紫稻细胞质雄性不育系叶片全蛋白双向电泳分析

通过对几种不育系叶片全蛋白双向电泳图谱分析证明;紫稻不育系具有不同于野败型,红莲型和马协型不育系的蛋白图谱,说明其可能是一种新的细胞质质源,同时,紫稻不育系与保持系蛋白图谱之间在三叶期和分蘖期时差异均不明显,但各材料本身蛋白图谱在两个不同时期之间差异很大,不育系与保持系图谱表现出的蛋白质（多肽）的差异,可能与不育系雄性不育有关。     

红莲型和野败型水稻细胞质雄性不育系线粒体DNA(mtDNA)的比较研究

以水稻红莲型和野败型细胞质雄性不育系为材料,用不连续蔗糖梯度离心法提纯线粒体,以饱和酚-氯仿-异戊醇法抽提mtDNA。用琼脂糖电泳和电镜观察比较不育系mtDNA差异,发现不育系中有小分子mtDNA(即m_2和m_3),不同细胞质间的小分子mtDNA存在差异。在不育系与保持系间名为m_1的大分子mtDNA无差异;本文还研究了mtDNAm_1的理化性质。     

高粱热激蛋白(HSPs)的电泳分析与雄性不育性

本文以高粱雄性不育系为材料进行了热激蛋白的研究。苗期单向电泳表明,保持系在热激(40℃)过程中可溶性蛋白有24条带,对照(28℃)22条,出现2条带差异,不育系在热激后有33条带,比对照28条多5条,且有4条加强带,共9条带产生差异。苗期双向电泳,不育系在热激后比对照有19点产生差异,保持系热激后与对照比较有6点产生差异。表明双向电泳揭示了热激后蛋白质在分子量和电性方面有差异。花粉母细胞期幼穗的热激蛋白,不育系对照可溶性蛋白仅有3条带,热激后有11条,8条带有差异。保持系对照有10条带,热激后有15条,1条加强,共6条带有差异。幼穗期不育系蛋白质突出的缺少19kd以上的大分子量的带。从个体发育看,不育系由苗期可溶性蛋白比保持系带数多,发育到花粉母细胞期比保持系突出地减少,尤其是大分子量的蛋白带消失,热激后明显地增加了与可育的保持系相同的蛋白成份,表明不育系有其特殊的基因调控。     

水稻线粒体基因组翻译产物与细胞质雄性不育性

通过线粒体离体翻译产物的电泳和放射性自显影分析,发现水稻BT型不育细胞质(农虎26A和丰锦A不育系)的线粒体基因产物比可育细胞质(农虎26B和丰锦B保持系)缺少一个22KD多肽。它反应了BT型不育细胞质线粒体基因组中有关育性的基因变异。不育系与恢复系杂交后,杂种F_1的育性虽然恢复,但是它的线粒体基因产物中仍然缺少22KD多肽。然而,在杂种F_1的线粒体蛋白质的电泳染色图谱中则显示出一个22KD多肽条带。杂种F_1中的这个22KD多肽是核基因产物。它弥补了线粒体基因组中育性基因的缺陷。     

水稻细胞质雄性不育系和保持系的线粒体COⅠ、COⅡ基因组织结构差异的分析

我们研究了水稻珍汕97不育系和保持系的线粒体DNA的PstⅠ、HindⅢ、BamHⅠ酶切电泳带型,发现了不育系和保持系的线粒体DNA分子结构的显著不同,而不育系和保持系的叶绿体DNA的HindⅢ酶切片段没有差异。以线粒体的细胞色素C氧化酶亚基Ⅰ、Ⅱ(COⅠ、COⅡ)基因为探针,与珍汕97不育系、保持系的叶绿体、线粒体DNA酶切片段进行分子杂交,证明叶绿体DNA没有COⅠ、COⅡ基因的同源顺序,发现不育系和保持系COⅠ、COⅡ基因有组织结构上的差异,但不能确证这些差异是否与细胞质雄性不育有关。     

普通小麦T型细胞质雄性不育系及其保持系线粒体多肽的电泳比较研究

应用单向SDS-PAGE和双向IEF-SDS电泳技术对两个品种的普通小麦(T.aestivum L.)T型细胞质雄性不育系及其保持系的线粒体多肽进行了比较研究,结论如下:1.黄化苗期不育系和保持系线粒体多肽在单向SDS-PAGE和双向IEF-SDS电泳行为上无明显差别;2.在孕穗期幼穗线粒体多肽的单向SDS-PAGE图谱上,两个不育系都缺少28Kd多肽带纹,因而不育系和保持系间表现出明显的差异。双向IEF-SDS凝胶电泳证实28Kd带纹实际上是分子量相同而等电点分别为5.58和5.65的两个多肽;3.线粒体基因的表达是具有时空性质的;4.线粒体与T型细胞质雄性不育可能存在着某种特定的关系。 本文还就T型细胞质雄性不育的分子机制进行了探索性讨论。     

