Salt tolerance conferred by over-expression ofOsNHX1 gene in Poplar 84K

OsNHX1 gene (Na⁺/H⁺ antiporter gene ofOryza sativa L.) was introduced into Poplar 84K withAgrobacterium tumefaciens- mediated transformation. PCR, Southern and Northern blot analysis showed thatOsNHX1 gene was incorporated successfully into the genome of Poplar 84K and expressed in these transgenic plants. Salt tolerance test showed that three lines of transgenic plants grew normally in the presence of 200 mmol/L NaCl, while the Na⁺ content in the leaves of the transgenic plants grown at 200 mmol/L NaCl was significantly higher than that in plants grown at 0 mmol/L NaCl. The osmotic potential in the transgenic plants with high salinity treatment was lower than that of control plants. Our results demonstrate the potential use of these transgenic plants for agricultural use in saline soils.     

OsNHX1基因转化84K杨的研究

采用农杆菌介导的方法开展了水稻Na /H 逆向转运蛋白基因OsNHX1转化84K杨的研究.建立了84K杨的叶外植体高频再生系统,经诱导不定芽及诱导生根阶段卡那霉素连续筛选,获得了大量卡那霉素抗性转化植株.PCR检测和Southern杂交检测表明,OsNHX1基因已成功整合到84K杨基因组.耐盐实验表明,转基因植株能在200 mmol NaCl条件下正常生长.     

转AlNHX1基因大豆后代遗传稳定性及耐盐性分析

为了获得耐盐性有所提高的转AlNHX1基因大豆后代材料,以已获得的转AlNHX1基因的6个株系中的3个株系后代为研究对象,通过分别对这3个株系转基因大豆各后代进行PCR分子检测并结合耐盐性鉴定,以分析外源基因在转基因大豆中遗传稳定性和耐盐性.结果表明:AlNHX1基因在转基因植株的后代中遗传;选取3个株系中部分阳性植株做耐盐性检测,结果表明:转基因大豆耐盐性状均好于野生型大豆.在150mmol/L NaCl胁迫下,转基因大豆叶片中维持了相对较高的K+/Na+比值,相对含水量较野生型提高了9%,而渗透势降低了39%,表明转基因大豆具有较好的吸水和保水能力;在盐胁迫下,超氧化物歧化酶(SOD)与过氧化物酶(POD)活性较野生型大豆分别提高了45%与69%.综上,通过耐盐筛选获得的转AlNHX1基因大豆具有较强的耐盐性.     

NaCl、Na2SO4对盐芥生长和生理胁迫效应的比较

分别用含有0、100、150、200mmol/L NaCl和Na2SO4的Hoagland培养液处理盐芥幼苗,处理一定时间后测定其鲜重、干重、离子含量、质膜透性、Na /K 比值、光合速率等的变化.结果表明:两种盐处理都能抑制盐芥的生长,并且Na2SO4处理比相同浓度的NaCl处理抑制更明显;盐芥肉质化程度降低;盐芥地上部及根部Na 含量远远高于对照,并且盐主要积累在地上部.这些结果说明盐芥不是稀盐盐生植物,也不是拒盐植物,推测其耐盐机制可能建立在基础生理生化的代谢上.     

野大麦在不同逆境下的生理生态特性

通过生态模拟,分析野大麦在碱性盐（Ｎａ２ＣＯ３,１０ｍｍｏｌ／Ｌ）,单纯碱（浓氨水,ｐＨ＝１０）中性盐（ＮａＣｌ,２０ｍｍｏｌ／Ｌ）磷酸中和碱性盐（ｐＨ＝７）聚乙二醇（ＭＷ４０００,３０ｍｍｏｌ／Ｌ）各处理下生物量,细胞膜透性,脯氨酸,柠檬酸,叶绿素,Ｋ＾＋,Ｎａ＾＋等胁变指标变化,结果表明,高ｐＨ是野大麦最大受害因素,其次才是钠离子效应,与前两者相比,渗透效应非常微弱,用磷酸中和降低Ｎａ２Ｏ３的     

CaCl2和Ca（NO3）2对降低玉米幼苗质膜透性的作用

以玉米为实验材料，分别用１５０ｍｍｏｌ／Ｌ，１５０ｍｍｏｌ／Ｌ，ＮａＣｌ＋１５ｍｍｏｌ／Ｌ ＣａＣｌ２及１５０ｍｍｏｌ／ＬＮａＣｌ＝１５ｍｍｏｌ／ＬＣａ（ＮＯ３）２的１／２浓度Ｈｏａｇｌａｎｄ溶液连续处理５天后，分别测定ＭＤＡ含量，质膜透性，植物鲜重，植物干重及植物体内Ｎａ＾＋含量。     

