适于SDS-PAGE分析的苹果叶片蛋白质提取方法

为探索苹果叶片蛋白质SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析的样品制备方法,比较了三氯乙酸(TCA)/丙酮沉淀法、改良的Tris-HC1法、酚-甲醇/醋酸铵沉淀法、二硫苏糖醇(DTT)/丙酮法及Tris-HC1法等5种蛋白质的提取方法.制备的样品经SDS-PAGE分离采用银染显色.结果表明,改良的Tris-HC1法在加大PVPP用量和蛋白质提取缓冲液的基础上,将丙酮沉淀蛋白质的时间由30 mln增加到1 h,使蛋白质的得率最高为3.243μg/μL,上样量5μL得到的蛋白质条带数量最多,条带最清晰为41条.利用这一改良方法对苹果实生树不同节位蛋白质的动态变化进行了研究,共发现6条蛋白质差异带,分子量分别为71.9,60.5,52.6,41.1,35.3,18.5 kDa,条带清楚,背景清晰.因此,改良的Tris-HC1法适用于苹果叶片的SDS-PAGE分析.     

苦瓜种子蛋白质组双向电泳技术条件的优化

通过对苦瓜种子蛋白质样品的提取方法、等电聚焦参数、SDS-PAGE分离胶浓度以及胶条pH范围的优化筛选,建立了适合苦瓜种子蛋白组分析的双向电泳体系.结果表明:使用TCA-丙酮提取方法提取苦瓜种子蛋白,更适用于双向电泳分析,结合使用pH=5~8 IPG胶条以及优化的聚焦程序,能获得分辨率较高、蛋白点清晰、重复性好的2-DE图谱.经银染显色,可检测约660个蛋白点,主要分布在p H=5~8;该体系适合用于分析苦瓜种子的蛋白质组.     

陆地棉花药蛋白质组分析中双向电泳技术体系的建立与优化

建立了适用于陆地棉花药蛋白质组研究的双向电泳技术.以陆地棉花药为材料,采用液氮研磨的方法破碎组织,然后用三氯乙酸(TCA)/丙酮沉淀法和苯酚抽提结合甲醇/醋酸铵沉淀法提取蛋白质.采取载体两性电解质pH梯度等电聚焦/SDS-PAGE和固相pH梯度等电聚焦/SDS-PAGE双向凝胶电泳,对陆地棉花药总蛋白质进行了分离.通过...     

茶树叶片蛋白的SDS-PAGE分析

以茶树鲜叶为材料,对4种常用的蛋白提取方法(TCA沉淀法、冷丙酮沉淀法、TCA-丙酮沉淀法、Tris-丙酮沉淀法)、SDS-PAGE条件及影响蛋白定量的因素进行了比较研究。确定了一套优化的适用于茶树叶片蛋白提取、定量、凝胶制备、电泳和染色的方法。利用这一优化方法对茶树鲜叶蛋白和不同蔗糖浓度处理的愈伤组织蛋白进行了研究,找出了差异条带。     

不同品种油葵水浸提液对3种药用甘草化感效应的研究

为探讨油葵对甘草潜在的化感作用,选择矮大头(567 DW)、矮早丰(BKZ 9806)、新葵4号以及新葵6号等不同品种油葵和3种药用甘草为研究对象.用油葵根、茎、叶等器官不同浓度的水浸提液分别处理乌拉尔甘草、胀果甘草和光果甘草的种子和幼苗,比较0,25,50 mg/mL 3个浓度的水浸提液对3种甘草种子萌发和幼苗生长影响格局的差异.结果表明,不同油葵品种、不同油葵器官的水浸提液的化感效应不尽相同,矮大头浸提液对胀果甘草的种子萌发和幼苗生长均无明显抑制作用,并且其根的提取液显著促进了根的生长,在25,50 mg/mL浓度下分剐比ck高出26％和37％(p＜0.05),RI分别为0.21和0.27.根据本实验结果,在生产实践中建议选用矮大头与胀果甘草进行套种,可望能提高种植收益.     

