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1.
为研究苹果属植物抗缺铁的分子生物学机理,从铁高效基因型小金海棠（Malus xiaojinensis）中克隆了Fe3+-还原酶基因MxFRO。MxFRO2的cDNA序列长2283bp,开放阅读框为2166bp,编码722个氨基酸。半定量RT-PCR结果表明MxFRO在小金海棠的根和叶中均受缺铁胁迫诱导。 将MxFRO转入野生型酵母中,结果表明转基因酵母的Fe3+-还原酶活性是对照的2.8倍,因此我们初步推测MxFRO基因编码的蛋白具有Fe3+-还原酶活性。 相似文献
2.
通过染色体步移法从小金海棠（Malus xiaojinensis）基因组中克隆了三价铁螯合还原酶（ferric chelate reductase, MxFRO2）基因翻译起始位点上游1 738 bp的启动子序列。生物信息学分析表明,该启动子片段中存在光响应元件G-box、生长素应答元件、铜元素响应元件、TATA-box、CAAT-box等顺式作用元件。从TAIR网站获得了拟南芥（Arabidopsis thaliana）中8个FRO基因上游的启动子序列,通过PlantCARE分析了其与MxFRO2启动子的异同。根据在线预测结果,克隆了翻译起始位点上游1 644 bp（MxFul）和259 bp（MxD1）两段序列,构建了其与GFP融合的瞬时表达载体,并将重组质粒通过PEG介导法分别转化拟南芥叶片原生质体,结果表明,小金海棠MxFul和MxD1两段启动子序列都能驱动GFP蛋白在拟南芥原生质体中的瞬时表达。 相似文献
3.
高pH值和铁素对毛白杜鹃和迎红杜鹃根系Fe~(3+)还原酶活性的影响 总被引：3，自引：0，他引：3
以耐碱性不同的两种杜鹃花(毛白杜鹃和迎红杜鹃)的组培苗为材料,对其根系Fe3+还原酶活性与铁素和pH值间的关系进行研究,结果表明:培养最初阶段高pH值(NaHCO3提供)与铁素共同诱导根系Fe3+还原酶活性增加;Fe3+对根系Fe3+还原酶活性的诱导作用具有类似于代谢途径中的底物诱导效应;在10d的缺铁处理中,迎红杜鹃根系Fe3+还原酶活性增加,而毛白杜鹃没有增加,说明缺铁对酶活性的诱导存在种间差异;杜鹃花根系Fe3+还原酶活性与植物耐碱性存在正相关。 相似文献
4.
小金海棠S-腺苷甲硫氨酸合成酶基因（MxSAMS）的克隆和表达分析 总被引：1，自引：1，他引：0
从本实验室建立的小金海棠缺铁差减文库中分离出一个cDNA片段,在GenBank中发现该片段与其它植物的S-腺苷甲硫氨酸合成酶(SAMS)基因具有90%的同源性。利用RACE技术成功地获得小金海棠（Malus xiaojinensis）SAMS基因(MxSAMS)的cDNA全长序列（GenBank登录号：EU639408）,该基因cDNA全长1 479 bp,最大开放阅读框1 176 bp,编码392个氨基酸,酶蛋白理论分子量为42.99 kD;与其它植物的蛋白质氨基酸序列同源性在88.1%~92.6%之间。推测的MxSAMS蛋白质三级结构包含3个保守结构域和一个保守的ATP结合位点GAGDQG。Southern 杂交显示,MxSAMS基因在小金海棠基因组中是单拷贝的。半定量RT-PCR结果表明,该基因在小金海棠的根和叶中均受缺铁胁迫诱导增强表达。 相似文献
5.
小金海棠MxYSL1基因的克隆与表达分析 总被引：2，自引：1，他引：1
小金海棠属于铁高效基因型。为了研究小金海棠的抗缺铁分子机理,根据苹果的EST库得到一个791 bp的YSL(yellow stripe 1-like protein)基因片段序列,设计特异引物,在苹果铁高效基因型小金海棠（Malus xiaojinenesis）中克隆到此基因片段。3'-RACE得到YSL基因的3'端序列,拼接后得到1 114 bp的3'端基因序列。预测该基因片段编码一个含7个跨膜区的蛋白多肽,与拟南芥AtYSL1同源性达74%,命名为MxYSL1。RT-PCR及Northern blot分析表明,该基因在小金海棠各个检测部位都有表达。在根、茎与成熟叶中,该基因在低铁(EDTA-NaFe,4 μmol/L)时减弱表达,在过量铁(EDTA-NaFe,320 μmol/L)供应时增强表达。MxYSL1基因在新叶中的表达趋势与以上器官中的表达趋势正好相反。 相似文献
6.
