中间偃麦草染色体组型研究

通过系统对比分析 ABD、AB、AG 等类型小麦与中间偃麦草杂交后 F_1的减数分裂资料确认中间偃麦草没有与小麦 B 组同源的染色体组。减数分裂染色体 N 带分析进一步证实了这一点。以目前研究来看,中间偃麦草染色体组型以有两组彼此部分同源,另一组与它们远缘为宜,建议以字母 NNiX 代之。     

Formation of unreduced gametes is impeded by homologous chromosome pairing in tetraploid <Emphasis Type="Italic">Triticum turgidum</Emphasis> × <Emphasis Type="Italic">Aegilops tauschii</Emphasis> hybrids

It is believed that unreduced gametes with somatic chromosome numbers play a predominant role in natural polyploidization. Allohexaploid bread wheat originated from spontaneous hybridization of Triticum turgidum L. with Aegilops tauschii Coss. Unreduced gametes originating via meiotic restitution, including first-division restitution (FDR) and single-division meiosis (SDM), are well documented in triploid F₁ hybrids of T. turgidum with diploid Ae. tauschii (genomic constitution ABD, usually with 21 univalents in meiotic metaphase I). In this study, two T. turgidum lines known to carry genes for meiotic restitution were crossed to tetraploid Ae. tauschii. The resulting F₁ hybrids (genomes ABDD), had seven pairs of homologous chromosomes and regularly formed 14 univalents and seven bivalents at metaphase I. Neither FDR nor SDM were observed. The distribution of chromosome numbers among progeny obtained by self pollination and a backcross to T. turgidum showed the absence of unreduced gametes. These results suggest that high homologous pairing interfered with meiotic restitution and the formation of unreduced gametes. This may be related to asynchronous movement during meiosis between paired and unpaired chromosomes or to uneven distribution of chromosomes in anaphases, resulting in nonviable gametes.     

Cytological stability of common wheats with leaf rust resistance derived from Triticum timopheevi Zhuk

Nine common wheat strains derived from crosses involving Triticum timopheevi were studied for cytological stability and resistance to individual physiologic races of leaf rust of wheat. One strain 53R-201-4 was resistant to all the eleven races used. Another five stable strains had resistance to some of the races and were susceptible to others. Three highly unstable strains had resistance to a few of the leaf rust races used. The six stable strains had normal meiosis, varying between the most stable Cheyenne and the less stable Minturki-the two checks. In general, the inheritance of resistance to leaf rust race 9 in crosses of three unstable and one stable strain was monogenic and regular. Metaphase I chromosome pairing and meiotic irregularities at anaphase-I, dyad and quartet stages in stable strain 55–2687 and in F₁ and F₂ plants from the cross strain 55–2687 × Cheyenne indicated only slight reduction in chromosome pairing or chiasma formation in the F₁ plants. The F₂ was meiotically less stable than the F₁. The possible reasons for the increased meiotic instability of the F₂ are discussed.     

温光敏核不育小麦C412S的育性转换及其APRT基因的表达

以选育的小麦(Triticum aestivum L.)温光敏核不育系C412S为试材,以C412S回交转育时的受体亲本C412为常规品系对照,通过分期播种试验研究了其育性转换特性。用涂抹压片法观察减数分裂和小孢子发育进程,用半定量RT-PCR技术分析不育与可育条件下不同发育时期的幼穗中腺嘌呤磷酸核糖基转移酶基因(APRT)的表达水平。结果表明,通过调整播种期改变雄性发育的温光条件,C412S表现出完全不育—高不育—半不育—正常可育的育性转换特性。C412S花粉败育的高峰在单核小孢子晚期,主要表现圆败型不育。C412S的育性敏感期是从花粉母细胞形成期到成熟花粉期,其中最敏感的时段是花粉母细胞形成期到减数分裂期。与对照相比,C412S的APRT1基因序列有个别碱基变异,但编码氨基酸序列没有变化。在花粉母细胞形成期至单核期,与晚播可育条件相比,早播不育的C412S幼穗中APRT基因转录水平下调,因此认为,其育性转换与幼穗中APRT基因转录水平有一定关系。     

