黄芩的花粉母细胞减数分裂及核型分析

采用压片法,对黄芩花粉母细胞减数分裂及核型进行了研究。结果表明:黄芩的大多数花粉母细胞减数分裂中染色体的行为正常,在终变期同源染色体配对后可形成9个二价体,后期Ⅰ染色体以9∶9的方式向细胞两极分离,其减数分裂为同时型;在少数花粉母细胞减数分裂中观察到落后染色体、染色体桥等异常行为;其花粉粒育性为76.49%。黄芩的染色体数目为2n=2X=18,核型公式为K(2n)=2X=18=16m+2 sm,染色体相对长度组成为2n=1 s+4M1+3M2+1L,其核型为"1A"型。     

二倍体矮牵牛花粉母细胞异常减数分裂的观察与核型分析

以夏季高温时期二倍体矮牵牛花蕾为材料,采用常规制片法,对花粉母细胞异常减数分裂进行观察并选取终变期进行染色体核型分析。结果发现:异常减数分裂主要表现为具有多核仁、二价体提前分离成单价体、赤道板外的染色体,姊妹染色单体提前分离、不均等分离,落后和丢失染色体,具有微核的三分体和四分体,中期Ⅱ纺锤体定位发生异常出现融合纺锤体和八字形纺锤体可导致2n花粉产生;矮牵牛终变期核型公式为K(2n)=2x=14=10m+4sm(2SAT),其中第1、4、5、6、7号为中部着丝粒染色体,第2号(具有随体)、3号为亚中部着丝粒染色体,染色体相对长度组成为2n=14=4M_1+10M_2,核型分类为2A型。研究表明,矮牵牛异常减数分裂可导致2n花粉和不育花粉的产生,利用终变期进行核型分析具有材料丰富、二价体形态清晰不需人为配对分析等优点,为矮牵牛细胞学研究提供了新方法。     

埃塞俄比亚芥与芥蓝杂交获得异源三倍体及其细胞学研究

以埃塞俄比亚芥(2n=4x=BBCC=34)和芥蓝(2n=2x=CC=18)为材料,通过相互杂交获得了异源三倍体(2n=3x=BCC=26)。该异源三倍体生长势较强;叶色等介于双亲之间;株型、花型和花大小偏向于埃塞俄比亚芥;花色与芥蓝的相同,为白花。减数分裂观察表明:在终变期,一般形成9个二价体和8个单价体(9Ⅱ+8Ⅰ),且B、C两组染色体表现出一定程度的分群现象;中期Ⅰ,CC基因组的9个二价体排列在赤道板上,而B组的8个单价体游离在赤道板周围;后期Ⅰ分到两极的染色体以13/13和12/14占多数,偶见落后的染色体。该BCC异源三倍体的获得为创建CC+B染色体的异附加系和研究B、C基因组间的亲缘关系奠定了基础。     

番木瓜核型和减数分裂研究

对番木瓜核型和花粉母细胞减数分裂行为的研究表明,番木瓜染色体数目为2n=18,由9对中部着丝粒染色体组成。核型公式为2n=2x=18m。花粉母细胞减数分裂正常,在终变期和中期Ⅰ观察到9个二价体,未观察到染色体结构变异和行为异常。     

中国产酸模性染色体的初步观察

中国产的酸模也有明显的性染色体,雌株:2n=12A+XX;雄株:2n=12A+X+Y_1Y_2。雄株的性染色体在减数分裂终变期时,形成 Y_1—X—Y_2的链状三价体,中期Ⅰ时形成“v”字形或环状,偶有形成 XY 二价体和一个 Y 单价体。最后形成6A+X 和6A+Y_1Y_2的小孢子。酸模中可出现雌雄同株而具22条染色体(2n=18A+XX+YY)的三倍体不育株。酸模中还可见到同一株内有不同染色体数目的细胞,这一株至死未见开花。     

