小麦高分子量谷蛋白亚基近等基因系的研究

以安农8455为母本,Pan555为父本组合杂交,并采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳法(SDS PAGE)对安农8455/Pan555的620粒F2籽粒的高分子量麦谷蛋白亚基(HMW GS)进行了测定,结果表明:F2籽粒中共出现了15种带型组合,可稳定出多对近等基因系;Glu A1a与Glu A1c分离比为1∶1;Glu B1b与Glu B1c分离比为9∶7;Glu D1d与Glu D1i分离比为1∶1;Glu B1b与或Glu B1c的不完全表达率为4 0%。     

家鸡LDH同工酶的发生遗传学分析

采用不同连续缓冲系统的聚丙烯酰胺凝胶电泳法，结合组织特异性染色和酶谱区带活性扫描技术，对家鸡胚胎期和生后期不同生长发育阶段各组织器官的乳酸脱氢酶（ＬＤＨ）同工酶进行了测定。结果表明，鸡胚胎期Ｂ型ＬＤＨ同工酶占优势（５１．４￣９９．８％），生后期则有３种不同表现：心、眼、脑组织继续保持Ｂ亚基的优势地位；肝、肾组织呈现Ｂ型酶含量的微弱上升；胸肌则发生Ｂ型酶到Ａ型的颠倒转换。ＬＤＨ同工酶的这种表达特征是     

聚丙烯酰胺凝胶电泳法鉴定禽呼肠孤病毒

从感染禽呼肠孤病毒的细胞中,以SDS和酚提取病毒核酸,通过聚丙烯酰胺凝胶电泳(PAGE)法将分节段的核酸分开,经银染色显示,呈3-3-1-3分布,此法可用于禽呼肠孤病毒的快速检测、鉴定。同样方法未能使传染性法氏囊病毒核酸显示。     

What proteomic analysis of the apoplast tells us about plant–pathogen interactions

Plant pathogens have developed different strategies during their evolution to infect and colonize their hosts. In the same way, plants have evolved different mechanisms acting against potential pathogens trying to infect and colonize their tissues. Regulation of a wide variety of proteins is required in order to perceive the pathogen and to activate the plant defence mechanisms. The apoplast is the first compartment where these recognition phenomena occur in most plant–pathogen interactions, allowing the exchange of different molecules and facilitating inter‐ and intracellular communication in plant cells. Proteomic analysis of the apoplast in recent years has found the initial biochemical responses involved in pathogen recognition and early defence responses. However, this proteomic approach requires some specific experimental conditions to obtain an extract free of cytoplasmic proteins and nonprotein contaminants that affect the subsequent stages of separation and quantification. Obtaining the highest proportion of proteins from the apoplastic space in infected tissues requires different steps such as extraction of apoplastic washing fluids and preparation of total secreted proteins (protein precipitation, solubilization, separation and digestion). Protein identification using mass spectrometry techniques and bioinformatics tools identifying peptides for the extracellular exportation is required to confirm the apoplastic location. This review compiles the most commonly used techniques for proteomic studies, focusing on the early biochemical changes occurring in the apoplast of plants infected by a wide range of pathogens. The scope of this approach to discover the molecular mechanisms involved in the plant–pathogen interaction is discussed.     

Variation in seed proteins from mutagen-treated cultivars and selected lines of Vigna unguiculata L. Walp

The storage protein profiles of the seeds of two IITA cowpea cultivars (‘IT84E–124’ and ‘Vita 7’) exposed to three mutagens—sodium azide (NaN₃), ethyl methane sulphonate (EMS) and ⁶⁰Co gamma rays—and those of 18 selected M₃ lines were investigated by sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis (PAGE). The total seed protein, globulin and albumin fractions showed differences in number and intensity of subunit bands, but the least differences were found in the albumin fraction. Both high and low molecular weight bands were observed, the highest being 94.5 kDa and the lowest 12.0 kDa.‘Vita T showed less variability compared with ‘IT84E–124’, as indicated by the relative similarity indices (S.I.) of the two cultivars. The lowest S.I. of 0.200 was found among ‘IT84E–124’ lines while the lowest S.I. among ‘Vita T lines was 0.4706. A number of lines with particular traits were found to be characterized by the presence of specific polypeptide bands. This study demonstrates that induced mutation could create additional variability to supplement existing germplasm and that SDS-PAGE is a useful tool for discriminating and estimating genetic similarities among selections.     

两优培九剑叶衰老过程光合膜功能及蛋白质复合物的变化

为探讨高产杂交稻两优培九在衰老过程中,剑叶光合膜蛋白质复合物的含量变化规律及其与光能吸收、转化、传递的关系,以大田栽培自然衰老剑叶为材料,利用活体叶绿素荧光动力学技术,并结合类囊体膜蛋白质复合物蓝绿温和胶电泳分析。结果表明,两优培九剑叶叶绿素含量、光合性能、类囊体膜蛋白稳定性等都在抽穗期达到顶峰,随后开始衰退,在扬花期、灌浆期尚保持较高水平,而进入籽粒成熟阶段衰退明显;随着衰老进程,光合膜蛋白质复合物有序非同步降解,稳定性为LHCII > PSIIcore > PSIcore > ATPase & Cyt b₆/f > LHCI;PSI和PSII蛋白和相应电子传递活性的稳定性及下降幅度差异较大;衰老过程叶绿素a/b的不断下降与相对于反应中心更稳定的捕光天线有关,剑叶生长后期LHCII维持高水平保持了叶片对光能的吸收,并可能在调节光系统间能量分布和协助过剩能量耗散中起重要作用。     

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.     

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     

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。