The role of salinity in seed maturation of the euhalophyte Suaeda salsa

Controlled conditions were used to investigate how salinity maintains the salt tolerance of seeds and seedlings of the euhalophyte Suaeda salsa. Seeds were harvested from S. salsa plants that had been treated with 1 or 500 mM NaCl for 113 days in a glasshouse. The results showed that high salinity (500 mM NaCl) increased chlorophyll concentration and oxygen production in embryos of maturing seeds. At 500 mM NaCl, the phosphatidylglycerol and sulfoquinovosyldiacylglycerol levels and the digalactosyldiacylglycerol/monogalactosyldiacylglycerol ratio were higher in young seedlings derived from seeds whose source plants were cultured in 500 mM rather than in 1 mM NaCl. When seeds were incubated with 600 mM NaCl, the conductivity and malondialdehyde concentration in the embryos was greater if the source plants had been cultured in 1 mM rather than in 500 mM NaCl. The opposite pattern was evident for seedling survival and shoot weight. In conclusion, salinity during seed maturation may increase the salt tolerance of seeds and seedlings by increasing the oxygen production in the embryos of the maturing seeds and by changing the lipid composition of membranes in the seedlings.     

Effects of salinity and nitrate on production and germination of dimorphic seeds applied both through the mother plant and exogenously during germination in Suaeda salsa

Salinity and nitrogen are two important environmental factors that affect the distribution of halophytes in their natural saline habitats. Seeds of the euhalophyte Suaeda salsa L. were harvested from plants that had been treated with 1 or 500 mm NaCl combined with 0.5 or 5 mm NO₃^?‐N (nitrate) for 115 days in a glasshouse. Germination was evaluated under different concentrations of NaCl and nitrate. Plants exposed to high salinity (500 mm ) and low nitrate (0.5 mm ) tended to produce heavy seeds. Either high salinity (500 mm ) or high nitrate (5 mm ) increased the brown/black seed ratio. The concentrations of Na⁺, K⁺, and Cl^? were higher in brown than in black seeds, and NO₃^? concentrations were higher in black than in brown seeds, regardless of NaCl and nitrate treatments during plant culture. Regardless of NaCl and nitrate concentrations during germination, seeds from plants grown with 0.5 mm nitrate generally germinated more rapidly than seeds from plants grown with 5 mm nitrate, and the difference was greater for black than for brown seeds. Exogenous nitrate during germination enhanced the germination of brown seeds less than that of black seeds. Producing more brown seeds and heavy black or brown seeds appears to be an adaptation of S. suaeda to saline environments. Producing more black seeds, which tend to remain dormant, should reduce competition for nitrogen and appears to be an adaptation to nitrogen‐limited environments. In conclusion, nitrate provided exogenously or by mother plants to black seeds may act as a signal molecule that enhances the germination of black S. suaeda seeds.     

盐旱互作对不同生境盐地碱蓬种子萌发和幼苗生长的影响

研究了盐旱互作对潮间带和盐碱地生境盐地碱蓬棕色种子萌发、地上部生长和离子积累的影响。不同盐浓度预处理后,未萌发的种子风干后复水,其萌发率与对照相比没有降低,说明两种生境盐地碱蓬种子萌发期间都耐干湿交替。400mmol/LNaCl溶液浇灌的潮间带生境盐地碱蓬幼苗在第3次干旱处理后萎蔫幼苗的百分比高于盐碱地生境的,而复水后正常幼苗的百分比却相反。400mmol/LNaCl溶液处理下,第3次干旱处理复水后,盐碱地生境盐地碱蓬幼苗地上部离子含量(主要是Na+和Cl-)高于潮间带生境的。表明在干旱情况下,盐碱地生境盐地碱蓬幼苗能积累更多的无机离子,降低渗透势,提高根系吸收水分的能力。上述结果说明,盐碱地生境盐地碱蓬幼苗较潮间带生境盐地碱蓬幼苗更耐盐与干旱的交互作用。     