Problems in breeding and cytology of sugar cane

In connection with the increase of chromosome number found in species hybrids of Saccharum the author studied embryo-sac development within several Saccharum froms from early stages of meiosis in the embryo-sac mother cells to stages where fertilization already had taken place.Normally by meiosis within E M C's a row of four haploid megaspores is formed from which the innermost, the chalazal megaspore, develops into an embryo-sac and the other three degenerate.Haploid embryo-sacs, thus formed, may occur in Saccharum. Frequently however, in Saccharum endo-duplication takes place, that is splitting of the chromosomes within the nucleus of the chalazal megaspore, through which the chromosome number is increased.It is described how within Saccharum by this phenomenon embryo-sacs may be formed with diploid and tetraploid chromosome numbers and with numbers between haploid and diploid.The increase of chromosome number, observed in the young megaspore after meiosis, could be confirmed by counting the chromosomes of dividing nuclei within young embryo-sacs.Chromosome numbers found in dividing zygotes and endosperm nuclei within fertilised embryo-sacs also pointed to the occurrence of egg-cells and polar nuclei that must be haploid, diploid and tetraploid.The significance of this research in relation to plant breeding will be discussed in Part V.     

普通小麦-簇毛麦易位系T4VS·4VL-4AL的选育与鉴定

利用染色体C-分带和基因组原位杂交分析,从普通小麦-簇毛麦4V染色体二体异附加系（DA4V）与普通小麦农林26-离果山羊草3C染色体二体异附加系（DA3C）杂种后代中选育出小麦-簇毛麦纯合易位系T4VS·4VL-4AL。SSR和RFLP标记分析表明,该易位染色体包括4VS、4VL近着丝粒部分区段和4AL顶端区段;该易位系具有良好的细胞学稳定性,结实正常,为杀配子染色体诱发形成的补偿型易位;易位系T4VS·4VL-4AL高抗梭条花叶病,是小麦抗病育种新种质。     

八倍体小偃麦与普通小麦杂种自交和回交后代染色体和性状分离的研究

以八倍体小偃麦小偃６９３和普通小麦烟农１５的六个不同杂种世代为材料，研究了自交和顺交对杂种后代染色体和性状分离的不同影响。结果表明，随自交和以烟农１５为轮回亲本回交世代的增加，染色体数目逐渐减少，但回交比自交能使后代中偃麦草染色体丢失更快；回交后代ＰＭＣＭＩ染色体构型较为简单，平均交叉结数减少，回交次数过多不利于偃麦草与普通小麦染色体发生遗传重组；自交和回交世代中小偃麦类型、中间类型和小麦类型出现     

Leaf rust and stripe rust resistance genes Lr54 and Yr37 transferred to wheat from Aegilops kotschyi

The tendency of unpaired meiotic chromosomes to undergo centric misdivision was exploited to translocate leaf rust and stripe rust resistance genes from an Aegilops kotschyi addition chromosome to a group 2 chromosome of wheat. Monosomic and telosomic analyses showed that the translocation occurred to wheat chromosome arm 2DL. The introgressed region did not pair with the corresponding wheat 2DL telosome during meiosis suggesting that a whole arm may have been transferred. Female transmission of the resistance was about 55% whereas male transmission was strongly preferential (96%). The symbols Lr54 and Yr37 are proposed to designate the new resistance genes.     

