Cytoplasm Influences the Photosynthetic Efficiency in Brassica carinata

Cytoplasmic effects on the photosynthesis was investigated in experimentally produced Brassica carinatas of reciprocal cytoplasmic origin. These are obtained by hybridizing B. nigra x B. oleracea var. italica ( B. carinata 226) and B. oleracea x B. nigra ( B. carinata 241). Natural B. carinata and the two parents viz. B. nigra and B. oleracea var. italica were also included in this study. It was observed that the rate of photosynthesis in B. carinata 226 with nigra cytoplasm, and natural B. carinata was higher than that of B. carinata 241 with oleracea cytoplasm. This was substantiated by the diurnal values of photosynthesis among these cultivars. The result indicated that the cytoplasm does affect the rate of photosynthesis in B. carinata and high photosynthetic efficiency in natural B. carinata was due to B. nigra maternal parent.     

Seed Color Inheritance in Brassica carinata A. Braun, Cultivar S-67

The inheritance of seed color was studied in the brown seeded Ethiopian mustard (Brassica carinata A. Braun), cultivar S-67. Seed color was controlled by a single gene pair. The heterozygous condition resulted in light yellow-brown set-d indicating incomplete dominance [semidominance] of brown over yellow. The homozygous recessive condition resulted in yellow seed. The significance of these findings in relation to seed color inheritance in other Brassica species is discussed.     

Intraspecific breeding for reduced glucosinolate content in Ethiopian mustard (Brassica carinata A. Braun)

A total of six Ethiopian mustard ( Brassica carinata A. Braun) lines showing reduced levels of total glucosinolate content were developed through two different approaches. In the first case, eight lines with good agronomic performance under southern-Spanish conditions were first evaluated during two years for glucosinolate content, and a pedigree selection was then performed during three further generations. The line N2-142, with an average glucosinolate content of 82 μmoles g-1 seed, was developed from the original line C-49, with an average glucosinolate content of 115 μmoles g-1 seed. In the second case, chemical mutagenesis (EMS 1% v/v) was applied to seeds from the line C- 101, with an average glucosinolate content of 125 μmoles g- 1 seed. Five mutant lines showing an average glucosinolate content between 20 and 30 μmoles g-1 seed lower than the wild line C-101 were isolated in the M_3 or M_4 generation. The reduced glucosinolate content of these mutants was confirmed by developing and analysing the M_5 generation. Previous results in B. juncea suggest that a further reduction of total glucosinolate content might be achieved through genetic recombination between the different lines developed in this work. This revised version was published online in July 2006 with corrections to the Cover Date.     

Selection for reduced linolenic acid content in Ethiopian mustard (Brassica carinata Braun)

Reducing linolenic acid content is one of the most important objectives for the development of Ethiopian mustard lines with high oil quality. This work was aimed at searching for variability of the fatty acid composition of oil within a germplasm collection of Ethiopian mustard. A total of 217 lines were analysed by gas-liquid chromatography (GLC) in 1991, and one was selected as having reduced values of both linolenic acid content (10.2% versus 14.0% of total fatty acids as the collection average) and linoleic acid desaturation ratio (LDR, 0.34 versus 0.45). After 3 years of pedigree selection for low linolenic acid content, this line showed, in 1995, average values of this fatty acid of 5.4% and 2.4% in two different environments, compared with 11.6% and 8.3%, respectively, in the control. The values of the LDR were 0.18 and 0.09, respectively, compared with 0.36 and 0.27 in the control line.     

Plant Regeneration from Callus Cultures of Brassica carinata A. Br. and its Implications to Improvement of Oil Seed Brassicas

Protocols of plant regeneration have been developed for Brassica carinata for creating somaclonal variation for plant type and adaptability, so that this species can fit into cropping systems in Indian agriculture. The response of cotyledonary and stem explants was assessed for callus induction and shoot regeneration on MS and B₅ basal media containing different combinations of auxin and cytokinin concentrations. MS medium supplemented with BA and NAA favoured callus induction. Supplementing MS with combinations of BA and IAA, as also with BA alone, regenerated shoots from the ex pi ants with a high frequency. The frequency of shoot regeneration and the mean number of shoots per explant were higher in cotyledons than in stem explants on identical growth regulator combinations. On B₅ medium, supplemented with BA (2 mg/l) and IBA (0.4 mg/l), compact callus was produced which regenerated shoots on transfer to medium containing BA (0.8 mg/l). Genotypic differences among carinata accessions for regeneration were also observed.     

