红菜薹雄性不育系花药败育的细胞形态学观察

采用石蜡切片技术,在光学显微镜下系统研究了红菜薹(Brassica campestris L.ssp．chinensis L.var．utilis TsenetLee．)波里马胞质雄性不育系(Polima CMS)、红菜薹萝卜胞质雄性不育系(Ogura CMS)及相应保持系花药发育过程的细胞形态学特征。观察结果表明：红菜薹Polima CMS花药发育受阻于孢原细胞阶段,不形成花粉,属无花粉型,此不育系花药不形成绒毡层和中层;而红菜薹Ogura CMS花药败育发生于小孢子母细胞期或四分体时期,表现为绒毡层细胞异常,挤压四分体,导致四分体和绒毡层同时解体而败育。     

甘蓝胞质雄性不育系和保持系POD同工酶分析

以育成稳定多代的甘蓝胞质雄性不育系及其保持系为试材,以盛花期的根、叶、花蕾和花以及9d和13d幼苗的叶和根为试样,采用聚丙烯酰胺凝胶垂直板电泳技术,分析了POD同工酶的谱带差异。结果表明,不育系和保持系：POD同工酶存在谱带增减,活性强弱变化和器官特异表达等差异;不育系和保持系间POD同工酶差异以花、大蕾和小蕾最大,叶和根间差异不明显。     

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。     

小麦胞质突变型雄性不育系85EA与其保持系线粒体DNA的比较研究

８５ＥＡ是通过电子束辐照获得的胞质突变型小麦不育要用ＲＦＬＰ和ＲＡＰＤ技术对８５ＥＡ及春保持系的线粒体ＤＮＡ进行了比较研究。ＲＦＬＰ分析表明８５ＥＡ线粒体基因组中ｃｏｘⅡ基因的位置结构与保持系发生了变化；ＲＡＰＤ分析中引物ＯＰＤ－１５扩增产物在不育系和保持系间有明显差异，不育系的扩增产物比保持系多１条分子量为０．６ｋｂ的特展览 要带，用Ｔ－ｅａｓｙ ｖｅｃｔｏｒ克隆该不育系特异条带并命名为ＯＰＤ－     

植物胞质雄性不育及育性恢复的分子机制研究进展(综述)

本文从与雄性不育有关的线粒体基因引起雄性不育的机理、雄性不育育性恢复机制以及育性恢复基因的克隆等方面,介绍国内外对植物细胞质雄性不育分子机理的研究进展,并对今后的研究进行讨论。     

二角型小麦雄性不育系花粉败育的细胞学研究

应用石蜡切片和醋酸洋红压片法,从细胞学角度对4个同质异核二角型小麦雄性不育系ms(bicor)-8222、ms(bicor)-80(6)、ms(bicor)-5-4及ms(bicor)-5-8的花粉败育过程进行了比较研究。结果表明：(1)4种不育系败育特点总趋势基本雷同,但也因特定核型亦存在一些差异;(2)二角型雄性不育系小孢子发生与其保持系相似,花粉败育就花药而言,中层有延迟退化、解体的趋势,就花粉粒而言,败育主要发生在二细胞花粉粒后期至三细胞花粉粒期;(3)4个不育系花粉败育异常,主要表现为二分体细胞内有微核出现,内缘壁周缘细胞均呈次生纤维状增生;维管束鞘细咆排列不规则,花粉大小不一;(4)花粉败育与营养供应无直接关系,细胞核对花粉育性及育性相关性状相对细胞质影响较小。     

辣椒胞质雄性不育保持基因的分子标记

利用RAPD标记技术,以辣椒胞质雄性不育系8A及其同型保持系8B为材料,对辣椒胞质雄性不育基因和保持基因进行比较分析.结果表明:引物H7只在保持基因池有1条稳定的特异扩增带,在不育基因池未扩增出此条带,该标记可能与辣椒胞质雄性不育保持基因相连锁,命名为H7-F850.序列分析结果表明,标记H7-F850序列全长857 bp,GenBank登录号为GU208822.1.核酸序列比对结果显示,H7-F850与其具有部分同源性的片段大小均小于340 bp,未发现与其具有较高同源性的DNA序列.该RAPD标记H7-F850已成功转化为SCAR标记SF640.     

