茶树总蛋白质提取方法及双向电泳体系的建立

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。     

三种甘蔗叶片蛋白质分离纯化方法比较及双向电泳体系优化

为了优化适用于甘蔗叶片蛋白质的分离纯化方法与双向凝胶电泳体系,比较了3种蛋白质分离纯化方法对甘蔗叶片蛋白质的SDS-PAGE电泳图谱、蛋白质提取率以及双向凝胶电泳的影响,同时优化了双向电泳条件。结果显示,SDS提取法的SDS-PAGE电泳图谱中条带较少,蛋白质沉淀杂质多,效果差;三氯乙酸/丙酮沉淀法蛋白质沉淀得率高,但沉淀复溶性差,蛋白质溶液浓度低,难于满足双向电泳大胶的高上样量要求;酚抽提法蛋白质沉淀得率虽低,但沉淀纯度高、复溶性好,蛋白质溶液浓度高,是适合双向电泳分离纯化甘蔗叶片蛋白质的理想方法。酚抽提法蛋白质双向凝胶电泳合适的胶条p H范围为4~7,17 cm大胶合适的上样量为800μg,平均可分离出954个蛋白点。     

槟榔叶片总蛋白质提取及双向电泳条件初探

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.     

橡胶草胶乳蛋白双向电泳方法的建立

采用Tris饱和酚法和TCA/丙酮沉淀法提取橡胶草(Taraxacum Kok-saghyz Rodin)胶乳蛋白质,并建立蛋白双向电泳方法.结果表明,TCA/丙酮沉淀法得到的双向电泳图谱横条纹较多,背景模糊,得到约446个蛋白质点;Tris饱和酚法提取样品双向电泳的聚焦效果较好,图谱显示约630个蛋白点,其中包括较丰富的低分子量蛋白.     

‘四季蜜’龙眼花芽总蛋白质提取方法及双向电泳体系的优化

利用TCA丙酮法、丙酮法、TCA酚抽提法和改良酚抽法等4种方法,提取‘四季蜜’龙眼花芽的总蛋白质,在蛋白产量、单向SDS-PAGE和2-DE图谱等方面进行比较,并对IPG胶条种类、蛋白质上样量及等电聚焦条件等进行优化。结果表明:采用改良酚抽法,选用24 cm、pH3~10的胶条,按120μg上样,在适宜的等电聚焦条件下,可获得背景清晰、重复性好、分辨率高的2-DE图谱,本研究为‘四季蜜’龙眼花芽蛋白质组学后续研究奠定了基础。     

对科研院所科技开发工作的思考

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。     

茶树蛋白质双向电泳样品制备技术研究

为探索一种适用于茶树蛋白质双向电泳的样品制备方法,研究比较了TCA/丙酮沉淀法、改良的Tris-HCl抽提法,以及酚-甲醇/醋酸铵沉淀法3种植物蛋白质样品制备方法。结果表明,改良的Tris-HCl抽提法较适合于茶树蛋白质双向电泳的样品制备。该改良方法制备的茶树蛋白质样品经双向电泳分离,可获得1117±9.89个蛋白质点,其中大部分蛋白质的等电点集中在pI5~7之间,相对分子质量集中在15.00~95.00kD之间,背景清晰且蛋白质得率较高,能满足茶树蛋白质组学研究的需要。     

萌发期大豆总蛋白提取及双向电泳体系的优化

蛋白质样品的制备和双向电泳条件是蛋白组学研究中很关键同时也是很棘手的问题。本文比较了TCA/丙酮法、酚抽提、苯酚/SDS法3种不同的蛋白提取方法,并对双向电泳的一些条件进行了优化,建立了适合萌发期大豆总蛋白的双向电泳体系,即采用17cm、p H4-7的IPG胶条,凝胶浓度为12%,蛋白质上样量为0.2mg。IEF程序为:250V 1h,500V 1h,3 000V 3h,8 000V 1h,10 000V 1h,10 000V 60 000Vh。本研究为萌发期大豆的蛋白组学研究提供了实验方法。     

