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1.

In the present study, wild orchardgrass ( Dactylis glomerata L.) collected from four different locations (Oltu, Olur, Uzundere and Uluba < ) of Erzurum (Turkey) were used to investigate various electrophoretic analyses within and between four ecotypes based on peroxidase (PRX), polyphenol oxidase (PPO) and malate dehydrogenase (MDH) enzymes. Four natural populations of Dactylis glomerata showed genetic variability within and among ecotypes based on similarities and polymorphism of isoenzyme bands. In addition, a dendogram analysis was made according to band patterns indicating that ecotypes could be grouped showing characteristics in relation to each other, notably Oltu-Uluba < and Olur-Uzundere. However, the degree of relationship of Oltu-Uluba < (6 units) was higher than Olur-Uzundere (12 units).  相似文献   

2.
Current tools used to assess the safety of food and feed derived from modern biotechnology emphasize the investigation of possible unintended effects caused directly by the expression of transgenes or indirectly by pleiotropy. These tools include extensive multisite and multiyear agronomic evaluations, compositional analyses, animal nutrition, and classical toxicology evaluations. Because analytical technologies are rapidly developing, proteome analysis based on two-dimensional gel electrophoresis (2DE) was investigated as a complementary tool to the existing technologies. A 2DE method was established for the qualitative and quantitative analysis of the seed proteome of Arabidopsis thaliana with the following validation parameters examined: (1) source and scope of variation; (2) repeatability; (3) sensitivity; and (4) linearity of the method. The 2DE method resolves proteins with isoelectric points between 4 and 9 and molecular masses (MM) of 6-120 kDa and is sensitive enough to detect protein levels in the low nanogram range. The separation of the proteins was demonstrated to be very reliable with relative position variations of 1.7 and 1.1% for the pI and MM directions, respectively. The mean coefficient of variation of 254 matched spot qualities was found to be 24.8% for the gel-to-gel and 26% for the overall variability. A linear relationship (R2 > 0.9) between protein amount and spot volume was demonstrated over a 100-fold range for the majority of selected proteins. Therefore, this method could be used to interrogate proteome alterations such as a novel protein, fusion protein, or any other change that affects molecular mass, isoelectric point, and/or quantity of a protein.  相似文献   

3.
采用蛋白质组双向电泳技术和已鉴定蜂王浆蛋白质组的比对,对王浆高产蜜蜂（浆蜂）和中华蜜蜂（中蜂）的蜂王浆蛋白质组进行了差异比较。结果表明,浆蜂蜂王浆的总蛋白质点数（93个）显著高于中蜂蜂王浆总蛋白质点数（70个）,它们的等电点主要集中在5-8之间,分子量在50-80kDa之间,其中共有蛋白质点为30个;通过与已鉴定的浆蜂蜂王浆蛋白质组比较,这些共有蛋白质点推断为蜂王浆主蛋白1、2、3和葡萄糖氧化酶。同时两蜂种蜂王浆的蛋白质丰度也存在较大差异,共有点中有4个蛋白质的丰度中蜂显著高于浆蜂,7个浆蜂显著高于中蜂中。结果表明浆蜂和中蜂蜂王浆蛋白质种类及丰度均有较大差异。  相似文献   

4.
为检测和分析0.5和3.0Gy 2种剂量重离子辐照绵羊精子差异表达的蛋白质,用二维凝胶电泳分离精子蛋白,凝胶用硝酸银染色,PDQuest8.0检测分析差异表达蛋白斑点并经高效液相色谱串联离子阱质谱鉴定,结果显示:2种剂量凝胶图谱分析后得到8个共同差异蛋白斑点,鉴定为3种表达上调的蛋白质(谷氧还蛋白1、转录因子AP-2α...  相似文献   

5.
The market of specialty eggs, such as omega-3-enriched eggs, organic eggs, and free-range eggs, is continuously growing. The nutritional composition of egg yolk can be manipulated by feed diet; however, it is not known if there is any difference in the composition of egg white proteins among different egg varieties. The purpose of the study was to compare the egg white proteins among six different egg varieties using proteomics analysis. Egg white proteins were analyzed using two-dimensional gel electrophoresis (2-DE), and 89 protein spots were subjected to LC-MS/MS. A total of 23 proteins, belonging to Gallus gallus , were identified from 72 detected protein spots. A quiescence-specific protein precursor in egg white was identified for the first time in this study. Significant differences in the abundant levels of 19 proteins (from 65 protein spots) were observed among six egg varieties. Four proteins, ovalbumin-related protein Y, cystatin, avidin, and albumin precursor, were not different among these six egg varieties. These findings suggest that the abundance, but not the composition, of egg white proteins varied among the egg varieties.  相似文献   