Transformation of the salt-tolerant gene of<Emphasis Type="Italic">Avicennia marina</Emphasis> into tobacco plants and cultivation of salt-tolerant lines

Salt-tolerant gene, CSRG1, which was isolated from a kind of salt-tolerant mangroves, Avicennia marina, constructed the transgenic plasmid, pGAM189/CSRG1. CSRG1, GUS, Kmr and Hyg^r could be transferred into tobacco genome by the ameliorated leaf discs method of agro-bacterium-mediate transformation. Thirteen stable resistant lines were obtained when fifty transgenic explants were selected through 50 mg/L hygromycin and 150 mg/L kanamycin. Assessments of PCR amplification, Southern blot analysis and GUS histochemical staining showed that CSRG1 has been integrated into the genome of the eleven transgenic lines (frequency of transformation was 22%). Northern bolt analysis revealed that CSRG1 had expressed in transgenic lines. The assessments of salt-tolerant ability and photosyn-thetic rates indicated that the survival rate of the transgenic lines is 80%—90% and the transgenic lines could increase by 30%—40% in plant height, even when they were cultivated in MS medium containing 2% NaCl and the total seawater (salinity 24). It is supposed that the special physiologic metabolic pathway formed by the products of CSRG1 can really endow the tobacco plants with the high salt-tolerant ability, not only to Na^ stress, but also to the comprehensive stress of various ions.     

转海蓬子Na+/H+逆向运输蛋白基因番茄的耐盐性研究

以转海蓬子Na /H 逆向运输蛋白基因(Nhap)的番茄(Lycopersicon esculentum Mill.)阳性植株和对照植株为材料,测定叶片相对含水量、质膜透性、K /Na 比值和叶绿素含量4项耐盐指标.结果表明:在盐胁迫条件下,转基因植株的相对含水量、 K /Na 比值和叶绿素含量明显高于对照植株,而叶片质膜透性明显低于对照植株,说明Nhap基因已经在番茄转化植株体内表达,提高了转基因植株的耐盐性.     

水分和盐分胁迫对海滨锦葵生长的效应

分别用含有0、100、200mmol/LNaCl和等渗PEG的Hoagland培养液处理海滨锦葵幼苗,10天后测定植株的生长、水分状况、Na、K离子含量、渗透调节能力及光合特性的变化.结果发现:NaCl和等渗PEG均引起生长的下降,但等渗PEG对海滨锦葵的影响比NaCl的影响大,说明海滨锦葵对盐胁迫的适应能力较强,而对渗透胁迫的适应能力较差.     

NaCl和KCl胁迫对碱蓬根和地上部分生长的效应

一定浓度的NaCl促进碱蓬的生长和地上部分的肉质化，而同浓度的和KCl无此作用，表明碱蓬对Na^ 离子的专一性需求。碱蓬根和地上部分对盐胁迫的反应不同：根比地上部分对高浓度NaCl和KCl胁迫（400mmol/L）反应敏感，受胁迫伤害的程度大。表明高浓度K^ 可能首先伤害根，继而对整株植物产生伤害。     

Molecular characterization of GmNHX2, a Na＋/H＋ antiporter gene homolog from soybean,and its heterolosous expression to improve salt tolerance in Arabidopsis

Na＋/H＋ antiporters have been well documented to enhance plant salt tolerance by regulating cellular ion homeostasis. Here, a putative Na＋/H＋ antiporter gene homolog GmNHX2 from soybean was cloned and predicted to encode a protein of 534 amino acids with 10 putative transmembrane domains. GmNHX2 was expressed in all soybean plant tissues but enriched in roots and its expression was induced by NaCI and polyethylene glycol （PEG） treatments. GmNHX2 exhibits greater sequence similarity with LeNHX2 and AtNHX6 than that of AtNHX1 and AtSOS1. Although phylogenetic analysis clustered GmNHX2 with organellar （tonoplast and vesicles） antiporters, the GmNHX2-EGFP （enhanced green fluorescent protein） fusion protein was possibly localized in the plasma membrane or organelle membrane of transgenic plant cells, Furthermore, transgenic Arabidopsis plants expressing GmNHX2 were more tolerant to high NaCl concentrations during germination and seedling stages when compared with wild-type plants. These results suggest that GmNHX2 is a membrane Na＋/H＋ antiporter and may function to regulate ion homeostasis under salt stress.     

