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。     

盐胁迫对翅碱蓬生长和渗透调节物质浓度的影响

盐胁迫环境抑制植物的生长,不同的盐分浓度对植物的抑制伤害存在差异.本文探讨了翅碱蓬(Suaeda heteroptera Kitagawa)对盐生环境的生理生态适应机制,研究其在不同盐浓度下,株高、根长、鲜重、干重等生长指标以及相对含水量、游离脯氨酸、可溶性糖和蛋白质含量的变化情况.结果表明:盐分对翅碱蓬幼苗的抑制作用...     

贮存时间对盐地碱蓬种子萌发及生长特征的影响

研究黄河三角洲滨海湿地不同贮存年限盐地碱蓬种子的萌发及植株生长特征,以期为利用土壤种子库进行滨海湿地盐地碱蓬群落的恢复提供科学数据。种子萌发实验结果显示：随着种子贮存年限的增加,盐地碱蓬种子活力呈明显下降趋势(P<0.05),表现为种子发芽率和发芽速度的下降;在同一盐度条件下,盐地碱蓬种子发芽率、发芽速度随贮存年限增加呈线性下降趋势;同一年限的种子随盐度的增加,发芽率呈线性下降趋势。植株生长实验结果显示：贮存时间短的种子出苗率高于贮存时间久的种子;随着贮存时间的增加,盐地碱蓬植株的密度呈下降趋势,但植株高度、含水量及单株鲜重及干重无明显差异,而单位面积生物总量差异显著(P<0.05)。在未来滨海湿地盐地碱蓬群落的修复过程中,应该根据滨海湿地的退化时间制定盐地碱蓬群落的恢复方案。研究结果可为滨海湿地生态恢复提供理论支持和科学参考,以实现滨海湿地恢复过程中降低恢复成本、提高恢复效率、加快恢复进程的目标。     

盐度、氮、磷含量对翅碱蓬生长的影响

目的：探明盐度、氮、磷含量对翅碱蓬生长的影响。方法：采用盆栽方法,控制土壤的盐度、氮、磷浓度,设计正交试验,研究在不同水平的盐度、氮含量和磷含量条件下翅碱蓬的发芽和生长状况。使用三因素方差法对试验结果进行显著性分析。结果：盐度显著影响翅碱蓬种子萌发和苗重;盐度和氮含量显著影响翅碱蓬苗高、叶绿素积累及对氮磷的吸收;磷含量影响不显著。结论：盐度和氮含量是影响翅碱蓬生长的关键因子,低盐环境有利于翅碱蓬生长,氮含量增加促进翅碱蓬的生长。磷含量对翅碱蓬生长影响不显著。     

盐生植物翅碱蓬的内生真菌多样性分析

【目的】对辽宁省盘锦市双台子河口国家自然保护区的盐生植物翅碱蓬(Suaedaheteroptera Kitag.)的内生真菌进行研究。【方法】利用传统分离方法对翅碱蓬的根、茎、叶组织进行分离和纯化,根据形态学和理化特征,并结合rDNA-ITS序列分析对菌种进行鉴定。【结果】共分离得到内生真菌49株,这些内生真菌分属于13个属,其中小丛壳菌属(Glomerella)、链格孢属(Alternaria)、毛盘孢属(Colletotrichum)和枝孢属(Cladosporium)为优势菌属。翅碱蓬的内生真菌分布具有一定的组织特异性,两种不同生境中的翅碱蓬内生真菌群落丰富度差异较小,相似度较高,但在某些种属上存在特异性。【结论】翅碱蓬体内含有丰富的内生真菌资源,其内生真菌具有很高的宿主特异性,而且其分布受生境影响。     

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...     

