Comparison of the isozyme variation in tall fescue parents and their somaclones

Summary This study was conducted using the isozymes ACP-1, ADH-1, GOT-2, GOT-3, MDH, 6-PGD-1 and PGI-2 to: a) compare isozyme banding patterns of tall fescue somaclones with parents and b) correlate tissue culture-induced chromosome abnormalities with variant banding patterns. The 174 somaclones were grouped into seven categories based on their meiotic analyses and time of regeneration from culture. Differences in isozyme frequency between categories compared by chi-square tests were greatest for MDH, 6-PGD-1 and PGI-2, and least for ACP-1. The most significant differences in frequency were found between somaclones and parents. In comparisons of somaclone categories, the most different isozyme distributions were between the early vs. late regenerated somaclones. No significant differences in isozyme frequencies were found between all 42-chromosome somaclones vs. aneuploid somaclones and the three somaclone groups (42-normal, 42-abnormal, aneuploid) compared to each other. This study suggests that culture-induced isozyme variation alters the distribution of the isozyme phenotypes, but is not directly correlated with chromosome abnormalities.     

Wheat x pearl millet hybridization: consequence and potential

Summary Eight grain pearl millet (2n=14) accessions were crossed as male to hexaploid spring wheat cv. Fukuho (2n=6x=42). An average of 80% wheat pistils showed pearl millet pollen tube entry in the ovules, compared to 56% in wheat x maize cv. Seneca 60 cross. Of the 15 embryos, obtained through in vitro immature seed culture from wheat x pearl millet crosses, 3 plantlets were produced and grown to maturity. These three were of the somatic chromosome constitution 2n=42, 21 and 22, respectively. Haploid wheat plant (2n=21) apparently originated from pearl millet chromosome elimination during embryogenesis. The 22 chromosome plant had retained a single pearl millet chromosome at tillering stage, but this chromosome was eliminated from pollen mother cells prior to and also during gamete formation. The significance and potential uses of this wide cross is discussed.     

Genomic Relationships of Triticum sharonense with Other S-Genome Diploid Triticum Species

Triticum sharonense was hybridized with autotetra-ploid T. speltoides, T. longissimum and T. bicorne. Meiotic analysis of these hybrids showed that T. sharonense is almost equally related to both T. speltoidcs and T. longissimum, while it is comparatively distant from T. bicorne. Therefore, this study does not support treating T. sharonense as a subspeies or variety of T. lottgissimum.     

The association of genes for purple coleoptile with chromosomes of the wheat variety Mironovskaya 808

Summary The association of genes for purple pigment in the coleoptile with the chromosomes of the winter wheat variety Mironovskaya 808 was investigated using monosomic F₂ analysis. The segregation ratio for F₂ hybrids of Chinese Spring monosomics x Mironovskya 808 seems to indicate that the purple colour of the coleoptile is determined by two dominant genes, Rc₃ and Rc₄, which are located on the chromosomes 7D and 6B respectively, and which reinforce each other. Apart from these two genes, suppressors found on the chromosomes 2A, 2B, 2D, 4B and 6A also play a role in the intensity of the purple colour.With the aid of a Chinese Spring telocentric chromosome marker it was observed that the Rc₃ gene is located on the chromosome arm 7DS, at a distance of 16±4.23 crossover units from the centromere.     

Homoeological relationships between the f chromosome of Brassica rapa and the e chromosome of Brassica oleracea

Eight plants of the putative double monosomic addition line (DMAL, 2n= 20) were developed by crossing a monosomic chromosome addition line of radish [f(A)‐type monosomic addition line (MAL) (2n= 19)] carrying the f chromosome of Brassica rapa (2n= 20, AA) with another [e(C)‐type MAL (2n= 19)] having the echromosome of Brassica oleracea (2n= 18, CC). The homoeological relationships between the two alien chromosomes were investigated by morphological, cytogenetic and random amplified polymorphic DNA (RAPD) analysis. Seventeen morphological traits that were not present in the radish cv. ‘Shogoin’ were observed in both MALs and these traits were substantially exhibited in DMAL plants. At the first metaphase of pollen mother cells (PMCs), the two parental MALs showed a chromosome configuration of 9II +1I, demonstrating impossibility of recombination between the R and the added chromosomes. The DMALs formed 10II in approximately 73% of PMCs, with one bivalent showing loose pairing between two chromosomes differing in size. In an attempt to identify the two MALs by RAPD‐specific markers using 26 selected random primers, 13 and 20 bands were specific for the f(A)‐type and the e(C)‐type MALs, respectively; 12 bands were common to both MALs (26.7%). In conclusion, the f chromosome of B. rapa is homoeologous to the e chromosome of B. oleracea. The genetic domain (genes) for 17 morphological traits are linked to each homoeologous chromosome bearing 27% of the corresponding RAPD markers.     

