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1.
槟榔叶片总蛋白质提取及双向电泳条件初探   总被引:4,自引:1,他引:3  
采用3种提取植物组织蛋白的方法(TCA/丙酮沉淀法、E-TCA法、酚法)提取槟榔叶片总蛋白,在蛋白产量,一维和二维电泳图谱等方面对3种方法的提取效果进行比较.结果表明,TCA/丙酮沉淀法效果较好,是槟榔叶片总蛋白提取的可选方法,同时优化了双向电泳的条件和上样量.  相似文献   

2.
油菜种子蛋白质提取方法研究   总被引:1,自引:0,他引:1  
通过对比改进的Tris-HCl法、酚提取-甲醇/醋酸铵沉淀法、TCA/丙酮沉淀法和DTT/丙酮法4种方法提取油菜未成熟种子蛋白质,以期找到合适的蛋白质提取方法。结果表明,TCA/丙酮沉淀法和DTT/丙酮法所得蛋白质沉淀较多,颜色亮白,效果较好。进一步通过定量分析及双向电泳图谱的比较分析表明,TCA/丙酮沉淀法在2-DE图谱上蛋白质点较多,分布均匀,图像清晰,而DTT/丙酮法相对于本实验中TCA/丙酮法检测到较少的蛋白质点外,2-DE图谱上有明显的纵条纹。因此TCA/丙酮沉淀法为提取油菜种子中蛋白质的最优方法。  相似文献   

3.
为探索适用于茶树叶片蛋白质组学研究的双向电泳技术体系(2-DE),比较三氯乙酸(TCA)/丙酮沉淀法和改良酚抽法2种不同总蛋白质提取方法、不同蛋白上样量对2-DE的影响。结果表明:改良酚抽法更适合茶树总蛋白质的提取;采用1.5 mg的蛋白质上样量,最终可获得蛋白质点较多、背景清晰、分辨率较高的双向电泳图谱,为进一步深入开展茶树蛋白质组学的研究奠定了基础。  相似文献   

4.
为探讨红麻细胞质雄性不育形成的分子遗传机理,以红麻细胞质不育系L23A和保持系L23B开花期叶片为材料,对其红麻蛋白质双向电泳体系进行研究,比较TCA/丙酮和酚抽提法提取红麻叶片总蛋白,同时优化上样量和样品制备方法,建立了红麻蛋白质的双向电泳技术体系和凝胶成像染色法,利用PDquest软件分析双向电泳凝胶的相关差异蛋白。结果表明:(1)TCA/丙酮法比酚抽提法更适合于ISO-DALT电泳系统;(2)TCA/丙酮提取的红麻叶片粗蛋白平均产量为31.67 mg/g,是酚抽提法的1.77倍, 但裂解效率为160 mg/g,是酚抽提法的36%;(3)每根胶条最佳上样量为300 μg;(4)考马斯亮蓝G-250比R-250以及银染色法效果更佳,在17 cm×17 cm的凝胶上平均可获得708个蛋白质点。本研究建立的红麻细胞质雄性不育系与保持系总蛋白提取方法及双向电泳体系,可为揭示红麻雄性不育分子机理奠定基础。  相似文献   

5.
大豆是我国重要的农作物之一,蛋白质组学技术是解析植物响应逆境胁迫的重要工具之一。为进一步推进对响应干旱胁迫的大豆叶片蛋白的鉴定工作,以大豆幼苗的叶片为研究对象,用甘露醇溶液模拟干旱胁迫,处理24 h后提取叶片蛋白粉进行双向电泳分离和图谱差异分析。结果表明:采取三氯醋酸(TCA)-丙酮沉淀法提取的叶片蛋白进行双向电泳分离能够得到稳定清晰的双向电泳蛋白图谱。采取PD-Quest软件对干旱胁迫下的大豆叶片蛋白表达图谱进行差异分析,共检测和匹配425个大豆叶片蛋白点,其中差异表达的蛋白点有101个,60个表达量上调,41个表达量下调。本研究结果表明有一部分大豆叶片蛋白响应了干旱胁迫。  相似文献   