6.
为建立一套适合于牡丹试管苗茎基部蛋白的双向电泳技术,以便更好地利用蛋白质组技术研究牡丹试管苗不定根的发生机理,本研究比较了三种不同蛋白质提取方法对双向电泳结果的影响,并在蛋白质上样量方面进行了比较。结果表明,乙酸铵/甲醇酚提取法所得2-DE图谱的蛋白点很少,仅检测到45个,且较模糊,有明显的拖尾现象,分辨率很低;乙醇/乙醚丙酮法所得的蛋白点也较少（101个）,较模糊,且横竖纹干扰较大;三氯乙酸/丙酮法所得蛋白点数较多,可检测到434个清晰的蛋白点,且形状规则,重复性好,适合后续分析,操作也较为简便。用三氯乙酸/丙酮法提取蛋白,采用800μg、1000μg和1200μg三个不同的上样量进行双向电泳,在上样量为1200μg时（IPGpH3～10,24cm）,蛋白质在12%SDS-PAGE胶上得到了较好的分离,在2-DE图谱上可分辨出562个蛋白点。因此,三氯乙酸/丙酮法是较适合于牡丹试管苗茎基部蛋白质提取的方法,1200μg是较为合适的上样量。  相似文献   

7.
Sixteen ecotypes of Corbarini small tomatoes were studied. The antioxidant activity was evaluated with the DMPD (N, N-dimethyl-p-phenylenediamine) method in the water-soluble fraction (S-AA) and with the ABTS [2, 2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)] method in the water-insoluble fraction (I-AA). S-AA and I-AA were strongly related to each other, varying to a large extent between the ecotypes considered. They were also significantly correlated with fruit weight, total solids, and refractive index. The combination of the latter variables (by multiple regression analysis) accounted for 36% and 44% of the total variability of S-AA and I-AA, respectively. Moreover, when the ecotypes were subdivided according to their shape (round, pear-shaped, oval, and long), S-AA and I-AA were both significantly higher in round tomatoes and lower in the long ones. In conclusion, the antioxidant activity varies to a considerable extent between different ecotypes of Corbarini small tomatoes. These differences are related to shape and some other fruit characteristics.  相似文献   

8.
Centuries of co-evolution between Castanea spp. biodiversity and human populations has resulted in the spread of rich and varied chestnut genetic diversity throughout most of the world, especially in mountainous and forested regions. Its plasticity and adaptability to different pedoclimates and the wide genetic variability of the species determined the spread of many different ecotypes and varieties in the wild. Throughout the centuries, man has used, selected and preserved these different genotypes, vegetatively propagating them by grafting, for many applications: fresh consumption, production of flour, animal nutrition, timber production, thereby actively contributing to the maintenance of the natural biodiversity of the species, and providing an excellent example of conservation horticulture. Nonetheless, currently the genetic variability of the species is critically endangered and hundreds of ecotypes and varieties are at risk of being lost due to a number of phytosanitary problems (canker blight, Chryphonectria parasitica; ink disease, Phytophthora spp.; gall wasp, Dryocosmus kuriphilus), and because of the many years of decline and abandonment of chestnut cultivation, which resulted in the loss of the binomial male chestnut. Recently, several research and experimentation programmes have attempted to develop strategies for the conservation of chestnut biodiversity. The purpose of this paper is to give an overview of the status of biodiversity conservation of the species and to present the results of a 7?year project aimed at the individuation and study of genetic diversity and conservation of Castanea spp. germplasm.  相似文献   