虉草种子破眠蛋白质双向电泳体系的建立

为了建立一套适合于虉草种子破眠蛋白质双向电泳的技术体系,以内蒙古民族大学农学院选育的"通选一号"虉草种子为试验材料,对其蛋白质提取方法、分离技术等进行了探讨。结果表明:采用三氯乙酸/丙酮沉淀法(TCA/A法)提取种子蛋白,结合使用p H 4~p H 7的18 cm IPG胶条,200μg的上样量,12%SDS-PAGE胶,硝酸银法染色,可得到清晰、丰富的蛋白质点。     

玉米叶片2种总蛋白质提取方法的比较

为比较不同方法提取的玉米叶片蛋白质对双向电泳效果的影响。采用TCA-丙酮沉淀和TRIzol试剂盒2种方法,提取国审玉米品种郑单958叶片全基因组蛋白质,并对提取的蛋白质样品进行双向电泳。结果表明:TCA-丙酮方法提取的蛋白质一向水平聚焦电压上升缓慢且达不到程序设置的电压,而TRIzol试剂盒方法实时监测图与程序设置图相吻合;对电泳胶图分析后可知,TRIzol试剂盒法分离出的蛋白点多于900个,低丰度蛋白质得到充分显现,大多数蛋白点分子量为20~80 k Da,p H值为4~7,蛋白点的形状较规则;TCA-丙酮沉淀法分离出的蛋白点仅有350个,表明样品中蛋白质分离种类较少,高丰度蛋白质过度显现,低丰度蛋白质检出率大大降低。     

适用于蛋白质组分析的粉红单端孢菌蛋白提取方法的建立

为了优化一种适用于粉红单端孢菌双向电泳的蛋白质提取方法,以期为蛋白质组学水平研究粉红单端孢菌的致病机制奠定基础。以分离纯化后的粉红单端孢菌为试验材料,分别采用超声-TCA-丙酮、超声-磷酸-TCA-丙酮、超声-SDS、超声-TCA/丙酮-酚/SDS联合抽提和TCA/丙酮-酚/SDS联合抽提等5种方法提取菌体蛋白质,通过对比分析蛋白质含量以及纯度筛选出2种较好的提取方法。在筛选聚丙烯酰胺电泳凝胶浓度的基础上,再通过双向电泳对比分析,得出最佳的蛋白质提取方法。结果表明,12%的聚丙烯酰胺凝胶浓度获得的单向电泳图谱背景清晰且无严重的拖尾现象,超声-TCA/丙酮-酚/SDS联合抽提法获得蛋白样品的质量浓度和含量分别为6.650 mg/mL和2.993 mg/g,该法提取蛋白通过SDS-PAGE分析条带清晰,双向电泳分析可获得1 238个独立清晰的蛋白点。由此可知,超声-TCA/丙酮-酚/SDS联合抽提法获得的粉红单端孢菌体蛋白适合于双向电泳分析及蛋白质组学研究。     

向日葵幼苗全蛋白提取及双向电泳技术体系的建立

本研究以向日葵幼苗为材料,比较TCA-丙酮提取法、Tris饱和酚提取法以及水溶法三种提取蛋白的方法,并且对双向电泳技术进行研究,初步建立向日葵幼苗蛋白的双向电泳体系。研究结果表明,用Tris饱和酚提取法提取蛋白含量最高,经蛋白定量后显示含量可达43.359 ug/u L;用不同浓度分离胶进行SDS-PAGE凝胶电泳,结果显示用8%分离胶可分离条带数较多,最多可分离20条蛋白条带;用不同聚焦时间以及不同上样量进行双向电泳,结果显示一向IEF聚焦时间为6 h左右,上样量为1 mg的条件较为适合,用Image Master5.0软件分析检测到蛋白点为102个。     

酸溶蛋白质电泳方法鉴定小麦种子纯度

本试验通过对小麦种子酸溶蛋白质的提取液浓度、提取时间和电泳凝胶组分浓度进行研究,建立了一种对小麦种子纯度进行鉴定的常规电泳技术方法.结果表明该方法具有以下特点:①样品提取时间短;②蛋白质组分多态性丰富;③电泳谱带清晰,分辨率高.     

Biological Activity and Quantification of Suspected Allelochemicals from Alfalfa Plant Parts

Autotoxicity restricts reseeding of alfalfa (Medicago sativa L.) after alfalfa until autotoxic chemical(s) breaks down or is dispersed into external environments. A series of aqueous extracts from leaves, stems, roots and seeds of alfalfa ‘Vernal’ were bioassayed against alfalfa seedlings of the same cultivar to determine their autotoxicity. The highest inhibition was found in the extracts from the leaves. Extracts at 40 g dry tissue l^?1 from alfalfa leaves were 15.4, 17.5 and 28.7 times more toxic to alfalfa root growth than were those from roots, stems and seeds, respectively. A high‐performance liquid chromatography (HPLC) analysis with nine standard compounds showed that the concentrations and compositions of allelopathic compounds depended on the plant parts. In leaf extracts that showed the most inhibitory effect on root growth, the highest amounts of allelochemicals were detected. Among nine phenolic compounds assayed for their phytotoxicity on root growth of alfalfa, coumarin, trans‐cinnamic acid and o‐coumaric acid at 10^?3 m were most inhibitory. The type and amount of causative allelochemicals found in alfalfa plant parts were highly correlated with the results of the bioassay, indicating that the autotoxic effects of alfalfa plant parts significantly differed.     

