缺硫对镉胁迫下水稻幼苗非蛋白巯基物质含量和谷胱甘肽硫转移酶活性的影响

通过设置缺硫(S)处理,研究了镉(Cd)胁迫下水稻生长情况、幼苗Cd和非蛋白巯基含量以及谷胱甘肽硫转移酶(GST)活性的动态变化.结果表明:Cd胁迫明显抑制了水稻生长,显著诱导了巯基物质[非蛋白巯基(NPT)、谷胱甘肽(GSH)、植物螯合肽(PC)]的合成,GST活性表现出先升后降的趋势.缺S处理下,尽管水稻根部对Cd的吸收和向地上部的转运都有所增加,但Cd胁迫程度并未明显增强,巯基物质含量明显降低,根部GST活性提高.表明巯基物质和GST在水稻抗Cd胁迫过程中互为补充,在一定程度上减轻了Cd的毒性效应.     

脱金属硫蛋白与镉离子的络合作用及构象研究

用圆二色（ＣＤ）谱地研究兔肝脱金属硫属蛋白的两个亚型与Ｃｄ＾２＋的络合作用及对重组ＭＴ构象的影响。观测了ａｐｏ－ＭＴ垢巯基在空气和室温下的稳定性。在ＰＨ４．７１,镉重组ＭＴ１的ＣＤ谱特征峰在２５７ｎｍ（＋）,２３８ｎｍ（－）,２２６ｎｍ（＋）与镉诱导的天然ＭＴ１相同。在空气存在和ＰＨ７．９０的ＣＤ谱只有２４３ｎｍ（＋）一个峰。向两亚型分别加入７ｅｑＣｄ＾２＋测定ＣＤ谱随ＰＨ值的变化,发现在ＰＨ２．     

镉胁迫对花生籽实品质的影响及响应机制

采用盆栽试验,研究了重金属镉(Cd)对花生籽实品质安全性的影响及花生籽实对镉胁迫的响应机制。结果表明:在供试镉处理范围内(≤10mg.kg-1),镉胁迫对花生籽实脂肪和蛋白质含量均有显著影响,但品种间存在差异;花生籽实中的镉含量随外源镉含量的增加而显著增加(P<0.05),且在土壤低镉浓度(≤1mg.kg-1)下籽实更易富集镉;受镉污染的花生籽实,其蛋白质是络合镉的主要营养部位,且其镉含量远高于食品中镉的限量值,而脂肪中镉的含量甚微,因此供试花生籽实不能作为人体植物蛋白来源,但可以作为人体食用油脂来源。     

水分胁迫下植物体内OH的产生与细胞的氧化损伤

干旱是植物组织的一种重要的胁迫因子。它能干扰植物细胞中活性氧产生与清除的平衡,导致植物细胞遭受氧化胁迫。过去十多年来,人们对水分胁迫下植物体内活性氧的产生、活性氧对植物的伤害及植物保护系统的作用进行了大量的研究,取得了明显的进展［１～４］。然而,水分...     

镉诱导家兔肝脏含锌金属硫蛋白的纯化及鉴定

预先给家兔皮下注射CdCl_2四次,取肝脏经匀浆、离心、乙醇沉淀后,通过Sephadex G-50凝胶过滤层析,DEAE Sepharose Fast Flow离子交换层析和Sephadex G-25层析柱脱盐,得到MT的两个“亚型”—MT-1和MT-2,对其进行鉴定,其紫外扫描图谱示在280和250nm处无吸收峰,而最大吸收在220nm处。MT-1和MT-2均不含镉和铜,而每分子结合6个锌。用简化巯基试剂测定,MT-1含18.8个巯基,MT-2含19.1个巯基;HPLC分析MT主要以单体存在,也有少量二聚体。氨基酸组成与Cd-MT一致。整个纯化过程采用示波极谱法和火焰原子吸收光谱法测锌含量。结果表明前者可代替后者。     

嗜酸热硫球菌(S-5)对硫化物的氧化

从中国一热泉区分离到一种新的嗜热菌——嗜酸热硫球菌(Sulfosphaerellus thermoarido-philum)S-5能直接氧化元素硫(S)和黄铁矿(FcS2)等硫化物,氧化元素硫的温度范围是55—80℃,最适70℃;pH范围是1．0—5．5,最适2．5。在最适条件下,含元素硫1％,静置10天,细菌氧化元素硫溶液中的硫酸根(soi一)使浓度从1．10 g／L增加到6．56g/L,pH从2．35降到1．00。还发现嗜酸热硫球菌与中温的氧化亚铁硫杆菌(Thiobacillus ferrooxidans)T一9以同样方式氧化黄铁矿,但是前者的氧化能力比后者的更强,而且温度范围也更广。当使用一200目黄铁矿矿粉、矿浆浓度10％、振荡氧化20天时,在接人嗜热菌的溶液中,可溶性铁浓度从1．79 g/L增加到16．40 g／L,pH从2．23降到0．85,而接人中温菌的溶液中可溶性铁浓度从1．23 g,L只增加到7．37 g,L,pH从2．23只降到1．20。     

