利用花粉辐射诱发普通小麦与大赖草染色体易位的研究

将小麦－大赖草Ｌｒ．２和Ｌｒ．１４染色体二体异附加系９４Ｇ１５和９４Ｇ４５即将成熟花粉经＾６０Ｃｏ－γ射线辐射处理,然后分别给普通小麦品种扬麦５号和绵阳１１授粉。辐射Ｍ１的ＰＭＣ ＭＩ期染色体Ｇｉｅｍｓａ Ｃ－分带和基因组ＤＮＡ荧光原位杂交处理后进行染色体配对分析,从１７株Ｍ１单株中筛选到５株ＰＭＣ ＭＩ期大赖草染色体与小麦配对的个体。根据这５株单株自交Ｍ２种子根尖细胞有丝分裂中期染色体Ｃ－分带和     

Inducement of chromosome translocation with small alien segments by irradiating mature female gametes of the whole arm translocation line

Haynaldia villosa Schur. (syn. Dasypyrum villosum Candargy, 2n=14, VV) has been proved to be an important genetic resource for wheat improvement. The development of translocation with small alien chromosome segments, especially interstitial translocation, will be helpful for better utilization of its useful genes. Up to now, most of the reported Triticum aestivum – H. villosa translocation lines are involved in a whole arm or large alien fragments. In this paper, we report a highly efficient approach for the creation of small chromosome segment translocation lines. Before flowering, the female gametes of wheat-H. villosa 6VS/6AL translocation line were irradiated by 60CO-γ ray at 160 Rad/M dosage rate and three dosages (1600, 1920, 2240 Rad). Anthers were removed from the irradiated florets on the same day and the florets were pollinated with normal fresh pollens of T. aestivum cv. Chinese Spring after 2-3 days. Genomic in situ hybridization (GISH) at mitosis metaphase of root-tip cell of M1 plants was used to detect the chromosome structural changes involving 6VS of H. villosa. Among the 534 M1 plants screened, 97 plants contained small segment chromosome structural changes of 6VS, including 80 interstitial translocation chromosomes, 57 terminal translocation chromosomes and 55 deletion chromosomes. For the 2240 Rad dosage treatment, the inducement frequencies of interstitial translo-cation, terminal translocation and deletion were 21.02%, 14.01%, and 14.65%, respectively, which were much higher than those previously reported. The M2 seeds were obtained by backcrossing of 74 M1 plants involving 146 chromosomes structural changes of 6VS, and it was found that the structural aberrations in the M1 plants could be transmitted to their progenies. Irradiating mature female gametes of whole arm translocation is a new and highly efficient approach for creation of small segment chromosome struc-tural changes, especially for interstitial translocations.     

Inducement of chromosome translocation with small alien segments by irradiating mature female gametes of the whole arm translocation line

Haynaldia villosa Schur. (syn. Dasypyrum villosum Candargy, 2n=14, VV) has been proved to be an important genetic resource for wheat improvement. The development of translocation with small alien chromosome segments, especially interstitial translocation, will be helpful for better utilization of its useful genes. Up to now, most of the reported Triticum aestivum-H. villosa translocation lines are involved in a whole arm or large alien fragments. In this paper, we report a highly efficient approach for the creation of small chromosome segment translocation lines. Before flowering, the female gametes of wheat-H. villosa 6VS/6AL translocation line were irradiated by ⁶⁰C_O-γ ray at 160 Rad/M dosage rate and three dosages (1600, 1920, 2240 Rad). Anthers were removed from the irradiated florets on the same day and the florets were pollinated with normal fresh pollens of T. aestivum cv. Chinese Spring after 2 — 3 days. Genomic in situ hybridization (GISH) at mitosis metaphase of root-tip cell of M₁ plants was used to detect the chromosome structural changes involving 6VS of H. villosa. Among the 534 M₁ plants screened, 97 plants contained small segment chromosome structural changes of 6VS, including 80 interstitial translocation chromosomes, 57 terminal translocation chromosomes and 55 deletion chromosomes. For the 2240 Rad dosage treatment, the inducement frequencies of interstitial translocation, terminal translocation and deletion were 21.02%, 14.01%, and 14.65%, respectively, which were much higher than those previously reported. The M₂ seeds were obtained by backcrossing of 74 M₁ plants involving 146 chromosomes structural changes of 6VS, and it was found that the structural aberrations in the M₁ plants could be transmitted to their progenies. Irradiating mature female gametes of whole arm translocation is a new and highly efficient approach for creation of small segment chromosome structural changes, especially for interstitial translocations.     

