小麦耐盐细胞系脯氨酸SOD酶核酸和蛋白质合成动态分析

对已筛选出的小麦耐盐细胞系的耐盐稳定性进行有关生理、生化分析。结果表明：耐盐细胞系脯氨酸含量不仅稳定,而且高于对照;ＳＯＤ活性在耐盐系增加,耐盐系对一定的盐浓度损伤ＤＮＡ修复能力大于对照系,耐盐系ＲＮＡ和蛋白质合成量分别是对照的５．４和４．５倍,并对小麦耐盐性的有关问题进行了讨论。     

玉米耐盐早期筛选体系的初步研究

耐盐性鉴定是植物耐盐分子育种的重要环节,为了建立玉米耐盐性鉴定的早期筛选体系,选用了丹340,Mo17和TL94B 3个自交系,设置了10个NaCl浓度梯度,并通过室内沙培盐胁迫处理实验,对出苗率、6天株高、10天株高、14天株高、地上部和根系鲜重等性状进行了考察.结果表明:丹340和TL94B的耐盐性较强,而Mo17的耐盐性较弱,且对盐胁迫敏感;出苗率与盐浓度无显著相关,不宜作为耐盐性筛选的定量指标;株高、地上部和根系鲜重都与盐浓度高度负相关,达极显著水平,可以作为玉米耐盐性早期筛选的指标.研究还发现:在沙培条件下,20 mmol.L-1左右的NaC l浓度对幼苗的生长有促进作用;进行玉米耐盐性鉴定的NaC l临界下限浓度是100~120 mmol.L-1.     

Transformation of japonica rice with RHL gene and salt tolerance of the transgenic rice plant

Overexpression of the yeast HAL2 gene increases salt tolerance of yeast and plant. Rice HAL2-like (RHL) gene was introduced into a japonica rice cultivar HJ19 with Agrobacterium tumefaciens-mediated transformation. Transgenic plants in R0 generation were selected on the principle of GUS-positive, RHL gene PCR-positive and normal growth. Hygromycin-resistant plants of some transgenic lines in R1 generation increased salt tolerance during the seedling and booting stage, being less damaged in the cytomembrane and stronger in leaf tissue viability under salt stress during booting period. Southern analysis of transgenic lines tolerant to salt in R1 generation showed that the RHL gene expression cassette had been successfully integrated into rice genome. Moreover, gene engineering breeding methodology and really salt-tolerant rice cultivar were discussed.     

盐胁迫、辐射条件下耐盐与不耐盐水稻POD同工酶、全蛋白变化的研究

耐盐水稻品种９３４３经盐胁迫后,根系内诱导出一条活性很强的ＰＯＤ３同工酶谱带,而对盐敏感的１１８７、中作３２１、中作９３盐胁迫后活性很强的ＰＯＤ１谱带消失。蛋白质的变化呈现出与ＰＯＤ变化相类似的趋势,耐盐品种９３４３盐胁迫后某些蛋白质谱带含量增加,出现分子量与调渗蛋白相类似的２６ＫＤ蛋白质,但对盐敏感的三个品种则蛋白质含量下降,没有检测到２６ＫＤ蛋白,辐射后经ＮａＣｌ胁迫,耐盐的９３４３中同工酶和     

小麦耐盐系筛选及稳定性研究

以小麦幼穗、幼胚为原始材料,进行耐盐突变体的筛选,并对其愈伤组织及再生植株后代进行耐盐稳定性的生理生化分析。结果表明：耐盐系在高盐浓度下其鲜重、干重明显高于对照系;耐盐系保持较高的Ｋ＾＋／Ｎａ＾＋比;与对照组相比,耐盐系种子醇溶蛋白电泳带有１４条,其中Ｂ２,Ｂ３,Ｂ４带为耐盐系所特有,Ｂ１带含量高于对照。但对照Ｂ５带合成量增加;耐盐系再生植株后代可溶蛋白电泳带耐盐系为２８条蛋白带,而对照系为２６条     

应用农杆菌介导法的多年生黑麦草遗传转化研究

以草坪型多年生黑麦草Lolium perenne L.成熟种子为外植体,经过愈伤组织诱导和植株再生研究,建立了该草种的遗传转化体系,并成功地应用农杆菌介导法将克隆自辽宁碱蓬的甜菜碱醛脱氢酶基因(BADH)转移进多年生黑麦草,获得的转基因株系Gsc-LP5通过PCR检测证明了目的基因的存在,通过叶片离体检测法证明了作为检测基因随同质粒一同转入的抗潮霉素基因的表达,通过盆栽耐盐试验证明了转基因植株具有较强的耐盐能力.     

