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1.
L Malysheva  T Sjakste  F Matzk  M R?der  M Ganal 《Génome》2003,46(2):314-322
In the present investigation, genomic in situ hybridization (GISH) and barley microsatellite markers were used to analyse the genome constitution of wheat-barley hybrids from two backcross generations (BC1 and BC2). Two BC1 plants carried 3 and 6 barley chromosomes, respectively, according to GISH data. Additional chromosomal fragments were detected using microsatellites. Five BC2 plants possessed complete barley chromosomes or chromosome segments and six BC2 plants did not preserve barley genetic material. Molecular markers revealed segments of the barley genome with the size of one marker only, which probably resulted from recombination between wheat and barley chromosomes. The screening of backcrossed populations from intergeneric hybrids could be effectively conducted using both genomic in situ hybridization and molecular microsatellite markers. GISH images presented a general overview of the genome constitution of the hybrid plants, while microsatellite analysis revealed the genetic identity of the alien chromosomes and chromosomal segments introgressed. These methods were complementary and provided comprehensive information about the genomic constitution of the plants produced.  相似文献   

2.
Individual plants from the BC1F5 and BC1F6 backcross progenies of barley--wheat (= H. geniculatum All.) (2n = 28) x T. aestivum L. (2n = 42)] and the BC1F6 progeny of their amphiploids were used to obtain alloplasmic euploid (2n = 42) lines L-28, L-29, and L-49 and alloplasmic telocentric addition (2n = 42 + 2t) lines L-37, L-38, and L-50. The lines were examined by genomic in situ hybridization (GISH), microsatellite analysis, chromosome C-banding, and PCR analysis of the mitochondrial 18S/5S repeat. Lines L-29 and L-49 were characterized by substitution of wild barley chromosome 7H1 for common wheat chromosome 7D. In line L-49, common wheat chromosomes 1B, 5D, and 7D were substituted with homeologous barley chromosomes. Lines L-37, L-38, and L-50 each contained a pair of telocentric chromosomes, which corresponded to barley chromosome arm 7H'L. All lines displayed heteroplasmy for the mitochondrial 18S/5S locus; i.e., both barley and wheat sequences were found.  相似文献   

3.
Genes for winter hardiness and frost tolerance were introgressed from Festuca arundinacea into winter-sensitive Lolium multiflorum. Two partly fertile, pentaploid (2n = 5x = 35) F(1) hybrids F. arundinacea (2n = 6x = 42) x L. multiflorum (2n = 4x = 28) were generated and backcrossed twice onto L. multiflorum (2x). The backcross 1 (BC(1)) and backcross 2 (BC(2)) plants were preselected for high vigor and good fertility, and subsequently, a total of 83 BC(2) plants were selected for winter hardiness after 2 Polish winters and by simulated freezing tests. Genomic in situ hybridization (GISH) was performed on 6 winter-hardy plants selected after the first winter and shown to be significantly (P < 0.05) more frost tolerant than the L. multiflorum control. Among the analyzed BC(2) winter survivors, only diploid (2n = 2x = 14) plants were found. Five plants carried 13 intact L. multiflorum chromosomes and 1 L. multiflorum chromosome with a single introgressed F. arundinacea terminal chromosome segment. The sixth BC(2) winter survivor appeared to be Lolium without any Festuca introgression capable of detection by GISH. A combined GISH and fluorescence in situ hybridization analysis with rDNA probes of the most winter-hardy (after 2 winters) and frost-tolerant BC(2) plant revealed the location of an F. arundinacea introgression on the nonsatellite arm of L. multiflorum chromosome 2, the same chromosome location reported previously as a site for frost tolerance genes in the diploid and winter-hardy species Festuca pratensis.  相似文献   

