河流域的建群种与生态恢复的指示器,是遏制土地荒漠化和保护群落稳定结构的天然屏障.仅依靠人工调查对荒漠河岸林进行监测已不能满足当下维系塔里木流域生态环境,高光谱遥感成为当下胡杨种群监测识别的重要手段.就沙漠干旱地区胡杨叶片色素高光谱遥感监测的主要研究成果做出了概括,并经过对有关论文内容的总结与数据的梳理,明...     

电导法评价核桃无融合生殖后代的抗寒性

为探讨核桃无融合生殖后代抗寒性的遗传变异情况,对核桃自然授粉后代和无融合生殖后代的1a生枝条的相对电导率、相对电导率变化倍数、半致死温度等进行了测定比较。结果表明：核桃自然授粉后代和无融合生殖后代1a生枝条的相对电导率、相对电导率变化倍数均随处理温度的降低而增加,表现出相似的变化规律,呈倒“S”型曲线;在低温胁迫下,除少数处理外,二者之间的相对电导率、相对电导率变化倍数差异不显著;结合Logistic方程,得到中林1号无融合生殖后代、绵核桃无融合生殖和自然授粉后代的半致死温度（LT50）分别为-22．81℃、-20．36℃和-23．71℃,变化幅度较小。由此可见,核桃无融合生殖后代的抗寒性没有发生改变。     

普鲁士兰法测定胡杨中植物多酚含量

首次采用普鲁士兰法对胡杨中植物多酚含量进行了研究.取胡杨枝粉碎后过40目筛,用丙酮水溶液提取液在45℃下浸提,提取液处理后用P-B法进行测定.首先从稳定性、最大吸收波长、线性关系、重复性和准确性等角度对P-B法进行了方法改进和验证.然后用改进的P-B法对胡杨枝提取液进行测定,测得结果是胡杨枝中总酚含量平均为1.191%、RSD为2.840%.     

人为干扰对额济纳胡杨林群落组成及更新的影响

额济纳绿洲是我国西北干旱地区的一道强大的生态防线。但由于水资源匮乏和人为干扰影响,绿洲中胡杨林已严重退化。我们认为人为干扰会对胡杨群落的组成和更新造成影响。应用双向指示种分析法(TWINSPAN)和典范对应分析(CCA),从人为干扰角度对额济纳不同地区胡杨群落进行了分类和排序。结果表明:人为干扰会影响群落林下物种多样性和根孽幼苗生长情况;林下物种多样性主要受旅游干扰影响,并与总干扰强度的关系遵循"中度干扰假说";根孽幼苗生长情况与放牧干扰呈负相关,当根孽幼苗数多的时候,一定程度上能够抵御外界干扰对其生长状况的影响;适度的人为干扰可以增加物种多样性,但要保证胡杨林的繁殖更新,需要以消除放牧干扰影响为前提。     

辽河三角洲盐碱地树种优化配置模式研究

在辽河三角洲盐碱地台田上,布置小胡杨纯林、白蜡纯林、小胡杨×刺槐混交林、白蜡×刺槐混交林4种树种配置模式,对不同配置模式下的树木生长和土壤水、盐、温度进行了研究。结果表明,小胡杨、白蜡、刺槐在辽河三角洲盐碱地上造林均能成活,成活率达92%～98%。混交配置模式对树木生长具有显著的促进作用,优于纯林,其中小胡杨×刺槐混交模式好于白蜡×刺槐模式。与CK相比,4种模式均能改善土壤水分状况,降低土壤盐分,调控土壤温度,防止盐分表聚,4种模式间比较顺序是小胡杨×刺槐混交林>白蜡×刺槐混交林>白蜡纯林>小胡杨纯林。可见,小胡杨×刺槐混交模式可作为该地区防护林营建的首选优化配置模式。