Chromosome doubling of mature haploid Brussels sprout plants by colchicine treatment

Summary The effect of colchicine concentration, and the mode and time of colchicine application in doubling the chromosomes of haploid Brussels sprout plants was assessed by scoring treated plants for the presence of diploid flowers and seed set after self-pollination.When colchicine treatments were applied after the plants had been vernalised, using 2 dose rates and 3 methods of application, only 38.1% became doubled and only 13.8% produced seed. Treatment with 0.1% colchicine with or without the addition of 2% dimethyl sulphoxide gave doubling rates similar to those reached with 0.05% colchicine, but resulted in more damage to the apices.When 0.05% colchicine solution was injected into the plants' apices at varying times during vernalisation, the rate of doubling was 71.2% on average and 50.7% of plants gave seed on selfing.Overall doubling rates of plants where the apex was easily accessible were 79.0%, while for plants with dense terminal heads they were 35.0% and for plants with visible buds 31.4%.     

Chromosome location of genes affecting polyphenol oxidase activity in seeds of common and durum wheat

This study used cytogenetic stocks to investigate the chromosomal location of genes responsible for polyphenol oxidase (PPO) activity in common and durum wheat seeds. Substitution lines of chromosome 2A of hexaploid varieties ‘Cheyenne’, ‘Thatcher’ and ‘Timstein’ in ‘Chinese Spring’ showed significantly higher PPO activity than all other substitution lines of the same variety, with the exception of substitutions of ‘Cheyenne’ chromosome 3A and ‘Thatcher’ chromosome 4B. Substitution lines of chromosome 2A of Triticum turgidum var. dicoccoides and of chromosome 2D of ‘Chinese Spring’ into the tetraploid variety ‘Langdon’ showed a significant increase in PPO activity relative to all other substitution lines in Langdon. The gene(s) responsible for high PPO activity in chromosome 2D from ‘Chinese Spring’ was mapped on the long arm within a deletion that represents 24% of the distal part of the arm. This study shows that genes located in homoeologous group 2 play a major role in the activity of PPO in wheat.     

Effects of substituted D genome chromosomes on photosynthetic rate of durum wheat (Triticum turgidum L. var. durum)

Summary A durum wheat cultivar Langdon (LDN) and fourteen disomic D genome chromosome substitution lines of Langdon, where A or B genome chromosomes were replaced with homoeologous D genome chromosomes of Chinese Spring (CS), were used to assess the compensatory effect of the D genome chromosomes on photosynthetic rates at tetraploid level. The LDN 1D(1B) and LDN 3D(3B) lines showed significantly higher photosynthetic rates than Langdon, whereas LDN 1D(1A) and LDN 3D(3A) lines were not greatly different from Langdon. It appears that chromosomes 1B and 3B decrease photosynthesis. This suggests the differentiation of the effects on the photosynthesis within the first and third homoeologous groups. Substitution with the 2D chromosomes did not compensate the effects of either 2A or 2B chromosomes as it reduced photosynthetic rate compared to plant with either chromosomes 2A or 2B. Tetra CS had a higher photosynthetic rate than CS and Penta CS. The photosynthetic rate of CS was similar to that of Penta CS, which lacked one set of D genome. The results suggest that it may be possible to increase photosynthesis, if both sets of the D genome were entirely removed from hexaploid wheat. However, it is difficult to conclude that the lower rate of photosynthesis of the hexaploids was mainly attributable to D genome chromosome effects, because we did not find a dose dependent effect of D genome. Homoeologous differentiation of chromosomes may be involved in photosynthesis.     

Somaclonal and chromosomal effects of genotype, ploidy and culture duration in Asparagus officinalis L.

Chromosome and morphological variations of embryogenic calli-derived plants of gynogenic haploid, diploid, triploid and tetraploid asparagus (Asparagus officinalis L.) were investigated. Artificial tetraploids were produced using colchicine treatment of seeds of diploid cultivar, ‘Poultom’. ‘Haidel’ (2X) was crossed with the artificial tetraploids, from which one gynogenic haploid, one diploid, 6 triploid, 3 mixoploids were obtained. Embryogenic calli were first obtained from crown buds, subsequently induced to form somatic embryos, and after 30 days, induced to germinate. Chromosome variation in embryogenic calli-derived plants increased with increasing duration of subculture, particularly when low ploidy levels of plants such as haploid and diploid were used as explants. Approximately 80% of haploid-derived plants showed morphological variations such as dwarfness and abnormal morphological characteristics, although no differences were observed in cladodes and stem characteristics between other polyploid-derived plants and their parents. The data presented here would supply important fundamental information for commercial mass-propagation using somatic embryogenesis. This revised version was published online in July 2006 with corrections to the Cover Date.     