4.
Bil'danova LL  Salina EA  Pershina LA 《Genetika》2003,39(12):1673-1679
The backcross progenies of the barley-wheat hybrids Hordeum vulgare L. (2n = 14) x Triticum aestivum L. (2n = 42) and two alloplasmic lines derived from them were studied using microsatellite markers of barley and wheat. The F1 hybrids and first backcross plants BC1 contained the genetic material of both cultivated barley and the cultivars of common wheat involved in developing of these hybrid genotypes. The genomes of BC3, BC4, and alloplasmic lines contained no microsatellite markers of the cultivated barley, whereas chromosomes of each homeologous group of common wheat were identified. In chromosomes of backcross progenies BC3, BC4, and alloplasmic lines yielded by backcrosses of hybrids and various common wheat cultivars, microsatellite markers of the parental wheat cultivars were shown to undergo recombination.  相似文献   

5.
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.  相似文献   

6.
The cytogenetics of the backcross generations and self-bred progenies (BC2F1, BC3F1, BC2F2, BC3F2 and BC2F3) of intergenetic hybrid of Triticum aestivum L. x Eremopyrum orientale (kedeb) Jaub. Et Spach were studied. The results showed that the plants BC3F1 (2n = 43 ) isolated from the backcross generations of the plants (2n = 44 ) accounted for 41.09%, but the plants (2n = 44) isolated was only 4. 11%, and the plants BC2F2 (2n = 44) isolated from self-cross generations of the plants BC2F1 (2n = 44) accounted for 13.21%. The number of univalents in pollen mother cells was higher in some plants (BC2F1) and the averages of univalents were negatively related to the backcross seed-setting and self-cross seed-setting with a related coefficient of - 0.6766* and -0.7429* respectively. The results of genomic in situ hybridization (GISH) showed that some plants of BC2Fs (2n=44) contained different number of alien chromosomes. All those indicated that alien chromosomes caused unusual pairing and segregation of wheat homologous chromosomes and also lowed the hereditary stability of wheat chromosomes.  相似文献   

7.
The karyotype and numeric changes in chromosomes among taxa of Lycoris (spider lilies) have been attributed to whole-arm rearrangements; however, the history of karyotype evolution of Lycoris is still ambiguous. In the natural habitat, one-third of Lycoris taxa are interspecific hybrids that are mainly sterile and extremely diverse in morphologies. Lycoris are geophytes with the reproductive stage initiated inside the bulbs during the storage period, which brings some inconveniences in collecting meiotic materials for studying chromosome pairing. The partial fertility of an artificial F1 interspecific hybrid between L. aurea (2n = 14) and L. radiata (2n = 22) provides an alternative option for tracing the meiotic process in F1 hybrids. The chromosome compositions of those functional gametes generated by the F1 hybrid could be recovered according to the chromosome complements of backcross progenies. We perform genomic in situ hybridization (GISH) analysis on somatic chromosomes of 34 BC1 plants (2n = 14–22) to reveal chromosomal divergences in number and composition of those functional gametes. GISH results also indicated a high homology between the MT- and A-genomes of Lycoris, reflecting on the partial fertility and frequently homoeologous recombination at meiosis of the F1 interspecific hybrids. The diverse chromosome complements and recombinant patterns presented in these functional gametes suggested that interspecific hybridization is an important force in driving diversification among Lycoris species. We suggest that the MT-karyotype genome may be the ancestral type in Lycoris, and some other chromosomal rearrangements in addition to centromeric fission may have played roles in the karyotype evolution of Lycoris.  相似文献   

8.
Chromosome association and chiasma formation were studied in pollen mother cells at metaphase I of four allotriplod BC1 plants (2n=3x=24) obtained from the backcross of the hybrid Alstroemeria aurea x A. inodora with its parent A. inodora. We distinguished the chromosomes of both parental species by genomic in situ hybridization (GISH), whereas the individual chromosomes were identified on the basis of their multicolour FISH banding patterns obtained after a second hybridization with two species-specific satellite repeats as probes. All the four BC1 plants possessed two genomes of A. inodora and one of A. aurea. Variable numbers of recombinant chromosomes, resulting from meiotic recombination in the interspecific hybrid, were present in these plants. The homologous A. inodora chromosomes generally formed bivalents, leaving the homoeologous A. aurea chromosomes unassociated. High frequencies of trivalents were observed for the chromosome sets that contained recombinant chromosomes, even when the recombinant segments were small. Chromosome associations in the trivalents were restricted to homologous segments. The implications of the absence of homoeologous chromosome pairing on gamete constitution and prospects for introgression in Alstroemeria are discussed.  相似文献   