几种甘蓝型油菜雄性不育胞质效应的研究

用共同的保持系陕３Ｂ将０７Ａ、２２Ａ、ＮｉｋＡ、陕２Ａ、ＰｏｌＡ５种不同的ＣＭＳ转育成同核异质不育系,研究不育胞质与可育胞质的差异和５种不育胞质之间的差异。     

甘蓝胞质雄性不育系及其保持系花药和叶片生化特性分析

对甘蓝胞质雄性不育系CMS158及其保持系花药激素、游离氨基酸和叶片叶绿素含量进行了比较研究。结果表明,不育系花药KT和IAA含量极显著低于保持系,GA含量不育系和保持系没有显著差异。游离氨基酸总量不育系花药极显著高于保持系,脯氨酸和胱氨酸含量不育系花药极显著低于保持系,赖氨酸含量两系无明显差异,苏氨酸在两系中均未检出,其余氨基酸含量均是不育系花药高于保持系;不育系花药中丝氨酸含量最高,保持系花药中脯氨酸含量最高,两系中天冬氨酸和脯氨酸含量的差异最大。叶片叶绿素含量不育系和保持系无显著差异。     

棉花晋A细胞质雄性不育系的细胞形态学观察

棉花晋A细胞质雄性不育系雄性败育的主要时期是在造孢细胞增殖--小孢子母细胞形成时期.造孢细胞和小孢子母细胞退化导致雄性不育的主要细胞学特征是造孢细胞不能进行正常的有丝分裂,胞内常含有n个微核,小孢子母细胞细胞质液泡化,并且认为绒毡层的退化与小孢子母细胞败育密切相关.     

青花菜雄性不育系选育研究初报

以具有细胞质雄性不育基因的青花菜不育材料CMS94008为不育源,通过杂交和连续回交的方法选育出不育性稳定、经济性状良好的一批青花菜胞质雄性不育系,并以CMS92100、CMS93-2、CMS9905、CMS95234等不育材料及其转育父本为代表,对不育系的主要特征及农艺性状进行系统的观察鉴定。     

Overcoming hybrid incompatibility between Nicotiana africana Merxm. and N. tabacum and development of cytoplasmically male sterile tobacco forms

The incompatibility between the wild species N. africana Merxm. and the cultivated species N. tabacum has been overcome by in vitro techniques. Underdeveloped F₀ seeds, placed on MS medium with supplements, produced plants which upon reaching the stage of anthesis proved to be completely sterile. Female sterility of F₁ hybrids was overcome by applying tissue culture methods. Explants of stem parenchyma were grown in vitro. In every passage investigations were made of their callus production, organogenesis and cell polyploidization. The regenerants showed a great diversity in their morphological and cytological characters. Pollination of the R₁ plants (N. africana × N. tabacum) with N. tabacum produced normally seeded capsules. BC₁ plants were male sterile. The male sterility of the first backcross generation was preserved in BC₂ and BC₃, proving its cytoplasmic origin.     

Differences on the microsporogenesis and tapetal development of male fertile and cytoplasmic male sterile pepper (Capsicum annuum L.)

To clarify the time and cause of pollen abortion, differences on the microsporogenesis and tapetum development in the anthers of male fertile maintainer line and cytoplasmic male sterile (CMS) line pepper were studied using transmission electron microscopy. The results showed that CMS line anthers appeared to have much greater variability in developmental pattern than male fertile maintainer line ones. The earliest deviation from normal anther development occurred in CMS line anthers at prophase I was cytomixis in some microspore mother cells (MMCs), and vacuolisation in tapetal cells. Then, MMCs in CMS line anthers developed asynchronously and a small part of ones at the different stage degenerated in advance appearing to have typical morphological features of programmed cell death (PCD). Most MMCs could complete the meiosis, but formed non-tetrahedral tetrad microspores with irregular shape and different size and uncertain number of nuclei, and some degenerated ahead of time as well. Tapetal cells in CMS line anther degenerated during meiosis, and were crushed at the tetrad stage, which paralleled the collapse of pollens. Pollen abortion in CMS line anthers happened by PCD themselves, and the premature PCD of tapetal cells were closely associated with male sterility.     

Timing of morphological and histological development in premeiotic anthers of Nicotiana tabacum cv. Xanthi (Solanaceae)

The tobacco stamen has been the object of many developmental studies, and the organ has more recently become a model for molecular genetic studies of anther differentiation. However, the spatial and temporal details of cellular differentiation of early anther development have never been thoroughly characterized. In the present study, the age of 15 tobacco flowers from plants grown under constant light and temperature was estimated using growth analysis. Prior to tissue fixation for light microscopy, moulds of stamen and anther primordia were made with a dental impression polymer so morphological and histological observations could be made on each tissue sample. Flower ages spanned an 8-d interval during which petal and stamen initiation occurred, and sporogenous cells reached the leptonema stage of meiosis. The initial development of the tetrasporangiate anther shape largely preceded periclinal division of archesporial initials. Anatomically, periclinal divisions in the hypodermal ∗∗∗(l₂) layer were observed before archesporial initials began to divide. These data indicate differences in the cellular basis of tobacco anther development compared to earlier clonal analyses of Datura. The pattern of mitotic cell division associated with microsporangial development suggested modal peaks in division over time. The ability to estimate developmental time in the tobacco anther has implications for future studies directed at understanding mechanisms of anther evolution via heterochrony.     