9.
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10.
Biochemical characteristics of patatin proteins purified by ion-exchange and affinity chromatography from tubers of 20 potato cultivars were studied to evaluate their genotype differences with respect to utility groups, table potato cultivars (TPCs) and processing potato cultivars (PPCs). Both groups of cultivars showed similar values of protein content in dry matter (3.98-7.39%) and of patatin relative abundance (5.40-35.40%). Three mass levels (～40.6, 41.8, and 42.9 kDa) of purified patatins were found by MALDI-TOF MS within all cultivars. Differences among mass levels corresponding with the mass of sugar antenna (～1.2 kDa) confirmed the previous concept of different glycosylation extentsin patatin proteins. It was showed that the individual types of patatin varying in their masses occur in the patatin family in a ratio specific for each of the cultivars, with the lowest mass type being the major one. Electrophoretic analyses demonstrated wide cultivar variability in number of patatin forms. Especially 2D-PAGE showed 17-23 detected protein spots independently on the utility group. Specific lipid acyl hydrolase (LAH) activity of purified patatins from the individual tested cultivars varied between 0.92 and 5.46 μmol/(min mg). Patatin samples within most of the TPCs exhibited higher values of specific LAH activity than samples of PPCs. It may be supposed that individual patatin forms do not have similar physiological roles.  相似文献   

11.
Proteome analysis was successfully applied to study the alterations in fish muscle proteins during ice storage. The processes occurring during post-mortem metabolism are known to lead to characteristic changes in the texture and taste of fish muscle. Endogenous proteases are anticipated to play the major role in these processes, although the exact mechanisms during fish meat tenderization have yet to be depicted. Protein changes in cod (Gadus morhua) muscle were followed during 8 days of storage. Within the partial proteome (pI 3.5-8.0, MW 13-35 kDa) significant changes were found in 11 protein spots. In nine protein spots the intensity increased, and for eight of these the increases were significant (p < 0.05) within the first 2 h post-mortem. In contrast, two protein spots decreasing in intensity showed significant (p < 0.03) changes after 8 days, thereby indicating that in the fish muscle different biochemical processes are involved in the protein changes observed post-mortem.  相似文献   

12.
Plant roots grow into the soil for efficient acquisition of various nutrients, such as inorganic nitrogen, ammonium, and nitrate. A previous study has revealed the genetic diversity of foliar functions of Arabidopsis thaliana ecotypes in an environment containing ammonium; however, the function of roots remains unclear. This work focuses on the root system architecture (RSA) of Arabidopsis ecotypes to investigate the genetic factors regulating ammonium-dependent RSA changes. Arabidopsis ecotypes were grown on vertical agar medium containing ammonium as a major nitrogen source, and root growth and RSA were determined. Arabidopsis ecotypes showed differential sensitivity to ammonium. The shoot dry weight of some ecotypes decreased, whereas that of other ecotypes increased in ammonium medium. The RSA changes also varied among the different ecotypes in response to ammonium. The total root length, measured as the sum of primary root and lateral root length, of some ecotypes was reduced, whereas that of other ecotypes showed no significant difference in ammonium medium. Compared with lateral roots, the primary roots showed a sharp response to ammonium supply. Notably, the RSA showed a partial correlation with shoot dry weight in ammonium medium. Because Col-4 and Ler-0 showed opposite RSA responses to ammonium supply, these two ecotypes were selected for further genetic analysis. Quantitative trait locus (QTL) analysis of recombinant inbred lines of Col and Ler showed the involvement of several genetic factors in ammonium-dependent RSA changes. Moreover, QTL analysis revealed that the primary structure of nitrogen-related enzymes do not account for changes in RSA in response to ammonium supply.  相似文献   

13.
Milicia species(M. excelsa andM. regia, Moraceae, theirokos) are among the most useful indigenous rain forest treespecies in Africa. They are not grown in plantations but extractedfrom the natural forest at an unsustainable rate. The majorconstraint on cultivation and afforestation is thegall-forming insect Phytolyma lata, whichdestroys seedlings through gall formation, causing dieback of theshoot. Genetic diversity studies should aid the design ofconservation and resistance-breeding strategies. This study,therefore, aimed to investigate the genetic variability inMilicia from three West African countries andthe power of bulk-family DNA in partitioning of geneticdiversity. Genetic variation within and between 5 natural populationsof Milicia from Ghana, Côte d'Ivoireand Sierra Leone was investigated using random amplified polymorphicDNA (RAPD) and non-coding chloroplast DNAsequencing. RAPD analysis of 41 DNA samples with 8 primers produced atotal of 74 bands with 40 bands (54.1%) beingpolymorphic. Dendrogram analysis produced 2 major clusters separatingdry and moist/wet ecotypes. Sub-clustering furtherseparated accessions by forest type and/or geographic region. Agreater proportion of the total genetic variability was attributed tovariation between populations (62.2%) than withinpopulations (37.8%). Sequencing of chloroplasttRNALeu intron (cptrnL) and intergenic spacer(cpIGS) between cptrnL and tRNAPhe(trnF) did not show any variation. The RAPD analysissuggests that trees from wet/moist sites are mostlyM. regia, while those fromdry sites are mostly M.excelsa. Sequencing results, however, suggestthat the two species are closely related. These results are discussedwith reference to conservation strategies.  相似文献   