Over-expression of an Arabi-dopsisd δ-OAT gene enhances salt and drought tolerance in transgenic rice

δ-OAT, ornithine-δ-aminotransferase, is the key enzyme involved in proline biosynthesis. In this study the Arabidopsis δ-OAT gene was transferred into rice (Oryza sativa L. ssp japonica cv. Zhongzuo 321), whose successful integration was demonstrated by PCR and Southern blot analysis. The over-expression of the gene in transgenic rice was also confirmed. Biochemical analysis showed that, under salt or drought stress conditions, proline contents in the leaves and roots in transgenic rice plants were 5- to 15-fold of those in non-transgenic controls. Under stress conditions, germinating rate of transgenic lines is higher than that of controls. Although the growth of rice plants tested were more and more retarded with the increasing of NaCI concentration, the transgenic plants grow faster compared to the controls under the same stress condition. Meanwhile, the resistance to KCl and MgSO4 stresses was also found enhanced in transgenie rice. Furthermore, the over-expression of δ-OAT also improved the yield of transgenic plants under stress conditions. The average yield per plant of transgenic lines increases about 12%--41% more than that of control line sunder 0.1 mol/L NaCI stress. These data indicated that the over-expression of δ-OAT, with the accumulation of proline, resulted in the enhancement of salt and drought tolerance and an increase of rice yield, which is of significance in agriculture.     

碱胁迫对宁夏枸杞叶中Na＋，K＋，Ca2＋动态变化的影响

摘要：为探讨宁夏枸杞叶中离子平衡与盐碱胁迫的关系,研究不同浓度的NaHCO3溶液胁迫下,枸杞叶中Na^＋,K^＋,Ca^2＋的浓度变化,同时采用非损伤微测技术研究了枸杞叶中Na^＋,K^＋,Ca^2＋的流速变化．结果表明,在同一时间内（7,14,21d）,Na^＋的浓度随NaHCO。浓度的升高总体呈升高趋势,K^＋和Ca^2＋的浓度总体呈下降趋势,c（Na^＋）／c（Ca^2＋）随NaHCO3浓度的升高而升高;随着时间的变化,各个处理下枸杞叶中Na^＋的浓度总体呈现先降后升的趋势,K^＋的浓度总体呈现下降趋势,Ca^2＋的浓度总体呈现先升后降的趋势,c（Na^＋）／c（K^＋）总体呈现升高趋势,c（Na^2＋）／c（Ca^2＋）总体呈现先降后升的趋势;NaHCO。溶液胁迫7d时,诱导了枸杞叶肉细胞中净Na^＋,K^＋,Ca^2＋外排的增加．碱胁迫下造成c（Na^＋）／c（K^＋）和f（Na^＋）／c（Ca^2＋）升高的原因为,叶片中K^＋和Ca^2＋外排和Na^＋大量积累,这也是枸杞不耐碱的原因之一．可为种植枸杞改良盐碱地提供参考．     

K^＋抑制盐生植物碱蓬生长机理的研究

碱蓬幼苗用０，２５０，５００ｍｍｏｌ／Ｌ的ＮａＣｌ和等浓度的ＫＣｌ处理后，测定其各项生理指标。结果发现，２５０和５００ｍｍｏｌ／Ｌ ＮａＣｌ处理后碱蓬幼苗鲜重和干重稍有下降，而含水量和肉质化程度升高；幼苗中Ｎａ＾＋含量上升而Ｋ＾＋，Ｃａ＾２＋含量下降；另外，碱蓬质膜透性稍有增大，渗透调节能力明显增加。     

植物根部的拒Na~+作用与叶片Na~+含量的相关性分析

在 10 0mmol/LNaCl条件下 ,伽蓝菜、碱蓬、白刺、滨藜、沙枣等 5种不同类型植物根部木质部液中Na+ 浓度都低于外界培养液中Na+ 浓度 ,说明 5种植物根部都有一定的拒Na+ 作用 ;但是叶片Na+ 含量与根部的拒Na+ 作用并没有明显的相关性 ,而与植物的相对蒸腾速率和相对生长速率呈显著的正相关 .     