Genome discrimination and chromosome pairing in the Hordeum chilense × Aegilops tauschii amphiploid

Tritordeum (X Tritordeum Ascherson et Graebner) is a synthetic amphiploid belonging to the Triticeae tribe, which resulted from crosses between Hordeum chilense and wheat. It presents useful agronomic traits that could be transferred to wheat, widening its genetic basis. In situ hybridisation with total genomic DNA from H. chilense and cloned, repetitive DNA sequences (pTa71 and pAs1) probes were used to discriminate the parental origin of all chromosomes, to analyse the chromosome pairing and to identify the chromosomes in pollen mother cells (PMCs) at metaphase I of the tritordeum line HT251 (H^chH^chDD, 2n = 4x = 28). The H. chilense total genomic DNA and the ribosomal sequence pTa71 probes, allowed the unequivocal discrimination of the 14 chromosomes of H^ch genome-origin and the 14 chromosomes of D genome-origin. Chromosome pairing analysis revealed meiotic irregularities such as reduced percentage of PMCs with complete homologous pairing, high frequency of univalents, most of H. chilense-origin and a reduced frequency of intragenomic multivalents from both genomes. The H. chilense genome revealed high meiotic instability. After individual chromosome identification at metaphase I with the pAs1 probe, we found the occurrence of pairing between chromosomes of different homoeology groups. The possible interest of the tetraploid tritordeum in the improvement of other Triticeae species is also discussed.     

高粱A2型细胞质雄性不育系小孢子发生的细胞学观察和减数分裂染色体行为分析

高粱A₂型细胞质雄性不育性（CMS）的细胞质来源于IS12662C, A₂细胞质杂交种目前已用于生产。本文以A₂/B₂V4为材料，对A₂ CMS小孢子败育过程作了细胞学观察，并对小孢子败育过程中减数分裂的染色体行为作了分析。研究发现，在A₂雄性不育系A₂V4的花药发育过程中，绒毡层细胞不形成或提前解体；绒毡层细胞畸形化；绒毡层细胞虽发育正常，但小孢子母细胞减数分裂行为异常；这些都导致小孢子退化。A₂细胞质雄性不育花粉母细胞减数分裂行为从后期Ⅰ开始出现异常，同源或姊妹染色体向两极分离时滞后或不分裂；染色体多倍化；一个细胞内出现多核和多核仁现象，最终导致小孢子败育。     

十个八倍体小偃麦的细胞学鉴定和染色体构成分析

培育新的八倍体小偃麦,对于利用偃麦草遗传物质进行小麦的遗传改良具有重要意义。本研究利用细胞学和基因组原位杂交技术,对从中间偃麦草与小麦品种烟农15杂交后代选育出的10个八倍体小偃麦山农TE256、山农TE259、山农TE261、山农TE262、山农TE263、山农TE265、山农TE266、山农TE267-1、山农TE270和山农TE274进行了细胞学鉴定和染色体构成分析。结果表明,10个八倍体小偃麦绝大多数单株根尖细胞的染色体数目为2n=56,个别单株含有54或55条染色体;大多数2n=56单株的花粉母细胞在减数分裂中期I的染色体构型为2n=28II,少数花粉母细胞存在单价体、三价体或四价体,后期I染色体可均等分向两极,仅有极少数细胞出现染色单体提前分离等现象;10个八倍体小偃麦均含有普通小麦的全套染色体和中间偃麦草的1个混合染色体基组,其中间偃麦草染色体是由来自中间偃麦草3个不同染色体基组的染色体构成的混合染色体基组,其染色体构成分别为2St+8J~S+2J+2J-St、2St+8J~S+4J、2St+8J~S+2J+2J-St、2St+8J~S+2J+2J-St、2St+8J~S+2J+2J-St、6St+4J~S+2J+2J-St、4St+6J~S+2J+2J-St、2St+8J~S+4J、2St+8J~S+4J和4St+6J~S+4J,与目前已报道的八倍体小偃麦均有所不同。研究结果可为这些新型八倍体小偃麦的研究和有效利用提供参考依据。     

Reduced chromosome association between the short arms of 5H homologues in Hordeum vulgare L. at metaphase I