辽宁盘锦三角洲翅碱蓬根系及内生细菌群落多样性

【目的】翅碱蓬(Suaeda heteroptera)是一种典型的盐碱指示物,对重金属和石油污染的盐碱土壤有一定的修复作用,但是关于翅碱蓬根系与根系内生微生物之间的关系、微生物的多样性以及根系微生物在生物修复中所起作用的研究较少。本文以盘锦"红海滩"的翅碱蓬为例,研究翅碱蓬根系及根系内生细菌菌群种类和结构。【方法】通过传统的培养方法和非培养的高通量测序方法对翅碱蓬根系土壤样品、根系附着物样品以及根系匀浆样品中微生物群落多样性进行分析。【结果】传统方法共分离得到67株细菌,选择代表菌28株,根据其形态特征和生理生化特征,结合16S r RNA基因序列比对进行鉴定,它们分别属于盐单胞菌属(Halomonas)、海细菌属(Marinobacterium)、芽孢杆菌属(Bacillus)、副球菌属(Paracoccus)、假单胞菌属(Pseudomonas)、游动球菌属(Planococlus)、沙雷氏菌属(Serratia)、刘志恒菌属(Zhihengliuella)等。利用高通量测序技术对样品进行多样性分析,其中根系附着物样品的菌群丰度和多样性最高,依次分别为根系土壤样品和根系匀浆样品。3个样品中有效序列群落结构可分为12个门,包括酸杆菌门(Acidobacteria)、放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)、绿菌门(Chlorobi)、绿弯菌门(Chloroflexi)、蓝菌门(Cyanobacteria)、厚壁菌门(Firmicutes)、芽单胞菌门(Gemmatimonadetes)、浮霉菌门(Planctomycetes)、变形菌门(Proteobacteria)、螺旋体门(Spirochaetes)和疣微菌门(Verrucomicrobia)。根系匀浆样品中蓝菌门为优势门类,占整个菌群的42%,变形菌门为次优势类群,占33%。变形菌门在根系附着物样品中为优势门类,占46%,拟杆菌门为次优势门类,占16%。根系土壤样品中拟杆菌门为优势门类,占整个菌群的37%,次优势类群为变形菌门,占20%。【结论】翅碱蓬根系和内生菌具有丰富的多样性,其根系微生物可能会在重金属和石油污染土壤的生物修复中起一定的修复作用。     

辽宁碱蓬SlCMO基因盐胁迫表达分析及盐诱导启动子鉴定

胆碱单加氧酶（choline monooxygenase, CMO）是合成甜菜碱的关键酶,甜菜碱在植物抵抗渗透胁迫中起着重要的作用。本研究室前期克隆了盐生植物辽宁碱蓬CMO（Suaeda liaotungensis CMO）基因及启动子。本研究对SlCMO基因在盐胁迫下的表达及盐诱导启动子进行分析。qRT-PCR分析SlCMO基因在辽宁碱蓬不同器官及盐胁迫下的表达,结果表明,SlCMO基因在根、茎、叶中均有表达,其中茎、叶中的表达量较高,SlCMO基因在根、茎、叶中的表达均受盐胁迫诱导。5′端缺失分析SlCMO启动子的盐诱导区段,结果表明,pC5（-267~+128 bp）是SlCMO启动子的盐诱导功能区段,推测pC5调控SlCMO 基因的盐诱导表达。本研究为SlCMO 基因表达调控研究奠定基础,也为植物抗盐基因工程提供可用的启动子。     

翅碱蓬氨基酸、蛋白质和脂肪酸成分的研究

翅碱蓬(Suaeda heteroptera)是广泛分布于我国北方滨海盐碱地的藜科植物。到目前为止,对翅碱蓬的有用化学成分和生理生化特性还没有进行过仔细的研究。仅报道过种子含油量测定的结果。因此对翅碱蓬的开发和利用缺少足够的资料。我们从分析种子和茎叶蛋白质、氨基酸和脂肪酸成分入手,探讨翅碱蓬种子作为人类食品,茎叶作为牲畜饲料的可能性。     

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。     

不同生境盐地碱蓬对氮饥饿的响应

为了探讨不同生境盐地碱蓬对低氮生境的适应机制,测定了盐渍环境下(200 mmol/L Na Cl)不同浓度硝态氮(0.3、5 mmol/L NO~-_3-N)预处理两种生境盐地碱蓬经氮饥饿后的NO~-_3含量、硝酸还原酶(NR)活性、光合特性及生长状况。结果表明,0.3和5 mmol/L NO~-_3-N处理以及进行氮饥饿时,潮间带生境盐地碱蓬叶片NO~-_3含量均高于内陆生境盐地碱蓬。与内陆生境盐地碱蓬相比,氮饥饿后,潮间带生境盐地碱蓬叶绿素含量、NR活性和光合放氧速率下降幅度均小于内陆生境盐地碱蓬,在0.3mmol/L NO~-_3-N预处理进行氮饥饿时趋势更加明显。0.3 mmol/L NO~-_3-N预处理后氮饥饿对潮间带生境盐地碱蓬根冠比没有影响,却降低内陆生境盐地碱蓬根冠比。上述结果表明,低氮条件下潮间带生境盐地碱蓬具有较高的NO~-_3储存能力,在环境持续氮素缺乏时具有较高的NO~-_3-N再利用能力,能更好地维持氮代谢以及光合性能。说明潮间带生境盐地碱蓬能更好地适应低氮生境。     