Changes in Seed Yield and Quality with Maturity in Onion (Allium cepa L., cv. 'Early Cream Gold')

Development of onion (Allium cepa L., cv. ‘Early Cream Gold’) seed under cool climate conditions in Tasmania, Australia occurred over a longer duration than previously reported, but similar patterns of change in yield components were recorded. In contrast to previous studies, umbel moisture content declined from 85 to 67 % over 57 days while seed moisture content decreased from 85 to 31 %. Seed yield continued to increase over the duration of crop development, with increasing seed weight compensating for seed loss resulting from capsule dehiscence in the later stages of maturation. Germination percentage was high and did not vary significantly from 53 to 77 days after full bloom (DAF), but mean germination time declined and uniformity of germination increased significantly over the same time period. The percentage abnormal seedlings declined with later harvest date, resulting in highest seed quality at 77 DAF. The results of this study suggest that the decision to harvest cool climate onion seed crops before capsule dehiscence will result in a loss of potential seed yield and quality.     

Location of a high-lysine gene and the DDT-resistance gene on barley chromosome 7

Summary The high-lysine gene in Risø mutant 1508 conditions an increased lysine content in the endosperm via a changed protein composition, a decreased seed size, and several other characters of the seed. The designation lys3a, lys3b, and lys3c, is proposed for the allelic high-lysine genes in three Risø mutants, nos 1508, 18, and 19. Linkage studies with translocations locate the lys3 locus in the centromere region of chromosome 7. A linkage study involving the loci lys3 and ddt (resistance to DDT) together with the marker loci fs (fragile stem), s (short rachilla hairs), and r (smooth awn) show that the order of the five loci on chromosome 7 from the long to the short chromosome arm is r, s, fs, lys3, ddt. The distance from locus r to locus ddt is about 100 centimorgans.     

Simultaneous Determination of Four Phenolic Acids in Radix Salviae Miltiorrhizae Formula Granules by QAMS

[Objectives]This study aimed to establish a QAMS(quantitative analysis of multi-components by single-marker)method for simultaneous determination of four phenol...     

Pollen and pollination experiments. III. The viability of apple and pear pollen as affected by irradiation and storage

Summary Apple and pear pollen was irradiated with doses of 0, 50, 100, 250 and 500 krad (gamma rays) and stored at 4°C and 0–10% r.h. From the in-vitro germination percentages an average LD 50 dose of about 220 krad was estimated. For both irradiated and untreated pollen a close and corresponding lineair relationship existed between germination percentage and pollen tube growth.Irradiated pollen was much more sensitive to dry storage conditions than untreated pollen, resulting in less germination and more bursting. Apparently, irradiation caused the pollen cell membrane to lose its flexibility faster than normal. Rehydration of dry-stored, irradiated pollen in water-saturated air restored germination percentages up to their initial levels. The importance of this procedure in germination trials is stressed.     

Water Extraction Process of Chinese Herbal Compound Man Gan Ning

[Objectives]To optimize the water extraction process of Chinese Herbal Compound Man Gan Ning and establish a method for its extraction and content determination...     

Present status and future strategy in breeding faba beans (Vicia Faba L.) for resistance to biotic and abiotic stresses

Progress is being made, mainly by ICARDA but also elsewhere, in breeding for resistance to Botrytis, AScochyta, Uromyces, and Orobanche; and some lines have resistance to more than one pathogen. The strategy is to extend multiple resistance but also to seek new and durable forms of resistance. Internationally coordinated programs are needed to maintain the momentum of this work.Tolerance of abiotic stresses leads to types suited to dry or cold environments rather than broad adaptability, but in this cross-pollinated species, the more hybrid vigor expressed by a cultivar, the more it is likely to tolerate various stresses.     

Optimization of Extraction Technology and Determination of Content of Total Phenols of Leaves in Acanthopanax giraldii Harms

[Objectives] To determine the optimum extraction technology for total phenols of leaves in Acanthopanax giraldii Harms.[Methods]The single factor test and ortho...     