Genetic diversity of Brassica carinata with emphasis on the interspecific crossability with B. rapa

Brassica carinata (BBCC, 2n = 34) possesses many good agronomic characters and, to provide more genetic information about this species and utilize it further, 110 accessions of B. carinata were tested for genetic variation by using 233 amplified fragment length polymorphism markers, so that a dendrogram could be constructed. To combine the good traits of B. carinata and B. rapa (AA, 2n = 20) with those of B. napus (AACC, 2n = 38) which is the major rapeseed variety in China, interspecific crosses between these two species have been made which have resulted in 276 doubled hybrids (AABBCC). These hexaploids combine positive characters and can be used for crossing with B. napus . Brassica carinata as the female parents exhibited extensive differences in interspecific crossability and 20 high crossability accessions were identified. The Poisson Regression Model analysis result (P < 0.0255) indicated that such differences were due to the genotype of the accessions. Accessions with high crossability could promote gene flow within the genus Brassica .     

Glucosinolates Determined by HPLC in the Seeds of Microspore-Derived Homozygous Lines of Rapeseed (Brassica napus L.)*

Microspore culture was employed to measure the relative efficiencies of anther culture and isolated microspore culture for the regeneration of embryoids and plants of Brassica napus. The yield of embryoids and plants was at least 10-fold greater from isolated microspores than from anther cultures. Approximately 1400 microspore-derived homozygous line's, the parental varieties and the corresponding F₂ plants were grown in a field trial. Important agricultural characteristics, such as morphological homogeneity, growth rate, onset of flowering and seed setting were evaluated subjectively and seed yield and glucosinolate content of individual plants were determined. The relative concentrations of up to S different glucosinolates in these seeds were measured via an automated high performance liquid chromatography (HPLC) system. The alkenyl and indole glucosinolates, the two most important categories of glucosinolates, were found in varying proportions and were independently determined in these line's. Our results do not support the previously suggested connection between low concentrations of glucosinolates and weak growth and/or poor seed yield. Additionally, no evidence was found that the lines derived from isolated microspore culture were subjected to unexpected selection pressures that might adversely affect the diversity of the lines obtained. These results demonstrate that microspore culture is a powerful tool not only for genetic analysis bur also for practical plant breeding.     

Ploidy of broccoli regenerated from microspore culture versus anther culture

The use of microspore or anther culture to generate doubled-haploids (DH) is an important adjunct to broccoli breeding) Regenerated populations from broccoli anther culture are usually mixtures of ploidy. However, ploidy composition of populations derived from microspore culture has not been reported. The purpose of the present study was to characterize regenerants derived from microspore culture, to evaluate factors influencing these characteristics and to compare results with those from anther culture. Eight populations, four from each culture method, were generated simultaneously using the same four F₁ hybrids as donor parents. The ploidy level of all regenerants was determined by DNA flow cytometry: the majority of them were diploid. As in anther culture, a mixture of ploidy was observed in all populations derived from microspore culture. Ploidy variation was more frequent among clonal families from anther culture (10%) than microspore culture (5%)‘Everest’ was the most productive donor parent with both methods, while ‘Greenbelt’ and ‘Major’ were least productive in anther and microspore culture, respectively. Genotype specificity for the total number of regenerated plants and ploidy composition occurred in both culture methods.     

Effect of donor plant age and inflorescence age on microspore culture of Brassica napus L.

Summary Effect of age of donor plants and age of inflorescence on embryogenesis in microspore culture of B. napus was examined. Microspores isolated from buds of older plants had a higher embryo yield than those of younger ones. The effect of the age of inflorescence showed a different pattern. In older plants, a higher embryogenesis response was observed in microspores isolated from buds of new inflorescences, while in young plants, microspores isolated from buds of old inflorescences showed high embryo yield. These different responses were considered to be attributable to a difference in the developmental stage of pollen at the time of microspore isolation. Our results indicated that microspores collected from older inflorescences and older plants have sufficient embryogenic potential when the optimum developmental stage of pollen was used. Frequency of embryo to plant conversion was influenced by the size of embryos subcultured, but not by donor plant age or the age of the inflorescence.     