9.
Brassica carinata, an allotetraploid with B and C genomes, has a number of traits that would be valuable to introgress into B. napus. Interspecific hybrids were created between B. carinata (BBCC) and B. napus (AACC), using an advanced backcross approach to identify and introgress traits of agronomic interest from the B. carinata genome and to study the genetic changes that occur during the introgression process. We mapped the B and C genomes of B. carinata with SSR markers and observed their introgression into B. napus through a number of backcross generations, focusing on a BC(3) and BC(3)S(1) sibling family. There was close colinearity between the C genomes of B. carinata and B. napus and we provide evidence that B. carinata C chromosomes pair and recombine normally with those of B. napus, suggesting that similar to other Brassica allotetraploids no major chromosomal rearrangements have taken place since the formation of B. carinata. There was no evidence of introgression of the B chromosomes into the A or C chromosomes of B. napus; instead they were inherited as whole linkage groups with the occasional loss of terminal segments and several of the B-genome chromosomes were retained across generations. Several BC(3)S(1) families were analyzed using SSR markers, genomic in situ hybridization (GISH) assays, and chromosome counts to study the inheritance of the B-genome chromosome(s) and their association with morphological traits. Our work provides an analysis of the behavior of chromosomes in an interspecific cross and reinforces the challenges of introgressing novel traits into crop plants.  相似文献   

10.
To increase the potential for establishing a complete series of tomato chromosome addition-sbstitution lines in a potato background, six new BC1 progeny were produced. All of them originated from crosses between three different hexaploid potato (+) tomato fusion hybrids. Three different somatic hybrids, viz., C31-17-5, C31-17-24, and C31-17-51, were used as female parents, and four different tetraploids, viz., Katahdin, Frieslander, 6704-1, and AM66.42 were used as male parents. A characterisation of the genomes of the three fusion hybrids and the six BC1 progenies (6739, 2001, 2002, 2003, 2004, and 2005) through genomic in situ hybridization and restriction fragment length polymorphism (RFLP) analysis indicated that there was preferential tomato chromosome elimination in the fusion hybrids. Similar analyses of the six BC1 progeny indicated that a variable number of the alien tomato chromosomes (6-11) were present in individual plants. RFLP analysis using chromosome specific DNA probes indicated that BC1 progenies had retained all 12 tomato chromosomes, albeit in different individual plants. This means that the available BC1 progenies have the potential for establishing a complete series of tomato chromosome addition-substitution lines in a potato background.  相似文献   

11.
Wheat (Triticum aestivum) and jointed goatgrass (Aegilops cylindrica) can cross with each other, and their self-fertile backcross progenies frequently have extra chromosomes and chromosome segments, presumably retained from wheat, raising the possibility that a herbicide resistance gene might transfer from wheat to jointed goatgrass. Genomic in situ hybridization (GISH) was used to clarify the origin of these extra chromosomes. By using T. durum DNA (AABB genome) as a probe and jointed goatgrass DNA (CCDD genome) as blocking DNA, one, two, and three A- or B-genome chromosomes were identified in three BC2S2 individuals where 2n = 29, 30, and 31 chromosomes, respectively. A translocation between wheat and jointed goatgrass chromosomes was also detected in an individual with 30 chromosomes. In pollen mother cells with meiotic configuration of 14 II + 2 I, the two univalents were identified as being retained from the A or B genome of wheat. By using Ae. markgrafii DNA (CC genome) as a probe and wheat DNA (AABBDD genome) as blocking DNA. 14 C-genome chromosomes were visualized in all BC2S2 individuals. The GISH procedure provides a powerful tool to detect the A or B-genome chromatin in a jointed goatgrass background, making it possible to assess the risk of transfer of herbicide resistance genes located on the A or B genome of wheat to jointed goatgrass.  相似文献   