Rod or ring bivalents at metaphase I (MI) reflect whether crossing-over has occurred on one or both pairs of chromosome arms, respectively, during meiotic prophase (pachytene). In cultivated barley ( Hordeum vulgare ) the highest frequencies of rod bivalents were found for satellite chromosomes (either 5H or 6H). To distinguish between chromosome 5H and 6H associations, a recombinant line derived from an H. vulgare × H. bulbosum hybrid with a hemizygous introgression from H. bulbosum on the short arm of chromosome 6H was probed with 45S rDNA and pSc119.2 using fluorescent in situ hybridization and compared with non-recombinant H. vulgare. The frequencies of rod bivalents, caused by failure of the short arms of 5H chromosomes (5HS) to associate during meiosis in the recombinant line, were similar to the frequencies of rod bivalents involving the satellite chromosomes in H. vulgare. Hence, rod bivalent formation in H. vulgare satellite chromosomes is probably based on reduced association of 5HS, corresponding to a low frequency of genetic recombination. 5HS is, therefore, more likely to show linkage disequilibrium than the other chromosome arms.     

Elucidation of intergenomic recombination and chromosome translocation: meiotic evidence from interspecific hybrids of Lilium through GISH analysis

Intergenomic exchange has been found to be a normal phenomenon in sexual polyploids. In order to distinguish whether such exchanges are derived from intergenomic recombination or translocation, 13 genotypes of an interspecific hybrid, which were previously used as parents to generate sexual polyploids, were selected for a detailed meiosis analysis. In all genotypes under study, variable numbers of bivalents (0–12) resulting from homoeologous pairing were observed. But in two genotypes (006001-6 and 006001-13), a multivalent which was either a quadri- or a trivalent, as well as a bivalent involving two Asiatic chromosomes, was observed. Two Asiatic chromosomes in 006001-6 which were non-homologous were always found to be associated together, and formed trivalent or quadrivalent with chromosomes from homoeologous genome. This indicated that there was a duplication between these two Asiatic chromosomes. Such a duplication might have resulted from the segregation of a chromosomal translocation between two non-homologous chromosomes in the Asiatic parent ‘Connecticut King’ which was transmitted to the progeny (006001-6). With the exception of two genotypes, in 11 genotypes that formed variable frequencies of bivalents, the homoeologous chromosome pairing and chiasma formation were similar to that between homologous chromosomes. After from the analysis of anaphase I stages it was evident that the expected types of chiasma formation involving non-sister chromatids gave rise to two strand single, two strand double, three strand double, four strand double and multiple exchanges. Whereas these events resulted from locus specific homoeologous exchanges, the translocations resulted from an aberrant form of non-homologous chromosomal exchange of segments. Elucidation of such differences is only possible through the analysis of meiosis using GISH.     

水稻减数分裂异常突变体osmd的表型分析及基因定位

减数分裂是有性生殖生物繁殖的基础,减数分裂和受精过程的正常进行保证了有性生殖生物配子体和合子体世代交替,并维持了遗传物质的稳定性。因此发现和克隆减数分裂相关基因可为进一步理解减数分裂调控过程提供研究基础。通过正向遗传学方法,筛选一个水稻减数分裂相关突变体并将其命名为osmd(Oryza sativa meiosis defect)。对其生长过程、花器官、花粉育性进行观察分析;通过DAPI,FISH染色对其进行染色体行为分析;并对该突变体进行遗传分析、基因定位和候选基因分析。结果表明,与野生型相比,osmd突变体营养生长过程正常,花器官以及雌蕊形态正常,但是成熟期花药不饱满,花粉粒不能被I2-KI染色。对突变体osmd花粉母细胞减数分裂过程观察发现,在粗线期染色体不能完全配对、终变期出现单价体、二分体时出现滞后染色体、四分体时期出现多个微核。通过FISH试验确定osmd突变体同源染色体不能正常配对。遗传学分析表明,osmd突变体不育表型由一个单核隐性基因控制;通过图位克隆将osmd的突变位点定位到10号染色体上的2个Indel分子标记Chr10-312和Chr10-1003-3之间,该区间共有900 kb,有133个开放阅读框。该区间内一个已知水稻减数分裂相关基因OsPAIR3编码区序列无变化。由此推测Os MD是一个未报道的参与水稻减数分裂同源染色体配对过程的蛋白。     