紫雨桦耐盐性及花色苷合成相关基因的表达特性

【目的】探讨紫雨桦的耐盐性与花色素苷合成相关基因的相关性,为揭示紫雨桦的耐盐机理提供参考。【方法】以紫雨桦和垂枝桦(对照)为试材,测定盐胁迫下其叶绿素荧光参数及生理指标的变化,同时采用qRT-PCR方法分析紫雨桦和垂枝桦中查耳酮合酶基因(CHS)、二氢黄酮醇还原酶基因(DFR)、花色素苷合成基因及MYB转录因子家族基因的表达特性。【结果】经质量分数0.8%NaCl胁迫12 d后,紫雨桦叶片花青素含量指数显著高于垂枝桦,是垂枝桦的3.4倍,其盐害指数显著低于垂枝桦,胁迫12 d的联苯胺蓝(DAB)和氮蓝四唑(NBT)染色显示,紫雨桦叶片中O^-₂和H₂O₂的累积少于垂枝桦; 在叶绿素荧光参数方面,NaCl胁迫12 d时,紫雨桦的F_v/F_m值仍在正常值范围内,与胁迫0 d比较,其qP和Φ_PSⅡ值的降幅也显著小于垂枝桦(P<0.05),紫雨桦的NPQ较0 d提高了282.29%,升幅显著高于垂枝桦(P<0.01)。 0.8%NaCl胁迫下CHS、DFR花色素苷合成基因及MYB转录因子家族基因的表达特性是:紫雨桦中的BpMYB10、BpCHS2和BpDFR5基因表达规律明显不同于垂枝桦,NaCl胁迫下上述3条基因持续高表达。【结论】 富含花青素的紫雨桦能够耐受一定浓度范围的NaCl胁迫,紫雨桦中的BpMYB10、BpCHS2和BpDFR5基因表达规律明显不同于垂枝桦,这些基因可能与花青素的合成密切相关,花色苷可增强紫雨桦清除活性氧自由基的能力,减缓膜质氧化,提高其耐盐性。     

小麦耐盐突变体筛选及生理生化特性分析

以小麦幼穗、幼胚为原料材料,进行耐盐突变体的筛选,并对其愈伤组织及再生植株后代进行耐盐稳定性的生理生化分析,结果表明 （１）耐盐系在高盐浓度下其鲜重、干重明显高于对照系;（２）耐盐系保持较高的Ｋ＾＋／Ｎａ＾＋比;（３）对照系种子有１１条醇溶蛋白电泳带,而耐盐系为１４条,与对照系相比Ｂ２、Ｂ３、Ｂ４带为耐盐所特有,Ｂ１带含量高于对照物,但Ｂ５带含量低于对照系;（４）耐盐系再生植株后代可溶蛋白电泳带为     

Transformation of japonica rice with<Emphasis Type="Italic">RHL</Emphasis> gene and salt tolerance of the transgenic rice plant

Overexpression of the yeastHAL2 gene increases salt tolerance of yeast and plant. RiceHAL2-like (RHL) gene was introduced into ajaponica rice cultivar HJ19 withAgrobacterium tumefaciens-mediated transformation. Transgenic plants in R₀ generation were selected on the principle of GUS-positive,RHL gene PCR-positive and normal growth. Hygromycin-resistant plants of some transgenic lines in R₁ generation increased salt tolerance during the seedling and booting stage, being less damaged in the cytomembrane and stronger in leaf tissue viability under salt stress during booting period. Southern analysis of transgenic lines tolerant to salt in R₁ generation showed that theRHL gene expression cassette had been successfully integrated into rice genome. Moreover, gene engineering breeding methodology and really salt-tolerant rice cultivar were discussed.     

野生番茄耐盐性研究及其利用

将带有二叶一心的小苗扦插于附加不同浓度NaCl的MS培养基中,研究不同番茄材料发根、植株生长以及POD酶酶谱及其活性的变化,并通过花粉管导入技术将野生番茄的总DNA导入到栽培种。研究结果表明,Lycopersicon Pennellii LA716和Lycopersicon pimpinellifolium LA2184始终具有相对高的生根率,在150mmol/L NaCl胁迫10d时,相对发根率分别为86.8％和100％。参谋工的4份野生种,在附加有75mmol/L NaCl的培养基上的株高和叶片数均高于对照（无NaCl）,随着NaCl浓度的不断提高,株高和叶片数教下降,Lycopersicon Pennellii LA716下降幅度最小。4份野生种的耐盐性明显优于2坐栽培种,当NaCl浓度提高到300mmol/L时,4份野生种的鲜重可达对照的0.90、1.07、1.60和1.53倍,而份栽培种则为对照的0.55和0.33倍。当NaCl浓度为75mmol/L时,Lycopersicon peruvianum LA111和Lycopersicon Pennellii LA716材料POD酶酶带数多于对照2条和1条,Lycopersicon cheesmanii LA166和Lycopersicon pimpinellifolium LA2184则少于对照1条和3条。利用花粉管导入技术可以将野生番茄的DNA导入到栽培番茄。     

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.     

玉米幼苗对NaCl胁迫的生理响应和耐受阈值分析

以水培华农5号玉米（Zea mays L.var Huanong 5）幼苗为材料,综合分析了NaCl胁迫条件下幼苗的生理反应变化及其不同组织和生理过程的胁迫耐受阈值.结果表明：玉米叶片对NaCl胁迫最为敏感,耐受阈值约为150 mmol/L,且主要影响因素是气孔限制,非气孔限制因素对NaCl耐受性较强.PSⅡ的耐盐阈值在200～300 mmol/L之间.     