6.
为建立一套适合茶树雌蕊总蛋白质分离的双向电泳体系,对茶树雌蕊总蛋白的提取方法、SDS-PAGE凝胶浓度、聚焦条件和IPG胶条pH范围等进行了比较优化。结果表明,TCA/丙酮-clean up kit法提取总蛋白,用17 cm pH 5~8 IPG胶条,13%SDS-聚丙烯酰胺凝胶进行分离,用考马斯亮蓝R250染色,可以获得清晰、分辨率高、重复性好的双向电泳图谱,共检测到约1200个蛋白点,主要分布在pH 5~8,相对分子量14~117 kD范围内,碱性蛋白得到有效分离,可以满足茶树雌蕊的蛋白组学分析和研究。  相似文献   

7.
为建立一套适合茶树雌蕊总蛋白质分离的双向电泳体系,对茶树雌蕊总蛋白的提取方法、SDS-PAGE凝胶浓度、聚焦条件和IPG胶条pH范围等进行了比较优化。结果表明,TCA/丙酮-clean up kit法提取总蛋白,用17 cm pH 5~8 IPG胶条,13%SDS-聚丙烯酰胺凝胶进行分离,用考马斯亮蓝R250染色,可以获得清晰、分辨率高、重复性好的双向电泳图谱,共检测到约1 200个蛋白点,主要分布在pH 5~8,相对分子量14~117 kD范围内,碱性蛋白得到有效分离,可以满足茶树雌蕊的蛋白组学分析和研究。  相似文献   

8.
为给小麦抗旱蛋白质组学研究提供技术支撑,以两个抗旱性不同的冬小麦品种周麦18和晋麦47三叶期幼苗为材料,比较分析了PEG胁迫处理后两种不同蛋白质提取方法TCA/丙酮法和磷酸钠缓冲液研磨法对IEF/SDSPAGE双向电泳的影响,并在IPG胶条pH范围、蛋白质上样量和SDSPAGE胶浓度等方面进行了探索与优化。结果表明,采用TCA/丙酮法提取叶片全蛋白,选用17 cm、pH 4~7的IPG线性胶条,在上样量为150 μg·胶条-1,以及12%SDSPAGE胶浓度下进行双向凝胶电泳,蛋白质能够更好地被分离,2DE图谱上可分辨出约400个蛋白点。利用该体系比较了PEG胁迫后两个品种双向电泳图谱的差异,周麦18有9个蛋白点存在差异,其中5个下调表达,4个上调表达(3个新诱导表达);晋麦47有5个蛋白点存在差异,均上调表达(1个新诱导表达)。  相似文献   

9.
以玉米(Zea mays L.)中糯二号为材料,采用改进的TCA(三氯乙酸)-丙酮沉淀法提取玉米叶片的蛋白质,并从蛋白样品纯化和上样量2个方面对玉米叶片双向电泳体系进行优化。结果表明:蛋白纯化后且上样量为150 μg时,能获得背景清晰,横条纹、竖条纹少,蛋白点多的双向电泳图谱。这为开展玉米中糯二号的蛋白质组学研究提供技术支持。  相似文献   

10.
以橡胶树无性系RRIM600为材料,采用TCA(三氯乙酸)-丙酮沉淀法提取胶乳全蛋白,17 cm pH 5~8固相pH梯度预制干胶条和快速银染法对提取的蛋白进行分离和染色,所得到的电泳图谱背景清晰,横条纹和竖条纹少,且蛋白点数量多。通过本研究获得了高分辨率的胶乳全蛋白双向电泳(2-DE)图谱的方法,为进一步开展橡胶树胶乳全蛋白质组学研究奠定了基础。  相似文献   

11.
12.
Summary

The efficiency of N fertilizers is usually poor; often less than 50% of the applied N is taken up by the crop. This review focuses on various N fertilizers with respect to the significance of different N loss pathways, namely (i) ammonia volatilization, (ii) dinitrogen and nitrogen oxide emissions, and (iii) nitrate leaching. Further, the significance of biological N immobilization, ammonium fixation and, finally, the impact of nitrate vs. ammonium uptake on crop yield are also discussed. The reviewed literature shows that N fertilizers may differ markedly in their susceptibility to losses. There is, however, considerable scope to improve N efficiency of each N source by proper N management practices.  相似文献   