The combined effect of salt stress and heat shock on proteome profiling in Suaeda salsa

Under natural conditions or in the field, plants are often subjected to a combination of different stresses such as salt stress and heat shock. Although salt stress and heat shock have been extensively studied, little is known about how their combination affects plants. We used proteomics, coupled with physiological measurements, to investigate the effect of salt stress, heat shock, and their combination on Suaeda salsa plants. A combination of salt stress and heat shock resulted in suppression of CO₂ assimilation and the photosystem II efficiency. Approximately 440 protein spots changed their expression levels upon salt stress, heat shock and their combination, and 57 proteins were identified by MS. These proteins were classified into several categories including disease/defense, photosynthesis, energy production, material transport, and signal transduction. Some proteins induced during salt stress, e.g. choline monooxygenase, chloroplastic ATP synthase subunit beta, and V-type proton ATPase catalytic subunit A, and some proteins induced during heat shock, e.g. heat shock 70 kDa protein, probable ion channel DMI1, and two component sensor histidine kinase, were either unchanged or suppressed during a combination of salt stress and heat shock. In contrast, the expression of some proteins, including nucleoside diphosphate kinase 1, chlorophyll a/b binding protein, and ABC transporter I family member 1, was specifically induced during a combination of salt stress and heat shock. The potential roles of the stress-responsive proteins are discussed.     

黄河口不同氮基质碱蓬种子萌发及幼苗生长对盐分及氮输入的响应

2014年4-11月,选择黄河入海口北部滨岸高潮滩的碱蓬湿地为研究对象,基于野外原位氮负荷增强模拟试验(N0,无额外氮输入;N1,低氮输入;N2,中氮输入;N3,高氮输入),获取相应的不同氮基质种子(S0,S1,S2和S3),以研究其发芽率以及幼苗生长状况对不同盐分胁迫和氮浓度交互作用的响应。结果表明,不同氮负荷影响下碱蓬成熟种子中的氮含量整体表现为S2S0S1S3,中氮输入更利于种子中氮养分的累积。盐分和氮交互作用下4种氮基质种子的发芽率总体表现为S2S1S0S3(P0.05),S2在不同盐分胁迫下的发芽率最高,幼苗的生长状况也最好。随着盐分的增加,4种氮基质种子的发芽率及幼苗生长状况均受到一定程度的抑制,但较低的盐分有助于其幼苗长度的增长,且随着氮输入量的增加这种抑制作用可得到一定程度缓解。盐分胁迫、氮浓度和种子类型作为单独因素出现时对碱蓬的发芽率、幼苗长度、鲜重和干重均产生显著影响,除幼苗长度受氮浓度和盐分胁迫交互作用的影响达到显著水平外(P0.05),其他因子交互作用对诸生态指标的影响并不明显。研究发现,不同氮输入处理不仅改变了原生环境碱蓬种子的氮含量,而且也使这些具备不同氮基质的种子对不同盐分胁迫与氮浓度环境具有不同的生态适应对策,中氮输入下的碱蓬种子(S2)无论在萌发率还是在幼苗生长状况上均优于其他氮基质的种子。未来,随着黄河口新生湿地氮养分供给的不断增加,当湿地氮养分达到中氮水平时,将更有利于碱蓬种子的萌发以及幼苗的生长,当氮养分达到更高水平时,碱蓬种子的萌发以及幼苗生长可能会受到一定程度的抑制。     

盐地碱蓬二型性种子的萌发和休眠及生态适应特性

二型性种子可能增强植物(尤其是盐生植物)应对难于预测环境变化的能力。该研究以运城盐湖盐地碱蓬(Suaeda salsa)二型性种子(黑色和棕色)为材料,采用室内观察和萌发方法对两种种子的形态、休眠和萌发特性进行比较,以揭示盐生植物二型性种子对盐渍化环境的适应对策。结果表明:(1)黑色种子体积小,种皮角质有光泽,而棕色种子体积大,种皮膜质无光泽。(2)新成熟的棕色种子在实验温度范围内萌发率均超过90%,而黑色种子仅在15/25℃下萌发最高(但低于85%),并且棕色种子萌发速度较黑色更快。(3)低盐(0.39mol/L)条件下,光照或黑暗对棕色种子萌发没有显著影响;高盐(0.39mol/L)条件下,棕色种子在黑暗中的萌发显著高于光照环境,而黑色种子在光照条件下的萌发率显著高于黑暗;所有温度和光照处理中,2种种子均随盐分浓度的升高而下降,但棕色种子在盐分浓度为0.78mol/L时萌发率仍能够达到30%以上,而黑色种子在盐分浓度大于0.59mol/L时几乎不萌发。(4)棕色种子为非休眠种子,而黑色种子具有非深度生理休眠特性;划破种皮、干储、低温层积和GA3处理均能够提高黑色种子的萌发率。研究发现,盐地碱蓬通过二型性种子休眠和萌发行为上的差异,采用"两头下注"对策提高其对盐渍化异质生境的适应性。     