应用原位杂交技术对小麦—黑麦代换系间杂交的细胞遗传研究

应用原位杂交技术结合染色体组型分析方法,对两个小麦-黑麦异源双代换系5R/5A和6R/6A杂交后代的遗传进行了研究,探讨同祖染色体配对的可能性并获得小麦-黑麦易位系。实验中对杂种F1代植株减数分裂各时期的花粉母细胞染色体行为进行分析,结果发现有22.91%的花粉母细胞中黑麦染色体与小麦染色体发生同祖配对。F2代通过C-分带、原位杂交鉴定,在45株中检测到9株易位,易位频率为20%,是目前报道易位频率最高的。染色体易位有的来源于同祖配对交换,有的来源于单价体错分裂或断裂的重建。     

Development of Triticum aestivum-Leymus racemosus translocation lines using gametocidal chromosomes

Maan[1] and Endo[2] et al. first reported that some chromosomes from Ae. longgissima, Ae. sharonensis and Ae. triuncialis showed preferential transmission when introduced into wheat background. The mechanism for this phenomenon rests with the fact that contrary to the normal fertility of gametes with these chromosomes, chromosome structural aberrations occur seriously in the gametes without these chromosomes, causing less compatibility in selective fertilization and resulting in semi-sterilit…     

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.     

Contribution of chromosomal imbalance to sperm selection and pre-implantation loss in translocation-heterozygous Chinese hamsters

Chinese hamster stocks with various structurally abnormal chromosomes have been produced by X irradiation. Among these stocks, 18 with various reciprocal translocations were used to investigate the participation of unbalanced gametes in fertilization and the development of unbalanced embryos. Among males as well as females heterozygous for the same translocation, there is no difference in the frequency of each disjunctional class. The participation of chromosomally unbalanced gametes in fertilization was investigated by chromosomal analysis of meiotic cells in heterozygotes for the 18 reciprocal translocations and pronuclei of fertilized ova obtained from crossing these heterozygotes. Compared with the expected frequencies from MII scoring, the frequencies of male pronuclei having a common deficiency of chromosome 1 (1q17-->1q42) or chromosome 3 (3p23-->3q31) decreased significantly in one-cell embryos. However, the frequencies of male pronuclei with other abnormalities were all consistent with those expected from MII scoring. In contrast, the frequencies of female pronuclei with any karyotype including the same abnormalities as those decreased in male pronuclei from the translocation heterozygotes were all consistent with those estimated from MII scoring. These results revealed clearly that most gametes with nullisomies as well as disomies for any chromosomal segments may participate in fertilization, whereas only male gametes nullisomic for certain segments of chromosomes 1 and 3 failed to participate in fertilization. The zygotic selection of chromosomal imbalance was also investigated by direct chromosomal and morphological analyses of preimplantation embryos from crosses between karyotypically normal females and male heterozygotes from the 18 stocks with various reciprocal translocations. These analyses revealed that some embryos were arrested in development at the two-cell stage. The karyotype of these two-cell embryos had a common deficiency in a segment of chromosome 1 or chromosome 2. Embryos with partial monosomy including chromosomes 1, 3, 4 and 5 showed arrested development at four- to eight-cell stages. Among day 4 embryos, some chromosomally unbalanced embryos, mainly with a deficiency of segments of chromosomes 1p, 1q, 2q, 5q, 7q and 8, had fewer blastomeres than karyotypically normal and balanced embryos. The homology between Chinese hamster and mouse chromosomes relating to abnormal embryogenesis at early stages has been partially confirmed from reported maps of chromosomes. The Chinese hamster is useful for further cytogenetic studies during the stages of meiosis and early embryogenesis.     