13.
Novel food and non-food uses for sorghum and millets   总被引:4,自引:3,他引:4  
Sorghum and millets have considerable potential in foods and beverages. As they are gluten-free they are suitable for coeliacs. Sorghum is also a potentially important source of nutraceuticals such antioxidant phenolics and cholesterol-lowering waxes. Cakes, cookies, pasta, a parboiled rice-like product and snack foods have been successfully produced from sorghum and, in some cases, millets. Wheat-free sorghum or millet bread remains the main challenge. Additives such as native and pre-gelatinised starches, hydrocolloids, fat, egg and rye pentosans improve bread quality. However, specific volumes are lower than those for wheat bread or gluten-free breads based on pure starches, and in many cases, breads tend to stale faster. Lager and stout beers with sorghum are brewed commercially. Sorghum's high-starch gelatinisation temperature and low beta-amylase activity remain problems with regard to complete substitution of barley malt with sorghum malt . The role of the sorghum endosperm matrix protein and cell wall components in limiting extract is a research focus. Brewing with millets is still at an experimental stage. Sorghum could be important for bioethanol and other bio-industrial products. Bioethanol research has focused on improving the economics of the process through cultivar selection, method development for low-quality grain and pre-processing to recover valuable by-products. Potential by-products such as the kafirin prolamin proteins and the pericarp wax have potential as bioplastic films and coatings for foods, primarily due to their hydrophobicity.  相似文献   

14.
Evolutionary aspects of the trade-off between seed size and number in crops   总被引:1,自引:0,他引:1  
Victor O. Sadras   《Field Crops Research》2007,100(2-3):125-138
Whereas the concept that availability of resources drives seed production is sound in principle, it is incomplete as there are many solutions to the allocation of resources that derive from the trade-off between number and size. This paper examines evolutionary aspects of this trade-off in annual grain crops. The analysis is centred on the working hypotheses that, for a given species and environment, allocation of resources to reproduction involves (H1) high plasticity in seed number, which allows for variable resource availability, and (H2) a relatively narrow range of seed size that results from evolutionary and agronomic selection. Comparisons between crops and fish are used to highlight common evolutionary elements in taxa where parents provide little or no care to their offspring, with the consequence that both number and early survival of offspring, hence fitness of parents, are partially related to embryo size and reserves.

The plasticity of seed number in relation to availability of resources is analysed against the established relationship between offspring number and parent growth rate during critical stages. The notion that seed size is under stabilising selection is analysed against three conditions: (1) mean seed size is conservative for a given species and environment, (2) seed size affects fitness, and (3) seed size is heritable. Databases from published papers were compiled to analyse the relative variability of seed size and number, and the heritability of seed size. Evidence for and against the link between seed size and parental fitness is revised using the Smith–Fretwell model as framework (Am. Nat., 108, 499–506).

The proposal of high plasticity of seed number and narrow variability of seed size resulting from stabilising natural selection is generally consistent with evolutionary and genetic considerations. Agronomic selection may have reinforced natural selection leading to relatively narrow seed size in species such as wheat and soybean, where cultivated types retained high plasticity for seed number. In contrast, selection for one or few inflorescences in crops like sunflower and maize may have morphologically reduced seed number plasticity and increased variability of seed size and its responsiveness to resource availability in relation to their wild ancestors.  相似文献   


15.
Flavonoids-enriched tissues of citrus such as peel, immature fruit and flower are consumed as culinary seasonings, tea ingredients in China for centuries. This HPLC quantitative study on the five citrus flavonoids, naringin, hesperidin, neohesperidin, sinensetin and nobiletin on a wide range of Chinese citrus fruits and several Traditional Chinese Medicinal food ingredients in East China, revealed a great diversity in flavonoid composition. Huyou peel (C. paradisi cv. Changshanhuyou) was found to be the best naringin (3.25%) and neohesperidin (2.76%) source; C. aurantium, a major ingredient of several citrus-related TCM, is also a suitable source of naringin and neohesperidin, and a good juice source for flavanone glycosides; the peel of Wenzhoumiju (C. unshiu) is one of the richest local species in hesperidin (up to 6.25%); Zaoju (C. subcompressa) has the highest content of nobiletin (0.59%), a polymethoxylated flavone. LC-ES-MS analysis of Zanthoxylum genus for flavonoids revealed for the first time the presence of significant amounts (0.74%) of hesperidin in the root of Liangmianzhen (Z. nitidum (Roxb.) DC), a relative of Sichuan pepper, which is a spice widely used in China.  相似文献   