芥蓝-菜心种间三倍体CCA的合成及细胞学研究

以菜心-芥蓝异源四倍体(AACC)为桥梁亲本,通过与亲本芥蓝(CC)回交和未成熟胚培养,在芸薹属蔬菜中首次获得基因组为CCA的异源三倍体(2n=3x=28).该异源三倍体生长势较强,开花期、株型、叶形、花色等略偏向于芥蓝;A、C染色体组间存在较高的同源性,减数分裂终变期不同花粉母细胞中形成的三价体数变动在1～8个,多为4～5个,后期Ⅰ染色体分离不均衡,有14/14、13/15、16/12、17/11和18/10等分离方式,与亲本芥蓝(CC)回交的结子率一般较低,蕾期人工授粉可达0.29%.     

麦蓝菜的花粉母细胞减数分裂及核型分析

采用压片法,对麦蓝菜的花粉母细胞减数分裂及体细胞核型进行了研究。结果表明,麦蓝菜的绝大多数花粉母细胞减数分裂中染色体的行为正常,在终变期同源染色体配对后可形成15个二价体;在少数花粉母细胞减数分裂中观察到落后染色体、染色体桥等异常行为;其减数分裂为同时型,其花粉粒育性为88.04%。麦蓝菜的染色体数目为2n=2X=30,核型公式为K(2n)=2X=30=22m(2SAT)+8 sm,染色体相对长度组成为2n=16M2+14M1,其核型为"1A"型。     

白菜型油菜(B. Chinensis)与海甘蓝(Crambe abyssinica)属间杂种的细胞学观察

白菜型油菜与海甘蓝属间杂种F₁代染色体数目早期为2n=55。经多代组培无性繁殖后,杂种染色体大量减少,数目变化于25～28条间,平均为26条。杂种PMC减数分裂中,平均配对构型为0.06Ⅲ+11.26Ⅱ+3.80Ⅰ,二价体数目变化于8～13之间,以10Ⅱ、11Ⅱ、12Ⅱ的细胞出现频率较高,分别为26.60%,23.91%和30.98%,单价体数目变化于0～8个之间。杂种染色体数目减少,二价体个数较高,这说明海甘蓝染色体在大量丢失的同时,白菜型油菜的染色体组发生了自然加倍。在后期Ⅱ观察到三分体、染色体落后、染色体桥等异常现象。      

芥蓝和结球甘蓝染色体组型及C-带带型的研究

本文用改进的染色体标本制片技术,研究了芥蓝和结球甘蓝的染色体组型和 C-带带型。两种植物的二倍体均由4对中着丝粒、5对亚中着丝粒染色体组成,其中一对为随体染色体。芥蓝和结球甘蓝具有统一的染色体组型公式:2n=18=8m+10sm(2SAT),但两者的某些染色体在编号顺序上有差异。在结球甘蓝中观察,到4种不同形态的随体。用 BSG C-带方法得到 C-带带型,带型公式,芥蓝为2n=18=CITS 型=10C+2CI_++4CT~++2CS;结球甘蓝为2n=18=CITS 型=8c+2CI_++6CT~++2CS。某些带纹具多态性和杂合性。本文从染色体水平上讨论了芥蓝与甘蓝的亲缘关系。     

玉米(Zea may)×四倍体多年生玉米(Zea perennis)可育三倍体形态学和细胞遗传学研究

通过将普通玉米与四倍体多年生玉米大量杂交,获得一株部分可育三倍体植株MT-71,为外源遗传物质在普通玉米的导入创造新种质。MT-71杂交结实率分别为3X/2X=7.84%,2X/3X=0.56%,3X/4X=0.52%,开放授粉结实率为1.18%;其花粉母细胞(PMC)染色体平均构型为2.30Ⅰ+3.72Ⅱ+6.49Ⅲ+0.23Ⅳ;3X/2X后代植株染色体几乎包括所有2n=20~30染色体数目,2X/3X和开放授粉后代中染色体数目仅有2n=20,21。结果表明,MT-71可产生有功能的雌雄配子,其中n=10,11的配子更易遗传;雌配子体对额外染色体的传递能力显著高于雄配子体,异源三倍体最有效的遗传方式为3X/2X。     