叶绿体类囊体膜多肽与细胞质雄性不育性

本实验利用单向SDS-PAGE及双向电泳技术,比较了玉米、甜菜和高粱三种作物的细胞质雄性不育系与其保持系之间叶绿体类囊体膜多肽的差异。结果表明,三种供试材料的不育系与其相应的保持系之间类囊体膜多肽的单向SDS-PAGE中,除个别条带染色深度有一些差异外,没有观察到明显的差异。但是,在双向电泳图式中,两系之间在33kd附近肽斑的大小、数量与分布方面显示出明显的差异,从而暗示,叶绿体类囊体膜多肽的组成与细胞质雄性不育性之间可能存在某种联系。此外,试验还表明,单向SDS-PAGE条带,几乎都是分子量相同而等电点不尽相同的一组多肽混合物;在双向电泳图谱上,它们可按等电点的差异分成若干个不同的多肽斑点。     

叶绿体基因组的翻译产物与细胞质雄性不育性

本实验通过对叶绿体基因组翻译产物的分析,发现高粱3197 A不育系及其核代换系白马丁A和遗3 A不育系均比保持系(3197 B)多1个52,000道尔顿的变异多肽。而细胞质来源不同的粒息A不育系除52,000道尔顿多肽外还有1个特有的80,000道尔顿变异多肽。小麦T型不育系具有1个54,000道尔顿的变异多肽。甜菜不育系与保持系无差异。这说明某些作物的细胞质雄性不育性与叶绿体遗传系统有关。在高粱恢复系和F_1叶绿体蛋白质中发现三种特有的多肽。它们是由核基因编码的,这可能是恢复基因的产物。由于恢复基因的作用,在F_1恢复育性的同时其叶绿体变异多肽的合成也受到了强烈的抑制。这进一步证明了不育性与叶绿体的联系。     

水稻线粒体atpA基因的克隆及其与细胞质雄性不育的关系

本研究以水稻BT型细胞质雄性不育系秋光和相应的保持系秋光为材料,提取线粒体DNA,用限制性内切酶完全酶解,以玉米线粒体atpA基因和波菜叶绿体atpA基因作为探针,进行分子杂交,将保持系线粒体atpA基因定位在3.5kb的Bam HI酶切片段上,并且以pBR322为载体,克隆了这一片段,另外,在Bam HI完全酶解普带的杂交结果中,不育系线粒体基因组中有两条阳性杂交带,分别是3.5kb和2.9kb,而保持系线粒体基因组中只有3.5kb一条阳性杂交带,因而认为水稻不育系线粒体基因组中可能有两个atpA基因拷贝,而相应的保持系线粒体基因组中只有一个atpA基因拷贝。     

水稻线粒体atpA基因的克隆及其与细胞质雄性不育的关系

本研究以水稻BT型细胞质雄性不育系秋光和相应的保持系秋光为材料，提取线粒体DNA，用限制性内切酶完全酶解，以玉米线粒体atpA基因和波菜叶绿体atpA基因作为探针，进行分子杂交，将保持系线粒体atpA基因定位在3.5kb的Bam HI酶切片段上，并且以pBR322为载体，克隆了这一片段，另外，在Bam HI完全酶解普带的杂交结果中，不育系线粒体基因组中有两条阳性杂交带，分别是3.5kb和2.9kb，而保持系线粒体基因组中只有3.5kb一条阳性杂交带，因而认为水稻不育系线粒体基因组中可能有两个atpA基因拷贝，而相应的保持系线粒体基因组中只有一个atpA基因拷贝。     

榨菜胞质雄性不育系与保持系间线粒体和叶绿体多肽比较

应用SIXS-PAGE技术对榨菜(Brassica juncea Coss.vat.tumida Tsen et Lee)胞质雄性不育系(CMS)和保持系(MF)不同发育时期(苗期、抽薹期、盛花期)的线粒体和叶绿体多肽进行了比较研究．结果表明：线粒体方面,各发育时期不育系比保持系多1条约37kD多肽带,另1条约35kD多肽仅出现于CMS盛花期的线粒体中,叶绿体方面,各发育时期保持系55kD多肽的表达量明显高于不育系叶绿体,其与Rubis CO大亚基分子量吻合。此外,还对植物胞质雄性不育系的分子机理进行了讨论。     

