Nicotiana tabacum L. as a source of cytoplasmic male sterility in interspecific cross with N. alata Link & Otto

A new source of cytoplasmic male sterility (CMS) in tobacco with interspecific origin is reported. In traditional selection wild tobacco species have been used as donors of cytoplasm. In the present study the cultivated species Nicotiana tabacum L. (n = 24) is a source of CMS. It was used as female parent and N. alata (n = 9) was involved as a pollinator. The F1 hybrid of this cross was completely sterile. Tissue culture method was applied to restore the female fertility. Regenerants obtained from the 5th passage were successfully pollinated with N. tabacum and seed-containing capsules were formed. All BC1P1 plants were male sterile. They possessed normally developed corollas, three-loculed or deformed pistils, and 1–2 stamens modified into secondary pistils. In some plants stamenless flowers were observed. Male sterility of BC1P1 was preserved in BC2P1–BC7P1 progenies confirming its cytoplasmic nature. This revised version was published online in July 2006 with corrections to the Cover Date.     

Variation in Nicotine Content of Cultured Cell Lines of Nicotiana Species and their Somatic and Sexual Hybrids

Several Nicotiana species, their intergeneric sexual and somatic hybrids were compared for their in vitro growth rate and nicotine content. Cell yield on a dry weight basis and HPLC analysis of their nicotine content revealed a very high variation within each cell line from parental, sexual and somatic hybrids for the two traits analyzed. The trigeneríc sexual hybrids (N. rustica × N. debneyi) × N. tabacum and only two of the N. rustica × N. tabacum somatic hybrid lines show a positive heterosis in nicotine content in vitro The in vitro growth rate and nicotine content did not demonstrate a statistically significant correlation.     

Enarthrocarpus lyratus-based cytoplasmic male sterility and fertility restorer system in Brassica rapa

Substituting the nuclear genome of Brassica rapa into the cytoplasmic background of Enarthrocarpus lyratus through backcross substitution helped in developing cytoplasmic male sterility (CMS). Alloplasmic male sterile plants had pale green leaves, small flowers with narrow petals and rudimentary anthers. Female fertility, low initially, improved considerably with advanced backcross generations. Male sterility expression was stable throughout the growing season. Except for EC 339014, all B. rapa accessions (38) evaluated were partial maintainers of the male sterility. Introgression of gene(s) for fertility restoration from the cytoplasm donor species was facilitated by homoeologous pairing between B. rapa and E. lyratus genomes, as was apparent from the very frequent occurrence of a trivalent in the monosomic addition plants (2n = 10 II + 1 I). Backcrossing of fertile monosomic addition plants with B. rapa led to the recovery of male fertile plants possessing the stable euploid chromosome number (2n = 20). These plants restored male fertility in crosses with different (lyr) CMS B. rapa genotypes, confirming the introgression of fertility restorer gene(s) from E. lyratus, the cytoplasm donor species.     

A new source of cytoplasmic male sterility in sunflower

Summary Interspecific substitutions of the nucleus of Helianthus annuus (2n=34) cv. Saturn into the cytoplasm of H. petiolaris (2n=34) by successive backcrossing, resulted in progenies with indehiscent anthers containing white, rather than normal yellow, pollen. Further backcrossing by cv. Saturn failed to restore pollen shed, suggesting that the male sterility was cytoplasmic. In vivo germination tests of pollen from 23 such plants from eight BC₅ lines, indicated complete pollen sterility for 14 plants, but normal seed set, suggesting that female fertility was not affected. Meiosis in all plants examined was normal.Crosses between seven sources of pollen-fertility restorer, one collection of wild H. annuus, and an existing source of cytoplasmic male sterility, resulted in a high frequency of plants with normal pollen shed in all F₁ progenies. However, no normal pollen shed was evident in F₁ progenies for similar crosses between BC₅ male-steriles and three of the seven restorer sources, nor for the single wild H. annuus evaluated. The foregoing suggests that the backcross substitution lines are a new source of cytoplasmic male sterility. The inheritance of restoration of pollen shed was complex and not fully elucidated. Some data suggested that two independent, complementary, dominant genes were required, but others indicated two to three independent, dominant genes.     