NaCl胁迫下外源NO供体硝普钠（SNP）对盐地碱蓬种子萌发的影响

用添加与不添加0．1mm01．L^-1NO供体硝普钠（sNP）的800mmol．L^-1NaCl溶液处理盐地碱蓬种子后,800mmol·L^-1NaCl处理下盐地碱蓬种子的萌发率、含水量和吸水速率显著增加,胚中脯氨酸的含量降低,但对Na^＋、K^＋和可溶性糖含量无显著影响。表明0．1mmol．L^-1SNP缓解800mmol．L^-1NaCl对盐地碱蓬种子萌发抑制的主要原因是盐地碱蓬种子含水量的提高,从而缓解了盐的渗透胁迫。     

盐胁迫下盐地碱蓬体内无机离子含量分布特点的研究

用不同浓度NaCl溶液处理盐地碱蓬（Suaeda salsa)植株后,测定并比较老叶、幼叶及根部的无机离子含量和对K的选择性,叶片及根部的Na^ 、Cl^-含量随盐度的增加而升高,且累积趋势相似,盐胁迫下根部Na^ 、Cl^-及总离子含量（K^ 、Na^ 、Ca^2 ,NO3^-,Cl^-)明显低于叶片,说明盐地碱蓬地盐胁迫下,以叶片优先积累大量离子（如Na^ ,Cl^-) 为其适应特征。NaCl处理下,叶片的K^ ,Ca^2 含量低于对照,但随盐度的增加保持相对稳定,而根部K^ 含量,K/Na比、对K的选择性则高于叶片,这对盐胁迫下地上部的K^ 亏缺有一定补偿作用。低盐度处理（100mmol/LNaCl)促进NO3^-的吸收,另外随盐度的增加,叶片渗透势下降,渗透调节能力增强,幼叶渗透势低于老叶,但渗透调节能力相同。     

重金属复合胁迫下碱蓬萌发生长及富集特征

为探究重金属复合胁迫对金川镍铜矿区广布植物碱蓬Suaeda salsa(L.)Pall.的影响,根据当地环境条件及预实验结果设置胁迫梯度,测定分析重金属胁迫下碱蓬种子萌发和芽期生理指标,并从野外站台取样研究碱蓬重金属富集能力。结果表明:无论是单一胁迫还是低浓度复合胁迫(Cu20和Ni20复合),发芽期碱蓬的生长均呈现"低促高抑"的趋势,即低浓度(≤40 mg/L)时促进碱蓬生长,高浓度(≥80 mg/L)时抑制碱蓬生长;高浓度复合胁迫(Cu320和Ni320复合)下均抑制碱蓬的生长。MDA(丙二醛)的含量随胁迫浓度的增加而增加;胁迫组可溶性蛋白和游离脯氨酸含量整体上高于对照组;单一胁迫下POD(过氧化物酶)活性随胁迫浓度增加而增加;复合胁迫下低浓度提高POD活性,高浓度抑制POD活性;碱蓬叶片的平均转移系数(TF)大于茎部,且平均转移系数大于1.00;碱蓬叶片的富集系数(BCF)大于根部大于茎部。碱蓬对Cu和Ni均具有很高的耐受性,但对于Cu的耐性强于Ni;低浓度时Cu、Ni复合胁迫对碱蓬生长的促进作用强于Cu、Ni单一胁迫,高浓度时则相反;碱蓬具有较高的重金属富集和转移能力;在当前矿区土壤环境背景下,碱蓬可以作为矿区生态恢复和重金属污染修复的备选植物。     

Ecological thresholds of Suaeda salsa to the environmental gradients of water table depth and soil salinity

The responses of Suaeda salsa to the environmental gradients of water table depth and soil salinity in the Yellow River Delta, China were analyzed from the aspect of ecological thresholds which were developed from the Gaussian model. Based on the correlation analysis of population biomass, density, height, coverage and abundance of Suaeda salsa, population biomass was selected as the population index for further analysis. The results indicated that the optimum water table depth for the growth of Suaeda salsa was about ?0.42 m, the ecological thresholds were from ?0.92 m to 0.08 m, and the optimum ecological thresholds were from ?0.67 m to ?0.17 m. To the soil salinity gradient, the optimum was about 12.71 g/kg, the ecological thresholds were from 5.17 g/kg to 20.25 g/kg, and the optimum ecological thresholds were from 8.94 g/kg to 16.48 g/kg. However, the effect of water-salinity interaction seemed to be important to the growth of Suaeda salsa, which was discussed through analyzing the water table depth-soil salinity relationship and their interactions. By using Ward cluster analysis and Gamma distances, 69 sampling sites were classified into 7 kinds of Suaeda salsa communities. It was found that there was a remarkable response of the community structure of Suaeda salsa to the water table depth and soil salinity gradients, which can be a switchover from xeromorphic and saline-alkali plants to limnophytes, and vice versa.     

