The production of inbred lines of brussels sprouts with dominant S-alleles

Summary The best sources of Brussels sprout inbred lines with both good agronomic characters and high self-incompatibility are likely to be cultivars of reasonably good agronomic type which have not been too intensively selected. Comparison of three cultivars of different agronomic quality showed that the cultivars of poor and moderate quality had about 55% of plants with a dominant S-allele, but the most highly selected cultivar had only 25% of such plants. A programme of S-allele screening is suggested which incorporates the minimum number of tests required to determine whether or not a particular plant has a dominant S-allele. A survey of S-alleles present in commercial F₁ hybrids showed that the frequency of dominant S-alleles was only 19% in hybrids released prior to the end of 1971, but was 50% in hybrids released since 1971.     

The effect of alternating temperatures on the self-incompatibility of some clones of Brussels sprouts (Brassica oleracea L. var. Gemmifera (DC.) Schulz)

Summary The influence of temperature on self-incompatibility of six S-homozygous clones of Brussels sprouts was studied. The clones were treated with constant temperatures of 14°, 17° and 20°C and alternating day and night temperatures of 17/14°, 20/14°, 23/14° and 26/14°C. To determine the degree of incompatibility the mean number of pollen tubes per style after selfing was calculated.The clone with the weak S-allele S-5 was less self-incompatible in the 23/14° and 26/14°C treatments than in the other treatments. The other clones with the weak S-alleles S-15 and S-45 remained sufficiently self-incompatible during all treatments. The clone with the rather strong S-39 showed the highest level of self-incompatibility at 26/14°C.In the two weakly self-incompatible clones with S-5 and S-39 a clear difference in self-incompatibility was found between young and old flowers. The strongly SI clones with S-39 showed no difference and in the two clones with S-15 the differences were small and significant in one case only.     

The role of pollen and stigma in determining the level of partial self-compatibility in brussels sprouts

Summary Brussels sprouts inbred plants, homozygous for the same recessive S-allele, were intercrossed in all combinations and the number of pollen tubes penetrating the stigmas of such crosses measured. The results of five such experiments showed that pollen as well as stigma is significant in determining the number of pollen tubes counted. It was also found that the capacity of a plant's stigmas to permit pollen tube penetration was independent of the capacity of its pollen to penetrate other plants' stigmas. Variation in pollen tube production between pollens of the plants tested was often greater than that between their stigmas.     

Increased atmospheric humidity post pollination: A possible aid to the production of inbred line seed from mature flowers in the Brussels sprout (Brassica oleracea var. Gemmifera)

Summary In two highly self incompatible inbred lines of Brussels sprouts the effect of increased atmospheric humidity post pollination was examined immediately following 1) hand pollination of green buds and open flowers, and 2) blowfly pollination of open flowers. Data were obtained for mean number of seeds set per pollination, mean number of fruits setting seed, and mean number of seeds produced per fruit which set for both varieties. Measured as number of seeds produced per minute spent pollinating, it was clear that open flower pollination followed by high humidity conditions was a much more efficient method of producing inbred line seed (46 seeds/minute) than green bud pollination (27 seeds/minute).     

Identification of S-alleles in Brassica oleracea

Summary S-alleles of Brassica oleracea were identified using a method which is based on the amplification of S-sequences from genomic DNA, followed by digestion of the PCR products with selected restriction enzymes (PCR-RFLP). A study was made in which the same S-allele was present in the homozygous state in a range of different crop types. This showed that, with minor exceptions, characteristic restriction patterns were obtained, and therefore that it was possible to identify the S-allele. To test whether the method was also suitable for the identification of both the S-alleles present in heterozygotes, a number of S-heterozygotes together with an F₂ population were screened. The results showed that the standard method was not very reliable for the identification of both of the S-alleles. This is because firstly, one of the S-alleles may be amplified preferentially, and secondly, the restriction patterns are not unique to a particular combination of S-alleles. Finally, although it is not possible to identify unequivocally both S-alleles of heterozygotes using a standard technique, the procedure can be modified for particular combinations of alleles to enable the identification to be made.     

Routine large-scale electrophoresis for plant breeding. An example with F1 seeds of Brussels sprouts (Brassica oleracea var.gemmifera)

Summary A routine method of large-scale electrophoresis is proposed for use in plant breeding. The method can be applied both with research and teaching and does not require sophisticated apparatus. A skilled laboratory technician can thus handle 768 samples a day. Within a series of electrophoretic investigations the individual electrophoresis can be stopped at any time, while the other investigations in the series continue.     

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.     

Inbred lines with dominant incompatibility alleles for use as parents of F1 hybrid brussels sprouts

Summary To improve the chances of obtaining highly self-incompatible inbred lines for use as parents of F₁ hybrid Brussels sprouts and to extend the range of mutually cross-compatible combinations available, new inbred lines are being produced from plants selected for the presence of dominant S-alleles. The material comprises 42 different inbred families representing 15 cultivars and contains 12 dominant S-alleles of uncommon occurrence in Brussels sprouts. Data on S-allele interactions in the material are presented.Tests showed that whilst many of the parent plants containing dominant S-alleles were highly self-incompatible, a few had only weak self-incompatibility. Although the chances of obtaining strongly self-incompatible inbreds may be increased by using material with dominant S-alleles, it remains necessary to test and select for strong self-incompatibility during the breeding programme.Production of such inbred lines requires the application of two independent selection procedures, one for agronomic type and one for S-allele constitution. Only a small proportion of the plants of a parent cultivar are acceptable on both counts. Thus large populations of the cultivar and large numbers of selections are necessary: this in turn involves much expensive S-allele screening. To alleviate this problem a wide-based panmix containing only dominant S-alleles is being produced. It is hoped that from this it will be possible to extract inbred lines which carry only dominant S-alleles, so avoiding the need to screen each parent plant for its S-allele content.     