14.
In 1999, natural populations of Dactylis glomerata L., Festuca arundinacea Schreb., Phalaris tuberosa L. and the macrotherm species Brachypodium rupestre (Host) R. et S., were collected as seed in Southern Italy (Apulia, Basilicata and Campania regions) to evaluate the potential as forage of native germplasm under dry conditions, using available commercial cultivars as controls. The persistence of accessions, biometric, phenologic, productive and qualitative parameters of forage have been studied for a 3-year period (2000–2002) in a typical Mediterranean environment. D. glomerata is widely present in the studied area with two subspecies (subsp. glomerata L., subsp. hispanica (Roth) Nyman). The research pointed out significant differences in plant size, earliness and single plant dry matter (DM) production. All the natural ecotypes belonging to this species showed higher persistence than the control cultivars. Some natural ecotypes appeared to have potential for improving DM yield (subsp. glomerata ecotypes) and reducing neutral detergent fibre (NDF) content (subsp. hispanica ecotypes). Natural ecotypes belonging to F. arundinacea, P. tuberosa, and B. rupestre are sporadically present in the studied area. Among them, two ecotypes of P. tuberosa, showing higher winter growth and earliness compared to cv. ‘Holdfast’, seem to be interesting for a future breeding programme.  相似文献   

15.
《Journal of plant nutrition》2013,36(9):1561-1573
Abstract

For this study, the diversity of some effects of salinity on rosette growth, seed production, and seed viability in Arabidopsis thaliana was explored. Plants from 5 accessions (“ecotypes”) were grown on a sand-peat mixture irrigated with nutritive medium. At the emergence of inflorescence axis, 35 mM NaCl was added to the solution. Several morphological parameters of the rosette were measured at the beginning of silique opening, irrigation was interrupted and the substrate was allowed to dry. The siliques and the mature seeds were harvested, and assayed for K+, Na+ and Cl?. Seeds were sown on pure water and germination kinetics were determined. There was a large inter-ecotype variability of the effect of salt on the rosette growth and on all factors contribution to reproduction (number of inflorescence axes, number of flower per axis, flower fertility, and seed viability). Among the five ecotypes, those with highest levels of Na+ and Cl? in the silique valves produced the less viable seeds. This comparison, (i) suggests that salt accumulation in the former organs has detrimental effect on seed maturation, and (ii) indicates that there is a natural variability for control of ion transport to fruits in A. thaliana.  相似文献   

16.
为了研究除草剂作用机理,用光系统Ⅱ抑制型除草剂阿特拉津（Atrazine）处理小麦幼苗,用2-DE技术和生物质谱方法,分析了叶绿体蛋白质组的变化。结果发现,在10mg·L-1浓度处理时,有7个叶绿体蛋白质斑点（斑点1,50kDa/PI8.1;斑点2,41kDa/PI8.4;斑点3,41kDa/PI7.6;斑点4,23kDa/PI7.1;斑点5,31kDa/PI5.0;斑点6,35kDa/PI8.9;斑点7,14kDa/PI8.1）丢失。对7个发生变化的斑点利用MALDI-MS方法,于NCBI进行数据查询,其中,有6个叶绿体蛋白质归属得到鉴别,它们是Calvin循环中,固定CO2的RuBPcase的激活酶（2个同工体和1个β型前体）,在H2O氧化裂解中起重要作用的23kDa氧释放蛋白（psbp protein）,在能量转贮中起重要作用的3-磷酸甘油酸激酶和催化HCO3--CO2水合作用可逆反应的碳酸酐酶。研究表明,叶绿体蛋白质组中丢失的6个蛋白质是Atrazine处理的相关蛋白。  相似文献   