F-box基因FOF2在拟南芥盐和冷胁迫响应中的功能分析

FOF2为F-box蛋白家族成员,其生物学功能尚不清楚.采用实时荧光定量PCR和生理学实验相结合的方法,对FOF2基因的表达模式及其在拟南芥抗盐和冷胁迫响应中的作用进行了分析.研究发现,FOF2在拟南芥根、茎生叶和果荚中表达较高,并且其表达受盐和冷胁迫诱导.FOF2过表达株系对盐胁迫敏感,与野生型相比种子萌发率低、幼苗主根较短;相反,fof2突变体对盐胁迫的敏感性则减弱.FOF2过表达和缺失突变体种子萌发对冷胁迫无响应,但其主根在冷处理中分别比野生型短或者长.盐处理下,FOF2过表达株系中盐胁迫反应相关基因的表达量显著降低,fof2突变体中则升高;冷处理下,FOF2过表达株系中冷胁迫反应相关基因的表达量显著升高,fof2突变体中则降低.结果表明,FOF2在植物抗盐胁迫响应中起负调控作用,在抗冷胁迫响应中则可能起正调控作用.     

小麦耐盐愈伤组织内源脯氨酸含量和细胞学特征

以小麦幼穗和幼胚为外植体，诱导出愈伤组织，通过在培养基中逐渐增加选择剂（ＮａＣｌ：Ｎａ２ＳＯ４＝４：１）方法，获得耐０．８％至２．０％选择剂的耐盐愈伤组织，其内源脯氨酸含量显著高于相同盐渍条件下的对照组．培养基中选择剂浓度达到１．６％耐盐细胞仍能保持细胞及细胞器正常形态，液泡化程度低，细胞核大，有的细胞中出现二或二个以上核仁．在培养基中选择剂超过１．２％，绝大多数细胞处于坏死或类坏死状态．耐盐愈伤组织在培养基中选择剂不超过２．０％，生长良好，可以再生植株，其后代在０．３％ＮａＣｌ土壤中仍表现耐盐性．     

耐盐稳定泡沫选择性堵水剂

耐盐稳定泡沫具有抗高温、堵水不堵油等优良特性,是很有发展前途的一种选择性堵剂．笔者研究了泡沫稳定的机理,优选了泡沫堵水的配方,选用具有支型结构的发泡剂和具有耐温耐盐结构的稳定剂．由耐盐活性剂（DM5512）、甜菜碱、烷基糖苷（APG）、十六烷基三甲基氯化铵（1631）？昆合制备成了离子型复合起泡剂（QP-1）,由羟丙基甲基纤维素（HPMC）、SMP—III、高分子聚合物（SP-8）混合制备成了稳定剂（WD-1）,使用时把QP-1和WD-1混合均匀,配成一定水溶液,然后发泡制备耐盐稳定泡沫．实验证明以上配方具有良好的泡沫稳定性和耐温耐盐性,并利用GE-5显微镜观察了泡沫的结构,讨论了可用作选择性堵7k剂的原因．     

耐盐细菌质粒的研究

本实验以1株耐盐细菌My23（耐18％NaCl）作为研究对象,采用碱裂解法对其进行质粒提取,结果表明可以提取得大小2个不同质粒;改变常规提取方法中的部分环节,如加入STE清洗菌体的培养基成分,溶菌酶的使用,以及加入碱裂解液Ⅰ后静置20min,对该耐盐菌质粒的提取有较好的作用。经SDS法将小质粒消除后的菌株与野生型菌株的耐盐功能比较,发现提取出的小质粒与耐盐无关。将大肠杆菌DH5α制备成感受态细胞,将耐盐菌的质粒进行转化,结果进一步表明耐盐性状与质粒没有关系。     

Unresponsiveness to a foreign antigen can be caused by self-tolerance

In mice, two sets of genes govern the immune response to the synthetic antigen GT. One maps to the major histocompatibility complex and behaves like a typical immune response gene. The second is a background gene encoding a cell surface structure found on B cells. Mice which express, and are therefore tolerant of, one form of this structure do not respond to GT. Thus, tolerance of self generates holes in the T-cell repertoire, partially crippling the immune system.     

Characterization of Arabidopsis calreticulin mutants in response to calcium and salinity stresses

As an important calcium-binding protein, calreticulin plays an important role in regulating calcium homeostasis in endoplasmic reticulum （ER） of plants. Here, we identified three loss-of-function mutants of calreticulin genes in Arabidopsis to demonstrate the function of calreticulin in response to calcium and salinity stresses. There are three genes encoding calreticulin in Arabidopsis, and they are named AtCRT1, 2, and 3, respectively. We found that both single mutant of crt3 and double mutant of crtl crt2 were more sensitive to low calcium environment than wild-type Arabidopsis. Moreover, crt3 mutant showed more sensitivity to salt treatment at germination stage, but tolerance to salt stress at later stage compared with wild-type plant. However, there was no obvious growth difference in the mutant crtl and crt2 compared with wild-type Arabidopsis under calcium and salt stresses. These results suggest that calreticulin functions in plant responses to calcium and salt stresses.