红麻GMS与CMS小孢子败育过程的细胞学及组织化学比较

以红麻细胞质雄性不育系(GMS)L23A、保持系L23B及从该保持系中发现的细胞核雄性不育系(CMS)L23BS为材料,采用石蜡显微制片技术,在光学显微镜下比较观察了三者花药发育过程中小孢子的发育过程及组织化学变化。结果表明,L23A的花药发育过程中,小孢子发育的不同阶段均出现败育现象,最早的败育表现为花粉母细胞在减数分裂之前的退化解体,最终形成空的花粉囊; 有的因花粉母细胞减数分裂过程异常而导致不能形成小孢子四分体;有的因小孢子在四分体中不能正常释放而败育,同时绒毡层细胞过度液泡化,并提早解体死亡; 花药发育早期含有少量蛋白质和淀粉,随着花药的发育逐渐变少。而L23BS小孢子败育的时期集中于四分体至单核花粉期间,表现为小孢子发育异常,有些小孢子不能从四分体里释放出来而影响其正常发育。在四分体以前与可育系类似,花药含丰富的蛋白质和淀粉。在败育的过程中,花药中的蛋白质和淀粉含量渐渐减少,但药隔组织中的在颗粒淀粉含量几乎不变。     

Cytological screening of rice TGMS lines

An investigation was undertaken to develop new stable thermo‐sensitive genic male sterility (TGMS) rice lines in intermated progenies (IMPs) of TGMS lines by using an anther clearing technique. The results indicate that both pre‐ and post‐meiotic genetic systems operate during anther development for the expression of sterility in TGMS lines. In all the TGMS lines, sterile anthers were small with empty pollen grains of irregular shape, except for TS 16, which showed pollen‐free anthers. This indicates that the sensitive stage of TS 16 is around stage IV (stamen and pistil primordia) of panicle development. Distinct differences were observed between sterile and fertile phases with respect to anther size, shape and colour of the pollen grains in TS 18 and TS 29. The pollen grains at the sterile phase were small and irregular in shape while in the fertile phase they were plump and larger with a yellow colour, establishing that the occurrence of sterility in TS 18 and TS 29 is post‐meiotic. Three distinct classes of pollen fertility percentage viz. <20%, 50‐70% and >90% were observed in IMPs. Anther clearing in IMPs showed distinct developmental patterns of pollen production with respect to distinct classes of pollen fertility. Less than 20% pollen fertility was observed in hybrids such as TS 15 × TS 16, TS 15 × Co 47, TS 18 × TS 16 and TS 18 × Co 47 which hold promise for developing new TGMS lines with a good plant type and acceptable quality.     

萝卜雄性不育小孢子发生的形态学研究

对萝卜雄性不育花进行形态学观察，发现两种类型的不育花，即花粉败育型和雄蕊萎缩型。比较研究了可育花及花种类型的不育花小孢子发育的细胞形态学特征，发现萝卜小孢子败育是个连续过程，在造孢细胞时期就已出现败育迹象，在小孢子发育过程中，绒毡层细胞出现液泡化、肥大、增生等异常现象。小孢子的败育与绒毡层细胞异常同步发生，小孢子一直信滞在单核期不再进一步发育，最终消失。另外，在正常可育小孢子发育过程中还偶尔观察到了不育花粉粒的存在。     

Electron-microscopy on anther tissue and pollen of male sterile and fertile wheat (Triticum aestivum L.)