Isolation of induced mutants in Ethiopian mustard (Brassica carinata Braun) with low levels of erucic acid

Ethiopian mustard (Brassica carinata Braun) is a potential oil crop for the rain-fed Mediterranean area. However, its usage is limited by the high erucic and high glucosinolate content of the oil and meal, respectively. In the course of a mutagenesis programme, an agronomically good line of Ethiopian mustard was treated with EMS in order to widen the natural variability of nutritional traits in this species. As a result of this programme several low erucic mutants were isolated; two of these mutants showed erucic acid values in the M4 generation in the range 5–10% of total fatty acids. Near-infrared reflectance spectroscopy (N1RS) was successfully applied as a rapid screening method for erucic acid in this breeding programme.     

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.     

Effect of incubation temperature regimes and culture medium on broccoli microspore culture embryogenesis

Conditions for reliable induction of embryogenesis from isolated microspores were studied in ten genotypes of broccoli. Embryo yields were significantly increased in almost all of the broccoli genotypes by the incubation at 32.5 ^°C for 1 day, than when the standard incubation at 30 ^°C for 2 days was used. Treatments of 48 hours at 32.5 ^°C produced less than optimal results suggesting that broccoli microspores are more sensitive to high temperatures than those of B. napus. The use of the ¹/₂ NLN-13 medium yielded greater number of embryos than the standard NLN-13. The magnitude of the response to the redution of the concentration of major salts by half in the NLN medium varied with the different genotypes. High embryogenic broccoli cultivars, such as ‘Shogun’, ‘SDB9’, and ‘Green Valiant’, presented a better response to the reduction of the concentration of major salts by half in NLN-13. Reduction never produced a detrimental effect on embryo yield and seems not to have any effect in the subsequent development of embryos in plants. This revised version was published online in July 2006 with corrections to the Cover Date.     

Different Behavior of Brassica juncea and B. carinata as Sources of Phoma lingam Resistance in Experiments of Interspecific Transfer to B. napus

With the aim to transfer Phoma lingam resistance into rape, successful interspecific crosses were made between three oilseed rape varieties (Brassica napus) and the resistant species B. carinata and B. carinata. Although both hybrid types B. napus×B. juncea and B. napus×B. carinata showed the same high level of resistance as the respective resistant parent, the resistance could be only transferred from juncea crosses. After three backcross generations, lines morphologically undistinguishable from rape, fertile, and with a high degree of resistance were obtained. The resistance of B. carinata was practically lost in the first backcross. A possible explanation of this different behavior could be a higher recombination between the genomes B and C (juncea crosses) than between B and A (carinata crosses). The: applied embryo culture increased the yield of hybrids and first backcross plants and reduced considerably the generation time.     

RAPD markers linked to microspore embryogenic ability in Brassica crops

In order to identify the markers linked to microspore embryogenic ability in Brassica crops, RAPD segregation analyses were performed in a microspore-derived (MD) population and a F₂ population derived from F₁between ‘Ho Mei’ (high responsive parent in microspore embryogenesis) and ‘269’ (low responsive parent) in Chinese cabbage, and between ‘Lisandra’ (high responsive parent) and ‘Kamikita’ (low responsive parent) in oil seed rape. After 230 and 143 primers were screened, a total of 148 and 52markers were detected to be polymorphic between the parents in Chinese cabbage and oilseed rape, respectively. Twenty-seven percent of the markers in the MD population showed a significant segregation distortion in both crops. Of the markers showing segregation distortion in the MD population, 71–75% of the markers followed the expected Mendelian segregation ratio in the F₂ population. When the relationships between such distorted markers and microspore embryogenesis of the F₂ population were examined, 7 and 3 markers were identified to be associated with embryogenic ability in Chinese cabbage and oilseed rape, respectively. These markers showed additive effects on embryo yields, and the plants having more alleles of the high responsive parent produced higher embryo yields. These markers maybe useful in marker-assisted selection for improving microspore responsiveness straits in Brassica crops. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparisons of Isozyme Patterns in Resynthesized Amphihaploid Rapeseed (Brassica napus) and Their Parental Species Brassica campestris and Brassica oleracea

Resynthesized rapeseed plants produced by embryo culture and the progeny of their selfed parents from Brassica oleracea and Brassica campestris were analyzed by starch gel electrophoresis staining of enzymes GPI, LAP and SDH. In at least one unequivocal case it could be proved for each enzyme system studied that the alleles of both parents arc independently expressed in the newly synthesized individuals and that in the pattern of the dimeric enzyme GPI interlocus bands of the homoeologous loci arc visible. Comparisons with enzyme patterns in cultivated B. napus leads to the conclusion that in general active enzyme loci of both parental genomes are present. This is of great significance in estimating slide frequencies, degree of heterozygosity and other genetic parameters of B. nupus populations/cultivars.     