盐地碱蓬过氧化氢酶基因的克隆及盐胁迫下的表达分析

过氧化氢酶是清除H2O2的重要酶类,从400mmol/LNaCl处理的盐地碱蓬（Suaeda salsa(L.)Pall)地上部分的cDNA文库中克隆了两个编码过氧化氢酶的cDNA(Sscat1和Sscat2)。其中Sscatl(1.7kb)是一个全长cDNA克隆,编码一个492个氨基酸的开放阅读框架。而Sscat2(1.1kb)是一个cDNA片段。据编码Sscatl3′端的287个氨基酸的cDNA序列与Sscat2的cDNA序列进行的BLAST同源性分析表明,Sscat1和Sscat2在核苷酸水平的一致性则为一个单拷贝基因。Northern杂交结果表明在盐胁迫条件下Sscat1和Sscat2的表达存在差异;400mmol/LNaCl处理48h的盐地碱蓬根中的Sscat1和Sscat2mRNA水平比对照显著提高,但是在叶中仅Sscat1受盐诱导表达,不同盐处理时间下的表达分析也证实。在盐地碱蓬叶中仅Sscat1受盐诱导表达。这说明Sscat1和Sscat2在盐地碱蓬中是差异调控的,生理分析表明过氧化氢酶的活性在盐胁迫条件下显著提高。     

盐地碱蓬过氧化氢酶基因的克隆及盐胁迫下的表达分析

过氧化氢酶是清除H2O2的重要酶类.从400 mmol/L NaCl处理的盐地碱蓬(Suaeda salsa(L.)Pall)地上部分的cDNA文库中克隆了两个编码过氧化氢酶的cDNA(Sscat1和Sscat2),其中Sscat1(1.7kb)是一个全长cDNA克隆,编码一个492个氨基酸的开放阅读框架,而Sscat2(1.1kb)是一个cDNA片段.据编码Sscat1 3＇端的287个氨基酸的cDNA序列与Sscat2的cDNA序列进行的BLAST同源性分析表明,Sscat1和Sscat2在核苷酸水平的一致性为71.9%,在氨基酸水平上的一致性为75%.Southem杂交表明,Sscat1在盐地碱蓬基因组中为多拷贝基因,Sscat2则为一个单拷贝基因.Northern杂交结果表明在盐胁迫条件下Sscat1和Sscat2的表达存在差异:400 mmol/L NaCl处理48h的盐地碱蓬根中的Sscat1和Sscat2 mRNA水平比对照显著提高,但是在叶中仅Sscatl受盐诱导表达.不同盐处理时间下的表达分析也证实,在盐地碱蓬叶中仅Sscat1受盐诱导表达.这说明Sscat1和Sscat2在盐地碱蓬中是差异调控的.生理分析表明过氧化氢酶的活性在盐胁迫条件下显著提高.     

水盐环境梯度下翅碱蓬(Suaeda salsa)的生态阈值

在对翅碱蓬(Suaeda salsa)生物量、密度、株高、盖度、多度相关性分析的基础上选取了生物量作为翅碱蓬生物指标,利用高斯模型分析了黄河三角洲翅碱蓬种群沿水深、土壤盐分的生态阈值,翅碱蓬最适水深为-0.42m,水深生态阈值区间为[-0 92,0.08](m),水深最适生态阈值区间为[-0.67,-0.17](m);最适土壤盐分为12.71 g/kg左右,其盐分生态阈值区间为[5 17,20.25](g/kg),盐分最适生态阈值区间为[8.94,16.48](g/kg).通过分析不同实验区的水盐关系及其交互作用,探讨了水盐交互作用对翅碱蓬生长的影响.最后,通过离差平方和聚类分析,将3个实验区69个样地划分为7类.随着水深和盐分的梯度变化,7类样地的翅碱蓬群落呈现明显的演替.     