野生大豆叶片蛋白质组双向电泳技术体系的优化研究

为给野生大豆蛋白质组学分析提供技术支撑,以耐盐碱野生大豆叶片为材料,对Tris-饱和酚法、Tris-HCl法和TCA-丙酮法3种蛋白质提取方法进行了比较,并对蛋白上样量、等电聚焦参数及凝胶浓度进行了优化。结果表明:使用Tris-饱和酚法提取蛋白质,上样量为2 000μg,等电聚焦90 000 V·h,凝胶浓度为10%时,可获得背景清晰、蛋白点数较多、重复性较好的蛋白双向电泳图谱。     

小麦穗突变体Sda1与其野生型叶片总蛋白双向电泳体系的建立

为探索适合小麦穗突变体Sda1叶片蛋白的双向电泳体系,寻找Sda1与其野生型叶片的差异蛋白,以Sda1与其野生型的抽穗期叶片为材料,从蛋白质提取、双向电泳条件等方面对适合Sda1及其野生型叶片蛋白的双向电泳体系进行探索。分析试验得到的双向电泳图谱,发现利用TCA/丙酮提取叶片总蛋白,用pH 4~7的17cm线性胶条,采用13%浓度的分离胶,上样量为900μg,利用改良的等电聚焦程序,能够得到背景清晰、分辨率高的电泳图谱;电泳图谱在低分子区蛋白点分布均匀,并且重复性好。利用PDQuest 8.0.1软件分析图谱,得到27个差异点蛋白,相比于野生型植株,Sda1突变体表现为蛋白表达量下调与缺失,发现6个缺失蛋白,21个表达下调蛋白。     

Sorghum and millet phenols and antioxidants

Sorghum is a good source of phenolic compounds with a variety of genetically dependent types and levels including phenolic acids, flavonoids, and condensed tannins. Most sorghums do not contain condensed tannins, but all contain phenolic acids. Pigmented sorghums contain unique anthocyanins that could be potential food colorants. Some sorghums have a prominent pigmented testa that contains condensed tannins composed of flavan-3-ols with variable length. Flavan-3-ols of up to 8–10 units have been separated and quantitatively analyzed. These tannin sorghums are excellent antioxidants, which slow hydrolysis in foods, produce naturally dark-colored products and increase the dietary fiber levels of food products. Sorghums have high concentration of 3-deoxyanthocyanins (i.e. luteolinidin and apigenidin) that give stable pigments at high pH. Pigmented and tannin sorghum varieties have high antioxidant levels that are comparable to fruits and vegetables. Finger millet has tannins in some varieties that contain a red testa. There are limited data on the phenolic compounds in millets; only phenolic acids and flavones have been identified.     

Tuber and starch quality of wild and cultivated potato species and cultivars

Summary Between 1993 and 1998 205 different potato cultivars and 1220 accessions/genotypes of wild and cultivated potato species from the IPK Genebank Gatersleben were evaluated. Parameters interesting for starch isolation and especially for the use of starch were determined. Altogether, there was a higher variability in wild potato species than in cultivated potatoes for all characteristics investigated: dry matter content, starch content, protein content, amylose content and mean particle diameter of starch granules.     

种子加工、检验理论与技术现状及思考

种子加工及检验是种子向市场流通的关键，是商业化育种中赋予种子商品属性不可或缺的环节。种子加工、检验理论与技术的研究是完善种业产业链、实现规模化商业育种、夯实“育繁推一体化”种业科学的重要一环。中国的种业科学技术体系正在形成与完善之中，受制于行业发展水平，种子加工及检验理论与技术相对薄弱。本文回顾了种子加工及检验理论与技术研究发展历程，提出了完善中国种业科学发展的策略与建议。围绕种业发展的需要，在做好品种优质化繁育的基础上，分析种子加工及检验理论与技术的新方向和新需求，加强种业应用性研究，建立先进的种子质量检验体系，研制适合现代种业发展需求的种子加工设备，打造种子加工产业标准化生产体系，完善种业科学技术学科建设，促进种业产业持续健康发展。     