12.
大白菜-结球甘蓝5号和8号单体异附加系的筛选和鉴定   总被引:3,自引:0,他引:3  
为建立成套的大白菜-结球甘蓝异附加系,以大白菜-结球甘蓝异源三倍体(AAC,2n=3x=29)与二倍体大白菜(AA,2n=2x=20)回交世代BC1为材料,对其进行细胞学观察并从中筛选单体异附加系。结果表明,BC1群体植株发生了染色体数目变异,变异范围为2n=20~29,其中以2n=23~24的非整倍体植株居多,占总观察株数的51.44%;对2个2n=21的植株进行核型分析,初步鉴定为大白菜-结球甘蓝5号和8号单体异附加系,分别命名为CO-5—1和CO-8—2。  相似文献   

13.
白菜与甘蓝之间体细胞杂交种获得与遗传特性鉴定   总被引:1,自引:0,他引:1       下载免费PDF全文
廉玉姬 《生物工程学报》2012,28(9):1080-1092
为拓宽白菜育种的基因资源,改良白菜品质,以白菜(Brassica campestris,2n=20,AA)和甘蓝(B.oleracea L.var.capitata,2n=18,CC)的子叶和下胚轴为材料分离、制备原生质体。采用40%聚乙二醇(Polyethylene glycol,PEG)进行原生质体融合。融合细胞在以0.3 mol/L蔗糖、0.3 mol/L葡萄糖为渗透稳定剂,附加1.0 mg/L 2,4-D+0.5 mg/L 6-苄氨基嘌呤(6-BA)+0.1 mg/L 1-萘乙酸(NAA)+1.0 mg/L激动素(Kinetin,Kin)的改良K8p培养基中培养并诱导细胞分裂。小愈伤组织经增殖培养后在MS+0.2 mg/L玉米素(Zeatin,ZEA)+1 mg/L 6-BA+0.5 mg/L Kin+0.4 mg/L NAA的固体分化培养基上诱导出不定芽。30 d后再转入MS基本培养基,获得完整的再生植株。将生根的植株转移到花盆,并对其杂种性质进行形态学、细胞学和分子生物学鉴定。结果表明,经细胞融合分裂出的320个愈伤组织中,获得了35棵再生植株,其再生率达10.94%。形态学观察显示,绝大多数再生植株的叶面积较大,株型和叶型为两种杂交亲本的中间型,部分植株的叶片浓绿、肥厚。染色体计数结果显示,36.4%的再生植株染色体数为2n=38;36.4%的再生植株的染色体数为2n=58 60;27.2%的再生植株的染色体数为2n=70 76,超过两个融合亲本的染色体数的总和。流式细胞仪测定DNA含量显示,再生植株DNA含量变化比较大,其结果与染色体鉴定结果相吻合。随机扩增多态性DNA(Randomamplified polymorphic DNA,RAPD)和基因组原位杂交(Genomic in situ hybridization,GISH)分析结果证明再生植株具有双亲基因组。体细胞杂种花粉育性比较低,杂交、回交后其育性逐渐获得恢复,与白菜回交后代逐渐恢复了育性。通过体细胞杂交和回交、杂交获得了形态变化广泛的个体,为白菜的品种育种提供多样的种质资源。  相似文献   

