Comparative analysis of different methods revealing the involvement of proteins during ageing of rice seeds

Seed longevity is a very important characteristic controlling seed quality. However, the mechanism underlying this characteristic is poorly understood. In this study, 'FH7185', a storable rice variety, the germination rate of which was 66.63%, much higher than the control after artificial ageing under relative humidity 88% and 42°C for 21 days. Different methods were applied to reveal the involvement of proteins during ageing of rice seeds. In total, 35 differential proteins were identified from 2D‐PAGE, and 3,719 proteins from iTRAQ analysis. A comparison of these two methods showed that not all protein types could be detected on the 2D‐PAGE gels, and the dynamic range was somewhat limited. So the comprehensive proteome map from the iTRAQ analysis was used to identify the quantitatively regulated proteins that played roles in seed longevity, and found that the most marked change was the increased abundance of many metabolic enzymes, especially the ones involved in α‐linolenic acid metabolism in the embryos during ageing. OsLOX2 and OsLOX3 had a negative effect on seed longevity, which were lower in FH7185 than the control. The dehydrogenase (LOC_Os07g44430) which played a major catalytic role in lipid peroxidation, also changed significantly during ageing. Therefore, OsLOX2, OsLOX3 and Loc_OS 07G44430 may be involved in the regulation of seed longevity with a synergistic effect.     

干旱胁迫对白桦实生幼苗叶片蛋白质的影响

The studies on leave protein of Betula platyphylla seedlings by the methods of SDS-PAGE electrophoresis and dimensional gel electrophoresis was concerned. The result indicated that some new proteins such as 29kD pI4.3, 29kD pI4.2, 14kD pI5.3 and 11.5kD pI5.16 protein point were induced by soil drought stress, and some protein disappeared,for instance four protein between 12.4 - 11.5kD pI5.8 - 6.44 in seedlings leaves of B. platyphylla during soil drought stress.     

偶氮类色素胭脂红对泥鳅组织酯酶同工酶表达的影响

采用肌肉注射法进行染毒,运用聚丙烯酰胺凝胶电泳法研究胭脂红在不同注射剂量、不同染毒时间对泥鳅肝脏、心脏、脾脏、肌肉酯酶同工酶表达的影响。试验结果表明,经一定剂量的胭脂红注射后,泥鳅酯酶同工酶在脾脏中表达最强,肝脏和心脏次之,肌肉最弱,其中脾脏的表达随时间、浓度呈减弱趋势,其余组织则呈先增强再减弱最后增强的趋势。     

籼稻体细胞胚胎发生特异性蛋白质研究

以籼稻Oryza sativa L.subsp. indica品种“广陆矮4号”幼穗和种子为外植体，在含有2，4-D2.0mg/L的MS培养基上，分别形成胚性和非胚性愈伤组织，建立体细胞胚胎发生实验系统。应用Native/SDS双向凝胶电泳分析，结果表明，非胚性愈伤组织分化培养前后均存在一种分子量为45kD的非胚性蛋白质N1，另一种非胚性蛋白质N2（54kD）仅     

Identification of cultivars of pasture legumes (Centrosema macrocarpum,C. pubescens and C. sp.n.) by acid polyacrylamide gel electrophoresis of cotyledons storage proteins

Summary An electrophoretic procedure was developed for storage proteins which can discriminate cultivars of forage legumes, Centrosema macrocarpum, C. pubescens and C. sp.n. Proteins extracted from cotyledons were separated in 10% polyacrylamide gel containing 10% sucrose. Electrophoretic patterns are presented to illustrate the results that can be obtained with the procedure described. Cultivars of all three species were distinguishable based upon the variation in their acid soluble seed proteins.Joint publiccation of the Department of Food Science, University of Manitoba (No 101) and Centro Internacional de Agricultura Tropical-CIAT.     

磷酰胺除草剂APM诱导洋葱根尖分生组织细胞异常有丝分裂和蛋白质组分变化

研究了不同浓度APM(4～6 μmol/L)和不同处理时间(4～6 h)对洋葱根尖细胞有丝分裂的影响.结果表明,当用5.6 μmol/L APM处理洋葱分生组织16 h后,中期细胞有丝分裂指数(Met.Ⅰ)从0.8(对照)分别提高至5.0、5.2.结果也同时表明,当APM浓度超过4 μmol/L时,严重影响细胞有丝分裂.在根尖细胞中,多极分裂细胞(尤其是3～4级),中期染色     

不含 SBTI-A_2的基因导入我国大豆的遗传表达

大豆 Glycine max(L.)Merrill 种子中的主要营养抑制因子是 Kunitz 胰蛋白酶抑制剂(SBTI—A_2)。经利用标准 SBTI 比较谱带的聚丙烯酰胺凝胶电泳(PAGE)分析方法,证实了我国栽培大豆中豆19、临沂81-551、通县元豆、中作84-C_(42)和中作84-C_(02)均属 Ti~aTi~a 型。并利用上述5个黄淮海夏大豆品种或品系与美国不含 SBTI-A_2(titi)品系 L_(81-4590)和 L_(83-4387)杂交,在7个组合的 F_2代中得到了 titi 型。其表型比例为3∶1,符合孟德尔单基因遗传规律。进一步的研究正在进行中。     

聚丙烯酰胺凝胶电泳的一种辅助辨型方法

聚丙烯酰胺凝胶电泳（PAGE）是实验室常用的基本鉴定方法之一。但由于做胶方式以及各实验室仪器设备良莠不齐，经过一系列纷繁复杂的操作，电泳结果未必尽如人意。其中凝胶带型的判断就是一个较为困惑的问题，一般的聚丙烯酰胺胶跑出的条带总体上都有弧度，越是靠近凝胶两侧越是严重。在本实验中，我们设计了一种较为简单易行的方案，即将难以区分的条带样品做成混合样marker来辅助辨型，通过调整电压和凝胶浓度来达到区分条带的目的。我们用该方案可以将相差0～3bp大小的条带区分开来。     

鸡毒支原体特异膜抗原的制备

采用 Ｓ Ｄ Ｓ聚丙烯酰胺凝胶电泳分离纯化鸡毒支原体膜蛋白。选择切取１１０ Ｋ Ｄ、９８ Ｋ Ｄ、６６～６２ Ｋ Ｄ 和５６ Ｋ Ｄ 特异蛋白条带，以 Ｐ Ｂ Ｓ（ｐ Ｈ７．４）浸泡过夜，浸出液置于透析袋封口，透析袋外散布 Ｐ Ｅ Ｇ（ Ｍ ＝ ６ ０００）浓缩膜抗原后获得特异膜抗原，为鸡毒支原体特异诊断方法的建立和单克隆抗体的制备奠定了基础。