Chromosome pairing in triploid intergeneric hybrids ofFestuca pratensis withLolium multiflorum, revealed by GISH

Genomic in situ hybridisation (GISH) was used to reveal chromosome pairing in two partly fertile, triploid (2n = 3x = 21) hybrids obtained by crossing the diploid (2n = 2x = 14) Festuca pratensis Huds. (designated FpFp), used as a female parent, with the autotetraploid (2n = 4x = 28) Lolium multiflorum Lam. (designated LmLmLmLm), used as a male parent. The pattern of chromosome pairing calculated on the basis of the mean values of chromosome configurations identified in all 100 PMCs analysed, was: 0.71I Lm + 2.24I Fp + 2.18II Lm/Lm + 0.54II Lm/Fp + 4.18III Lm/Lm/Fp. A relatively high number of Lm/Lm bivalents and Fp univalents, and a low number of Lm/Fp bivalents and Lm univalents indicated that the pairing was preferential between L. multiflorum chromosomes. Other observations regarding chromosome pairing within the Lm/Lm/Fp trivalents also confirmed this preferential pairing in the analysed triploids, as the Fp chromosome was not randomly located in the chain- and frying-pan-shaped trivalents. The similarities and differences in chromosome pairing at metaphase I and the level of preferential pairing between Lolium chromosomes in the different triploid Lolium-Festuca hybrids are discussed.     

Genome discrimination in progeny of interspecific hybrids between Brassica napus and Raphanus raphanistrum.

Genomic in situ hybridization (GISH) applied to the F1 interspecific hybrid between oilseed rape (Brassica napus, AACC, 2n = 38) and wild radish (Raphanus raphanistrum, RrRr, 2n = 18) showed the predicted 19 chromosomes from B. napus and 9 chromosomes from R. raphanistrum. The very low female fertility of these interspecific hybrids when backcrossed to R. raphanistrum led to only two descendants. Their chromosome number varied between 45 and 48. Both of these progenies showed only 9 chromosomes from R. raphanistrum and 36-39 chromosomes from B. napus. These results indicate the efficiency and limits of GISH as a suitable tool to assess and interpret the behavior of chromosomes after such interspecific crosses. The unexpected chromosome combination is discussed.     

Production and characterization of intertribal somatic hybrids of Raphanus sativus and Brassica rapa with dye and medicinal plant Isatis indigotica

Intertribal somatic hybrids of Raphanus sativus (2n = 18, RR) and Brassica rapa spp. chinensis (2n = 20, AA) with the dye and medicinal plant Isatis indigotica (2n = 14, I I) were firstly obtained by polyethylene glycol-induced symmetric fusions of mesophyll protoplasts. One mature hybrid with R. sativus established in field had intermediate morphology but was totally sterile. It had the expected chromosome number (2n = 32, RRI I) and parental chromosomes were distinguished by genomic in situ hybridization (GISH) analysis, and these chromosomes were paired as 16 bivalents in pollen mother cells (PMCs) at diakinesis and mainly segregated equally as 16:16 at anaphase I (A I), but the meiotic disturbance in second division was obvious. Five mature hybrids with B. rapa established in field were morphologically intermediate but showed some differences in phenotypic traits and fertility, two were partially fertile. Cytological and GISH investigations revealed that these hybrids had 2n = 48 with AAIIII complement and their PMCs showed normal pairing of 24 bivalents and mainly equal segregation 24:24, but meiotic abnormalities of lagging chromosomes and micronuclei appeared frequently during second divisions. AFLP analysis showed that all of these hybrids had mainly the DNA banding pattern from the addition of two parents plus some alterations. Some hybrids should be used for the genetic improvement of crops and the dye and medicinal plant.     