The role of the seed coat in adaptation of dimorphic seeds of the euhalophyte Suaeda salsa to salinity

The role of the seed coat in adaptation of dimorphic seeds of the euhalophyte Suaeda salsa to salinity was investigated during germination and early seedling growth. Black and brown seeds were treated with chloroform for 1 min before the extract was used to analyze waxes and the seeds to investigate the protective role of the seed coat under saline conditions. Waxes in black seed coats were more abundant than those in brown seed coats. Salinity (500 mM NaCl) increased the concentration of Na⁺ and decreased the concentration of K⁺ in both black and brown seeds regardless of chloroform treatment. Chloroform treatment alone (in the absence of NaCl) had no effect on the concentration of Na⁺ or K⁺ in black or brown seeds and in the presence of 500 mM NaCl had no effect on the concentration of Na⁺ or K⁺ in brown seeds. However, chloroform treatment increased Na⁺ and decreased K⁺ in black seeds with 500 mM NaCl. A change of MDA (malondialdehyde) concentration in black and brown seeds treated with or without chloroform was similar to the change of Na⁺ concentration. High salinity (1500 mM NaCl) pretreatment for 40 days had a less adverse effect on germination of black seeds compared with brown seeds after they were transferred to fresh water regardless of chloroform treatment. Similar results were found for seedling emergence. In conclusion, a black seed coat may be more protective than a brown seed coat, probably by shielding the embryo from ion toxicity, because of its higher content of waxes. Thus black seeds can better maintain seed viability than brown seeds for extended periods under hypersaline conditions.     

盐地碱蓬(Suaeda salsa)APX 基因的克隆及盐胁迫下的表达

从盐地碱蓬 (Suaedasalsa)中克隆了抗坏血酸过氧化物酶 (ascorbateperoxidase ,APX)的全长cDNA(SsAPX) ,基因注册号为AY0 34 893。SsAPX全长 1.1kb ,推导的氨基酸序列长为 2 5 0个氨基酸残基。BLAST同源性分析表明 ,该cDNA与已报告的菠菜(Spinaciaoleracea)细胞质抗坏血酸过氧化物酶基因同源性最高 ,在核苷酸水平上一致性为 87% ,在氨基酸水平上一致性为 89%。Southern杂交表明APX基因在盐地碱蓬基因组中只有 1个拷贝。盐 (NaCl 40 0mmol/L)处理不同时间后的Northern杂交分析表明盐地碱蓬中SsAPX基因在盐胁迫下表达量增加 ,而且在盐胁迫下抗坏血酸过氧化物酶的活性也显著地增加 ,说明该基因受盐诱导。推测抗坏血酸过氧化物酶可能在保护盐地碱蓬免受氧化损伤的过程中起到一定作用     

盐地碱蓬(Suaeda salsa)中SsINPS基因的分子克隆及表达特性分析

肌醇 1 磷酸 (I 1 P)合成酶 (EC5 .5 .1 .4,INPS)是肌醇生物合成中的关键酶 ,催化葡萄糖 6 磷酸 (G 6 P)到I 1 P的反应。从该实验室已构建的NaCl40 0mmol/L处理的盐地碱蓬 (Suaedasal sa)cDNA文库中克隆了肌醇 1 磷酸合成酶的全长cDNA (S .salsamyo inositol 1 phosphatesynthase,SsINPS) ,基因注册号为AF43 3 879。SsINPS全长约 1 986bp ,含有开放式阅读框架 1 5 3 0bp ,3′和 5′的非翻译区分别为 1 3 9bp和 3 1 7bp ;推导的氨基酸序列全长 5 1 0个氨基酸残基 ,分子量约为 5 6 .7kD ,pI值为 5 .3 5。BLAST同源性分析表明 ,该cDNA与已报告的冰叶日中花 (Mesembryanthemumcrys tallinum)的INPS基因同源性最高 ,其中 ,核苷酸水平的同源性为 91 % ,氨基酸水平上的同源性为84%。以SsINPS全长cDNA为探针进行的South ern杂交结果表明 ,SsINPS基因在盐地碱蓬基因组中只有一个拷贝 ;Northern结果表明 ,在盐处理(40 0mmol/L的NaCl)下 ,SsINPS在叶中的表达量有显著的增加。从而说明SsINPS在盐胁迫下是上升调节的     