植物质膜H+-ATPase能把质子泵出根外酸化土壤,增加铁的溶解度,利于植物对铁的吸收.小金海棠是一个苹果铁高效基因型砧木,为研究质膜H+-ATPase在小金海棠铁吸收过程中的功能,本研究通过同源克隆的方法从小金海棠(Malus xiaojinensis)中克隆到了一条全长2865 bp含有完整编码区的质膜H+-ATPase基因(MxHA2),编码954个氨基酸(GenBank登录号:JQ867095).系统进化分析结果表明MxHA2蛋白与碧桃(Prunus persica)的PPA2蛋白亲缘关系较近.基因枪法在洋葱(Allium cepa)表皮内进行亚细胞定位分析,结果表明,MxHA2与GFP的融合蛋白定位于细胞膜上.荧光定量PCR分析表明,缺铁胁迫后,MxHA2基因在根和叶中均有不同程度的增强.把MxHA2基因转化到酿酒酵母(Saccharomyces cerevisiae)(BJ2168)后,转MxHA2的酿酒酵母H+-ATPase活性极显著增强,且缺铁处理后其生长量持续高于对照,表明该基因编码的蛋白具有H+-ATPase活性,其过量表达能提高酵母菌株的耐缺铁性.结果提示,MxHA2可能在小金海棠的耐缺铁胁迫过程中起重要作用,为进一步研究小金海棠的铁高效机理提供了依据,在果树耐缺铁分子育种上有重要的应用前景. 相似文献
7.
本试验从小金海棠(Malus xiaojinensisensis)中克隆得到2个具有 CDS全长的ferritin基因(1182bp和940bp)和4个具有CDS片段的ferritin基因(934bp、594bp、1070bp和834bp)。序列分析表明它们均具有真核生物铁蛋白保守区(Euk_Ferritin)这一典型的植物铁蛋白结构特征。预测蛋白质相对分子量在21.58kDa和34.32kDa之间。进化树分析表明小金海棠的ferritin基因与梨的ferritin基因在同一进化分支上。根据qPCR和半定量RT-PCR结果,可将小金海棠ferritin基因的时间表达模式划分为早期表达型(MxFer1)、晚期表达型(MxFer2)、组成表达型(MxFer6)和波动表达型(MxFer3-5);就组织部位而言,MxFer1的表达具有组织特异性,只在叶中特异表达,而MxFer2-6在根和叶中均有表达。 相似文献
8.
为探明小金海棠(Mdus xiaojinensis)类黄色条纹蛋白基因(MxYSL5)的表达和调控机制,应用Tail-PCR技术从小金海棠中克隆了翻译起始位点上游891 bp的启动子序列.生物信息学分析表明,该启动子片段中存在光响应元件、乙烯应答元件、赤霉素响应元件、TATA-box、CAAT-box等顺式作用元件.以GFP为报告基因,构建了含MxYSL5基因启动子的植物表达载体MxYP5-GFP.将MxYP5-GFP用基因枪法转化洋葱(Allium cepa)表皮细胞,瞬时表达结果表明,该启动子能够驱动GFP在洋葱表皮细胞中的表达,可以用于MxYSL5的表达示踪. 相似文献
9.
试管微嫁接早期鉴定小金海棠与苹果品种的亲和性* 总被引：8，自引：0，他引：8
用试管微嫁接法鉴定嫁接亲和性的结果表明,苹果(Malus pumila)富士／小金海棠(M.xiaojinensis)和苹果新红星／小金海棠两种砧穗组合的嫁接成活率均在80％以上;嫁接后10d左右,接穗新叶开始生长;嫁接后2个月,嫁接苗移栽成活率达70％左右。移栽1个月后,嫁接口已完全愈合;移栽3个月后,富士／小金海棠和新红星／小金海棠两种嫁接苗的茎杆粗细均匀,无大小脚现象,嫁接苗的平均株高分别为8．08cm和7．34cm,平均干周分别为1．30cm和1．32cm,长势与同时移栽的富士、新红星组培苗一致。从早期的鉴定结果可以看出,小金海棠与富士、新红星具良好的嫁接亲和性。 相似文献
10.