Indeterminate meiotic restitution (IMR): a novel type of meiotic nuclear restitution mechanism detected in interspecific lily hybrids by GISH

A detailed analysis of microsporogenesis was carried out in three diploid lily cultivars (2n=2x=24) and three diploid interspecific hybrids (2n=2x=24) using DNA in situ hybridisation methods (GISH and FISH). In cvs. Gelria (Lilium longiflorum; L genome), Connecticut King and Mont Blanc (both Asiatic hybrids; Agenome) meiosis was regular and only haploid gametes were formed while the three interspecific hybrids between L. longiflorum×Asiatic hybrid (LA) showed a variable frequency of meiotic nuclear restitution and stainable 2n-pollen formation ranging from 3% to 30%. An analysis of meiotic chromosome behaviour of the LA hybrids through GISH and FISH revealed that: (1) the parental chromosomes could be clearly discriminated into univalents, half-bivalents and bivalents in the PMCs; (2) in some of the PMCs the entire complement was present either as univalents or half-bivalents which had the potential to divide equationally (following centromere division) during the first division leading to first division restitution (FDR) gametes; (3) more frequently, however, in one and the same PMC the univalents and half-bivalents divided equationally whereas the bivalents disjoined reductionally at the same time giving rise to 2n-gametes that could vary from the well-known FDR or SDR 2n-gametes. We indicate this novel type of restitution mechanism as Indeterminate Meiotic Restitution (IMR). In order to confirm the occurrence of IMR gametes, the chromosome constitutions of eight triploid BC₁ progenies derived from backcrossing the 2n-gamete producing the LAhybrids to the Asiatic hybrid parents were analysed through in situ hybridisation. The results indicated that there were seven BC₁ plants in which FDR 2n-gametes, with or without homoeologous recombinations, were functional, whereas in one case the 2n-gamete resulting from IMR was functional. In the latter, there was evidence for the occurrence of genetic recombination through homoeologous crossing-over as well as through the assortment of homoeologous chromosomes. A singular feature of the IMR 2n-gamete was that although it transmitted a euploid number of 24 chromosomes to the BC₁ progeny, the number of chromosomes transmitted from the two parental species was dissimilar: 9 L-genome chromosomes and 15 A-genome chromosomes instead of 12 of each. Received: 15 May 2000 / Accepted: 4 December 2000     

Cytology of higher polysomics in Coix gigantea (Poaceae): Pentasomy and hexasomy

Among a natural population of Coix gigantea a pentasomic (2n+3; 2n=23) and a hexasomic (2n+4; 2n=24) were isolated and studied cytologically for the first time. The extra homologous chromosomes in the pentasomic plant did not disturb the behaviour of the rest of the bivalents through meiosis except for the rare occurrence of a few univalents. The majority of PMCs at diakinesis showed nine bivalents and a pentavalent. The various anaphase segregations led to the formation of sub- and super-haploid male gametes with from n–1 (n=9) to n+4 (n=14) chromosomal constitutions. The four extra homologous chromosomes in the hexasomic plant, however, disturbed meiosis leading to sticky chromosomal configurations and sterility. At diakinesis, the majority of the PMCs showed nine bivalents and a hexavalent.The origin of these higher polysomics is considered to be through fusion of male and female super-haploid gametes produced by the trisomics that occurred in large numbers in the population. The occurrence and dominance of nullisomics together with other aneuploids in the diploid population of C. gigantea make it quite certain that the 2n=20 form of the species is at a tetraploid level.     