Chromosomal location of genes influencing plant characters and evapotranspiration efficiency in bread wheat

Little is known of genes that influence root development and drought resistance in bread wheat. The evapotranspiration efficiency (ETE = ratio of vegetative dry weight to total water used) of spring bread wheat tall landrace ‘Chinese Spring’ is relatively high. We used 42 ditelosomic and dimonotelosomic lines of Chinese Spring to identify chromosome arms that influence plant characters and ETE. Multiple regression analyses indicated that 96% of the variation observed in ETE was explained by variation in vegetative dry weight and total water used. Variation in plant height, number of spikes (tillers), root dry weight and shoot dry weight (excluding grains) together explained 88% of the variation observed in plant vegetative dry weight. Chromosome arms involved in expression of days to heading and maturity, plant height, number of spikes, root dry weight, shoot dry weight, number of grains, grain weight, and carbon isotope discrimination (Δ) were identified. Specifically, both arms of chromosome 2A, the long arm of chromosome 2B, and the short arm of chromosome 2D might carry genes with positive effects on number of spikes, root dry weight, and shoot dry weight. None of the aneuploids produced grain yield greater than Chinese Spring. The short arms of chromosomes 6A and 4D might carry genes that suppress Δ. Chromosome 1D might carry genes that increase relative water loss. The chromosome arms belonging to homoeologous group 2 might carry genes with positive effects on ETE. The genetic basis of ETE in modern wheats could be broadened by substituting specific chromosome arms of landrace wheats carrying desired characters into their genomes. This revised version was published online in July 2006 with corrections to the Cover Date.     

The production of aneudihaploids in Solanum tuberosum L. group Tuberosum (the common potato)

Summary Trisomic cytotypes were produced in dihaploid (diploid) plants of Solanum tuberosum L. Group Tuberosum, the common potato, according to two methods. Firstly, the aberrant types were selected, through chromosome counting, from parthenogenic dihaploid offspring of tetraploid-diploid crosses. In dihaploid populations from twelve tetraploid potato varieties the frequencies of aneuploids ranged from 3.5 to 11.0%. About 95% of these aneuploids had only one, and the others not more than two extra chromosomes. Secondly, the aneuploids were produced from triploid-diploid crosses. Seedset strongly depended on the crossability of the parental plant material, and the best results were obtained when the motherplants were grafted onto tomato. On avarage the three most successful cross combinations resulted in approximately 0.7 berries per pollination and 6 seeds per berry. With regard to seedsize the seed could be divided in two groups, viz, normal and small seeds. Half of the seed did not germinate or produced inviable seedlings, especially among the small seed group. About 93% of the plants was neuploid, with 25, 26 and 27 being the predominant chromosome number. It was concluded that the production of trisomics was the most successful through triploid-diploid crosses. The results were discussed with reference to the chromosomal behaviour in the meiosis in triploid plants (Lange & Wagenvoort, 1973a). It thus was possible to relate the low seedset to the distribution of chromosomes in the meiotic anaphases. On the one side this distribution resulted in a limitation of the avalability of gametes with monohaploid and near-monohaploid chromosome numbers, while on the other side the macrospores with higher chromosome numbers seemed to be inviable. Finally a crossing scheme was presented for transmitting the trisomic condition into a genetic background with better homogeneity and more homozygosity.Extended version of a lecture presented at a Eucapia meeting about Potato Dihaploids, held at Wageningen (Netherlands). See Lange and Wagenvoort, 1973b.     