The inheritance of partial self-compatibility in Brassica oleracea L.: Results from a half diallel homozygous for a moderately recessive S-allele

Summary In a study of partial self-compatibility in Brassica oleracea, flower number, seeded siliqua and seed production were recorded on self-and cross-pollinated inflorescences of the progenies of a half diallel between six in bred Brussels sprout plants homozygous for the same moderately recessive incompatibility allele S₄₅.On both self-and cross-pollinated inflorescences significant amounts of additively controlled genetic variation were found for seed set per flower. For cross-pollinated inflorescences this was also the case for the two components of seed set, seeded siliquae per flower and seeds per seeded siliquae, but for self-pollinated ones only seeded siliquae production showed significant additive variation. Considerable heterosis and gene interaction were always present and a simple additive dominance model did not explain the variation.Two of the parents transmitted lower levels of partial self-compatibility to their progenies and, in one of these, dominant genes appeared to be responsible. The most important feature determining the production of self seeds was found to be the number of flowering sites at which the incompatibility mechanism failed rather than the number of seeds produced at each site.     

Breeding system,taxonomy, and breeding strategy in cauliflower,Brassica oleracea var. botrytis L.)

Summary Cauliflowers representing those maturing in Britain in autumn, winter and spring were assessed for self-compatibility by self-pollen tube growth. A tendency existed for groups taking longer to mature to be more self-incompatible, but a range of incompatibilities was apparent in all groups. These results complement those of other workers.It is argued that despite demonstrable heterosis within many groups of cauliflowers, radical advances are probably best made by selecting self-compatible material from annual × biennial hybrid populations.     

Genotype environment interactions and sib content in F1 hybrid —brussels sprouts

Summary Sib content of F₁ hybrid seed lots obtained from crossing four highly self incompatible brussels sprout lines was examined at two locations for four years. Parental combinations accounted for by far the greatest proportion of the total variation observed. Significant second and third order interactions terms suggest complex interactions between parental genotype, years, and locations. Attempts to interpret these data in terms of the regression model of Finlay & Wilkenson (1963), were unsuccessful.     

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.     

Carbon dioxide treatment as an effective aid to the production of selfed seed in kale and Brussels sprouts

Summary The use of carbon dioxide to overcome the self-incompatibility mechanisms in marrow-stem kale and Brussels sprouts (Brassica oleracea L.) is discussed. Results are presented to show the effectiveness of the treatment, and a method is described of applying it as a routine procedure for the production of selfed seed in quantily.     

Inheritance of stalk rot resistance in cauliflower (Brassica oleracea var. Botrytis L.)

Summary The inheritance of resistance in cauliflower to stalk rot (Sclerotinia sclerotiorum (Lib.) de Bary) was investigated in population from six generations of six crosses. Disease incidence was recorded on 4 parents, 6 F^s ₁, 6 F^s ₂ and 12 back-crosses in a screenhouse under artificially created epiphytotic conditions. Resistance to stalk rot in this set of parents was found to be polygenic and under the control of recessive genes and due primarily to additive gene action. A breeding strategy emphasizing recurrent selection should lead to improvement in resistance.     

The inheritance of days to flowering in broccoli (Brassica oleracea var. italica)

Summary Crosses between an early flowering inbred broccoli (Brassica oleracea L. var. italica) HS140 and four later maturing inbred lines, S301, S310, s318, and S258, were studied to determine the inheritance of earliness as expressed by days to first open flower. Mean days to first flower for F₁ and F₂ were almost identical, and were close to the mean of the two parents in three crosses, indicating additive inheritance. In the cross HS140×S258, for which no F₁ plants were available, the F₂ mean was closer to the mean of the early parent, but this was likely due to a distortion of the data caused by the very late maturation of S258, the latest maturing parent in the study. Frequency distribution for parents and progenies supported the conclusions from parent and progeny mean values and indicated that days to maturity is a quantitative character, with mostly additive inheritance.Oregon Agricultural Experiment Station Technical Paper No. 7578.     

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.     

Inheritance of resistance to black rot (Xanthomonas campestris pv. Campestris) in cauliflower (Brassica oleracea var. Botrytis)

Summary Black rot disease caused by Xanthomonas campestris pv. campestris is a limiting factor in the commercial production of the cauliflower crop. Crosses were attempted between SN 445, a mid season cultivar resistant to black rot and two highly susceptible commercial cultivars (Pusa Snowball-1 and K-1). Studies of the F₁'s, F₂'s and back crosses indicated that SN 445, carries a dominant gene imparting resistance to black rot.     

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.     

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.     

Influence of timing of cross-versus self-pollination on self-incompatibility in Brussels sprout inbreds

Summary The impact of cross pollination 30 h, 4 h, and immediately prior to self-pollination, and self-pollination 2, 4, 8, 12, 24 and 30 h prior to cross-pollination was assessed for pollen tube number per style, seed number per siliqua, and proportion of self-seed (sibs) per siliqua, in two inbred lines of brussels sprout. Pollination procedure had a marked effect on the amount of sib-and hybrid seed produced. Cross pollination 30 h prior to self-pollination produced significantly greater numbers of sib progeny, as did self-pollination 8–12 h before cross pollination. Prior application of self pollen reduces the number of pollen tubes in the style. The results of this experiment suggest that self-incompatibility in brussels sprout could better be assessed by self-pollination with a subsequent cross pollination 8 h later, and the determination of amounts of sib and F₁ hybrid seed per siliqua.