部分烟草种质亲缘关系的AFLP分析

利用AFLP技术,对48份不同类型和地域来源的烟草种质的亲缘关系进行了分析。选用4对AFLP引物共检测到323条扩增带,其中174条具有多态性,平均多态检出率为55.72%。对AFLP扩增结果采用UPGMA法进行聚类分析,可以将48份烟草种质分为2大类群,即黄花烟草群和普通烟草群,后者又可进一步分为4组;48份材料的欧氏距离为1.41~10.78。初步研究表明,AFLP标记技术能较好地从分子水平揭示烟草种质资源的遗传背景和亲缘关系。     

Development of a new cytoplasmic-nuclear male-sterility line of soybean and inheritance of its male-fertility restorability

Based on previous work, a new cytoplasmic‐nuclear male‐sterile line NJCMS2A along with its maintainer NJCMS2B of soybean was released through four episodes of nuclear substitution backcrossing with male‐sterile plants of (N8855 × N1628)F₂ as donor parent and N1628 as recurrent parent. The male sterility of NJCMS2A was complete and female fertility was normal. A series of restorers from different sources were screened out and evaluated. It was found that from the crosses between NJCMS2A and its restorers, two pairs of duplicate dominant genes, allelic to those of NJCMS1A, controlled the inheritance of fertility restorability of NJCMS2A.     

Plasmons and male sterility types in Solanum verrucosum and its interspecific hybrid derivatives

Summary Solanum verrucosum (2n=24) is easily crossable with many diploid species and haploids, but usually as a female partner only. All interspecific hybrids studied appeared male sterile due to interaction of S. verrucosum plasmons with genes from male parents. Seven male sterility types have been discovered in the investigations made on 21 S. verrucosum introductions and their interspecific hybrid derivatives. These are briefly discussed. S. verrucosum proves to possess at least four plasmons denoted as [Tr^s], [Ps^s], [Sv^s] and [ns^s]. Each produces a diagnostic type of male sterility in the presence of the specific plasmon-sensitive genes which are widely spread. The role of hybrid sterility as a barrier to gene exchange between populations is discussed.     

应用SRAP与ISSR分析烟草种质资源遗传多样性及遗传演化关系

应用ISSR与SRAP两种分子标记,研究国内外96份烟草种质的遗传多样性及不同栽培类型种质的遗传演化关系。表明烟属种间具有丰富的遗传多样性,种间的遗传相似性(GS)在0.28~0.58之间,遗传分化系数(Gst)为0.83。普通栽培种品种间遗传多样性水平较低,品种间的遗传相似性在0.61~0.99之间,栽培种内的遗传多样性为烤烟>晒晾烟>白肋烟>香料烟。当相似系数在0.67作切割线时,基于2种标记的96份烟草种质资源的聚类结果为,(1)普通烟草栽培品种材料91份聚在同一大类,而黄花烟、黏烟草、浅波烟草、哥西氏烟草、香甜烟草5个种也分别为单独的个类,同普通烟草栽培种类群完全区别开来;(2)从进化上看,烤烟和晒晾烟间的遗传进化关系最近,香料烟和黄花烟之间的亲缘关系较远;普通烟草栽培种中国内外来源的烟草品种亲缘关系极其相近,遗传分化现象甚微;(3)2种分子标记虽然原理不同,但分析结果趋势相近(r=0.68,P=1.000)。     

四种细胞质来源的烟草不育系线粒体SSR位点差异

采用线粒体SSR(mtSSR)方法对普通烟草4种不育类型的不育系、不育胞质来源的烟草野生种和保持系进行线粒体位点差异分析,探讨mtSSR位点差异与胞质不育的关系。在烟草线粒体基因组上检索到24个能重复稳定扩增出片段的mtSSR,对烟草属植物的这些mtSSR分析结果显示,普通烟草种内mtSSR较为保守,多态性比率仅10.5%;4种不育类型的各自不育系与保持系、不育胞质上存在多个差异位点,在Nta(sua.)S类型中差异位点在线粒体编码基因orf138a、orf138c以及orf102~orf210的基因间区;在Nat(gla.)S类型中,差异位点在线粒体orf138a、orf138c两个编码基因以及nad2~sdh3、rps12~orf125b、orf116~orf101b、orf102~orf210的4个基因间区;在Nta(rep.)S类型中,差异位点在线粒体编码基因orf190以及orf104c~orf202的基因间区;在Nta(rus.)S类型中,差异位点在线粒体编码基因orf137以及rps12~orf125b的基因间区。这些位点差异可能与各自的雄性不育有关。另外,mtSSR引物TMS20在Nat(sua.)S、Nat(gla.)S、Nat(rep.)S和Nat(rus.)S类型的雄性不育系中分别扩增出了184、172、212和225 bp的片段,表明可用该引物区分4种烟草CMS类型。     