Summary 1. Anther tissue and pollen grains of cytoplasmic male sterile and fertile wheat were studied with the aid of an electron-microscope for differences in cytoplasmic structures.2. Clear and consistently occurring cytoplasmic differences were not observed, although unidentifiable particles were discovered in both the sterile and the fertile material.3. Male sterility finds expression in the small number of organelles in the sterile pollen grains which consequently are little or hardly physiologically active and thus subject to a rather rapid degeneration. On the contrary the fertile pollen grains are full of cell organelles and therefore well equipped for a complete development.4. The fertile pollen grain contains a clear intine which is thicker than the exine. Only a very thin intine was observed in the sterile material.5. Male sterility has no visible influence on exine formation. Ubisch bodies occur in the anthers of both male sterile and fertile plants. There is no sign of any contribution to exine formation on the part of the Ubisch bodies. There was more evidence of exine formation from within than from without. The normality of exine formation of sterile pollen grains need not indicate exine formation from without; it suggests rather that exine is largely formed before degeneration.     

New insights into genotypic thermodependency of cytoplasmic male sterility for hybrid barley breeding

The cytoplasmic male sterility (CMS) system msm1 in barley is known to be thermosensitive, sometimes resulting in spontaneous fertility restoration in the absence of the corresponding restorer gene Rfm1. Here, we investigated genotypic differences concerning temperature sensitivity and the plant developmental stage at which elevated temperature induces spontaneous fertility restoration in three CMS mother lines. While one line stayed completely male sterile, a significantly higher fertility was observed in two lines after treatment from growth stage DC 41 until maturation. Microscopic analysis revealed that sterile anthers contained neither intact pollen, nor remains of aborted pollen grains, whereas pollen was visible in anthers of potentially fertile plants. We conclude that the barley CMS system affects anther and pollen development prior to meiosis. Elevated temperature during heading and flowering can lead to a spontaneous fertility restoration by reactivating pollen growth. Nevertheless, genotypic variation exists enabling the selection for stable CMS mother lines and the development of F₁ hybrids with high hybridity. As spontaneous fertility restoration due to environmental effects is difficult to phenotype, further investigations will focus on the development of molecular markers for marker‐assisted selection.     

雄性不育野生茶树开花生物学特性

为观察野生茶树雄性不育株开花生物学特性,探讨野生茶树雄性不育表型特征。以野生茶树雄性不育株及可育株为试材,田间观察了野生茶树的开花物候期、单花发育进程和花器形态特征,并采用离体萌发法、染色法测定其花粉活力和柱头可授性。结果表明：野生茶树雄性不育株开花物候期为9月上旬—10月上旬,全花期持续约29~33天,花期与对照材料相近。雄性不育花从花芽分化至花瓣凋谢平均历时约107天,比对照长14天。雄性不育花器结构具有典型的雄不育特征,表现为花冠开展度小,花丝弯曲畸形,花药彼此粘连,皱缩干瘪,不裂药,花药内无花粉或有微量败育花粉。雄性不育花雌性器官发育正常,柱头从开花前1天至开花后4天具有可授性,最佳授粉时间为开花后1天。所调查的野生茶树雄性不育类型属于花药败育型和花粉败育型。     

温敏核不育小麦可育和不育花药的细胞化学观察

温敏核不育小麦BNS可育花药发育中,脂类物质一直很少。在小孢子分裂前,花药中的淀粉粒也不多。小孢子分裂形成二胞花粉后,伴随着营养细胞中大液泡的消失和细胞质内含物的增加,细胞质中出现淀粉粒并持续增加。即将开花的成熟花粉中积累大量淀粉粒,是其营养物质积累特征。不育花药在小孢子分裂以前与可育花药相似,小孢子未显示结构差异。小孢子分裂形成二胞花粉后,不育花粉营养细胞中的大液泡不消失,细胞质内含物也不持续增加,淀粉粒的积累终止,最终导致其没有内含物的积累。这种淀粉代谢的异常与花粉败育有关。     