为了研究不同铁效率基因型苹果砧木铁吸收利用的分子机理,本研究以铁低效基因型山定子(Malus baccata Borkh)为试材,根据实验室从铁高效基因型小金海棠(Malus xiaojinensis)克隆到与铁运输相关的基因柠檬酸合成酶基因(MxCS1)的全长序列设计特异引物,通过RT-PCR方法从山定子cDNA中克隆到柠檬酸合成酶基因CS,基因全长为1422bp,与金冠(Malus domestica Borkh cv.Golden Delicious)、小金海棠中的CS基因具有较高的同源性,将该基因命名为MbCS1(GenBank登录号:JQ898346)。利用生物信息学软件对山定子柠檬酸合成酶基因(MbCS1)进行预测分析,结果显示该基因预测编码473个氨基酸,相对分子量为54.26kD,理论等电点为8.95。亚细胞定位显示MbCS1蛋白定位在细胞膜上。半定量RT-PCR及Real-time PCR分析均表明,正常供铁时,该基因在山定子的根、茎、新叶中都有表达;缺铁处理(EDTA-NaFe,4μmol/L)时,该基因在根、茎和新叶中的表达加强,第9天达到最高值,之后开始下降;但各检测器官中表达增强的程度不同,其中茎中受缺铁诱导表达最明显。与小金海棠中MxCS1基因的表达趋势有明显的差别。本研究为高等植物抗性机理的深入研究以及铁低效资源型砧木资源的改良提供了基础资料。 相似文献
11.
《Journal of plant nutrition》2013,36(8):1381-1393
Abstract Root and leaf ferric chelate reductase (FCR) activity in Annona glabra L. (pond apple), native to subtropical wetland habitats and Annona muricata L. (soursop), native to nonwetland tropical habitats, was determined under iron (Fe)-sufficient and Fe-deficient conditions. One-year-old seedlings of each species were grown with 2, 22.5, or 45 µM Fe in a nutrient solution. The degree of tolerance of Fe deficiency was evaluated by determining root and leaf FCR activity, leaf chlorophyll index, Fe concentration in recently mature leaves, and plant growth. Root FCR activity was generally lower in soursop than in pond apple. Eighty days after plants were put in nutrient solutions, leaf FCR activity of each species was lower in plants grown with low Fe concentrations (2 µM) than in plants grown with high (22.5 or 45 µM) Fe concentrations in the nutrient solution. Leaves of pond apple grown without Fe became chlorotic within 6 weeks. The Fe level in the nutrient solution had no effect on fresh and dry weights of soursop. Lack of Fe decreased the leaf chlorophyll index and Fe concentration in recently matured leaves less in soursop than in pond apple. The rapid development of leaf chlorosis in low Fe conditions and low root and leaf FCR activities of pond apple are probably related to its native origin in wetland areas, where there is sufficient soluble Fe for adequate plant growth and development. The higher leaf FCR activity and slower growth rate of soursop compared to pond apple may explain why soursop did not exhibit leaf chlorosis even under low Fe conditions. 相似文献
12.
This work studied the behavior of different iron (Fe)-chelates as substrates of ferric chelate reductase (FCR) and their ability as Fe suppliers for mildly chlorotic plants. FCR activity and Fe concentration in xylem sap were determined in green stressed cucumber plants with different stress levels using different synthetic chelates as substrates. Both reduction and Fe concentration in the xylem sap were higher for the less-stable Fe chelates, except for Fe-EDTA, which presented a relatively low Fe concentration in sap. It was concluded that a high stability of the chelate in the nutrient solution reduces the Fe reduction, but other factors, such as the complexation of the Fe(II) by the chelating agents, should be considered when the complete process of Fe uptake is studied. The use of both indexes together, i.e., FCR determination and xylem sap concentration, is useful for understanding the Fe uptake from different Fe chelates. 相似文献
13.
Jos�� Sebasti��n D��vila Costa Virginia Helena Albarrac��n Carlos Mauricio Abate 《Water, air, and soil pollution》2011,216(1-4):527-535
Amycolatopsis tucumanensis, a recently recognized novel species showed remarkable copper resistance as well as efficient Specific Cupric Reductase Activity (SRACu) in both, copper adapted and non-adapted cells, under different temperatures of incubation. Its copper resistance strength was highlighted against other metal-resistant actinobacteria (Streptomyces sp. AB5A) and sensitive strains (Amycolatopsis eurytherma and Streptomyces coelicolor). Pre-adapted cells of A. tucumanensis displayed values of SRACu, on average, 65% higher than those obtained from non-adapted cells. In addition, preadaptation of A. tucumanensis improved the rate of Cu(II) reduction which was approximately, two-, seven- and ninefold higher than pre-adapted cells from Streptomyces sp. AB5A, A. eurytherma and S. coelicolor, respectively. A. tucumanensis showed the highest levels of SRACu at all temperatures and also the highest copper resistance profile, suggesting that these two abilities may be in close relationship. This ostensible versatility, related to the temperature, of adapted cells from A. tucumanensis might support the application of this strain under different bioremediation conditions. To our knowledge this is the first time that cupric reductase activity was demonstrated within the genus Amycolatopsis. 相似文献
14.