14.
Using 23 F1 hybrids, 14 BC1 and 32 BC2 progenies, the genome composition of Darwin hybrid tulips was analysed through genomic in situ hybridisation (GISH) of somatic chromosomes. All plants were diploids (2n = 2x = 24) with the exception of one tetraploid BC1 (2n = 4x = 48) and one aneuploid BC2 (2n = 2x + 1 = 25) hybrid. Morphometric analysis in F1 hybrids revealed a difference in the total length of chromosomes representing genomes of T. gesneriana and T. fosteriana, where the percentage of each genome equaled 55.18 ± 0.8 and 44.92 ± 0.6% respectively. GISH distinguished chromosomes from both parent genomes although there was a lack of consistent chromosome labelling in some cases. In both T. gesneriana and T. fosteriana chromosomes some segments of heterochromatin in the telomeric and intercalary regions exhibited a higher intensity of fluorescence. In situ hybridisation with 5S rDNA and 45S rDNA probes to metaphase chromosomes of F1 hybrids showed that these regions are rich in rDNA. A notable feature was that, despite genome differences, there was a considerable amount of intergenomic recombination between the parental chromosomes of the two species as estimated in both BC1 and BC2 offspring. The number of recombinant chromosomes ranged from 3 to 8 in BC1 and from 1 to 7 in BC2 progenies. All recombinant chromosomes possessed mostly a single recombinant segment derived from either a single crossover event or in a few cases double crossover events. This explains the fact that, unlike the situation in most F1 hybrids of other plant species, certain genotypes of Darwin hybrid tulips behave like normal diploid plants producing haploid gametes and give rise to mostly diploid sporophytes.  相似文献   

15.
GISH has been a particularly useful technique for studying the Lolium-Festuca species complex of forage grasses. The reason for this utility is two-fold: (i) the complex is unique amongst crop plants in which fertile hybrids, and backcross progenies, can be produced which recombine genomes and promiscuously exchange their genes through homoeologous recombination; (ii) dispersed repetitive DNAs differ between species, and this allows tracking of the identity of chromosomes and chromosome segments. This tracking property has enabled several fruitful lines of research to produce a harvest of new information for both fundamental and practical purposes. We review this first decade of GISH (genomic in situ hybridization) in Lolium-Festuca, and discuss and summarize the achievements which have accrued.  相似文献   

16.
The first backcross breeding programme for the transfer of freezing-tolerance genes from winter hardy Festuca pratensis to winter-sensitive Lolium multiflorum is described. A partly fertile, triploid F(1) hybrid F. pratensis (2n=2x=14) x L. multiflorum (2n=4x=28) was employed initially, and after two backcrosses to L. multiflorum (2x) a total of 242 backcross two (BC(2)) plants were generated. Genomic in situ hybridisation (GISH) was performed on 61 BC(2) plants selected for their good growth and winter survival characters in the spring following one Polish winter (2000-2001). Among the winter survivors, diploid chromosome numbers were present in 80% of plants. An appropriate single Festuca introgression in an otherwise undisturbed Lolium genome could provide increased freezing tolerance without compromise to the good growth and plant vigour found in Lolium. Among all the diploids, a total of 20 individuals were identified, each with a single F. pratensis chromosome segment. Another diploid plant contained 13 Lolium chromosomes and a large metacentric F. pratensis chromosome, identified as chromosome 4, with two large distal Lolium introgressions on each chromosome arm. Three of the diploid BC(2), including the genotype with Festuca chromosome 4 DNA sequences, were found to have freezing tolerance in excess of that of L. multiflorum, and in one case in excess of the F. pratensis used as control. A detailed cytological analysis combining GISH and fluorescence in situ hybridisation analyses with rDNA probes revealed that the other two freezing-tolerant genotypes carried a Festuca chromosome segment at the same terminal location on the non-satellite arm of Lolium chromosome 2.  相似文献   

17.
We used polymerase chain reaction (PCR) and fluorescence in situ hybridization (FISH) techniques to demonstrate the presence of Brassica B-genome chromosomes and putative B-genome introgressions in B. napus x B. juncea interspecific progeny. The B-genome--specific repeat sequence pBNBH35 was used to generate PCR products and FISH probes. The highest frequencies of viable progeny were obtained when B. napus was the maternal parent of the interspecific hybrid and the first backcross. B-genome--positive PCR assays were found in 34/51 fertile F2 progeny (67%), which was more than double the proportion found in fertile BC(1) progeny. Four B-genome--positive F(2)-derived families and 1 BC(1)-derived family were fixed or segregating for B. juncea morphology in the F(4) and BC(1)S(2), respectively, but in only 2 of these families did B. juncea-type plants exhibit B. juncea chromosome count (2n = 36) and typical B-genome FISH signals on 16 chromosomes. The remaining B. juncea-type plants had B. napus chromosome count (2n = 38) and no B-genome FISH signals, except for 1 exceptional F(4)-derived line that exhibited isolated and weak B-genome FISH signals on 11 chromosomes and typical A-genome FISH signals. B. juncea morphology was associated with B-genome--positive PCR signals but not necessarily with 16 intact B-genome chromosomes as detected by FISH. B-genome chromosomes tend to be eliminated during selfing or backcrossing after crossing B. juncea with B. napus, and selection of lines containing B-genome chromatin during early generations would be promoted by use of this B-genome repetitive marker.  相似文献   