厚轴茶雄性不育株花药败育的生物学特性和细胞学研究

为明确厚轴茶(Camellia crassocolumnaH. T. Chang)雄性不育株花器发育形态、花药和花粉败育时期及兵细胞学特征,利用体视显微镜、石蜡切片技术、染色体制片和DAPI染色法,对厚轴茶雄性不育株和可育株开花迚程、花器形态、花药发育过程、花粉母细胞减数分裂及小孢子发育过程比较观察。结果显示,厚轴茶花属于完全花,花药其四室、呈蝶形,花药壁发育为基本型,绒毡层细胞其双核,于四分体时期形成分泌型细胞,单核花粉期开始降解,花粉母细胞经过减数分裂Ⅰ、减数分裂Ⅱ和胞质分裂后形成四面体型四分体,小孢子呈三角形,成熟花粉为二细胞型花粉。花蕾发育早期,不育株雄蕊发育正常,与可育株无明显差异。花蕾发育后期,不育株花丝弯曲,花药粘连、干瘪、褐化、坏死,不裂药。不育株减数分裂期绒毡层细胞异常增生、排列混乱,单核至双核花粉期绒毡层延迟降解。不育株花粉母细胞减数分裂过程中存在环状单价体、滞后染色体、染色体桥、染色体缺失、不均等分离、微核和多分体等异常现象。不育株小孢子胞质紊乱,单核期花粉粒相互粘附,花粉壁皱缩变形,花粉细胞质和细胞核模糊不清,成熟花粉细胞空瘪凹陷。研究结果表明,厚轴茶雄性不育花...     

芝麻核雄性不育系ms86-1小孢子败育过程的超微结构

运用透射电子显微镜对芝麻核雄性不育系ms86-1的可育和不育花药进行了超微结构的比较观察。根据小孢子的细胞学形态特征,将芝麻花粉发育过程划分为小孢子母细胞形成期、减数分裂期、四分体期、单核小孢子早期、单核小孢子中期、单核小孢子晚期、花粉成熟期7个时期。对比观察表明芝麻核雄性不育的败育迹象起始于小孢子母细胞形成期,并伴随着进一步发育,败育现象逐渐明显,小孢子母细胞形成期小孢子母细胞壁形状不规则;减数分裂期小孢子母细胞壁严重扭曲变形,质膜外缺少早期外壁成分--原基粒棒;四分体期胼胝质壁外沉积物异常,呈绒毛状;四分体解体后形成畸形小孢子,孢子外壁不健全,绒毡层异常肥厚、降解延迟,释放极少量的畸形乌氏体;随后小孢子愈发皱缩,胞质凝集,内含物减少并逐渐凝聚成一团电子致密物质,最终走向完全败育。本研究揭示了不育小孢子的败育过程和败育特征,为深入研究芝麻核雄性不育败育机理奠定了基础。     

Male sterility in some cole crops

Some male sterile plants were found in seed fields of cauliflower, Brussels sprouts and white cabbage. In all 3 crops the male sterile flowers were characterized by shorter stamens and shrivelled anthers. Male sterility did not seem to be greatly influenced by the environment.In cauliflower male sterility is determined by 1 recessive factor. In Brussels sprouts this characteristic also depends on recessive factors. In this crop, however, the mode of inheritance is much more complicated. In addition to completely male sterile plants there were also partially male sterile plants of Brussels sprouts.Two plants of white cabbage were found to be male and female sterile.     

通过组织培养获得棉花不育株

棉花（ＧｏｓｓｐｉｕｍｈｉｒｓｕｔｕｍＬ．）体细胞培养获得的再生植株中存在很大差异,其中育性变异表现最突出,在１９９０～１９９１年培养再生的植株中发现９个不育株,其中１株雌雄全不育,形态上表现高大紧凑,花器小,畸形,柱头四裂,电镜观察表明其它粉粒空瘪,内容物少,另２株为雄性不育,电镜观察发现其花粉粒外壁发育不良。用鄂荆１号做父本与其杂交,Ｆ１代值株均不育,鄂荆１号回交ＢＣ１不育株与可育株呈１：１分     

Identification and inheritance of male sterility in groundnut (Arachis hypogaea L.)