Diel methane emissions in stands of Spartina alterniflora and Suaeda salsa from a coastal salt marsh

In this study, we aimed to understand the influence of plant type on the monthly variations of diel CH₄ fluxes from Spartina alterniflora and Suaeda salsa of coastal salt marshes at three growth stages (July, August and September). Dissolved CH₄ concentrations in porewater and sediment redox potentials were monitored, as were aboveground plant biomass and stem densities. CH₄ fluxes exhibited clear monthly variations and peaked in September in the S. alterniflora and S. salsa mesocosms. However, no discernible diel variation was observed in the CH₄ flux in the S. salsa mesocosm, probably due to its weak gas transport capacity. By contrast, notable diel variations of CH₄ flux with the peak of 1.42 and 3.67 mg CH₄ m⁻² h⁻¹ at 12:00 and the lowest of 0.75 and 2.11 mg CH₄ m⁻² h⁻¹ at 3:00 or 6:00 were observed in the S. alterniflora mesocosm on 11 August and 11 September, respectively, but not in July mainly due to low plant biomass masking diel variations in the porewater CH₄ concentration. The ratios of the maximum flux to minimum flux over the course of the day in the S. alterniflora mesocosm on 10 July, 11 August and 11 September were 1.28, 1.89 and 1.76, respectively, and corresponding values for porewater CH₄ concentration were 1.31, 1.39 and 1.17, respectively. CH₄ flux significantly correlated with CH₄ concentration in porewater, and both were significantly related to air temperature. These findings indicate that CH₄ production and CH₄ flux at the middle growth stage (August) exhibited greater responses to changes in air temperature, which in turn induced the higher diel variation. The higher diel cycle for CH₄ flux in August than in September was likely due to the higher proportion of CH₄ oxidized during diffusion within the aerenchyma system. Our results suggest that the extent of diel variations in CH₄ flux may have depended on the gas transport capacity of plants, and the highest diel variation occurred at the middle growth stage.     

The effect of plant growth regulators, nitric oxide, nitrate, nitrite and light on the germination of dimorphic seeds of Suaeda salsa under saline conditions

Suaeda salsa, a leaf succulent shrub in the family Chenopodiaceae, is one of the most important halophytes in China. Suaeda salsa produces dimorphic seeds (soft brown seeds and hard black seeds). Seeds of S. salsa were collected from the coastal salt flats near Huanghua City, China. Experiments were conducted to determine the salinity-alleviating effect of plant growth regulators, nitric oxide, nitrate, nitrite and light on the germination of dimorphic seeds of S. salsa. Brown seeds had a higher germination rate than black seeds in all experiments. Black seeds were more sensitive to salt in the absence of light in comparison to brown seeds. Brown seeds absorbed water more quickly in comparison to black seeds and were found to be more tolerant of salt stress. Our results showed that 1-aminocyclopropane-1-carboxylate (ACC, the immediate precursor of ethylene), nitrite, GA₄ and BA improved seed germination in the presence of salt. However, nitrate, GA₁, GA₃ failed to alleviate salt stress. ABA inhibited seed germination and seedling growth. Possible mechanisms involved in the alleviation of salt stress in S. salsa seeds and the ecological adaptation of the seeds to the environment are discussed.     

Specific regulation of SOD isoforms by NaCl and osmotic stress in leaves of the C3 halophyte Suaeda salsa L

The halophyte Suaeda salsa L., exposed to different NaCl concentrations (100 and 400 mmol/L) and polyethylene glycol (isoosomotic to 100 mmol/L NaCl) containing nutrient solutions under normal or K+-deficient conditions for 7 days, was used to study effects of NaCl salinity and osmotic stress on chlorophyll content, chlorophyll fluorescence characteristics, malonedialdehyde (MDA) content, and superoxide dismutase (SOD) isoform activities. Photosynthetic capacity was not decreased by NaCl treatment, indicating that S. salsa possesses an effective antioxidative response system for avoiding oxidative damage. Seven SOD activity bands were detected in S. salsa leaf extracts, including an Mn-SOD and several isoforms of Fe-SOD and CuZn-SOD. It turned out that NaCl salinity and osmotic stress lead to a differential regulation of distinct SOD isoenzymes. This differential regulation is suggested to play a major role in stress tolerance of S. salsa.     