八倍体小黑麦×普通小麦杂种后代群体中的染色体易位

用改良的Giemsa C-带技术以单株为基础分析了八倍体小黑麦×普通小麦的杂种BC_1,F_(?)和F_(?)代植株的核型。在鉴定了C-带核型的1098株杂种后代植株中,发现了78条小麦-黑麦和277条黑麦-黑麦易位染色体。在不同的世代和株系中,小麦-黑麦染色体易位率变化在4.35—14.07%之间,平均7.10%;黑麦-黑麦染色体易位率在0.48—52.78%之间,平均25.23%。鉴定的小麦-黑麦易位染色体涉及了黑麦的14条不同的染色体臂和小麦的A、B和D组染色体。易位的48.57%发生在小麦和黑麦的部分同源染色体之间,51.43%发生在非部分同源染色体之间。不同的黑麦染色体臂参与易位的频率不同。小麦-黑麦染色体易位主要发生在杂种的早期世代,使用适当的选择技术在F_3获得了纯合的易位植株。文中讨论了快速选育易位系的技术和它们在小麦育种中的应用问题。     

利用phlb突变体创造普通小麦-簇毛麦6VS端二体代换系

用中国春phlb突变体（C.Sphlbphlb)与普通小麦柎-簇毛麦6V（6A）异代换系（Sub.6V)杂交,再用phlb突变体与F     

Maize tertiary trisomic stocks derived from B-A translocations

Reciprocal translocations between supernumerary B chromosomes and the basic complement of A chromosomes in maize have resulted in a powerful set of tools to manipulate the dosage of chromosomal segments. From 15 B-A reciprocal translocation stocks that have the B-A chromosome genetically marked we have developed tertiary trisomic stocks. Tertiary trisomics are 2n + 1 aneuploids where the extra chromosome is a translocation element, in this case a B-A chromosome. Whereas B-A translocations produce aneuploidy in the sperm, the tertiary trisomic plant efficiently transmits hyperploid gametes maternally. Because the B-A tertiary trisomic stocks and the B-A translocation stocks from which they were derived are introgressed into the W22 inbred line, the effects of maternally and paternally transmitted trisomic B-A chromosomes can be compared. Data are presented on both the male and female transmission rates of the B-A chromosomes in the tertiary trisomic stocks.     

Disomic and ditelosomic alien chromosome additions in beet (Beta vulgaris), carrying an extra chromosome of B. procumbens or telosome of B. patellaris.

Alien chromosome transmission through the female germ line as well as meiosis in pollen mother cells were studied in disomic and ditelosomic alien chromosome additions of beet. Beta vulgaris, carrying an extra pair of chromosomes or telosomes of B. procumbens or B. patellaris, respectively. The alien chromosomes carried genes for resistance to the beet cyst nematode, Heterodera schachtii, and screening for this resistance was used to select plants with or without the alien chromosomes. A great variation for alien chromosome transmission was recorded and plants carrying two extra alien chromosomes were recovered in the backcross progenies of the disomic or ditelosomic additions. However, in these progenies the average frequencies of plants without alien chromosomes (86%) did not clearly differ from that in similar progenies of the original monosomic or monotelosomic chromosome additions, indicating that doubling the number of the alien chromosome did not enlarge their transmission to the next generation. The alien chromosomes fully paired at pachytene and desynapsed again before diakinesis, indicating decreased chiasma formation. At second metaphase nearly 60% of the cells had one extra chromosome, and the remaining cells carried two or no extra chromosomes in about equal proportions. The tetrads looked fully normal. The expected relation between the average number of alien chromosomes in the germ cells and in the plants of the progenies did not show up, indicating a strong selection favouring the female gametes without alien chromosomes.     

普通小麦—簇毛麦异附加系和异代换系的C—分带鉴定

用改良的C-分带技术鉴定南京农业大学细胞遗传研究室获得的普通小麦的簇毛麦V_2、V_3、V_4、V_6、V_7染色体异附加系和V_2、V_5异代换系,得到与N-分带和染色体配对分析一致的结果,并且由于C-分带可同时鉴别小麦全部21对染色体,鉴定出V_2异代换系中被代换掉的小麦染色体为1A。     

Failure of chromosomally abnormal sperm to participate in fertilization in the Chinese hamster.