小麦-黑麦代换系5A/5R与6A/6R杂交诱导同祖染色体配对与易位的研究

利用两个小麦-黑麦异源双代换系DS 5A/5R与DS 6A/6R杂交,探讨同祖染色体配对的可能性与创制小麦黑麦异源易位系.在方法上对杂种F1的减数分裂行为进行研究,观察5R与5A、6R与6A配对频率,探讨同祖染色体配对规律.实验结果看到杂交F1减数分裂中有22.91%的花粉母细胞有小麦染色体(ABD组)与黑麦染色体(R组)发生同祖配对.在F2及以后世代,通过染色体C分带、原位杂交检测,选择小麦-黑麦易位系.在F2代的45株中检测到9株有易位,易位频率为20%,是目前小麦-黑麦染色体易位频率最高的.染色体易位有的来源于同祖配对的交换,有的来源于单价体错分裂或断裂的重建.     

Cytogenetic studies on two F1 hybrids of autotetraploid rice varieties showing extremely high level of heterosis

Mechanisms of two F1 hybrids (D46A × DTP-4 and D46A × Dminghui63) of autotetraploid rice (2n = 4x = 48) showing extremely high pollen fertility 87.40% and 85.97%, respectively, seed set 82.00% and 79.00%, respectively and extremely high level of heterosis were analyzed cytologically. The chromosome pairing of D46A × DTP-4 and D46A × Dminghui63 was normal at metaphase I(MI), and had almost no I or III, with an average of 0.020I +14.36 II 6.44rod+7.91ring) +0.01III + 4.80 IV + 0.01VIII and 0.06 I + 17.67 II (11.01rod + 6.67ring)] + 0.06 III +3.10IV+0.01VI, respectively. The most frequent chromosome configurations were 10II+7IV and 12II+bIV. The bivalent frequency was less frequent in hybrids than that in restoring parents, and the same results were gained from univalents, trivalent and multivalents. However, the quadrivalent frequency was significantly higher in hybrids than that in restoring parents at MI. The other meiotic phases progressed normally, except for low percentages of PMCs with lagging chromosomes at AI and low percentages of PMCs with micronuclei at telophaseI (TI) and telophaseII (TII). PMCs with lagging chromosomes at AI and PMCs with micronuclei at TI and TII showed negative correlation between pollen fertility and seed set. Above 90% of the PMCs could form normal microspores, which resulted in the production of viable pollen grains, abnormal microspores were observed including penta-fission and hexa-fission. Based on these results we suggest that the two F1 hybrids had better behaviors of chromosome pairing and genetic stability than autotetraploid rice and other autotetraploid plants ever studied.     

Parental genome separation and elimination of cells and chromosomes revealed by AFLP and GISH analyses in a Brassica carinata x Orychophragmus violaceus cross

BACKGROUND AND AIMS: The phenomenon of parental genome separation during the mitotic divisions of hybrid cells was proposed to occur under genetic control in intergeneric hybrids between cultivated Brassica species and Orychophragmus violaceus (2n = 24). To elucidate further the cytological and molecular mechanisms behind parental genome separation, Brassica carinata (2n = 34) x O. violaceus hybrids were resynthesized and their chromosome/genomic complements analysed. METHODS: F(1) hybrids of the cross were obtained following embryo rescue, and were investigated for their cytological behaviour and subjected to genomic in situ hybridization (GISH) and amplified fragment length polymorphism (AFLP) to determine the contribution of parental genomes. KEY RESULTS: All the F(1) plants with high fertility closely resembled B. carinata in morphological attributes. These were mixoploids with 2n chromosome numbers ranging from 17 to 35; however, 34, the same number as in B. carinata, was the most frequent number of chromosomes in ovary and pollen mother cells (PMCs). GISH clearly identified 16 chromosomes of B. nigra in ovary cells and PMCs with 2n = 34 and 35. However, no O. violaceus chromosome was detected, indicating the presence of the intact B. carinata genome and elimination of the entire O. violaceus genome. However, some AFLP bands specific for O. violaceus and novel for the two parents were detected in the leaves. Cells with fewer than 34 chromosomes had lost some B. oleracea chromosomes. F(2) plants were predominantly like B. carinata, but some contained O. violaceus characters. CONCLUSIONS: The cytological mechanism for the results involves complete and partial genome separation at mitosis in embryos of F(1) plants followed by chromosome doubling, elimination of cells with O. violaceus chromosomes and some introgression of O. violaceus genetic information.     