不同外源物质对镉砷复合污染胁迫下油菜生理指标和镉砷积累的影响

为缓解重金属对作物的胁迫,降低重金属对作物的毒害作用,通过叶面喷施比较不同外源物质茉莉酸(JA)、褪黑素(MT)、亚精胺(SPD)和2,4-表油菜素内酯(EBL)对Cd、As胁迫下油菜生理指标及吸收积累Cd、As的影响。结果表明:喷施200μmol·L^-1的JA和MT均显著提高油菜叶片的叶绿素SPAD值,而喷施20μmol·L^-1的EBL显著降低油菜叶片的叶绿素SPAD值;与CK处理相比,喷施500μmol·L^-1SPD和20μmol·L^-1EBL分别降低油菜叶片中MDA含量41.0%和52.3%,同时EBL处理提高油菜叶片SOD活性88.2%;喷施200μmol·L^-1MT导致油菜地上部Cd含量比CK处理降低27.8%,而喷施200μmol·L^-1的JA和20μmol·L^-1的EBL却分别提高油菜地上部As含量159.8%和136.8%; JA、MT、SPD和EBL在某种程度上均缓解了Cd、As复合污染对油菜的胁迫,为重金属污染农田...     

脱金属硫蛋白与二价汞离子的络合作用及构象研究

用圆二色谱研究兔肝锐金属硫蛋白的两个亚型ａｐｏ－ＭＴ１、ａｐｏ－ＭＴ２与Ｈｇ＾２＋在不同分子数比和ｐＨ值下的络合规律。发现：（１）在ｐＨ２下与Ｈｇ＾２＋重组ＭＴｓ的ＣＤ谱特征峰是３０４ｎｍ（＋）,２６０ｎｍ（－）,２５０ｎｍ（＋）。络合Ｈｇ＾２＋的数目ｎ远远超过７。任何ｐＨ值下过量Ｈｇ＾２＋的存在都将破坏ＭＴｓ的正常构象。ｐＨ值对形成和维持ＭＴｓ稳定构象的影响很大,当ｐＨ≥４．９时,ａｐｏ－ＭＴｓ     

油菜叶片总蛋白质双向电泳样品制备方法的改进

以甘蓝型油菜"扬油6号"的叶片为试验材料,分别采用传统的TCA/Acetone(三氯乙酸/丙酮沉淀法)和改进的PEG(polyethylene glycol)分步提取法提取叶片可溶性总蛋白,并利用条件一致的蛋白质双向电泳体系进行比较。TCA/Acetone法提取的蛋白质双向电泳图谱背景中由于高丰度"housekeeping"结构蛋白的存在,特别是叶片中参与光合作用的Rubisco蛋白的干扰,图谱中低丰度调控蛋白受到了高度覆盖和遮蔽现象,影响双向电泳图谱的质量。而PEG分步提取法提取的蛋白质样品,可以剔除Rubisco蛋白,使获得的双向电泳图谱清晰,无斑点间的遮蔽现象,为油菜叶片蛋白质组定量和定性分析提供了丰富的信息。     

镉诱导黄瓜金属硫蛋白抗体的制备及纯化

用镉诱导黄瓜叶片组织产生金属硫蛋白（MT）,经分离纯化,血清白蛋白偶联后作为抗原。采用背部皮内、皮下和耳缘静脉注射免疫家兔,制备出效价为1：16的免疫血清。用饱和硫酸铵盐析法和Sephadex G-200柱层析分离纯化IgG。经测定纯化后的IgG蛋白含量为0．9364mg&#183;ml^-1。用纯化的IgG建立黄瓜Cd-MT的斑点免疫试验检测方法可检测黄瓜Cd-MT的最低含量为200Pg&#183;ml^-1。     