由小麦×玉米获得的小麦DH系花粉母细胞减数分裂观察

研究了由小麦×玉米远缘杂交获得的小麦双单倍体DH2代花粉母细胞减数分裂过程。结果表明，大多数双单倍体小麦与异源六倍体普通小麦染色体数目相同，这是染色体经秋水仙素成功加倍的结果。同时在少数细胞中观察到了染色体的“块状移动”，在这些“块”中存在少数“染色体落后”现象，这可能是由于在染色体加倍过程中部分细胞染色体数目发生变异造成的。     

以流式核型分析小麦根尖细胞的有丝分裂同步化过程

构建染色体特异文库是简化小麦基因组测序的有效途径,而通过同步化处理提高根尖细胞有丝分裂指数,使根尖细胞中富集高比例的中期染色体是实现染色体分选成功的前提。采用双阻断法用50 µmol L^-1羟基脲和100 µmol L^-1氟乐灵对普通小麦根尖细胞进行同步化处理,然后用流式核型分析法观测各周期细胞在不同时间点上的比例变化。结果显示,最佳方案为羟基脲处理10 h,恢复7 h,氟乐灵处理4 h。以此方案处理中国春小麦根尖可获得最高的有丝分裂指数,达70.1%。附加冰水处理(0°C)有助于减少染色体碎片和增加流式核型的分辨率,但对富集更多的中期染色体没有效果。     

A comparative study of the cytological and reproductive behaviour of wheat and triticale subjected to two temperature regimes

Summary A comparison was made of the meiotic stability and reproductive behaviour ofTriticale hexaploide cv. Rosner andT. aestivum cv. Pitic in response to two temperature regimes (15° and 30°C) imposed continuously from premeiosis to maturity. Mean frequencies of MI open bivalents, univalents, and lagging chromosomes at AI were significantly higher in Rosner than in Pitic. Such irregularity would appear to arise from the asynchronous disjunction of chromosomes between Metaphase I and Anaphase I, resulting in meiotic disturbances and a high frequency of abnormal microspores. Despite such abnormalities, floret fertility was equivalent to that observed in Pitic, and the proportion of aneuploids recovered in the progeny, although greater than expected in wheat, was much lower than would be predicted. It is suggested that there is a strong certation effect in the transmission of aneuploid microspores and that the irregularities observed during microsporogenesis do not apply directly to megasporogenesis.No appreciable effect of temperature on meiotic chromosome behaviour during microsporogenesis was observed in either wheat or Triticale. In Triticale, high temperatures (30°C) substantially reduced spikelet fertility, and in contrast, low temperature (15°C) increased the frequency of aneuploids.Contribution No 232, Department of Plant Science, University of Manitoba, Winnipeg 19, Manitoba, Canada     

Dosage Response of Rye Genes in a Wheat Background

Wheat (Triticum aestivum L.) breeders often utilize alien sources to supply new genetic variation to their breeding programs. However, the alien gene complexes have not always behaved as desired when placed into a wheat background. The introgressed genes of interest may be linked to undesirable genes, expressed at low levels or not at all. The short arm of rye (Secale cereale L.) chromosome one (1RS) contains many valuable genes for wheat improvement. In order to study rye gene response to varying copy number, wheat lines were constructed which contained zero, two or four doses of 1RS. The meiotic behavior of rye chromosome 1R, and wheat/rye translocation chromosomes, 1AL/1RS and 1BL/1RS was studied in the F₁ hybrids between wheat lines carrying 1R or the translocation chromosomes. The IRS arm was transmitted at a very high frequency; 98 % of the F₂ plants had at least one of the chromosomes with a IRS arm. In addition, 44 % of the F₂ plants received at least one copy of the chromosomes from each parent. Analysis of the meiotic behavior of the IRS arm suggested that few euploid wheat gametes were formed. Therefore, most of the pollen must have contained IRS. It is unknown whether the lack of euploid wheat pollen could account for the high transmission frequency of the rye chromosomes. There may have been differential survival of the embryos receiving the rye chromosome as well.