The selection of chromosomally abnormal gametes was investigated in the Chinese hamster by direct chromosome analysis of meiotic cells and one-cell embryos obtained from crossing heterozygotes for two reciprocal translocations, T(1;3)7Idr and T(1;3)8Idr. Expected frequencies of male and female gametes with different chromosome constitutions were estimated by scoring of secondary meiotic metaphase (MII) cells in the translocation heterozygotes. The frequency of gametes with each karyotype that participated in fertilization was investigated in pronuclei from translocation heterozygotes in one-cell embryos obtained from crossing the heterozygotes with karyo-typically normal animals. Compared with the expected frequencies from MII scoring, the frequencies of male pronuclei having some karyotypes in one-cell embryos decreased significantly. The karyotypes of male pronuclei showing a decreased frequency were commonly characterized by a deficiency of the long-arm segment of chromosome 1 (q13----qter) or by a deficiency of almost the whole arms of chromosome 3. On the other hand, the frequencies of female pronuclei with the same karyotypes were all consistent with those estimated from MII scoring. These results suggest that sperm nullisomic for certain segments of some chromosomes may fail to participate in fertilization.     

A strategy for detecting chromosome-specific rearrangements in rye.

To obtain translocations involving specific chromosomes in rye, a line in which chromosome 1R has large C-bands on its two telomeres but which lacks C-bands (or has very small ones) on the telomeres of the remaining chromosomes was used. About 6% of the plants produced using pollen from irradiated (1.2 krad (1 rad = 10 mGy)) spikes of this line possessed structural changes involving the labeled chromosome. These aberrations included translocations, ring chromosomes, isochromosomes, and telocentrics. It is concluded (i) that all nonlabeled chromosomes have the same probability of participating in reciprocal translocations with the labeled chromosome, 1R, and (ii) that most induced reciprocal translocations involved exchanges of chromosome segments of approximately equal length. The use of lines having the appropriate combination of telomeric C-bands improves the efficiency of obtaining reciprocal translocations involving specific chromosomes that could be used in the construction of detailed physical maps.     

Directed Synthesis of a Segmental Chromosomal Transposition: An Approach to the Study of Chromosomes Lethal to the Gametophyte Generation of Maize

Because of the haploid nature of the gametophyte generation of plants, most mutations that are lethal or detrimental to the gametophytes cannot be recovered. Our laboratory is currently developing several techniques to overcome this situation. In this paper, a procedure is described to generate directed segmental chromosomal transpositions. The method involves recovery of recombinants between reciprocal translocation overlaps such that one region of the genome is inserted into a nonhomologous chromosome in a predetermined and directed manner. This duplicated segment then could serve to cover deficiencies or mutations, lethal to the gametophytes, in the region from whence it originated. The manipulation of segmental chromosomal transpositions for analyzing mutants lethal or detrimental to the gametophyte generation is discussed. The procedure to generate transpositions, the translocations between normal A and supernumerary B chromosomes that generate deficiencies in the male gametes, the r-X1 chromosome that generates deficiencies in the female gametes and other techniques available in maize form a system to analyze gametophyte lethal mutations.     

八倍体小滨麦与缺体小麦杂交的细胞遗传学研究

八倍体小滨麦与缺体小麦杂交和回交,其后代ＢＣ１Ｆ１与Ｆ２相比较,染色体分离范围小,有利于４１条染色体类型的分离,若用异源双单体附加作父本与缺体回交,４１条染色体类型的分离率还会提高２倍左右;单体代换在自交世代的传递率为３１．９１％,二体代换的分离率为１９．３７％,异染色体的丢失率为２９．３４％;二体代换在自交世代的传递率为８５．２６％,异染色体的丢失率为９．２１％;ＰＭＣＭＩ染色体构型为２０．７６”＋０．３１’＋０．０３＂＋０．０１””,相对紊乱系数为０．０１,２ｎ＝２１”的细胞占８６．０９％。选育的二体代换系,不同程度地表现出大穗、多花、优质、抗多种病害等滨麦的优良性状。     