硫腺苷甲硫氨酸合成酶基因的克隆及其在盐胁迫条件下的不同表达

硫腺苷甲硫氨酸作为甲基供体在转甲基反应中起到重要作用.为了解硫腺苷甲硫氨酸在盐地碱蓬(Suaedasalsa (L.)Pall)耐盐中的作用,我们对可能编码硫腺苷甲硫氨酸合成酶的基因(SsSAMS2)进行了分析.该基因在经400 mmol/L NaCl处理的盐地碱蓬地上部分的λ-Zap cDNA文库中克隆到,其插入片段全长1 531 bp,包含一个395个氨基酸的开放阅读框架,该基因推断的分子量约为43 kD.SsSAMS2与长春花(Catharanthus roseus)的SAMS2在氨基酸水平上的一致性为93%.Southern杂交显示,SsSAMS2在盐地碱蓬基因组中可能是两个拷贝.Northern分析显示硫腺苷甲硫氨酸合成酶基因受NaCl等胁迫的正调控.酶活性检测表明,NaCl胁迫条件下该酶活性增强.     

盐地碱蓬GST基因的克隆、序列分析及其表达特征

从盐地碱蓬 (Suaedasalsa)幼苗的cDNA文库中克隆到一个 0 .9kb的全长cDNA ,同源性分析表明该全长cDNA与已报告的大豆 (Glycinemax)GST基因相应序列的同源性达 5 5 % ,可能编码由 2 35个氨基酸组成的谷胱甘肽转移酶 (glutathioneS transferase ,GST)。Southern杂交结果证明GST基因在碱蓬基因组中可能有至少两个以上的拷贝 ;Northern杂交结果表明 ,4 0 0mmol/L的NaCl处理 4 8h ,幼叶中GSTmRNA的表达量是对照的 2～ 3倍 ,说明碱蓬中GST基因受盐诱导     

黄河口滨岸潮滩不同生境下翅碱蓬硫元素的季节变化

2008年5-11月, 对黄河口滨岸潮滩不同生境下翅碱蓬(Suaeda salsa)硫(S)的季节变化特征进行了研究。研究表明: 中潮滩翅碱蓬(JP1)和低潮滩翅碱蓬(JP2)各器官生物量均具有明显的季节变化特征, 总体表现为JP1 > JP2; JP1和JP2地上与地下部分比值的变化较为一致, 整体表现为JP2 > JP1; 二者枯落物量呈递增变化; JP1和JP2叶、茎和枯落物中的全硫(TS)含量在生长季波动变化明显, 整体呈先增后减变化, 而根中的TS含量在生长季呈递减变化, 符合指数衰减模型; 二者不同器官及枯落物的TS累积量和S累积速率(V_S)季节变化明显, JP1地上部分的TS累积量和V_S明显高于JP2, 且二者地上部分的TS累积量和V_S均明显高于地下; JP1和JP2不同部分的S分配比差异明显, 其中叶的分配比最高, 分别为(38.34 ± 16.19)%和(66.27 ± 12.09)%, 说明叶是翅碱蓬重要的S累积器官。结果显示, 翅碱蓬的生态学特性和其所处生境的水盐状况对JP1和JP2生物量、TS含量、累积量、累积速率、分配比均具有重要影响。     