Efficient production of doubled haploid wheat plants by in vitro treatment of microspores with trifluralin or APM

Isolated microspore cultures from two doubled haploid (DH) lines of wheat, Triticum aestivum L., were used to develop an in vitro chromosome-doubling protocol. During the initial 24 h or 48 h of culture the microspores were treated with either of the two antimicrotubule herbicides trifluralin or amiprophos-methyl (APM) in concentrations ranging from 0.1 μM to 10μM. Untreated control cultures yielded 209 embryos per 100000 microspores, which is the equivalent of one spike. Among the regenerated plantlets 67% were green, and 15% of the flowering plants were spontaneously chromosome doubled. Treatments with both the herbicides had a significant effect on chromosome doubling, measured as the percentage of fertile regenerants. With the best combination of treatment duration (48 h) and herbicide concentration (10/μM) the percentage of fertile plants among regenerants could be increased up to 74% with APM and up to 65% with trifluralin. The largest numbers of DH plants per spike could be obtained with herbicide concentrations at 1–3 μM. Treatments with either herbicide at these concentrations resulted in an estimated average between the two genotypes of 27 DH plants per 100 000 microspores. These results demonstrate the high potential of APM and trifluralin as chromosome-doubling agents in isolated microspore cultures. The in vitro treatment integrated into tissue culture procedures will constitute an efficient method for chromosome doubling in future wheat breeding     

Role of chromosome 3A in stomatal resistance of winter wheat

Summary Leaf stomatal resistance, through transpiration and photosynthesis control, constitutes a major factor of productivity and adaptation in wheat. The aim of the investigations reported here was to identify chromosomal effects on the expression of the maximum stomatal resistance, determined under optimum conditions of irradiance and water supply. Leaf stomatal resistance was measured, on wheat grown in pots under natural and well-watered conditions, using a LI-COR LI-6200 portable photosynthesis system under a saturating light>1400 mol m^-1 s^-2. Reciprocal sets of chromosome substitution lines between two hard red winter wheat cultivars, Wichita and Cheyenne, were used to identify the chromosomes involved in the expression of this trait. The two parental cultivars were significantly different for the parameter investigated. Chromosome 3A appeared to be involved in the expression of the stomatal resistance value under optimum conditions. Taking into account the relationships previously established between this parameter and some agronomic traits, chromosome 3A might be involved directly in productivity-determining processes or in the adaptation to water conditions, in wheat.     

Genetic and environmental effects on chromosome doubling of sugarbeet (Beta vulgaris L.) haploids

Summary Chromosome doubling has been a limiting factor for production of doubled-haploids, a means of obtaining fruly homozygous individuals, and a time-saving alternative to repeated selfing for the creation of inbred lines. The existence of genetic, environmental and genotype × environment interaction effects on chromosome doubling ability of sugarbeet (Beta vulgaris L.) haploids was investigated. Haploids were derived from four distinct, highly heterozygous diploid populations through in vitro culture of unpollinated ovules. Ovule-derived plants were treated with colchicine to double their chromosome complement. Environmental effects were determined from replication of the experiment at seven dates. Both genetic and environmental factors were found to affect chromosome doubling ability of sugarbeet haploids. No significant interaction between genotype and environment was identified. The presence of genetic effects on chromosome doubling ability is discussed with regard to its implications on breeding programs.     

1B/1R易位型小麦雄性不育系孤雌生殖性的细胞遗传学研究

以偏、粘、易型 5个 1 B/1 R小麦雄性不育系为基本材料 ,用中国春二体 ,1 B°1 D 缺四体和 1 B重双端体作父本与其杂交 ,以研究这几类异质 1 B/1 R不育系产生单倍体的特性 ;同时利用细胞遗传学方法分析了关键性染色体 1 B/1 R的特征。结果表明 :5个不育系的 1 BL/1 RS易位染色体的易位断点均发生在着丝点附近 ;1 B/1 R型小麦雄性不育系单倍体频率的变化是三种细胞质与不同的 1 B/1 R不育系互作的结果 ;来源不同的异质 1 B/1 R型小麦雄性不育系及其 F1产生单倍体频率的差异及稳定性与 1 B/1 R染色体上 1 RS片段的长度有对应性关系。     

A reevaluation of karyotype in cucumber (Cucumis sativus L.)

A karyotype analysis was conducted on the mitotic chromosomes of cucumber (Cucumis sativus L.; 2n = 2 × = 14), using improved chromosome preparation and C-banding techniques. Chromosome pairs were characterized on the basis of length and arm ratio value. The length of metaphase chromosomes ranged from 1.48 to 2.31 m, and the total length of the haploid chromosome complement was 13.06 m. The karyotype of this species was symmetric, and consisted of six metacentric and one submetacentric chromosomes. All chromosomes were defined by their unique C-banding patterns. The C-banded karyotype included centromeric, telomeric, and intercalary bands depending on chromosome pair leading to a precise identification of each pair. The C-banding patterns were distinctly different from observations made in previous study.     

牛囊胚染色体制备的研究

本研究采用传统的细胞遗传学方法，从牛体外受精后产生的6～8d囊胚中制备染色体，以探讨适合牛胚胎染色体制备的方法。将牛的6～8d回收的囊胚分别在含0．1μg／mL、lμg／mL和10μg／mL秋水仙酰胺(Colcemid)的TCMl99液中继续培养4h或6h，以抑制细胞分裂停留在中期。经处理后的囊胚移入10g／L柠檬酸钠的低渗液中，室温下处理15～20min.将低渗后的单个胚胎转移至载玻片上，经固定液I和固定液Ⅱ固定后，用体积分数为25％的Giemsa溶液染色10min，蒸馏水冲洗脱色，于室温自然干燥。显微镜下计算胚胎细胞数和有丝分裂指数以及观察染色体的伸展情况。结果显示，1μg／mL秋水仙酰胺处理6h后染色体制备的结果较为理想。