Breeding a genie and cytoplasmic double-MS line of rapeseed (Brassica napus L.)

All rapeseed lines with Polima male sterility (MS) that are applied in hybrid cultivars have the problem that their sterility varies with temperature. To overcome this problem, two double-MS lines with genie (GMS) as well as cytoplasmic male sterility (CMS) genes were synthesized through seven generations of breeding, based on a systematic study of changes in fertility of the genie and cytoplasmic male sterility lines. The fertility of the new sterility lines was determined by observation of the floral organs and by pollen staining. The results showed that, in the double-MS lines, half the plants maintained the features of the Polima CMS line, while the other half behaved like the GMS line. The GMS genes were correctly expressed in the Pol cytoplasm, but there was little interaction between the GMS and CMS genetic systems.     

甘蓝型双低油菜胞质雄性不育系384A不育性的表现与遗传

对甘蓝型双低油菜细胞质雄性不育系３８４Ａ的不育性表现、遗传方式等方面的研究结果表明,该不育系属败药型不育,其败育彻底,育性稳定,花瓣开张正常,不育性的遗传受不育细胞质（Ｓ）和一对隐性核不育主效基因（ｒｒ）共同控制,属孢子体不育,基因型为Ｓ（ｒｒ）,同时也受若干位点上微效基因的影响。     

利用分子标记鉴定白菜细胞质雄性不育类型的研究

通过对9份白菜细胞质雄性不育材料的不育胞质类型进行分子标记鉴定,以期为今后更好的利用分子标记来进行辅助育种奠定基础。通过CTAB法提取白菜的基因组DNA,根据GenBank中orf138基因保守序列设计2对特异引物,对9份白菜基因组DNA进行PCR扩增,将扩增出条带的PCR产物进行测序,在NCBI中进行同源性比对,最终确定其不育胞质类型。结果表明：2对特异引物PI/PII和PIII/PIV只在4份白菜雄性不育基因中均扩增出条带,在另外5份材料中未扩增出条带,与田间育性鉴定相符,获得4份白菜Ogura胞质雄性不育的分子标记;在GenBank中用BLAST进行同源性比对分析,发现L1-CI、L3-CI、L3-F1 3个不育材料的特异片段与已报道的青花菜Ogu CMS所具有的Ogu orf138基因（Genbank登录号：HQ149728）同源度高达100%,L1-F1的同源度达99%,出现了一个变异位点,另外5份材料的胞质不育类型有待进一步研究。通过该方法鉴定出9份材料中有4份材料是萝卜胞质雄性不育的材料,得到了4条分子标记,该结果可为白菜细胞质雄性不育的分子鉴定提供了新的标记,也为今后利用分子标记辅助育种奠定了基础。     

Effect of temperature on the expression of cytoplasmic male sterility in cultivated barley (Hordeum vulgare L.)

Twenty‐three cytoplasmic male sterile BC₁F₁ barley lines were exposed to varying temperature treatments (TTs) to examine the effect of temperature on the stability of the expression of cytoplasmic male sterility (cms). The TTs used for this test were: (i) controlled low‐temperature treatment of 16°C (CL), (ii) controlled medium‐temperature treatment of 21–24/16–17°C day/night (CM) and (iii) ambient glasshouse‐temperature treatment of 24–41/16–17°C day/night (AG). The expression of cms was found to be variably influenced by temperature and by the genetic background of the cms recipient lines. Ten cms lines exhibited consistently complete male sterility over TTs, indicating that these lines are completely under cms genes control, while other lines showed partial revision to fertility across different TTs with profound influence of warm temperatures (CM and AG) in breaking down the cms in barley.     