硫腺苷甲硫氨酸合成酶基因的克隆及其在盐胁迫条件下的不同表达

硫腺苷甲硫氨酸作为甲基供体在转甲基反应中起到重要作用。为了解硫腺苷甲硫氨酸在盐地碱蓬(Suaeda salsa (L.) Pall)耐盐中的作用,我们对可能编码硫腺苷甲硫氨酸合成酶的基因(SsSAMS2)进行了分析.该基因在经400mmol/L NaCl处理的盐地碱蓬地上部分的λ-Zap cDNA文库中克隆到,其播入片段全长1531bp,包含一个395个氨基酸的开放阅读框架,该基因推断的分子量约为43kD.SsSAMS2与长春花(Catharanthus roseus)的SAMS2在氨基酸水平上的一致性为93%.Southern杂交显示,SsSAMS2在盐地碱蓬基因组中可能是两个拷贝.Northern分析显示硫腺苷甲硫氨酸合成酶基因受NaCl等胁迫的正调控.酶活性检测表明,NaCl胁迫条件下该酶活性增强.     

NaCl胁迫对2种表型盐地碱蓬种子萌发的渗透效应和离子效应研究

采用高低2个浓度的NaCl、LiCl及等渗甘露醇溶液处理紫红色表型(紫色型)和绿色表型(绿色型)盐地碱蓬种子,通过测定它们的种子萌发率、吸胀速率和胚内离子含量,研究NaCl胁迫对2种表型种子萌发的离子效应和渗透效应.结果表明:(1)2种表型盐地碱蓬种子萌发率在高浓度(300 mmol/L)和低浓度(100 mmol/L)NaCl处理下均显著降低,紫色型种子萌发率在低浓度下显著低于绿色型,而在高浓度下却显著高于绿色型;绿色型种子萌发率在高浓度(30 mmol/L)和低浓度(10 mmol/L)LiCl处理下均未受到显著影响,但紫色型种子萌发率却均极显著降低;2种表型盐地碱蓬种子萌发率在低浓度等渗甘露醇处理下均极显著低于低浓度NaCl处理,而高浓度等渗甘露醇处理却均与高浓度NaCl处理无显著差异.(2)2表型种盐地碱蓬种子的吸胀速率在低浓度NaCl处理下没有受到显著影响,但高浓度NaCl处理及与之等渗的高浓度甘露醇处理下都显著降低,而且紫色型种子的吸胀速率在等渗甘露醇处理时显著高于绿色型.(3)2种表型盐地碱蓬种子胚中的Na 含量和Na /K 在对照和低浓度NaCl处理下无显著差异,但紫色型种子胚中的Na 、K 含量在高浓度NaCl处理时都显著高于对照,且K 含量增加的幅度远大于Na 含量,导致紫色型种子胚中的Na /K 显著低于绿色型.研究发现,盐地碱蓬种子萌发在低浓度NaCl胁迫下主要受离子效应抑制,而高浓度NaCl胁迫下则主要受渗透效应抑制,紫色型种子萌发率在高浓度NaCl胁迫下高于绿色型的原因之一是前者能维持更低的Na /K 比.     

盐胁迫下盐地碱蓬叶片液泡膜H+-ATPase H 亚基的克隆与表达分析

利用EST随机挑取克隆测序的方法从盐地碱蓬叶片cDNA文库中分离得到了盐地碱蓬V-H -ATPase H亚基cDNA序列,并进行了H、c亚基基因表达及V-H -ATPase活性分析.结果表明,H亚基基因全长1 969 bp,包括100 bp的5′-非编码区和471 bp的3′-非编码区.开放阅读框为1 398 bp,编码465个氨基酸残基,分子量约52.8 kD.N orthern杂交分析表明盐胁迫明显诱导了H亚基表达,而且盐胁迫下H、c亚基及V-H -ATPase活性存在协同作用.这些结果表明盐胁迫下H和c亚基基因上调及V-H -ATPase活性的增加为N a 区隔化到液泡中提供了质子驱动力.     

翅碱蓬对镉的耐性及吸收特性的研究

重金属Cd具有高毒性,对生物体危害较大.盘锦红海滩优势物种翅碱蓬能够吸收一定量的重金属,因此可用于湿地滩涂污染的原位修复.通过水培的方法,以翅碱蓬为研究对象,测定不同浓度Cd胁迫下翅碱蓬的根长、苗高、鲜重、叶绿素含量、及根、茎、叶内Cd的含量,探讨Cd对于翅碱蓬生长发育的影响以及对Cd的吸收规律.结果表明:低浓度Cd对...