Inheritance of head splitting in cabbage (Brassica oleracea L. var. capitata L.)

Summary The inheritance of head splitting was probed by studying two moderate inbred lines of early-splitting and late-splitting cabbages, their F₁, F₂ and backcross progenies under field condition. It was concluded that there were at least 3 gene pairs for controlling head splitting. Gene action was mostly additive but partial dominance for early splitting was detected. Narrow sense heritability was estimated as 47%.     

Plant traits associated with resistance to Thrips tabaci in cabbage (Brassica oleracea var capitata)

Thrips tabaci is a major problem in the cultivation of cabbage for storage, as this pest causes symptoms that necessitate the removal of affected leaves from the product. Between cabbage varieties large differences in susceptibility occur. This study aimed to identify plant traits associated with these differences, in field experiments with natural infestation in 2005 and 2006. One factor affecting the amount of thrips damage was the timing of the development of the head. In an experiment with different planting dates especially the early maturing, more susceptible varieties were shown to benefit from later planting. In comparisons of multiple varieties in both years, regression studies showed that more advanced plant development in August and early September increased thrips damage at the final harvest. However, no single plant trait explained more than 25% (2005, Brix) or 48% (2006, compactness) of the variation in thrips damage. Optimal regression models, explaining up to 75% of the variation in thrips damage included Brix and leaf surface wax late in the season, as well as an indicator of plant development earlier in the season, and in 2005 also leaf thickness. The possible role of these plant traits in relation to thrips is discussed.     

结球甘蓝的离体快繁技术研究

以结球甘蓝无菌苗的胚轴和子叶为材料，几乎不生长愈伤组织，分化出不定芽。不定芽在添加6-BA的MS培养基上增殖很快，适宜的生根培养基为1/2MS+IBA0.5mg/L+ NAA0.1mg/L。通过增加培养基中糖和琼脂粉的用量，提高培养时的光照强度等有效控制了玻璃苗的产生。     

The somatic karyotype of cabbage (Brassica oleracea ssp. capitata)

Summary The individual chromosomes of cabbage are identifiable in nucellar cells found in the ovaries of immature pistils by the combination of their relative length, arm ratios, secondary constrictions, chromatic and achromatic regions and the presence of satellites.     

The allelic relationship between A IP(internal pigmentation) and A RC (red cabbage) in Brassica oleracea var. capitata L.

Summary Internal anthocyanin pigmentation (IP), previously reported to be controlled by a single gene dominant to aa of normal green cabbage, was studied further to determine its relationship to A ^RC A ^RC of red cabbage. When IP line R52 was crossed with an early red cabbage line. F₁ heads were pigmented throughout, but pigment intensity was intermediate. Subjective classification of F₂ plants by pigmentation intensity and distribution scores gave a ratio of 1 intense red throughout (red cabbage):2 medium red throughout:1 medium to light red, restricted to the central portion of the head (IP). The genotypes A ^RC A ^RC: A ^RC A ^IP: A ^IP A ^IP. respectively, are proposed to explain these 3 phenotypic classes. F₂ progenies contained no normal green plants, supporting the conclusion that A ^IP and A ^RC are alleles.Technical Paper 7052, Oregon Agricultural Experiment Station.     

Efficient doubled haploid production in microspore culture of loose-curd cauliflower (Brassica oleracea var. botrytis)

We present an improved protocol for highly efficient production of doubled haploid loose-curd cauliflower plants (Brassica oleracea var. botrytis) via microspore culture. Our experiment explored factors such as donor plant treatment, flower bud pretreatment, embryo germination medium, and ploidy characterization of regenerated plants. Our technique efficiently produced embryos from both tight- and loose-curd donor plants, although the embryo yields were genotype dependent. We achieved a germination rate of around 30 % by employing a hormone combination of zeatin, indole-3-acetic acid, and 6-benzylaminopurine pretreatment culture. We also used 1–4 days of cold pretreatment of the flower buds, which were submerged into NLN-13 medium, to induce microspore embryogenesis. Analysis using an FCM Ploidy Analyzer showed that more than 50 % of regenerated plants were spontaneously doubled haploids, more than 25 % were tetraploids, and fewer than 7 % were haploid. Visual examination of plants in the field revealed that they had distinct phenotypic characteristics relating to their ploidy level. The efficient production of double haploids using our improved microspore culture technique is a promising approach that can be applied in loose-curd cauliflower breeding programmes and genetic research.     