水稻配子体,孢子体细胞质雄性不育系线粒体体外翻译产物的分析

对水稻配子体细胞质雄性不育系粤泰Ａ,保持系粤泰Ｂ,Ｆ１代泰优２号和孢子体细胞质雄性不育系珍汕９７Ａ,保持系珍汕９７Ｂ,Ｆ１代汕优６３及另一种孢子体细胞质雄性不育系马协Ａ,保持系马协Ｂ,Ｆ１代马协６３的黄化苗线粒体离体翻译产物进行ＳＤＳ－ＰＡＧＥ分析。结果表明：粤泰Ａ的线粒体比粤泰Ｂ,泰优２号少合成２条多肽,Ｂ与Ｆ１的带型相近,Ａ特异合成４０．７ｋＤ多肽;     

Proteins of Axial Organs of Dormant and Germinating Horse Chestnut Seeds: 1. General Characterization

This is the first characterization of proteins from axial organs of recalcitrant horse chestnut seeds during deep dormancy, dormancy release, and germination. We demonstrated that, during the entire period of cold stratification, axial organs were enriched in easily soluble albumin-like proteins and almost devoid of globulins. About 80% of the total protein was found in the cytosol. Approximately one third of cytosolic proteins were heat-stable polypeptides, which were major components of total proteins. Heat-stable proteins comprised three groups of polypeptides with mol wts of 52–54, 24–25, and 6–12 kD with a predominance of low-molecular-weight proteins. The polypeptide patterns of heat-stable and thermolabile proteins differed strikingly. Heat-stable proteins accumulated in axes during the late seed maturation, comprising more than 30% of the total protein in axes of mature seeds. The polypeptide patterns of the total protein of axial organs and its particular fractions did not change in the course of seed dormancy and release. At early germination, the content of heat-stable proteins in axes decreased and their polypeptide pattern changed both in the cytosol and cell structures. We believe that at least some heat-stable proteins can function as storage proteins in the axes. Localization of storage proteins in the cells of axial organs and the role of heat-stable proteins in recalcitrant seeds are discussed.     

水稻细胞质雄性不育系及其保持系幼穗发育过程中内源激素的变化

利用酶联免疫测定技术研究了水稻细胞质雄性不育系珍汕97A及其保持系珍汕97B在幼穗发育过程中叶片、幼穗和花药中内源IAA、GA1＋4、ABA和iPAs含量的动态变化。结果表明,1）从幼穗发育的雌雄蕊形成期到三核花粉期,保持系叶片中IAA水平高于不育系,并在二核花粉期最高;在幼穗和花药中也以保持系为高。2）保持系与不育系叶片中GA1＋4含量变化趋势相似,均为先升后降,但从单核到三核花粉期以保持系为高;在幼穗和花药中也都以保持系为高。3）不育系叶片中ABA水平在幼穗发育早期明显高于保持系,中期与保持系相近,后期又高于保持系;在幼穗和花药中也都以不育系为高。4）保持系叶片、幼穗和花药中iPAs含量始终显著高于不育系。5）保持系叶片中IAA＋GA1＋4＋iPAs与ABA之比值也始终高于不育系。提示不育系叶片、幼穗和花药中IAA、GA1＋4、iPAs和ABA含量出现了异常,且IAA、GA1＋4和iPAs亏缺以及ABA盈积可能与水稻细胞质雄性不育发生有关。     

不同倍性不结球白菜Pol CMS及保持系生理生化特性比较

对二、四倍体不结球白菜Pol CMS及其保持系花蕾和薹叶进行生理生化特性比较分析,结果表明:不育系花蕾中可溶性蛋白、可溶性糖、脯氨酸含量均显著低于保持系,且各指标在不育系中四倍体低于二倍体,而在保持系中则四倍体高于二倍体;不育系MDA含量,POD、SOD、CAT活性均高于保持系,不育系中四倍体均高于二倍体.而保持系中四倍体均低于二倍体;薹叶中MDA含量,POD、SOD、CAT活性均为不育系高于保持系,不育系中四倍体高于二倍体;POD、EST同工酶显示不育系与保持系间均具特异酶带,但不同倍性间并无差异.     