The influence of the siderophore produced by plant growth‐promoting Pseudomonas putida strain WCS358, pseudobactin 358 (PSB358), on chlorophyll synthesis and iron(III) reduction in carnation cultivars Lena and Pallas grown in hydroponics was studied. Ferric pseudobactin 358 (4 μM) stimulated chlorophyll synthesis in cv. Lena, but not in cv. Pallas. FeEDDHA stimulated chlorophyll synthesis in both cultivars. Differences between the two carnation cultivars in the utilization of FePSB358 as an iron source are correlated with differences in iron efficiency reactions of the two cultivars: Fe‐deficient plants of cv. Lena produced more and longer root hairs than Fe‐deficient plants of cv. Pallas, and the ferric reducing activity of cv. Lena was higher than that of cv. Pallas. 相似文献
15.
Camino García-Balboa Dina Cautivo M. L. Blázquez F. González J. A. Muñoz A. Ballester 《Water, air, and soil pollution》2010,207(1-4):213-226
In the present work the kinetics of ferric reduction was investigated using dissimilatory ferric- and sulphate-reducing bacterial cultures. The effect of sulphate reduction on Fe(III) reduction was also studied. The study is an attempt to improve the biological reduction rate of Fe(III) as an alternative biotechnological way to the reduction step in steelmaking processing operations. The results obtained show that the reduction of ferric iron and sulphate took place in a successive way and none synergetic effect was detected. The simultaneous action of both metabolic activities did not enhance the process but slowed down the kinetics of ferric reduction. The reduction process of 3 g/L of soluble ferric and 3 g/L of sulphate lasted 25 days. Ferric iron was the first electron acceptor to be reduced in the first 15 days followed by the sulphate reduction in the following 10 days. That result suggests that ferric reduction is a preferential metabolic process over sulphate reduction when both electron acceptors coexist. None improvement in the kinetics was observed using an electron donor concentration in excess. In contrast, the total reduction of ferric ion (3 g/L) with adapted bacterial cultures was achieved in only 36 h. The presence of sulphate had no effect on the ferric reduction. Finally, an improved culture medium for ferric-reducing bacteria is also proposed. 相似文献
16.
The reduction capacity for ferric chelates of Geum urbanum L. showed a marked increase when plants were grown under conditions of iron‐shortage. Ferric ethylenediaminetetraacetate (FeEDTA) was reduced with a pH optimum between 5 and 6. The reaction exhibited a low substrate affinity with a Km much higher than the expected concentration range of soluble iron in the soil. Analysis of the saturation plots conform to Michaelis‐Menten kinetics. Both Vmax and Km values varied to a broad extent with changing assay and plant culturing conditions. Ferricyanide was reduced at significantly higher rates than FeEDTA and inhibited the reduction of FeEDTA compete‐tively. The kinetic characteristics of iron reduction by plants will be discussed in terms of ecological significance as part of an adaptation to the soil conditions. 相似文献
17.
We studied the spatial distribution of the amount and types of Fe oxides in a Ferric Acrisol having a red-yellow mottled horizon. Red mottles contain hematite and goethite; yellow mottles have goethite but in amounts markedly smaller than those of the contiguous red mottles. We think that reductive dissolution caused the disappearance of the hematite and part of the goethite in the yellow mottles. This is supported by the experimental dissolution of the iron oxides of the clay fractions: hematite dissolved, on the average, at a rate twice of that of goethite. 相似文献
18.
水稻土裂缝的演变及其还原酶活性的变化研究 总被引：2，自引：1，他引：2
在温室中对3种水稻土进行土柱培养,加水使土壤含水量充分饱和,然后土壤自然干燥。在该过程中,粘粒含量高的青紫泥和黄斑田产生了裂缝,而粘粒含量低的小粉土没有开裂。青紫泥和黄斑田在裂缝演变过程中,单位面积的总长度、最大宽度以及最大深度都遵循着一定的演变规律。裂缝的产生对于硝酸还原酶和羟胺还原酶活性的变化影响显著,但是亚硝酸还原酶活性的变化与裂缝的产生没有多大关系。在裂缝产生到稳定过程中,青紫泥和黄斑田的硝酸还原酶、亚硝酸还原酶和羟胺还原酶活性的演变规律相似。 相似文献