16.
Summary The viability of five pathogens was decreased by treatment with hot water when tested in vitro.Polyscytalum pustulans was most sensitive andRhizoctonia solani least sensitive. Potato tubers were exposed to 55°C for 5 min in a commercial continuous hot water treatment plant using naturally contaminated seed tubers and tubers which had been inoculated by dipping in comminuted cultures. The frequency of eyes colonised byP. pustulans, Helminthosporium solani, andR. solani was reduced to virtually zero and the effect persisted on tubers subsequently stored at 4°C and at 15°C for up to 16 wk. Results withColletotrichum coccodes were inconclusive. Treatment suppressedPenicillium spp. which, however, rapidly recolonised the eyes during storage, leading to higher contamination levels in the treated than in the untreated tubers. With tubers inoculated withPhoma foveata, good control was achieved when the incubation period before treatment was 10 d but not when the period was 42 d.  相似文献   

17.
Summary

Heterocyclic 1,4-benzoxazin-3-ones and related benzox-azolinones are important natural products of the cereal crops maize, wheat and rye. Much research has focused on the role of these compounds as defensive agents against plant diseases and pests. Studies of a wide variety of biological effects on herbivores and nematodes, plant pathogenic fungi and bacteria are reviewed in this article. Phytotoxic activity of the compounds is also considered with respect to allelopathic interaction with higher plants. Recent investigations of molecular aspects of interactions of cyclic hydroxamic acids and related benzoxazo-linones with acceptor organisms have demonstrated different effective detoxification strategies and tolerance mechanisms. This research has greatly advanced our knowledge about the multiple roles that these allelochemicals play in ecological systems.  相似文献   

18.
Wheat bran is an undervalued by-product of white flour and has great nutritional potential due to its high content in fibres and bioactive compounds. Micronized bran could be used as a food ingredient to improve the nutritional potential of cereal products, or be used as a starting material for other processes (bioactive compound extraction or bran fractionation). The aim of this work was to find a way to efficiently decrease the particle size of bran. The influence of the grinding temperature (ambient or cryogenic grinding) on the granulometric distribution of particles, their composition, and their microstructure was studied, at lab-scale and pilot-scale. It showed that the intrinsic characteristics of bran (glass transition within intermediate layers at −46 °C) had more influence on its grinding behaviour than the type of grinding device used: the particles size distributions obtained after grinding at lab-scale and pilot-scale were very similar. At both scales, the granulometric curves were narrow for cryogenic grinding, while for ambient grinding they were spread over the whole particle size range. Ultrafine particles were obtained in both ambient and cryogenic conditions. Negative temperatures, by increasing the material’s brittleness, favoured a fast fragmentation of bran: one step of cryogenic grinding allowed a median particle size of nearly 50 μm to be reached, whereas three successive steps of ambient grinding were needed for the same result. On the other hand, ambient temperature favoured the dissociation of the different constituent layers of wheat bran, and produced less composite particles than cryogenic grinding.  相似文献   

19.
The technique of spraying plants with inorganic forms of selenium can be employed for phytochemical production of organic selenium compounds. Fractionation of the plant material makes it possible to produce a highly concentrated and well defined selenium supplement with potential use in animal and human nutrition. The fractionation also gives opportunities to combine production of organic selenium compounds with other products, for example plant fibres. Multiple use of plants can contribute to a more efficient utilization of land area (in comparison to monocultures solely adapted to food production). It also gives the opportunity to develop systems suitable for long term fixation of carbon, as long as the plant material is not reoxidised to carbon dioxide. Plant fibres could provide raw material for the production of paper or building materials in combination with the production of organic selenium compounds preferentially accumulated in another fraction of the processed plant.  相似文献   

20.
Summary Macrosiphum euphorbiae, collected in the field from potato plants infected with potato leafroll virus (PLRV), transmitted the virus to fewer potato plants in a field trial than did laboratory-rearedMyzus persicae. In the laboratory,M. persicae was the only efficient vector of PLRV fromPhysalis floridana seedlings, potato sprouts or excised leaves toP. floridana. Two clones ofM. euphorbiae and one clone ofAulacorthum solani transmitted PLRV from infected potato plants toNicotiana clevelandii as effeciently asM. persicae but a clone ofAphis gossypii was an inefficient PLRV vector. An isolate of PLRV, whichM. persicae transmitted inefficiently from potato toN. clevelandii, was also transmitted inefficiently byM. euphorbiae andA. solani.  相似文献   

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