Summary Some plants without pods but with gynophores were observed in two F₄ progenies of two crosses of goundnut (Arachis hypogaea L.). The flowers on these plants had translucent white anthers with no or a few sterile pollen grains. Three such plants in the succeeding generation were hand pollinated with pollen from a short-duration Indian cv. JL 24. The resulting F₁ hybrid plants (male sterile x JL 24) were normal. Chi-square tests for segregation for male fertile and male sterile plants in F₂ and F₃ generations indicated that the male sterility in these crosses of groundnut is governed by two recessive genes. We designate these genes as ms₁ and ms₂ with ms₁ms₁ms₂ms₂ being a male sterile genotype.Submitted as ICRISAT J. A. No. 1812.     

Relationship Between Phytohormones and Male Sterility in Thermo-Photo-Sensitive Genic Male Sterile (TGMS) Wheat

Summary Fertility alternation normally occurs in a novel type of thermo-photo-sensitive genic male sterile (TGMS) wheat whose recessive genes in the nucleus are expressed as virtually complete sterility under lower temperature and shorter daylight conditions, but as normal fertility under higher temperature and longer daylight conditions during the developmental period from meiosis of pollen mother cells (PMCs) to the early mononucleate stage of microsporogenesis. In order to provide more information about the mechanism(s) of fertility alternation in TGMS wheat, the relationship between fertility alternation and levels of phytohormones was analyzed by endogenous hormone quantification and exogenous hormone treatments in the present research. Sown at an earlier date with a relatively low temperature and short daylight, TGMS wheat was male sterile phenotypically. Pollen abortion mainly occurred at the mononucleate to bi-nucleate stages of microsporogenesis and pollen abortion was mainly of the spherical type. Levels of endogenous indole-3-acetic acid (IAA), abscisic acid (ABA) and gibberellic acid (GA₃) in the anthers were at lower levels, and zeatin riboside (ZR) was higher. Sown at a later date, when the temperature during the critical period for fertility alternation was relatively high and the daylight was long, TGMS wheat was almost as fertile as the normal genotype and the contents of endogenous IAA, ABA and GA₃ in anthers were at higher levels, and the content of ZR was at a lower level. At the critical period for fertility alternation, treatments with exogenous IAA, ABA, GA₃ or aminoethoxy vinylglycine (AVG), an inhibitor of ethylene biosynthesis, decreased the frequency of sterile pollen grains and increased the frequency of self-pollinated fertile florets. Treatment with exogenous 2-chloroethyl phosphonic acid (CEPA), a releaser of ethylene, increased the frequency of sterile pollen grains and decreased the frequency of self-pollinated fertile florets. It was thus suggested that the fertility alternation of TGMS wheat was closely related to levels of phytohormones in the anthers, and changes of endogenous hormone levels were among the important factors responsible for the fertility alternation of TGMS wheat.     

洋葱细胞质雄性不育系及保持系小孢子发生的细胞学观察

为揭示洋葱细胞质雄性不育系小孢子的败育时期,利用石蜡切片技术对洋葱细胞质雄性不育系8A及其保持系8B的小孢子发育过程进行观察和比较。结果表明：石蜡切片洋葱细胞质雄性不育系8A和保持系8B造孢组织时期小孢子的生长发育没有明显的差异;洋葱不育系8A花药绒毡层在花粉母细胞时期就与中层完全分离,并逐渐膨大挤压花粉母细胞,最后逐渐降解。其降解后的残余物侵入药室,与小孢子混合粘连在一起,占据药室的一部分空间,并且不能供给小孢子生长发育所需要的营养物质,最终小孢子降解,花药败育。试验明确了洋葱细胞质雄性不育系8A的小孢子败育时期为花粉母细胞时期。因此,在洋葱雄性不育系8A的不育机理研究中,应重点在花粉母细胞时期进行深入研究。     