抗草丁膦和抗草甘膦转基因油菜的抗性基因向野芥菜的流动

通过人工去雄授粉和田间隔行种植试验,研究了抗草丁膦和抗草甘膦转基因油菜(Brassica napus)中的bar基因和EPSPS基因向野芥菜(B. juncea var. gracilis)流动的可能性。结果表明在人工授粉的情况下,以野芥菜为母本,分别以两种转基因油菜为父本,亲和性指数都很高,达13以上,与野芥菜自交或开放授粉条件下的亲和性指数没有明显差异,说明两种转基因油菜和野芥菜的亲和性较好。经两次除草剂筛选,人工杂交获得的所有F₁对相应的除草剂都表现出了明显的抗性,且经PCR检测扩增出了各自的特异性条带,说明人工杂交获得的所有F₁都携带了相应的抗性基因。F₁的适合度研究表明,两种F₁种子萌发率和母本都没有明显差异,营养生长明显好于母本。但花粉活力和结实率明显下降,携带抗草丁膦基因F₁的花粉活力和每角果粒数分别是32.4%和0.59粒,携带抗草甘膦基因F₁的花粉活力和每角果粒数分别是35.1%和0.58粒。经两次除草剂筛选和PCR检测,表明野芥菜和抗草丁膦油菜或与抗草甘膦油菜田间隔行种植分别能产生0.02%和0.014%的携带抗性基因的F₁杂种。以上结果表明抗除草剂转基因油菜的抗性基因具有向野芥菜流动的可能性,且bar和EPSPS基因向野芥菜流动的可能性类似,但对其可能引起的环境后果需要做进一步地深入研究。     

Molecular markers for low-glucosinolate alleles in oilseed rape (Brassica napus L.)

Two recent studies have mapped QTLs associated with the level of seed glucosinolates in oilseed rape (Brassica napus L.). It was likely that the two most significant QTLs identified in each study were the same, as they were linked to RFLP alleles identified by common DNA probes. To investigate the utility of these probes in breeding programmes, they were used to study RFLPs in a range of low- and high-glucosinolate cultivars and breeding lines. It was shown that all low glucosinolate spring and winter cultivars possessed a specific RFLP fragment identified by probe wg3f7 which is linked to theGSL-1 QTL, and all high-glucosinolate cultivars possessed a specific RFLP fragment identified by probe wg7a8, which is linked to theGSL-2 QTL. Cultivar Ariana, which has intermediate levels of glucosinolates possessed the low-glucosinolate fragment atGSL-1 but the high-glucosinolate fragment atGSL-2. A similar result was found with the cvs. Martina and Bronowski which have intermediate and variable levels of glucosinolates. There were no other RFLP fragments identified by other DNA probes which were specific to either the low- or high-glucosinolate phenotypes. The use of probes wg3f7 and wg7a8 in selection of low-glucosinolate lines in breeding programmes is discussed.     

Glucosinolate changes in developing pods of single and double low varieties of autumn-sown oilseed rape (B. napus)

Single and double low varieties of oilseed rape were grown in the 1987/88 and 1988/89 seasons to study changes in the concentrations of total and individual glucosinolates within pods during development. Total glucosinolate concentration in seeds of all varieties increased during development when expressed on a fresh weight basis. The levels of the major alkenyl glucosinolates present in the seed; 2–hydroxy-3–butenyl, 3–butenyl and 4–pentenyl had been reduced in the transition from single to double low varieties. The major indole glucosinolates in the seed, 4–hydroxy-3–indolylmethyl and 3–indolylmethyl were present in the same amounts in single and double low varieties but in the latter represented a greater proportion of the total seed glucosinolate content. A decline in the total glucosinolate concentration in the pod walls with time together with the analogous profile of individual glucosinolates in the seeds and pod walls suggests that the pod wall is a major site of seed glucosinolate synthesis. Other plant parts may also have an important role to play in provision of intact glucosinolates or precursors to the pod walls for glucosinolate biosynthesis.     

Chloroplast Transformation in Oilseed Rape

The chloroplast transformation vector pNRAB carries two expression cassettes for the spectinomycin resistance gene aadA and the insect resistance gene cry1Aa10. The two cassettes are sited between the rps7 and ndhB targeting fragments. Biolistic delivery of the vector DNA, followed by spectinomycin selection, yielded chloroplast transformants at a frequency of four in 1000 bombarded cotyledon petioles. PCR analysis and Southern blot of PCR products confirmed the site-specific integration of aadA and cry1Aa10 into the chloroplast genomes of transgenic oilseed rape. When transgenic oilseed rape leaves were fed to second instar Plutella xylostera larvae, 47% mortality was observed against this insect and the surviving larvae had significantly lower weight than the control. This is the first report of chloroplast transformation in oilseed rape and the introduction of novel genes between the rps7 and ndhB genes in the chloroplast genome. This offers an opportunity for improvement of oilseed rape by chloroplast genetic engineering.     