Microspore culture of white cabbage, Brassica oleracea var. capitata L.: Genetic improvement of non-responsive cultivars and effect of genome doubling agents

For the efficient application of haploid induction procedures in cabbage breeding, a sufficient number of regenerants should be achieved in a broad spectrum of genotypes. However, the majority of genotypes are somewhat recalcitrant. The efficiency of microspore culture was tested by crossing a responsive (28.7 embryos per Petri dish) and a non- responsive (0.1 embryo) cabbage cultivar. The embryo yield of one progeny was intermediate (18.9) while two were superior to the best parent cultivar (52.9 and 64.0 embryos). Thus, genes for haploid embryogenesis, present in responsive lines, can be effectively transmitted to responsive × non-responsive hybrids. Abscisic acid-induced desiccation of embryos was used for the efficient regeneration of plants. High germination percentages (54.7-70.6%) followed by normal plantlet development were achieved. Spontaneous genome doubling measured at the plantlet stage differed markedly in untreated genotypes. The percentage of diploids ranged from 21 to 67%. The effects of two antimitotic drugs applied to freshly isolated microspores were determined in two experiments. In the first experiment, trifluralin (0.5 and 1.0 mg:l) had no effect on embryo induction while oryzalin partly (0.125-0.25mg/l) or completely (0.5.mg/l) inhibited the formation of embryos. In the second experiment, higher concentrations of trifluralin increased the proportion of diploidized plants. Application of anti-mitotic drugs to microspores did generally not improve the overall production of haploid plants, which was higher in an untreated control.     

An S-allele survey of cabbage (Brassica oleracea var. Capitata)

Summary A total of 31 S-alleles was found in a survey of 197 cabbage plants representing 11 cultivars of diverse type. Most of these S-alleles also occurred in either kale or Brussels sprouts, but five of them have not been found previously and apparently occur only in cabbage. A more detailed study of five cultivars of spring cabbage showed only 12 S-alleles in all, with 6–10 S-alleles in four older cultivars and only 3 S-alleles in the newer more highly selected cultivar. S2 was by far the commonest S-allele, as it is in B. oleracea as a whole. The highly recessive alleles S5 and S15 were not particularly common in cabbage and this may partly explain why the sib problem in F₁ hybrids is apparently less in cabbage than in Brussels sprouts. Three cases were found in which an S-allele was completely recessive in both the stigma and the pollen. The problems for the breeder created by this rather unusual situation are discussed.     

Effect of genotype,environment and carbohydrate on anther culture response in head cabbage (Brassica oleracea L. convar. capitata (L.) Alef.)

Summary The effect of genotype, growing conditions for donor plants and type and concentration of carbohydrate in the culture medium was investigated for anther culture of head cabbage (white cabbage, savoy cabbage, pointed-headed cabbage). Strong genotypic effects on embryo formation from the cultured anthers were shown as well as superior embryo formation from anthers of field grown donor plants compared to plants grown in the greenhouse. When comparing 7, 10 and 13% sucrose in the medium, embryo response increased with increasing sucrose concentration. With maltose, which was generally inferior to sucrose as carbohydrate source for anther culture, the embryo response did not increase with maltose concentration above 10 per cent.     

Inheritance of internal pigmentation in green cabbage (Brassica oleracea var. Capitata L.)

Summary Internal anthocyanin pigmentation (IP) in otherwise normally green cabbage occurs in a number of Oregon State University breeding lines. Extracted pigment, tested for spectral absorption and for color reactions with lead acetate and aluminium chloride, was similar but not necessarily identical to pigment extracted from red cabbage cultivar Redman. When IP line R52 was crossed with normal green line C70, the F₁, F₂ and backcross progenies indicated that IP at the intensity found in R52 was determined by a single factor in homozygous condition, with intermediate levels of IP expressed by the heterozygous genotypes. Modifying factors also appear to influence the level of IP. In the cross R52 (IP)×R51 (normal green), expression of IP in the F₁ was much reduced. The F₂ failed to fit the expected 3 IP: 1 green ratio due to an excess of green plants, but instead, closely fit a 9:7 ratio. This may have resulted from incomplete expression of IP because of modifiers, rather than from the effects of a second major gene. An allele at the A (anthocyanin) locus of B. oleracea is tentatively proposed and designated A ^IP or a ^IP pending further identification.Oregon Agricultural Experiment Station Technical Paper No. 4690.     