The leaf tonoplast V-H(+)-Atpase activity of a C3 halophyte Suaeda salsa is enhanced by salt stress in a Ca-dependent mode

Suaeda salsa seedlings grown in Hoagland nutrient solution were treated with different concentrations of NaCl combined with two levels of Ca2+ (0 and 20 mmol/L) to study the effect of Ca2+ nutrition on the growth and activity of leaf tonoplast V-H(+)-ATPase. Increase of Ca2+ concentration in the solution markedly increased the relative growth quantity of S. salsa seedlings and Ca2+ and K+ concentration in the leaf cell sap under NaCl stress. The leaf V-H(+)-ATPase activity was significantly increased with increasing NaCl concentration under high Ca2+ application (20 mmol/L), but little changed under Ca2+ starvation (0 mmol/L). Western blot analysis showed that the leaf V-H(+)-ATPase of S. salsa was at least composed of A, B, D and c subunits, and their protein amounts were not affected by NaCl treatments under Ca2+ starvation (0 mmol/ L) with an exception of 100 mmol/L NaCl, but increased under high Ca2+ application (20 mmol/L). There was a positive correlation between activity of V-H(+)-ATPase and the protein amounts of the subunits. The results suggest that Ca2+ nutrition played an important role in the salt tolerance of S. salsa, and that enhancement of V-H(+)-ATPase activity under salt stress was Ca2(+)-dependent.     

Seed germination of the halophyteSuaeda japonica under salt stress

Suaeda japonica Makino belonging to the family Chenopodiaceae, is a halophyte and grows at the shore of Ariake sea in Japan. This plant presumably possesses high salt resistant nature, thus, we examined the mechanisms of seed germination under salt stress. The seeds maintained 80% germination rates on the medium containing 0.7 M NaCl. Germination rates varied depending on salt type; the germination rates under NaCl or KCI exhibited relatively lower values than ones under sodium gluconate or potassium gluconate. This different responses for salts seemed to be as a result of the presence of Cl⁻ ions. Although very high levels of betaine (compatible solute), were kept in the seedlings grown under no salt stress, the contents gradually increased as concentration of NaCl increased. Betaine is a factor present in plants that works to alleviate the effects of excessive soil salts. It is synthesized in leaves from betaine aldehyde, and this process is catabolized by betaine aldehyde dehydrogenase (BADH). When the seedlings were cultivated on the medium without NaCl, relatively high level of BADH activity was found. The activity increased 5-fold in the seedlings grown under 0.5 M NaCl stress. Increases in betaine content and BADH activity were found during seed germination. InS. japonica, the salt stress promoted BADH activity, subsequently endogenous betaine contents were increased, and increased betaine seemed to secure seed germination under salt stress.     

Comparative effects of salt and alkali stresses on growth, osmotic adjustment and ionic balance of an alkali-resistant halophyte Suaeda glauca (Bge.)

Effects of salt and alkali stresses on growth, osmotic adjustment and ionic balance of Suaeda glauca (Bge.), an alkali-resistant succulent halophyte, were compared. The results showed that alkali stress clearly inhibited the growth of S. glauca. Moreover, the concentrations of Na⁺ and K⁺ both increased with increasing salinity under both stresses, suggesting no competitive inhibition between absorptions of Na⁺ and K⁺. The mechanism underlying osmotic adjustment during salt stress was similar to alkali stress in shoots. The shared essential features were that organic acids, betaine and inorganic ions (dominated by Na⁺) mostly accumulated. On the other hand, the mechanisms governing ionic balance under both stresses were different. Under salt stress, S. glauca accumulated organic acids and inorganic anions to maintain the intracellular ionic equilibrium, but the anion contribution of inorganic ions was greater than that of organic acids. However, the concentrations of inorganic anions under alkali stress were significantly lower than those under salt stress of the same intensity, suggesting that alkali stress might inhibit uptake of anions, such as NO₃ ^– and H₂PO₄ ^–. Under alkali stress, organic acids were the dominant factor in maintaining ionic equilibrium. The contribution of organic acids to anions was 74.1%, while that of inorganic anions was only 25.9%. S. glauca enhanced the synthesis of organic acids, dominated by oxalic acid, to compensate for the shortage of inorganic anions.