HPLC分析油菜籽油中维生素E的组成与含量

本文利用ＨＰＬＣ法分析了５０份遗传背景丰富的白菜型油菜、甘蓝型油菜、芥菜型油菜和芸芥种子油中维生素E的组成与含量,研究结果显示,油菜种子油中主要含α-生育酚和? -生育酚,且α-生育酚、? -生育酚和维生素E总量均存在明显的基因型差异,甘蓝型油菜种子油中维生素E含量总体水平最高,平均总量较高,为123.11 mg/100g油, 维生素E含量最高的Omega,总量为144.73 mg/100g油, α/ ? -生育酚比值最高可达0.77。α-生育酚、? -生育酚和维生素E总量与类胡萝卜素含量均呈现显著以上负相关,种子油中α-生育酚与含油量呈现显著正相关,α-生育酚、? -生育酚和维生素E总量与生育期均呈现显著或极显著正相关,α-生育酚和维生素E总量与株高均呈现显著正相关,维生素E总量与千粒重呈显著正相关,而α-生育酚、? -生育酚和维生素E总量与全株角果数和每角粒数相关不显著。     

基于高通量测序分析大豆和油菜根际微生物群落结构的差异

根肿病是由芸薹根肿菌侵染引起的专性寄生性土传病害,严重制约着油菜等十字花科作物的可持续生产.前期研究发现,大豆作为前茬作物可以显著降低后茬油菜根肿病的发生和危害,\"豆-油轮作\"模式是一种值得探索和应用的根肿病防治新途径.为了解开大豆作为前茬防治根肿病发生的机理,本研究基于扩增子测序技术探究大豆与油菜根际土壤微生物的群落结构差异.结果表明:大豆和油菜根际土壤微生物类群在门水平的优势类群相同,包括变形菌门、拟杆菌门、酸杆菌门、放线菌门、子囊菌门、接合菌门、担子菌门和壶菌门等丰度都较高.但相比于油菜根际土壤,大豆根际土壤富含具有生防作用和促进植物生长的微生物,如黄杆菌属、鞘脂单胞菌属、芽孢杆菌属、链霉菌属、假单胞菌属、木霉属和盾壳霉属等;而一些植物病原细菌(如肠杆菌、黄单胞菌)和真菌(炭疽菌和尾孢菌)含量则低于油菜根际土壤;另外,大豆根际土壤中还富含具有固氮功能的根瘤菌属、慢生根瘤菌属和丛枝菌根真菌(如球囊霉属).可见,大豆根际土壤利于有益微生物生长并可抑制病原菌繁殖.大豆和油菜根际微生物组差异为大豆-油菜轮作防治根肿病提供了理论依据,并为根肿病的防治提供了一些潜在的生物防治资源.     

Effects of bromoethylamine on antioxidant capacity,lipid peroxidation,and morphological characteristics of rat liver

Administration of bromoethylamine (BEA, 1.2 mmol/kg) to fed rats induced a significant diminution in the activity of hepatic superoxide dismutase (at 1 h after treatment), catalase, and glutathione peroxidase and in the content of nonprotein sulfhydryls (at 15 h after treatment). The content of thiobarbituric acid reactants by the liver was enhanced by 1.9 times over control values (at 3 h). Light microscopy studies revealed that BEA (72 h after treatment) induced periportal fatty accumulation, focal liver cell necrosis, and diffuse inflammatory infiltrates, in addition to hypertrophic Kupffer cells and mitotic hepatocytes. Also, hypertrophic middle tunic or hypertrophic smooth muscle layers of arterioles was observed in the periportal space, with dilated sinusoidal capillaries and free macrophage infiltration. It is concluded that BEA induces a derangement in the antioxidant status of the liver with the consequent lipid peroxidation response, which may constitute a significant hepatotoxic mechanism of the haloaklylamine. © 1998 John Wiley & Sons, Inc. J Biochem Mol Toxicol 13: 47–52, 1999     

Polyethylene glycol and electric field treatment of plant protoplasts: characterization of some membrane properties

Petiolar protoplasts of a dihaploid line of winter oilseed rape Brassica napus L. ssp. oleifera were exposed to fusogenic polyethylene glycol (PEG) and electric field treatments. The surface properties and stability of membrane components of the treated protoplasts were investigated by contact angle measurements in aqueous two-phase systems and differential scanning calorimetry, respectively. The leakage of intracellular components was estimated with respect to amino acids, proteins and DNA. Both fusogenic treatments resulted in the same apparent changes in membrane surface hydrophobicity and the same destabilization of membrane components. However, the PEG-treated protoplasts were more leaky than both the control and the electric field-treated protoplasts. The results indicate that the molecular mechanisms of PEG- and electrical field-induced fusion are similar. However, the effects of the latter appear to be less harmful presumably because the parameters for electric field treatment are more easily controlled.