甘蓝型油菜萝卜质雄性不育恢复系的选育

为了获得甘蓝型油菜萝卜细胞质雄性不育(Cytoplasmic male sterility,CMS)高含油量双低品质恢复系,以Ougra-CMS不育胞质杂交种Oug-F1(S(Rfrf))为不育基因和恢复基因供体,高含油量双低亲本D89-44(N(rfrf)为受体,通过杂交、多代同交以及最后自交对恢复基因进行了定向转育...     

向日葵细胞质雄性不育性的机制与利用

向日葵杂种生产采用三系配套系统完成。因此,向日葵细胞质雄性不育性的研究备受关注。其中研究较为清楚的是PET1细胞质,其不育机理是线粒体DNA重组形成嵌合基因,编码新的多肽,导致小孢子的败育。然而,这些基因如何影响小孢子发育的生理机制仍不清楚,有待进一步深入研究。丰富多样的向日葵CMS会大大降低因遗传单一性给大规模生产带来的风险;未发现恢复系的CMS可以用于无花粉观赏向日葵杂交种的生产。     

中国甘蓝型油菜细胞质雄性不育的主要类型及在育种实践中的应用

我国甘蓝型油菜细胞质雄性不育的类型主要有：ogura CMS、nap CMS、pol CMS、Shan 2A CMS、Hau CMS、NCa CMS、Nsa CMS和NEA CMS。pol CMS和Shan 2A CMS在我国油菜杂种优势利用中发挥着重要的作用。新型不育胞质的育成以及在育种上的应用,不仅丰富了细胞质雄性不育种质资源,而且为我国油菜产业可持续发展提供了保证。     

Intergeneric symmetric and asymmetric somatic hybridization in Festuca and Lolium

Summary Intergeneric symmetric and asymmetric somatic hybrids have been obtained by fusion of metabolically inactivated protoplasts from embryogenic suspension cultures of tall fescue (Festuca arundinacea Schreb.) and unirradiated or 10–500 Gy-irradiated protoplasts from non-morphogenic cell suspensions of Italian ryegrass (Lolium multiflorum Lam.). Genotypically and phenotypically different somatic hybrid Festulolium mature flowering plants were regenerated.Species-specific sequences from F. arundinacea and L. multiflorum being dispersed and evenly-represented in the corresponding genomes were isolated and used for the molecular characterization of the nuclear make-up of the intergeneric, somatic Festulolium plants recovered. The irradiation of Italian ryegrass protoplasts with 250 Gy X-rays prior to fusogenic treatment favoured the unidirectional elimination of most or part of the donor chromosomes. Irradiation of L. multiflorum protoplasts with 500 Gy produced highly asymmetric (over 80% donor genome elimination) nuclear hybrids and clones showing a complete loss of donor chromosomes.The RFLP analysis of the organellar composition in symmetric and asymmetric tall fescue (+) Italian ryegrass regenerants confirmed their somatic hybrid character and revealed a bias towards recipient-type organelles when extensive donor nuclear genome elimination had occurred.Approaches aimed at improving persistence of ryegrasses based on asymmetric somatic hybridization with largely sexually-incompatible grass species (F. rubra and Alopecurus pratensis), and at transferring the cytoplasmic male sterility trait by intra- and inter-specific hybridization in L. multiflorum and L. perenne, have been undertaken.Abbreviations cpDNA chloroplast DNA - CMS cytoplasmic male sterility - 2,4-D 2,4-dichlorophenoxy-acetic acid - IOA iodoacetamide - mtDNA mitochondrial DNA - RFLP restriction fragment length polymorphism     