甘蓝型油菜温敏细胞核雄性不育系160S花药败育的形态学特征和细胞学研究

明确甘蓝型油菜温敏核雄性不育系160S花器形态变化、花药败育的时期和细胞学特征,初步探究败育的原因,为深入研究不育系160S的内在分子调控机制提供理论基础,也对其在油菜两系杂交育种中的实际应用具有指导意义。本研究在15℃和28℃条件下培养试验材料160S,利用体式显微镜分别观察花发育形态特征;采用醋酸洋红染色方法观察各时期小孢子发育形态;通过石蜡切片和苏木精-伊红染色对可育植株(MaleFertile/160S-MF)和不育植株(MaleSterile/160S-MS)花药细胞学特征进行显微观察;TUNEL染色法检测花药发育各时期绒毡层细胞凋亡情况。160S-MF在15℃表现为可育,雄蕊正常发育,成熟的花药呈黄色,形态饱满,正常开裂,表面一层有活性的花粉附着在上面;28℃条件下, 160S-MS花朵的雌蕊、萼片与160S-MF花朵无差异,但花瓣变小,花丝变短,雄蕊明显退化,花药干瘪呈黄褐色,无花粉粒附着在花药上,表现出雄性完全不育。160S-MF的小孢子能正常发育为成熟有活力的花粉。而160S-MS由于雄蕊完全败育,未观察到小孢子和花粉粒。160S-MS花药在造孢时期和花粉母细胞时期与160S-MF无明显差异,但在减数分裂期,160S-MS花药绒毡层形态和结构出现异常,绒毡层细胞排列不整齐,细胞空泡化,伴随提前解体。同时花粉母细胞发育受阻,无四分体结构形成,最终在减数分裂期完成前形成空的花粉囊。TUNEL检测发现, 160S-MS花药绒毡层细胞在减数分裂期开始凋亡。本研究结果表明, 160S属花粉母细胞败育型不育系,败育时期发生在减数分裂期,绒毡层异常降解,绒毡层未向腺质型转化,不能提供花粉母细胞发育所需要的营养物质,致使花粉母细胞发育受阻无法形成四分体结构,从而导致小孢子无法形成,花药形成空的花粉囊,产生雄性不育。     

Chromosome 1D as a possible location of a gene (s) controlling variation between wheat (Triticum aestivum L.) varieties for carbon isotope discrimination (Δ) under water-stress conditions

Cytoplasmic male-sterility (CMS) in pigeonpea has been reported when some wild relatives of pigeonpea were crossed as the female parent with cultivated types as the male parent. In this paper we report a new source of CMS developed by using the cultivated pigeonpea as the female parent and one of its wild relative Cajanus acutifolius as the pollen donor. This is the first report in pigeonpea where CMS has been developed using the cytoplasm of cultivated pigeonpea. Several pure line cultivars of pigeonpea restored pollen fertility whereas cv. HPL 24 partially maintained male-sterility. The wild species C. acutifolius used as one of the parents, maintained complete sterility. Cytological analysis revealed that both in male-sterile as well as the fertile floral buds, meiosis proceeded normally till the tetrad stage. However in the male-sterile genotypes during the formation of tetrads, the pollen mother cell (PMC) wall did not dissolve to release the tetrads unlike in the fertile genotypes and this major event was found to be responsible for male-sterility.     

绿豆雄性不育突变体msm2015-1的遗传学与细胞学分析

单产水平低一直是我国绿豆存在的产业问题。绿豆杂种优势显著是提高产量的有效途径,而雄性不育材料是实现杂种利用的宝贵资源。本研究利用~(60)Co-γ辐射诱变技术首次在绿豆中获得雄性不育突变体,并命名为‘不育绿豆2015-1’(male sterile mungbean2015-1,简写为msm2015-1)。msm2015-1营养生长期表型与‘苏绿1号’没有明显差异。花器官形态正常,但花药发白且不能正常开裂散粉,成熟期座荚率几乎为0。遗传分析显示,育性分离群体中可育与不育分离比符合3∶1,表明不育性状由单个隐性核基因控制。花粉发育的细胞学分析发现, msm2015-1的盛开花朵中花粉粒几乎未在柱头附着,体外亦未萌发。花粉细胞质未被Alexander染液正常着色, I_2-KI染色出现典败、圆败和染败3种类型。DAPI染色显示, msm2015-1花粉细胞核发育异常。醋酸洋红染色表明,败育发生在花粉发育早期,花粉母细胞在四分体时期减数分裂的不对称和异常多分裂是导致花粉败育的主要原因。     