空心莲子草雄蕊和雌化雄蕊发育的比较研究

采用形态解剖和石蜡切片法,对空心莲子草雄蕊和雌化雄蕊的形态和结构的发育过程进行了观察.结果显示:(1)在形态上,空心莲子草两性花雄蕊由聚药雄蕊和无药雄蕊构成,二者相间而生成一轮;而空心莲子草雌化雄蕊花的雌化雄蕊由雌化雄蕊和无药雄蕊构成,雌化雄蕊的外形如雌蕊状,其外轮为无药雄蕊,二者前后对应而生,成二轮;(2)在结构上,...     

Restoration of fertility in cytoplasmic male sterile (CMS) Nicotiana Sylvestris by fusion with X-irradiated N. tabacum protoplasts

Summary Restoration of male fertility was achieved by fusing protoplasts from male sterile (CMS) Nicotiana sylvestris plants with X-irradiated protoplasts derived from fertile N. tabacum plants. The CMS N. sylvestris plants were derived from a previous somatic hybridization experiment and contained alien (Line 92) cytoplasm. About one quarter of the regenerated plants were found to be cybrids. i.e. they consisted of N. sylvestris nuclei combined with all or some components of N. tabacum cytoplasm. In one half of these cybrids male fertility was restored to different levels. The chloroplasts of the two parental donors differ in respect to tentoxin sensitivity: chloroplasts of CMS N. sylvestris are sensitive while those of N. tabacum are insensitive. It could therefore be demonstrated that there was an independent segregation of chloroplast type and male fertility/sterility: several somatic cybrids were male fertile but tentoxin sensitive and others were tentoxin insensitive yet they were male sterile. Only in about one half of the somatic cybrids was male fertility restored together with restoration to tentoxin insensitivity.     

Mismatch-specific DNA breakdown in nuclear extract from tobacco (Nicotiana tabacum) callus

Mismatch-specific enzymatic activity was sought for in nuclei from normal and transformed plant cells originating from tobacco (Nicotiana tabacum) callus and crown gall tumor induced by Agrobacterium tumefaciens. The specific enzymatic activity was assayed with substrates derived from synthetic oligonucleotides (19-mer sequences corresponding to the human K-ras gene). Single-base changes in the middle of the sequence were the basis for creating heteroduplexes with all eight mismatches. Homo- and heteroduplexes were ligated in a size ladder and used as substrates. We detected mismatch-specific DNA breakdown and determined basic requirements for the reations. Kinetic analysis indicates the following reactivity order of preference: C : A=C : C=C : T>G : TA : AG : AG : GT : T>>G : C. It can be said now that specific mismatch recognition and repair activities have been detected in all kingdoms of living species.     

玉米新选细胞质雄性不育系小孢子发育的细胞学观察及DNA甲基化分析

细胞质雄性不育在高等植物中普遍存在,是杂种优势利用的重要工具,为推动植物杂种优势的利用发挥了重要作用。文章以本课题组前期新选的玉米细胞质雄性不育系A1、A2及保持系18(红)为材料,利用石蜡切片技术对不育材料小孢子发育过程进行细胞学观察,采用高效液相色谱法(HPLC)对不同发育时期的叶片及不同发育时期的雄穗DNA进行甲基化分析,从细胞学和表观遗传学角度了解不育系A1、A2的败育机制。结果表明：不育材料A1、A2小孢子发生败育的主要时期为四分体时期至单核小孢子中期。在不育系A2中还存在另一种败育方式,即在花粉母细胞时期表现出败育特征。甲基化分析结果表明,保持系18(红)的叶片DNA甲基化水平从苗期到拔节期迅速上升,而不育系A1、A2叶片DNA甲基化水平基本保持不变;保持系雄穗DNA甲基化水平表现为从花粉母细胞时期到双核期逐渐升高,而不育材料A1、A2从花粉母细胞时期到双核期的雄穗DNA甲基化水平表现为先上升后下降的趋势,达到最高峰的时期均出现在小孢子发育的四分体时期。从小孢子发育的细胞学观察结果可以发现,小孢子败育的主要时期往往具有较高的甲基化水平,推测DNA甲基化水平变化可能与不育材料A1、A2的花粉败育有关。