18.
Genomic in situ hybridization (GISH) is a useful tool to analyse natural polyploids, hybrid plants, and their backcross progenies as to their origin, genomic composition, and intergenomic rearrangements. However, in angiosperms with very small genomes (<0.6 pg/1 C), often only heterochromatic regions were found to be labeled. We have modified the GISH technique to label entire mitotic and meiotic chromosomes of Arabidopsis thaliana (2n = 10) and closely related species with very small genomes by using high concentrations of DNA (7.5-15 microg per probe per slide) or 5 microg of probe and long hybridization times (>60 h). According to our GISH data, Cardaminopsis carpatica (2n = 16) is most likely the diploid ancestor of the autotetraploid Arabidopsis arenosa (2n = 32). Furthermore, within the allotetraploid species Arabidopsis suecica (2n = 26), it was possible to elucidate the origin of chromosomes contributed by the parental species A. thaliana and A. arenosa for a specimen with 2n = 26 or a deviating chromosome number.  相似文献   

19.
In a previous study, we developed cytoplasmic male sterile lines of Allium fistulosum possessing the cytoplasm of A. galanthum, a wild species, by continuous backcrossing. Furthermore, we reported the presence of a pollen fertility-restoring gene (Rf) for cytoplasmic male sterility (CMS) in A. fistulosum from segregation of pollen fertility of backcross progenies. In the present study, genomic in situ hybridization (GISH), using genomic DNA of A. galanthum as the probe DNA and that of A. fistulosum as the blocking DNA, was applied to F1 hybrids between both species and backcross progenies to determine the chromosomal location of the Rf locus. By means of GISH, eight chromosomes from A. galanthum were clearly discriminated from those of A. fistulosum in the F1 hybrids, and chromosome substitution process through continuous backcrossing was visualized. Interestingly, the chromosome region from A. galanthum, specific to male fertile plants, was detected in one chromosome of BC4 to BC7 generations. Based on the karyotype analysis of the male fertile plants, the chromosome was identified as the 5F chromosome. Our results confirm that the Rf locus is located on the 5F chromosome of the male fertile plants. This is the first report that identified the chromosomal location of the pollen fertility-restoring gene in A. fistulosum.  相似文献   

20.
对十倍体长穗偃麦草(Thinopyrum ponticum)与普通小麦杂交F1及其与普通小麦回交BC1F1的形态学和细胞学特性进行了分析。结果表明,长穗偃麦草与普通小麦‘兰考矮早八’衍生F1(‘兰考小偃麦’)的根尖细胞染色体数为56条;花粉母细胞减数分裂中期Ⅰ染色体构型平均值为19.81Ⅰ+15.78Ⅱ+0.75Ⅲ+0.59Ⅳ;基因组荧光原位杂交(GISH)显示,兰考小偃麦中含有35条完整的长穗偃麦草和21条小麦染色体。‘兰考小偃麦’/‘科育818’和‘兰考小偃麦’/‘Cp02-3-5-5’杂交F1的根尖细胞染色体数及其所遗传的长穗偃麦草染色体数分别为50~52和16~22条,且存在染色体易位;花粉母细胞减数分裂中期Ⅰ平均染色体构型为14.54Ⅰ+17.40Ⅱ+0.55Ⅲ+0.14Ⅳ,平均49.4%的细胞出现多价体(三价体或四价体)。这些材料为创造小麦-长穗偃麦草新种质奠定了基础。  相似文献   

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