Interactive Effects of Elevated CO2 and Moisture Stress on the Photosynthesis, Water Relation and Growth of Brassica Species

Interactive effects of elevated CO2 and moisture stress on photosynthesis, growth and water relation of Brassica species were studied using open top chamber technology. Brassica species responded to the elevated CO2 significantly under moisture stress condition. The adverse effect of moisture stress on the photosynthesis and plant water components were minimized by elevated levels of CO2. Drought susceptible species of B. campestris and B. nigra responded better to elevated CO2 compared to drought tolerant Brassica species such as B. carinata and B. juncea. The plant water potential significantly improved by elevated CO2 coupled with higher stomatal resistance and root growth.     

High frequency spontaneous production of doubled haploid plants in microspore cultures of Brassica rapa ssp. chinensis

Brassica rapa (syn. Brassica campestris) ssp. chinensis is an important vegetable crop, but it is relatively recalcitrant to microspore culture. One genotype each of B. rapa ssp. chinensis var. communisand var. utilis were used formicrospore culture. Embryo production of3.8–42.4 embryos/bud was obtained. A high rate of plant regeneration directly from microspore-derived embryos without subculture was achieved by an improved protocol involving replacement of culture media and reduction of sucrose concentrations after 48 h of induction,among other modifications. More than 70%of regenerated plants were spontaneous diploids. Some spontaneous tetraploid plants were also obtained from isolated microspores of both genotypes tested. These tetraploids may be directly exploited a snew varieties in a Brassica rapabreeding programme. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetics of the Erucic Acid Content in Interspecific Hybrids of Ethiopian Mustard (Brassies carinata Braun) and Rapeseed (B. napus L.)

Ethiopian mustard (Brassica carinata Braun) is a potential oil crop in which genes for low erucic acid content of the seed oil have not yet been found. In order to solve this problem the potential of rapeseed (B. napus L.) varieties as a source of these genes has been tested. Reciprocal F₁ hybrids between B. carinata and a low erucic acid variety of B. napus, F₂, and backcrosses with B. carinata were obtained. The fatty acid composition was determined in half seeds of F₁ and segregating generations from reciprocal interspecific crosses. The genetic analysis indicated that the erucic acid content of the seed oil of B. carinata is controlled by two genes with no dominance and additive in action.     

A Survey of the Anther Culture Response of Brassica oleracea L. Cultivars Grown Under Field Conditions

Anther culture studies were conducted on a wide variety of different vegetable Brassica oleracea L. cultivars grown under field conditions. Representative selections were included from different varieties including: broccoli, var. italica; heading broccoli, var. botrytis; Brussels sprouts, var. gemnifera; Chinese Kale, var. alboglabra; Kales, vars. acephala, medullosa, ramosa, selensia; cauliflower, var. botrytis; kohlrabi, var. gongylodes; and cabbage, vars. capitata and sabanda. At least some microspore derived embryos were recovered from the majority of cultivars tested; however, rates of embryo formation were sometimes lower than those obtained under controlled environmental conditions. In general the anther culture response was greatest for cabbage and other vegetables that required vernalization for flowering. Results were lower for materials exposed to high temperature stresses before culture. Nevertheless, it is clear that the use of anther culture procedures in breeding programs is possible without access to extensive controlled environmental facilities.     

Microspore culture is successful in most crop types of Brassica oleracea L.

Summary Microspore culture was shown to be applicable to a broad range of accessions belonging to six horticulturally important crop types of Brassica oleracea: broccoli, white cabbage, cauliflower, savoy cabbage, Brussels sprouts and curly kale. Of 64 accessions tested 86% were responsive. Large genotypic differences were found in number of embryos produced per flower bud, and in frequency and mode of regeneration of plants from embryos. B. oleracea was characterized by a strong asynchrony of microspore development within single buds. Microspore populations optimal for culture contained a large proportion (10–40%) of binucleate pollen. An initial high temperature treatment was essential for microspore embryogenesis. Growth conditions of the donor plants during inflorescence formation were less critical.