A male sterile line with dominant gene (Ms) in cabbage (Brassica oleracea var. capitata) and its utilization for hybrid seed production

A male sterile plant, 79-399-3, was identified from a spring cabbage line 79-399 in 1979. Light-microscopic studies with paraffin section indicated that meiotic division stopped at the tetrad stage. The ratio of male sterile plants to fertile plants in the progenies of test crosses was 1:1 in five experiments conducted in 1982 and 1991–1994. However, some male sterile plants were sensitive and developed a very low number of viable pollen grains. When sensitive male sterile plants were selfed, the progenies segregated into male sterile and fertile plants at a ratio of 3:1. The male sterile plants from selfing of sensitive male sterile plants were propagated by tissue culture and crossed with different inbred lines. From the progenies of the crosses, populations with 100% male sterile plants were observed. The results indicated that homozygous dominant male sterile (MsMs) plants segregated from the selfed progenies. Populations with 100% male sterile plants and stable male sterility were developed as male sterile lines. Several favorable combinations were also selected by crossing the male sterile lines with inbred lines with high combining ability for desirable horticultural characters. This revised version was published online in July 2006 with corrections to the Cover Date.     

结球甘蓝产量性状的配合力与杂种优势分析

结球甘蓝全株重和叶球重是其产量性状的重要构成因素.研究其配合力和杂种优势表现,对甘蓝杂交育种和创制高产结球甘蓝杂交种具有重要意义.本研究通过6×7不完全双列杂交设计,对结球甘蓝产量性状的配合力效应和F,代杂种优势表现进行了分析,并基于SSR标记对13个亲本进行了聚类分析和遗传距离计算.结果表明,亲本880014、739...     

自花授粉诱导的甘蓝功能基因BoSPI的克隆与表达分析

自交不亲和性是甘蓝在长期进化过程中形成的防止自交衰败、促进杂交优势的一种复杂而完善的遗传机制。克隆自交不亲和性相关基因对甘蓝自交不亲和性的深入研究和利用有重要意义。本研究通过挖掘0~60 min自花和异花授粉的甘蓝柱头转录组数据, 筛选到一个受自花授粉诱导上调表达的基因, 命名为BoSPI。BoSPI开放阅读框534 bp, 编码177个氨基酸, 理论等电点为4.21, 不包含信号肽和跨膜区, 含有4个保守的EF-hand结构域。BoSPI基因起始密码子上游2000 bp的核苷酸序列中含有真菌诱导响应、代谢调节以及器官形成等应答元件。BoSPI基因在大肠杆菌中可诱导表达为17 kD的蛋白。BoSPI在柱头中表达量最高, 在花瓣、萼片、叶片、雄蕊表达量较低。BoSPI蛋白被定位在细胞膜和细胞质。自花授粉30 min后对BoSPI基因的诱导表达显著增强。表明BoSPI参与了柱头响应自花花粉刺激的分子过程, 可能是实现甘蓝自交不亲和性相关的某种新功能基因。     

结球甘蓝花粉类钙调素蛋白基因BoCML49的克隆及表达分析

以结球甘蓝E1为材料,提取花粉萌发前和萌发后的混合花粉总蛋白,总蛋白双向电泳后通过MALDI-TOF-MS鉴定分析差异点,根据差异点分析结果,利用同源克隆得到甘蓝BoCML49基因707 bp的cDNA片段。通过对BoCML49基因的qPCR分析表明其表达量在花粉萌发前约为萌发后的2.73倍,表明BoCML49基因在花粉萌发后表达下调。通过5¢-Race和3¢-Race分别得到550 bp和721 bp大小cDNA片段,最终得到结球甘蓝BoCML49全长cDNA序列,开放阅读框位于125~
1 078 bp处,加尾信号(AATAAA)位于第1 222 bp处。通过cDNA推导得到的氨基酸序列分析表明,BoCML49编码317个氨基酸残基,预测分子量为33.51 kD,pI为6.93,经Smart-embl预测其含2个重要的EF-hand结构,分别位于序列第150~178和第216~244位氨基酸残基处,预测结果显示其含有7个α-螺旋和10个β-折叠结构。系统发育树表明结球甘蓝BoCML49与拟南芥AtCML49和AtCML50的亲缘关系较近。BoCML49基因的原核表达得到37.55 kD的融合蛋白。     

Effect of activated charcoal on Brassica oleracea microspore culture embryogenesis

The effect of the addition of a 0.1 ml drop of activated charcoal (AC) on microspore culture embryogenesis was studied in nine morphotypes of Brassica oleracea. Embryo yields were significantly increased in all of the morphotypes by the addition of the 0.1 ml drop of AC to the microspore culture media. The magnitude of the response to the addition of AC varied with the different plants and morphotypes. The addition of AC never produced a detrimental effect. A qualitative improvement of the subsequent development of embryos to plants was also observed with the addition of AC. Data suggest that the addition of AC to microspore culture media promoted embryo production in different B. oleracea morphotypes. This revised version was published online in July 2006 with corrections to the Cover Date.     