用分子原位杂交（GISH）鉴定小麦－簇毛麦双倍体、附加系、代换系和易位系

用生物素标记的簇毛麦（Ｈａｙｎａｌｄｉａｖｉｌｌｏｓａ）染色体组ＤＮＡ（ｔｏｔａｌｇｅｎｏｍｉｃＤＮＡ）作探针,以普通小麦染色体组ＤＮＡ作遮盖（用量１：２００左右）,进行有丝分裂中期和减数分裂中期Ｉ染色体的分子原位杂交（ＧＩＳＨ）,经抗生物素蛋白－辣根过氧化物酶复合物（ｂｉｏ－ｓｔｒｅｐｔａｖｉｄｉｎ－ｈｏｒｓｅｒａｄｉｓｈｐｅｒｏｘｉｄａｓｅ）和联苯胺四盐酸（ＤＡＢ）检测显色后,小麦－簇毛麦双倍体、附加系、代换系和易位系中的簇毛麦染色体及染色体片段显棕色,与显浅蓝色的小麦染色体可明显区分。用ＧＩＳＨ不仅可以检测导入小麦中的簇毛麦染色质,而且可以清楚地显示出易位染色体断裂点的确切位置。将ＧＩＳＨ用于减数分裂期染色体配对分析,还可以清晰形象地显示出同源和非同源染色体之间的配对和分离情况。     

Transmission of chromosomal abnormalities: participation of chromosomally unbalanced gametes in fertilization and early development of unbalanced embryos in the Chinese hamster

Using 14 Chinese hamster stocks with various reciprocal translocations, chromosomally unbalanced gametes were produced and used to investigate the participation of the unbalanced gametes in fertilization and the development of unbalanced embryos. The selection of chromosomally abnormal gametes during fertilization was investigated by the chromosomal analysis of meiotic cells in heterozygotes for the 14 reciprocal translocations and pronuclei of fertilized ova obtained from crossing these heterozygotes. Compared with the expected frequencies from meiotic metaphase II (MII) scoring, the frequencies of male pronuclei having commonly a deficiency of chromosome 1 (q14-->q42) or chromosome 3 (p23-->q31) in one-cell embryos decreased significantly. However, the frequencies of male pronuclei with other abnormalities were all consistent with those expected from MII scoring. In contrast, the frequencies of female pronuclei with any karyotype including the same ones, as those decreased in male pronuclei from the translocation heterozygotes were all consistent with those estimated from MII scoring. These results suggest that gametes with nullisomies as well as disomies for any chromosomal segments may mostly participate in fertilization, whereas some sperm nullisomic for the specific segments of chromosomes 1 and 3 may fail to fertilize. On the other hand, the zygotic selection of chromosomal imbalance was investigated by direct analyses of pre-implantation embryos from crosses between chromosomally normal females and male heterozygotes from the 14 stocks with various reciprocal translocations. The chromosomal and morphological analysis revealed that some embryos were arrested in development at the two-cell stage and their common abnormality was partial monosomy for chromosome 1 or 2. Embryos with partial monosomy including chromosomes 1, 3 and 4 showed arrested development at four-eight-cell stages. Among day 4 embryos, some chromosomally unbalanced embryos, mainly with a deficiency of other segments, such as chromosomes 1p, 2q, 5q and 8, had fewer blastomeres than karyotypically normal and balanced embryos. The homology between the mouse and the Chinese hamster chromosomes relating to the developmental abnormalities at early stages was partially confirmed.     

Radiation-induced translocations with reduced Haynaldia villosa chromatin at the Pm21 locus for powdery mildew resistance in wheat

Haynaldia villosa Schur. (syn. Dasypyrum villosum Candargy, 2n = 2x = 14, genome VV), a species related to wheat, is highly resistant to powdery mildew. The powdery mildew resistance gene Pm21 from H. villosa was introduced into common wheat by means of a translocation line T6VS·6AL, where the 6VS chromosome arm of H. villosa was joined at the centromere with wheat chromosome arm 6AL. To develop small alien translocations, especially interstitial translocations of small alien chromosome segments, we irradiated mature female gametes of a T6VS·6AL translocation line with gamma rays. More than 20 new translocations and deletions of 6V chromatin were obtained and subsequently used to map Pm21. Pm21 was located in a small region (FL 0.45–0.58) by genomic in situ hybridization, molecular marker analysis, and powdery mildew response. Two homozygous translocation lines with small H. villosa chromosome fragments carrying Pm21 were identified by fluorescence in situ hybridization and molecular marker analysis: an interstitial translocation in which a small fragment of 6VS is inserted into chromosome 4B and a terminal translocation with a small fragment of 6VS inserted into 1A. These small alien translocations are being transferred into an adapted elite wheat background by backcrossing to allow their easy use in breeding programs.