17.
A combined proteomic approach was applied for the separation, identification, and comparison of two major storage proteins, beta-conglycinin and glycinin, in wild (Glycine soja) and cultivated (Glycine max) soybean seeds. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) with three different immobilized pH gradient (IPG) strips was an effective method to separate a large number of abundant and less-abundant storage proteins. Most of the subunits of beta-conglycinin were well-separated in the pH range 3.0-10.0, while acidic and basic glycinin polypeptides were well-separated in pH ranges 4.0-7.0 and 6.0-11.0, respectively. Although the overall distribution pattern of the protein spots was similar in both genotypes using pH 3.0-10.0, variations in number and intensity of protein spots were better resolved using a combination of pH 4.0-7.0 and pH 6.0-11.0. The total number of storage protein spots detected in wild and cultivated genotypes was approximately 44 and 34, respectively. This is the first study reporting the comparison of protein profiles of wild and cultivated genotypes of soybean seeds using proteomic tools.  相似文献   

18.
猪肌肉组织双向电泳分离条件的建立及常见问题分析   总被引：1，自引：0，他引：1  
为建立和优化猪肌肉组织蛋白质双向电泳技术,从提取方法、加样量、IPG胶条的选择、SDS-PAGE胶浓度、染色方法等多个方面对双向电泳分离效果进行比较研究.结果显示,液氮研磨+超声破碎法提取猪肌肉组织蛋白效果优于液氮研磨法,前者细胞破碎彻底蛋白质溶解性好且核酸污染少,图谱质量较好;18cm pH 4～7的IPG胶条分离效果比pH 3～10非线性IPG胶条好;对于银染,18 cm pH 4～7的IPG胶条150μg上样量比较适宜;同一肌肉样品的三块胶的重复性可达70％以上;同时,对双向电泳过程中的典型问题作了详尽分析.研究结果表明,通过优化的双向电泳条件获得了较高分辨率和较好重复性的猪肌肉组织双向电泳图谱,可用于后续蛋白质组学分析.  相似文献   

19.
The content of organosulfur compounds was determined in selected garlic cultivars grown at four locations in Andalusia, Spain. The organosulfur compounds studied were three γ-glutamyl peptides, namely, γ-l-glutamyl-S-(2-propenyl)-l-cysteine (GSAC), γ-l-glutamyl-S-(trans-1-propenyl)-l-cysteine (GSPC), and γ-l-glutamyl-S-methyl-l-cysteine (GSMC), and four cysteine sulfoxides (alliin, isoalliin, methiin, and cycloalliin). There was a significant effect of the location, cultivar, and garlic ecotype on individual organosulfur compound contents. Purple-type cultivars showed on average the highest contents of GSMC, GSAC, alliin, and methiin but the lowest isoalliin content. The impact of genotype was relatively high for GSAC, whereas this factor hardly contributed to the total variability in alliin and isoalliin content. Planting date had a significant effect on the content of alliin and isoalliin. Discriminant analysis evidenced the ability of organosulfur compounds to distinguish among garlic bulbs from different locations or ecotypes with 81 or 86% accuracy, respectively.  相似文献   

20.
This study evaluated effects of drought stress on the concentrations of mineral elements in the leaves of different ecotypes of castor plant. The experimental design was a split-plot randomized complete block with three replications. It was carried out at Fozveh Agricultural Research Station located 20 km from Esfahan, Iran in 2014. The main plot was four levels of water stress (30%, 45%, 60%, and 75% moisture content) and the sub-plot was six castor plant ecotypes (Esfahan, Ardestan, Arak, Naein, Yazd, and Ahwaz). The potassium (K), magnesium (Mg), calcium (Ca), sodium (Na), iron (Fe), copper (Cu), and zinc (Zn) contents were measured and found to be significantly affected by drought stress. Drought stress increased K, Ca and Na contents and decreased Fe, Cu, Zn, and Mg contents of the plants. Various ecotypes of castor plant showed different reactions to drought stress and the Fe concentration varied significantly by the castor plant ecotypes. Interaction between water stress and ecotypes was significant only for Cu and Zn. Understanding the element concentrations under drought stress can be useful for predicting growth and development of castor plants.  相似文献   

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