Cytoplasmic male sterility,resistance to gall mite and mildew,and season of leafing out in black currants

Summary Cytoplasmic male sterility (cms) is described in the F₁ hybrids Ribes × carrierei (R. glutinosum albidum × R. nigrum) and R. sanguineum × R. nigrum. In backcrosses to R. nigrum, progenies with R. glutinosum cytoplasm were either all male sterile, or segregated for full male fertility (F) and complete (S) and partial (I) male sterility. Ratios of F:I+S suggested that two linked genes controlled cms, F plants being dominant for one (Rf ₁) and recessive for the other (Rf ₂).Segregation for cms in relation to three linded genes, Ce (resistance to the gall mite, Cecidophyopsis ribes), Sph ₃(resistance to American gooseberry mildew, Sphaerotheca mors-uvae) and Lf ₁(one of two dominant additive genes controlling early season leafing out) indicated that Rf ₁and Rf ₂were in this linkage group. The gene order and approximate crossover values appeared to be: % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXafv3ySLgzGmvETj2BSbqef0uAJj3BZ9Mz0bYu% H52CGmvzYLMzaerbd9wDYLwzYbItLDharqqr1ngBPrgifHhDYfgasa% acOqpw0xe9v8qqaqFD0xXdHaVhbbf9v8qqaqFr0xc9pk0xbba9q8Wq% Ffea0-yr0RYxir-Jbba9q8aq0-yq-He9q8qqQ8frFve9Fve9Ff0dme% aabaqaciGacaGaamqadaabaeaafaaakeaacaWGdbGaamyzamaamaaa% baGaaiiiaiaacccacaGGWaGaaiOlaiaacgdacaGG0aGaaiiiaiaacc% caaaGaaiiiaiaacccacaGGGaGaamOuaiaadAgaliaaigdakmaamaaa% baGaaiiiaiaacccacaGGGaGaaiiiaiaaccdacaGGUaGaaiOmaiaacs% dacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaaaacaWGsbGaamOzaSGa% aGOmaOWaaWaaaeaacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaiaacc% cacaGGGaGaaiiiaiaacccaaaGaamitaiaadAgaliaaigdakmaamaaa% baGaaiiiaiaacccacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaiaacc% cacaGGGaGaaiiiaiaacccacaGGGaaaaiaadofacaWGWbGaamiAaSGa% aG4maaaa!6E4D!\[Ce\underline { 0.14 } Rf1\underline { 0.24 } Rf2\underline { } Lf1\underline { } Sph3\]. Crossover values of 0.36 for Ce-Lf ₁, and 0.15 for Lf ₁-Sph ₃were estimated from the relative mean differences in season of leafing out between seedlings dominant and recessive for Ce and Sph ₃.It is suggested that competitive disadvantage of lf ₁-carrying gametes and/or zygotes at low temperatures may be implicated in the almost invariable deficit of plants dominant for the closely linked mildew resistance allele Sph ₃. Poor performance of lf ₁- (and possibly lf ₂-) carrying gametes and young zygotes during periods of low temperature at flowering might also account for the liability of some late season cultivars and selections to premature fruit drop (running off).     

Screening techniques and sources of resistance to parasitic angiosperms

Parasitic angiosperms cause great losses in many important crops under different climatic conditions and soil types. The most widespread and important parasitic angiosperms belong to the genera Orobanche, Striga, and Cuscuta. The most important economical hosts belong to the Poaceae, Asteraceae, Solanaceae, Cucurbitaceae, and Fabaceae. Although some resistant cultivars have been identified in several crops, great gaps exist in our knowledge of the parasites and the genetic basis of the resistance, as well as the availability of in vitro screening techniques. Screening techniques are based on reactions of the host root or foliage. In vitro or greenhouse screening methods based on the reaction of root and/or foliar tissues are usually superior to field screenings and can be used with many species. To utilize them in plant breeding, it is necessary to demonstrate a strong correlation between in vitro and field data. The correlation should be calculated for every environment in which selection is practiced. Using biochemical analysis as a screening technique has had limited success. The reason seems to be the complex host-parasite interactions which lead to germination, rhizotropism, infection, and growth of the parasite. Germination results from chemicals produced by the host. Resistance is only available in a small group of crops. Resistance has been found in cultivated, primitive and wild forms, depending on the specific host-parasite system. An additional problem is the existence of pathotypes in the parasites. Inheritance of host resistance is usually polygenic and its transfer is slow and tedious. Molecular techniques have yet to be used to locate resistance to parasitic angiosperms. While intensifying the search for genes that control resistance to specific parasitic angiosperms, the best strategy to screen for resistance is to improve the already existing in vitro or greenhouse screening techniques.