Proteins of cotyledons of mature horse chestnut seeds

This is the first characterization of proteins from storage parenchyma of cotyledons of mature dormant recalcitrant horse chestnut (Aesculus hippocastanum L.) seeds and evaluation the cell protein-synthesizing capacity. It was established that the content of protein in cotyledons did not exceed 0.5% of tissue fresh weight. Soluble proteins (the proteins of the postmitochondrial supernatant or cytosol) comprised the bulk (up to 90%) of total proteins. Protein of subcellular structures (20000 g-pellet) comprised 5–7% of total protein. Cotyledon proteins were heterogenous in their charges and molecular weights of subunits. Cotyledon protein was easily extracted with a salt (1 M NaCl); they comprised 90% of water-soluble albumin-like proteins. The proportion of globulins was insignificant; it did not exceed 5%. Most water-soluble proteins (more than 80%) were tolerant to heat denaturing. Among these heat-stable proteins, two major groups of polypeptides dominated: an electrophoretically homogeneous component with a mol wt of 24–25 kD and a complex group from three to five polypeptides with mol wts in the range between 6 and 12 kD. Native heat-stable proteins had disulfide bonds. Four fractions of heat-stable proteins were obtained by ammonium sulfate fractionation; three of them were alike in their polypeptide composition and contained major components with mol wts of 24–25 and 5–12 kD. It was established that the active translational machinery functioned in the cells of storage parenchyma in cotyledons of mature dormant horse chestnut seeds. During each stage of stratification, cotyledon fragments incorporated ³⁵S-methionine into TCA-insoluble material more actively than axial organs. We discuss cotyledon protein composition, their function as a storage organ, and a possible role of heat-stable proteins.     

Analysis of chloroplast differences in leaves of rice isonuclear alloplasmic lines

The chloroplast being an important organelle of plant cells could possibly be associated with plant cytoplasmic male sterility (CMS). To better understand the correlation between (CMS) and chloroplast, we presented a comprehensive analysis based on the changes of photosynthetic parameters, chloroplasts ultrastructure, soluble sugar and starch content, the relative expression of sugar and starch metabolism genes, and chloroplast genome in rice isonuclear alloplasmic CMS lines at the flowering stage. Leaf gas exchange parameters did not affect by CMS lines (M2BS and M2A), although intercellular CO₂ concentration (C _i) was influenced in both M2BS and M2A. Ultrastructural observation results indicated that many starch granules were observed in the chloroplast of CMS lines, especially bigger size in M2BS, while few ones in M2B. Only the chloroplasts of M2A contained some additional number of lipoids compared with those of the other two lines (M2B and M2BS). Soluble sugar and starch contents in CMS lines (M2BS and M2A) were significantly higher than those in maintainer line (M2B) (p?<?0.01). The relative expression of sugar and starch metabolism genes indicated the imbalance of starch and sugar synthesis and decomposition may lead to accumulation of starch granules and demonstrated the presence of cytoplasmic effects. Moreover, chloroplast genome sequencing results showed similarity in both CMS lines, which revealed different single nucleotide polymorphisms (SNPs) and insertion/deletion (InDels) models compared with their maintainer line. Those models were located in psbD, rpoC2, rpl33, psbB, ndhA, ndhH, and intergenic regions. These findings, aligned with the possible association of CMS characteristics with cpDNA and genetically close relationship among both CMS lines, may contribute for future research.     

WA型,BT型和D型水稻雄性不育系细胞质基因组翻译产物分析和研究

通过对ＷＡ型、ＢＴ型和Ｄ型水稻雄性不育系细胞质基因即线粒体基因组和叶绿体基因组翻译产物的分析,初步寻找出与雄性不育和育性恢复有关的调控基因产物。根据实验结果,讨论了水稻细胞质雄性不育及育性恢复机制,并提出了两种育性恢复假说。     

Glutamine synthetase genes of pea encode distinct polypeptides which are differentially expressed in leaves, roots and nodules.

We have characterized the distinct polypeptides, primary translation products and mRNAs encoding glutamine synthetase (GS) in the various organs of pea. Western blot analysis of soluble protein has identified five distinct GS polypeptides which are expressed at different relative levels in leaves, roots and nodules of pea. Of the two GS polypeptides in leaves (44 and 38 kd), the 44-kd GS polypeptide is predominant and is localized to the chloroplast stroma. In roots, the predominant GS polypeptide is 38 kd. Upon Rhizobium infection of roots, three 37-kd GS polypeptides increase in abundance in the nodules relative to uninfected roots. cDNA clones encoding three different GS mRNAs have been characterized. Hybrid-select translation has identified three different GS primary translation products (49, 38 and 37 kd). Two cDNA clones (pGS134 and pGS341) are homologous to GS mRNAs most abundant in nodules which encode the 38- and 37-kd GS primary translation products. A third cDNA (pGS197) corresponds to a larger GS mRNA species specific to leaf poly(A) RNA, which encodes a 49-kd putative precursor to the mature chloroplast GS polypeptide. cDNA sequence analysis and Southern blot analysis of pea nuclear DNA identifies at least three genes encoding GS in pea which are related but distinct in structure and in vivo pattern of expression.