Role of male-sterile cytoplasm in resistance to barley yellow mosaic virus and Fusarium head blight in barley

Two types of male‐sterile cytoplasm, designated msm1 and msm2, in barley were investigated to determine whether these cytoplasms confer resistance to barley yellow mosaic virus (Ba YMV) and Fusarium head blight (FHB). Alloplasmic lines and isogenic lines of two cultivars showed the same reaction to each Ba YMV as that of their euplasmic lines. This demonstrates that the barley male‐sterile cytoplasms msm1 and msm2 have no effect on resistance to BaYMV. No significant difference in reactions to FHB was recognized among fertile alloplasmic lines of ‘Adorra’, but the difference in reactions to FHB between fertile and sterile isogenic lines of ‘Adorra’ was significant. The damage caused by FHB in the male‐sterile lines that produced sterile pollen was significantly greater than the damage in a sterile line that did not produce pollen. These results suggest that pollen or anthers are important factors in infection with or spread of FHB. For production of hybrid seeds, male‐sterile lines with no pollen production, such as those with msm1 male‐sterile cytoplasm, would reduce FHB infestation.     

Genetic and histological characterization of a novel recessive genic male sterile line of <Emphasis Type="Italic">Brassica napus</Emphasis> derived from a cross with <Emphasis Type="Italic">Capsella bursa</Emphasis>-<Emphasis Type="Italic">pastoris</Emphasis>

In a previously made cross Brassica napus cv. Oro (2n = 38) × Capsella bursa-pastoris (2n = 4x = 32), one F₁ hybrid with 2n = 38 was totally male sterile. The hybrid contained no complete chromosomes from C. bursa-pastoris, but some specific AFLP (amplified fragment length polymorphism) bands of C. bursa-pastoris were detected. The hybrid was morphologically quite similar to ‘Oro’ except for smaller flowers with rudimentary stamens but normal pistils, and showed good seed-set after pollination by ‘Oro’ and other B. napus cultivars. The fertility segregation ratios (3:1, 1:1) in its progenies indicated that the male sterility was controlled by a single recessive gene. In the pollen mother cells of the male sterile hybrid, chromosome pairing and segregation were normal. Histological sectioning of its anthers showed that the tapetum was multiple layers and was hypertrophic from the stage of sporogenic cells, and that the tetrads were compressed by the vacuolated and disaggregated tapetum and no mature pollen grains were formed in anther sacs, thus resulting in male sterility. The possible mechanisms for the production of the male sterile hybrid and its potential in breeding are discussed.     

T型细胞质雄性不育小麦T763A的败育特点及育性恢复

为了明确T型细胞质雄性不育小麦T763A败育的形态特征和细胞学特点及对T763A恢复系的选用提供依据,以不育系T763A,保持系763B,恢复系Tm3315B、Tm504B和TP731B为供试材料,进行外部形态特征观察和花粉粒制片(醋酸洋红、I2-KI和DAPI);并以中国春和黑麦为对照试材,对所有供试材料进行核型鉴定。结果表明:T763A败育类型为典败和圆败,成熟花粉粒皱缩无规则,内含物少,花粉败育,败育主要发生在单核晚期到二核期;所有供试材料均为非1B/1R类型;3个恢复系(Tm3315B、Tm504B和TP731B)恢复能力均较强,其中以Tm504B对T763A的恢复能力相对最好,这可能与T763A的胞质类型及与恢复系所含的恢复基因数量有关。