Effects of Organic and Chemical Fertilizers and Arbuscular Mycorrhizal Inoculation on Growth and Quality of Cucumber and Lettuce

Summary

Arbuscular mycorrhizae were inoculated into phosphorus-deficient soil fertilized with either organic or chemical fertilizer with cucumber (Cucumis sativus L.) as the first crop and lettuce (Lactu-ca sativa L.) as the second crop but without additional fertilization and AM inoculation. AM increased dry matter and fruit yield of cucumber significantly in the unfertilized, organic-fertilized and P-deficient plants compared with the fully chemical-fertilized plants. AM inoculation increased the available phosphorus in plant and soil by around 30% for all treatments except for those chemically-fertilized. The rate of AM infection did not differ significantly among the fertilization treatments, but the infection intensity was higher in unfertilized, organic-fertilized and phosphorus-deficient treatments than chemical-fertilized treatment. The residual effects of AM-inoculated to cucumber were evident for lettuce in all pre-treatments that were unfertilized and un-inoculated for the second cropping. Without P-fertilization, neither crop could grow optimally even when the soil was inoculated with AM, suggesting that AM could not serve as a substitute for phosphorus fertilizer. However, the other beneficial effects of AM on crop growth and yield could not be fulfilled with phosphorus fertilizer.     

Present Situation and Consideration of Seed Processing and Testing Theory and Technology

Seed processing and testing hold the key to the market circulation of seeds, and are an indispensable link in commercial breeding to endow seeds with commodity attributes. The research of seed processing and testing theory and technology is an important link in improving the seed industry chain, realizing large-scale commercial breeding and consolidating the seed industry science of ‘integration of breeding, propagation and promotion’. The scientific and technological system of seed industry in China is being formed and perfected, which is subject to the development level of the industry. The theory and technology of seed processing and testing are relatively weak. We reviewed the development of seed processing and testing theory and technology, and put forward strategies and suggestions to improve the sound development of China’s seed industry. In order to meet the needs of seed industry development and on the basis of high quality breeding of varieties, the new directions and demands of seed processing and testing theory and technology were analyzed. We will work to strengthen the applied research of seed industry, establish advanced seed quality inspection system, improve seed processing equipment suitable for the development needs of modern seed industry, establish standardized production system of seed processing industry, develop the scientific and technological disciplines of seed industry, and promote the sustainable and healthy development of seed industry.     

种子加工、检验理论与技术现状及思考

种子加工及检验是种子向市场流通的关键,是商业化育种中赋予种子商品属性不可或缺的环节。种子加工、检验理论与技术的研究是完善种业产业链、实现规模化商业育种、夯实"育繁推一体化"种业科学的重要一环。中国的种业科学技术体系正在形成与完善之中,受制于行业发展水平,种子加工及检验理论与技术相对薄弱。本文回顾了种子加工及检验理论与技术研究发展历程,提出了完善中国种业科学发展的策略与建议。围绕种业发展的需要,在做好品种优质化繁育的基础上,分析种子加工及检验理论与技术的新方向和新需求,加强种业应用性研究,建立先进的种子质量检验体系,研制适合现代种业发展需求的种子加工设备,打造种子加工产业标准化生产体系,完善种业科学技术学科建设,促进种业产业持续健康发展。     

芸薹属多倍体与二倍体作物 花柱及角果的发育差异

通过比较分析芸薹属多倍体复合种与其祖先二倍体基本种在花柱、柱头大小、角果长度、宽度、种子直径 及种子干重变化上的异同,发现多倍体复合种在这些发育性状上表现出处于两个祖先二倍体亲本之间,或只偏向 于其中一个祖先亲本。同一个多倍体品种在不同发育性状上具有不同的表现,即在某一发育性状上偏向于其祖先 亲本之一,而在另一发育性状上偏向其另一祖先亲本。     