Preliminary Study of the Characteristics of Several Glossy Cabbage(Brassica oleracea var. capitata L.) Mutants

To determine the characteristics and potential practical applications of glossy cabbage(Brassica oleracea var. capitata L.) mutants, five different glossy mutants were studied. The amount of epicuticular wax covering the mutant leaves was only approximately 30% that of the wild-type(WT) leaves. The wax crystals of WT plants were columnar and linear, while they were granular and rod-shaped in the mutants. Additionally, in WT cabbage, the primary wax components were alkanes, alcohols, fatty acids, ketones, and aldehydes. There was a significant decrease in the abundance of alkanes and ketones in the wax of the mutants. The glossy-green trait of the mutants may be the result of an inhibited alkane-forming pathway. Higher rates of chlorophyll leaching and water loss demonstrate that the mutant leaves were more permeable and sensitive to drought stress than the WT leaves. Growth curve results indicated that the growth rate of mutant-1 and mutant-3 was slower than that of the corresponding WT cabbage, resulting in shorter plants. However, the growth rate of mutant-2 was not influenced by the lack of coating wax. An investigation of the agronomic traits and heterosis of the glossy cabbage mutants indicated that all five mutants had glossy-green leaves, which was a favorable characteristic. The F1 plants derived from crosses involving mutant-2 exhibited obvious heterosis, suggesting the observed glossy-green trait is controlled by a dominant gene. Therefore, mutant-2 may be useful as a source of genetic material for future cabbage breeding experiments.     

Inheritance of internode length,plant form,and annual habit in a cross of cabbage and broccoli (Brassica oleracea var. capitata L. and var. italica Plenck.)

Summary When an inbred line of cabbage, Brassica oleracea L. var. capitata L., was crossed with an inbred line of broccoli B. oleraceae var. italica, the F₁ progeny were vigorous late annuals. All F₁ × broccoli backcross plants and 92% of the 3260 F₂ plants were annuals, while 40% of the F₁ × cabbage backcross plants were biennials. Annual habit is thus dominant and controlled by more than a single gene. Number of days to bud appearance in annuals varied continuously, and was primarily additive in inheritance. F₁ data suggested partial dominance for lateness but this was not supported by the F₂. Internode length was also continuous in distribution and primarily additive in inheritance, but with some dominance for short internodes in the F₁. Cabbage head forming ability was recessive and multigenic, with 2% of the F₂ plants forming heads, of which none were of commercial type and about half bolted as annuals. There was a significant chi square association between biennial habit and tendency for cabbage head formation. Clasping habit of terminal leaves was recessive to open leaves, multigenic, and associated with both cabbage heading and biennial habit.Technical Paper 4836, Oregon Agricultural Experiment Station; from an M.S. thesis by the senior author.     

An extended random primer amplified region (ERPAR) marker linked to a dominant male sterility gene in cabbage (Brassica oleracea var. capitata)

Similar to SCAR, an extended random primer amplified region (ERPAR) marker is a PCR amplified genomic DNA fragment at a single genetically defined locus. However, ERPAR uses specific primer pairs derived from RAPD primers by adding bases sequentially to their 3′-ends. As an example, an ERPAR marker was derived from a RAPD marker (OT11₉₀₀) linked to a dominant male sterility gene in cabbage (Brassica oleracea var. capitata). After two cycles of base adding and primer pair screening, a primer pair (5′-TTCCCCGCGACT-3′and 5′-TTCCCCGCGAGA-3′) amplified a single intense band with the same size as OT11₉₀₀. The identity of the new marker and OT11₉₀₀ was verified by segregation analysis. The new marker amplified by this extended primer pair was named as EPT11₉₀₀. The development of ERPAR exploits the importance of 3′-end bases of primers in PCR ERPAR shares advantages of SCAR, but eliminates the need for cloning and sequencing. It is a fast and universal way of converting RAPD markers into stable markers. This revised version was published online in July 2006 with corrections to the Cover Date.