Cytoplasmic and cytogenetic relationships among tetraploid Triticum species

Summary The F₁ hybrids from crosses of 59 accessions of wild and cultivated Triticum types including amphidiploids T. boeoticum-Ae. squarrosa, T. timopheevi-Ae. squarrosa, T. timopheevi-T. monococcum, T. boeoticum (4n), T. macha, and T. Zhukovskyi with T. durum Sel. 56-1 and/or T. aestivum were examined for male sterility and chromosome pairing at metaphase I of meiosis in pollen mother cells. Those hybrids which produced male-sterile F₁'s were recurrently backrossed with pollen from T. durum or T. aestivum to study segregation for male sterility and/or to confirm cytoplasmic male sterility.All T. timopheevi and T. araraticum accessions and several T. dicoccoides types, including T. dicoccoides var. nudiglumis from the Turkey-Iran-Iraq area, had male sterility inducing cytoplasm. The chromosome pairing in the F₁ hybrids indicated that all tetraploid Triticum accessions with male sterility inducing cytoplasm had genome AAGG. T. dicoccoides Körn types from the Turkey-Iran-Iraq area had genomes AABB and did not have male sterility inducing cytoplasm. Therefore, T. dicoccoides Körn and the T. timopheevi complex differ from each other cytoplasmically and cytogenetically and occur sympatrically in the Turkey-Iran-Iraq area.Possibly, the cytoplasm of the emmers was not derived from the putative diploid progenitors, T. boeoticum, Ae. speltoides, or Ae. bicornis as indicated by their nucleo-cytoplasmic and cytogenetic relationships with the tetraploid Triticum species. The cytoplasmic differences among Ae. speltoides, T. araraticum and T. timopheevi are of a relatively smaller magnitude than the cytoplasmic differences among T. timopheevi, T. boeoticum, and the emmers. A complete analysis of nucleo-cytoplasmic relationships among Triticum and Aegilops species may indicate the cytoplasmic donor(s) to the two tetraploid Triticum species complexes.Authorized for publication 19 July, 1972 as Paper No 397 in the Journal Series of the North Dakota Agricultural Experiment Stations.     

Modified complementation test of male sterility mutants in pepper (Capsicum annuum L.): preliminary study to convert male sterility system from GMS to CMS

The male sterility system in hybrid seed production can eliminate the cost of emasculation and ensure seed hybridity through avoidance of self pollination. GMS and CMS are two types of male sterility system that currently employed in pepper breeding. Conversion from GMS to CMS will increase the male sterility proportion of female parent from 50 to 100%. In this study, segregation analysis of four male sterile mutants consisting of one CMS mutant (CA1) and three GMS mutants (GA1, GA3 and GA4) showed that each had single recessive gene inheritance. A modified complementation test was performed by replacing male sterile mutants with their maintainer line as male parent. The nuclear restorer gene for CMS was independent of all nuclear restorer genes for GMS and all nuclear restorer genes for GMS were independent each other. Further observation on CMS and GMS male sterility loci revealed that GA1 and GA3 had mutated in both nuclear restorer genes for CMS and GMS, while CA1 and GA4 each carried mutation in single male sterility system of nuclear restorer gene for CMS and GMS, respectively. Conversion from GMS to CMS in the case of lines carried mutations in both sterility systems required only S-type cytoplasm donor, while lines carried mutation in single nuclear restorer gene for GMS required not only S-type cytoplasm but also rf allele donors. The important finding is the broader function of maintainer line in certain male sterility system that can be used as a maintainer or restorer line for other male sterility systems. We also confirmed that line CC1 is the general restorer for both CMS and GMS systems.     

Barriers to hybridization of Solanum bulbocastanum Dun. and S. Verrucosum Schlechtd. and structural hybridity in their F1 plants

Summary Results of reciprocal crosses between Solanum verrucosum (2n=2x=24) and S. bulbocastanum (2n=2x=24) are described in terms of pollen tube behaviour in styles, of berry and seed set, of fertility and of meiotic behaviour of the F₁ hybrids. Pollen tube growth of S. verrucosum is strongly inhibited in styles of S. bulbocastanum, whereas no inhibition is observed in the reciprocal cross. Therefore S. bulbocastanum x S. verrucosum fails to set berries or seeds, whereas the reciprocal cross produces both berries (54.4% berry set) and seeds (0.3 per berry). Only 14.6% of the seeds germinate. Both the diploid and corresponding tetraploid hybrid plants are vigorous, flower abundantly, have a rather regular meiosis (mainly rod bivalents), but show a high degree of cytoplasmic-genic male sterility. Crossability of the diploid hybrid plants is nil when used as pollen parents and near to zero when used as pistillate parents. The barriers to hybridization of the parent species (unilateral inhibition of pollen tube growth, somatoplastic sterility, cytoplasmic-genic male sterility and structural differences of the parental chromosomes) are discussed and methods are suggested to overcome these barriers.