The extent and position of homoeologous recombination in a distant hybrid of Alstroemeria: a molecular cytogenetic assessment of first generation backcross progenies

To estimate the extent and position of homoeologous recombination during meiosis in an interspecific hybrid between two distantly related Alstroemeria species, the chromosome constitution of six first generation backcross (BC₁) plants was analysed using sequential fluorescent in situ hybridization (FISH) and genomic in situ hybridization (GISH) analysis. Four different probes were used for the FISH analysis: two species-specific and two rDNA probes. The six BC₁ plants were obtained from crosses between the hybrid A. aurea×A. inodora with its parent A. inodora. GISH clearly identified all chromosomes of both parental genomes as well as recombinant chromosomes. The sequential GISH and FISH analysis enabled the accurate identification of all individual chromosomes in the BC₁ plants, resulting in the construction of detailed karyotypes of the plants. The identification of the recombinant chromosomes provided evidence which chromosomes of the two species are homoeologous. Two of the BC₁ plants were aneuploid (2n=2x+1=17) and four triploid (2n=3x=24), indicating that both n and 2n gametes were functional in the F₁ hybrid. Using GISH, it was possible to estimate homeologous recombination in two different types of gametes in the F₁ hyrid. The positions of the crossover points ranged from highly proximal to distal and the maximum number of crossover points per chromosome arm was three. Compared with the aneuploid plants, the triploid plants (which received 2n gametes) clearly possessed fewer crossovers per chromosome, indicating reduced chromosome pairing/recombination prior to the formation of the 2n gametes. Besides homeologous recombination, evidence was found for the presence of structural rearrangements (inversion and translocation) between the chromosomes of the parental species. The presence of the ancient translocation was confirmed through FISH analysis of mitotic and meiotic chromosomes. Received: 7 October 1998; in revised form: 4 December 1998 / Accepted: 10 December 1998     

High efficiency production and genomic<Emphasis Type="Italic">in situ</Emphasis> hybridization analysis of<Emphasis Type="Italic">Brassica</Emphasis> aneuploids and homozygous plants

Interspecific and intergeneric hybridizations have been widely used in plant genetics and breeding to construct stocks for genetic analysis and to introduce into crops the desirable traits and genes from their relatives. The intergeneric crosses between Brassica juncea (L.) Czern. & Coss., B. carinata A. Braun and Orychophragmus violaceus (L.) O. E. Schulz were made and the plants produced were subjected to genomic in situ hybridization analysis. The mixoploids from the cross with B. juncea were divided into three groups. The partially fertile mixoploids in the first group (2n = 36-42) mainly contained the somatic cells and pollen mother cells (PMCs) with the 36 chromosomes of B. juncea and additional chromosomes of O. violaceus. The mixoploids (2n = 30-36) in the second and third groups were morphologically quite similar to the mother plants B. juncea and showed nearly normal fertility. The plants in the second group produced the majority of PMCs (2n = 36) with their chromosomes paired and segregated normally, but 1-4 pairs of the O. violaceus chromosomes were included in some PMCs. The plants in the third group produced only PMCs with the 36 B. juncea chromosomes, which were paired and segregated normally. The mixoploids (2n = 29-34) from the cross with B. carinata produced the majority of PMCs (2n = 34) with normal chromosome pairing and segregation, but some plants had some PMCs with 1-3 pairs of chromosomes from O. violaceus and other plants had only PMCs with the B. carinata chromosomes. The Brassica homozygous plants and aneuploids with complete or partial chromosome complements of Brassica parents and various numbers of O. violaceus chromosomes were derived from these progeny plants. The results in this study provided the molecular cytogenetic evidence for the separation of parental genomes which was previously proposed to occur in the hybridizations of these two genera.