Combined nitrogen and sulphur fertilisation and its effect on wheat quality and protein composition measured by SE-FPLC and proteomics

Soil sulphur deficiency, which is increasingly prevalent in Western Europe, lowers wheat yields, and also affects the gluten quality of the flour. Differences in S availability may change the proportion of S-poor to S-rich gliadins and glutenin subunits. This may cause unpredictable and unwanted variations in wheat quality. The combined effects of nitrogen (N) and sulphur (S) fertilisers and split application of S and N on wheat gluten quality and composition were investigated. The results revealed effects of S fertilisation on dough quality. At high N fertilisation levels significant responses to S fertilisation were found which emphasised the need for precision application of S in intensive wheat production systems. Protein fractionation by SE-FPLC showed that quality differences were associated with changing proportions of high M_r polymeric proteins. Changes in protein composition of salt soluble proteins were also confirmed by proteomics. Glyceraldehyde-3-phosphate dehydrogenase and one of the serpin protein spots increased at high N, combined with the lower S level. The enzymes also increased in samples with increased S fertilisation combined with low N, but was not changed at higher N levels. Furthermore, at high N the serpin protein spot, and also a 27 K protein and one unidentified protein spot decreased with increasing S.     

我国麻类标准现状与分析

介绍了我国麻类标准的现状。通过分析,指出了存在的问题,主要表现在标准体系不完善、标准更新不及时,协调性差及贯彻执行不力等。提出了我国麻类标准应采取的对策。     

Effects of crop density and tillage system on grain yield and N uptake from soil and atmosphere of sole and intercropped pea and oat

Pea (Pisum sativum L.) and oat (Avena sativa L.) were grown as sole and mixed crops in various densities under two different tillage systems on a loess soil near Göttingen/Germany in a 2-year field experiment (2002/2003). In the conventional tillage system a mouldboard plough (CT) was used and in the minimum tillage system a rotary harrow (MT) was employed. The effect of crop density and tillage system on the grain dry matter and grain N yields, N₂ fixation and soil N uptake were determined to address the following questions: (i) which mixture compositions exhibit the highest grain yields compared to the sole crops, (ii) which mixture compositions also fix a high level of N₂ and leave low levels of residual inorganic soil N after harvest, and (iii) whether the intercrop advantage is influenced by the tillage system. For (i) the result in 2002 showed that the highest grain yields of both sole cropped pea and oat and intercropped pea and oat were achieved at the highest densities. In 2003, when the inorganic soil N content was higher and weather conditions were warmer and drier, grain yields were significantly higher than in 2002, but sole as well as intercropped pea and oat gave their highest grain yields at lower densities. For both years and tillage systems, the highest intercrop advantages were achieved in mixtures with densities above the optimal sole crop densities. The result for (ii) was that a distinctly higher proportion of nitrogen was derived from the atmosphere (Ndfa) by intercropped pea than by sole cropped pea. However, the uptake of soil N by intercropped pea and oat was not reduced in comparison with that of sole cropped oat as the decrease in the uptake of N from the soil by oat at lower oat densities in the mixture was compensated for by the soil N uptake of pea. Additionally, the N_min-N content of the soil following the mixtures and sole cropped oat did not differ, especially in the deeper soil layers because oat in mixture was forced to take up more soil N from deeper layers. Therefore, the risk of soil N losses through leaching after mixtures was lower compared to sole cropped pea. The tillage system (iii) had no significant influence on grain yield and soil N uptake, but N₂ fixation and the competitive ability of intercropped pea were higher under CT than with MT. An additional result was that intercropping led to a significantly increased grain N content of both pea and oat compared to the sole crops. The increase in grain N content from sole to intercrop was from 3.30 to 3.42% for pea and from 1.73 to 1.96% for oat as a mean for both years and tillage systems. The present study confirms that growing pea and oat as intercrops highlights potential economic and environmental benefits which still need to be understood in more detail in order to exploit intercropping to a greater extent.