盐清土壤环境对羊草生长与养分吸收的效应

通过室内模拟试验研究了不同盐度胁迫对羊草生长与营养吸收的影响,结果表明：土壤总盐0.172%-0.690%条件下,羊草植株生长受到抑制,生物量减少10.13%-33.20%。蒸散量和净光合速率随土壤盐度的增加而降低,尤其是日量大净光合速率受盐胁迫的影响显著,高盐处理羊草的日量大净光合速度较对照约减少1/3。盐分抑制羊草对N、P、K的吸收,尤其K的吸收受盐分抑制显著,不同盐度处理羊草植株茎叶与根系K含量均显著降低。羊草对Na的吸收累积随土壤盐度的增加而上升。     

转MT工程菌的构建及其对重金属的抗性研究

利用重叠延伸PCR技术克隆金属硫蛋白(MT)和绿色荧光蛋白(GFP)基因片段,并将两基因融合连接构建重组表达载体,采用氯化锂法转化毕赤酵母,获得工程菌株.荧光显微镜观察发现,工程菌在蓝光激发下发出绿色荧光,说明GFP基因被正确表达.在培养基中加入一定浓度的铜(1.0 mmol/L,1.5 mmol/L)、铬(150 μmol/L,200 μmol/L)、镉(120 μmol/L,140 μmol/L)、砷(40 μmol/L,60 μmol/L)化合物后,对照菌生长抑制,转基因菌株长势明显好于对照菌,表现出对金属离子的耐受性,说明工程菌过表达MT能够增强宿主对重金属离子的耐受性,提高菌株耐污能力,在微生物法净化重金属废水中具有一定优势.     

盐胁迫下甜高粱光能利用能力的变化

以甜高粱的一个优良品种---绿能一号为试验材料,对盆栽幼苗进行盐胁迫（0、50、100、200 mmol·L -1）处理。通过光合气体交换参数和叶绿素荧光猝灭动力学研究了盐胁迫对甜高粱幼苗碳同化能力和光系统Ⅱ光化学效率的影响。结果表明,在50 mmol·L -1盐处理下甜高粱的光能利用能力和对照相比无显著差异,随盐处理浓度的升高,甜高粱的光合利用能力受到明显的抑制,在100和200 mmol·L -1盐处理条件下,甜高粱的φPSII、qP 和 Pn 等指标均显著降低,在200 mmol·L -1的盐处理下有最小值。     

NaCl胁迫对黄瓜幼苗生长及光合作用特征的影响

以碧春黄瓜为材料,研究了不同浓度NaCl处理对黄瓜幼苗生长、叶片光合作用特性、以及脯氨酸和丙二醛含量的影响。结果表明:10 mmol/L NaCl处理促进了黄瓜幼苗生长,但高浓度NaCl(≥25 mmol/L)处理下,黄瓜幼苗的生物量、净光合速率(Pn)、气孔导度(Gs)、胞间CO2浓度(Ci)、光系统II电子传递量子效率(ΦPSⅡ)和叶绿素含量均随着浓度增加下降,气孔限制值(LS)、MDA和脯氨酸含量则随着浓度增加而上升;但NaCl处理对光化学效率(Fv/Fm)影响不大。由此说明,低浓度NaCl处理促进了黄瓜幼苗的生长,且NaCl浓度低于100 mmol/L时也未引起黄瓜幼苗叶片光抑制的发生,其光合作用下降的主要原因是叶绿素含量下降以及气孔限制,并且脯氨酸含量的增加也起到增强其抗性的作用。     

盐胁迫对甜高粱幼苗抗氧化酶活性的影响

研究了不同浓度NaCl溶液(含量0、50、100、150 mmol/L)处理对4种甜高粱幼苗超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性以及脂质过氧化产物丙二醛含量的影响。结果表明,与对照相比(即NaCl浓度为0),50 mmol/L NaCl处理显著增加了SOD、POD和CAT活性,而100和150 mmol/L NaCl处理则抑制了3种抗氧化酶活性。NaCl胁迫导致了脂质过氧化发生,具体表现为随着NaCl处理的浓度增加,丙二醛也增加。4种供试甜高粱对NaCl胁迫的应答有差异,其中辽甜1号在所有供试浓度的NaCl胁迫下,3种抗氧化酶活性均高于其他3个品种,而丙二醛含量最低,表明辽甜1号对NaCl的耐受性是最强的。