The effect of colchicine on triticale anther-derived plants: Microspore pre-treatment and haploid-plant treatment using a hydroponic recovery system

In cereals, chromosome doubling of microspore-derived haploid plants is a critical step in producing doubled haploid plants. This investigation was undertaken to study the effect of incorporation of colchicine in the induction medium for anther culture, and the effect of colchicine on anther culture-derived plants of triticale grown under controlled greenhouse conditions. In the latter case, chromosome doubling of adult sterile plants derived from anther culture of fourteen triticale populations was attempted, where androgenetic plants with non-dehiscent anthers were cloned and subjected to the colchicine treatment, and then grown with the aid of hydroponics. The hydroponic system provided optimal conditions for recovery of the affected haploids from the toxic effects of colchicine treatment and all colchicine-treated plants survived. A topcross-F₁ (TC₁F₁) population with timopheevii cytoplasm produced the highest percentage of plants with seed-set either due to chromosome doubling by colchicine (98%) or spontaneous doubling of chromosome number (15%). Colchicine-treated anthers performed inferior than control in both induction and regeneration phases. One of the key observation of this study was the reversal from reproductive stage back to the vegetative stage which in turn enabled further cloning of haploid plants under hydroponic conditions once they were identified as sterile. The one hundred percent survival rate of in vitro-derived plants, 100% survival rate of colchicine treated haploid plants and the high chromosome doubling success rate (X = 82.3) observed in this study imply that a temperature-controlled greenhouse with an hydroponic system provides an efficient environment for inducing chromosome doubling of haploid plants in cereals. This revised version was published online in August 2006 with corrections to the Cover Date.     

DNA甲基化酶抑制剂对体细胞染色体的影响

在动物的体细胞克隆过程中,供体细胞的细胞核要由高度甲基化状态转变为胚胎细胞的非甲基化状态,从而实现全能性的重新获得。试验利用DNA甲基化酶（5-aza-dc）抑制剂处理供体细胞,尝试建立一种既不影响染色体结构和数目,又能降低供体细胞细胞核的甲基化状态的最适浓度和方法,从而提供一种提高体细胞克隆动物效率的方法。在试验中,用不同浓度的甲基化酶抑制剂（5-aza-dc）处理奶牛成纤维细胞72 h,通过常规染色体核型分析方法制备染色体分裂相,检测其染色体数目、结构等的改变,结果显示,低浓度的甲基化酶抑制剂（0.005和0.01 μmol/L）处理,染色体没有发生明显的畸变（P＞0.05）,染色体能够保持正常的形态,高浓度（0.03与0.05 μmol/L）处理导致染色体畸变率显著增加（P＜0.05）,而0.1 μmol/L的5-aza-dc处理后染色体畸形率增加更为明显（P＜0.01）。总之,此试验的结果说明在对供体细胞做甲基化处理时用0.005和0.01 μmol/L的5-aza-dc处理不会影响细胞染色体结构和数目,但是高浓度处理则会导致染色体畸变率显著增加,不能用于体细胞克隆动物生产。     

川辐系列小麦品种品系的染色体结构变异研究

采用~(32)P内照射川育5号,选育出了川辐1号。采用~(60)Co丁射线辐照杂种F_2种子,选育出了川辐2号、川辐3号和川辐4号,以及多抗新材料4037和2270。川辐1号与常规品种绵阳19的染色体结构间差异较小,川辐2号与川辐1号之间有1对染色体结构存在差异。川辐2号与绵阳19号也有1对染色体结构存在差异。4037与绵阳19的杂种F_1,大量形成1个四价体,4037与川辐2号的杂种F_2,形成四价体、三价体、2个单价体、4个单价体的细胞都有,4037是一个典型的易位系,其基因组成与绵阳19更接近。2882的染色体结构与几个品种之间都存在着差异,其染色体结构值得进一步研究。4037的抗白粉病基因由主基因控制,表现为加性效应,为单基因遗传,是由~(60)Co丁射线诱变产生。     

Cytogenetic mechanism for reversion of triploids to heteroploid mosaics in Crassostrea gigas (Thunberg) and Crassostrea ariakensis

Triploid oyster induction has been very successful. However, during field culture, a certain proportion of triploids apparently loses one set of chromosomes, ‘reverting’ to diploid–triploid mosaics. The revertants will lose their aquaculture advantages due to the recovery of reproductive ability. Mitotic chromosomes were observed in mosaics of Crassostrea gigas and Crassostrea Ariakensis to elucidate the cytogenetic mechanism of reversion. Most individuals showed a modal number of 30 in triploid cells and varying numbers of chromosomes in other cells. From all 15 mosaics of C. gigas and 16 mosaics of C. ariakensis individuals, high proportions of aneuploid cells with various numbers of chromosomes were observed. Most of the aneuploid cells were hypotriploid cells with 26–29 chromosomes. Notably, in 10 out of 15 C. gigas and 9 out of 16 C. ariakensis, some cells consisted of 20–26 well‐spread chromosomes and 3–10 chromosome ‘clumps’. Individuals with more chromosome clumps tended to have higher percentages of hypotriploid cells. The correlation between these two categories of cells was significant (P<0.05) in C. gigas and highly significant (P<0.01) in C. ariakensis. Thus, we conclude that some chromosomes are eliminated during mitotic divisions because they clump together and are thus unable to undergo normal segregation.     

苏丹草与高粱染色体核型比较研究

采用去壁低渗火焰干燥法对4个苏丹草和6个高粱品种的核型进行比较研究。结果表明,苏丹草和高粱体细胞染色体数均为20(2n=20);苏丹草品种均为1A核型,并具有1对随体染色体,其中1、2号品种的第2对染色体为近中间着丝点染色体,3、4号品种为中间着丝点染色体;高粱品种除5号为1A核型、中间着丝点染色体外,其余的5个品种为2A或2B核型,且都有1或2对近中间着丝点染色体,9号品种还出现1对近顶端着丝点染色体;高粱8、9号品种各观察到1对随体染色体。染色体数量分析表明,第1到第10对染色体长短臂的绝对长度、相对长度以及绝对全长在苏丹草和高粱2类间的差异均不显著(P>0.05)。因此,苏丹草和高粱的遗传差异不在染色体长度上。