Transfer of powdery mildew resistance from Brassica carinata to Brassica oleracea through embryo rescue

Interspecific hybrid plants and backcross 1 (BC₁) progeny were produced through sexual crosses and embryo rescue between Brassica carinata accession PI 360883 and B. oleracea cvs Titleist’and‘Cecile’to transfer resistance to powdery mildew to B. oleracea. Four interspecific hybrids were obtained through application of embryo rescue from crosses with B. carinata as the maternal parent, and their interspecific nature confirmed through plant morphology and random amplified polymorphic DNA (RAPD) analysis. Twenty‐one BC₁ plants were obtained through sexual crosses and embryo rescue although embryo rescue was not necessary to produce first backcross generation plants between interspecific hybrids and B. oleracea. All interspecific hybrids and eight of the BC₁ plants were resistant to powdery mildew.     

Interspecific hybridization between Brassica carinata and Brassica rapa

The crossability between Brassica carinata (BBCC, 2n=34) and Brassica rapa (AA, 2n=20), and the cytomorphology of their F₁ hybrids were studied. Hybrids between these two species were only obtained when B. carinata was used as the female parent. The hybrid plants exhibited intermediate leaf and flower morphology, and were found to be free from white rust and Alternaria blight diseases. One of the four F₁ plants was completely male sterile, while the remaining plants had 4.8, 8.6, and 10.9% stainable pollen, respectively. No seed was produced on hybrid plants under self pollination or in backcrosses; but seed was obtained from open pollination. The occurrence of the maximum of 11 bivalents as well as up to 44.8%) of cells with multivalent associations in the form of trivalents (0‐2) and a quadrivalent (0‐1) in the trigenomic triploid hybrid (ABC, 2n = 27) revealed intergenomic homoeology among the A, B and C genomes. Meiotic analysis of F₁ hybrids indicated that traits of economic importance, such as disease resistance, could be transferred from B. carinata to B. rapa through interspecific crosses.     

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.     

Glucosinolate content in interspecific crosses of Brassica carinata with B. juncea and B. napus

Brassica carinata A. Braun is a highly productive oilseed crop in the Ethiopian highlands, but the seed has a high 2-propenyl glucosinolate content, which is undesirable. The objective of this study was to introgress, through interspecific crosses, genes for low 2-propenyl glucosinolate content from the B genome of B. juncea and C genome of B. napus into the B. carinata B and C genomes and thus develop low glucosinolate B. carinata. The cross [(B. carinata×B. juncea) ×B. carinata] yielded plants that contained only ～ 20 μmoles of 2-propenyl glucosinolate, which was an 85% reduction compared with levels in B. carinata seed. Plants of the [(B. carinata×B. napus) ×B. carinata] cross had normal high concentrations of 2-propenyl glucosinolate. Backcross plants of both interspecific crosses also contained 3-butenyl and 2-hydroxy-3-butenyl glucosinolates. The results of these crosses suggested that genes for glucosinolate synthesis were located on B genome chromosomes of B. carinata because B. napus C genome introgressions did not result in reductions of total glucosinolate contents. The total alkenyl glucosinolate content of one F₃ family of the B. juncea backcross was similar to that of the B. juncea parent. It was concluded that through further selection in this family, B. carinata plants could be identified that would be basically free of 2-propenyl glucosinolate, and have a low total alkenyl glucosinolate content.     

埃塞俄比亚芥与白菜型油菜间六倍体杂种的获得及其生物学特性研究

通过杂交和染色体加倍人工合成了埃塞俄比亚芥×白菜型油菜间的三倍体和六倍体杂种，并对其进行了细胞学鉴定、气孔观察以及一些生理生化特性的分析。在染色体记数、原位杂交和分子标记印证的基础上，比较了两种不同染色体加倍方法的效率；生理生化分析显示，六倍体杂种植株叶片的游离脯氨酸含量、质膜相对透性、Vc含量和叶绿素含量与三倍体杂种植株相比都有明显的差别。     

埃塞俄比亚芥研究进展

埃塞俄比亚芥(Brassica carinata)拥有很多优良的农艺性状:耐热,耐干旱,抗倒伏,抗黑胫病、霜霉病等多种病害,抗蚜虫、跳甲等多种害虫;并且其种质资源非常丰富。介绍了埃塞俄比亚芥的品质改良、遗传多样性分析、可交配性研究以及种质资源利用四个方面的研究进展,希望这些信息能为埃塞俄比亚芥的遗传改良以及油菜育种中如何高效利用埃塞俄比亚芥的优良性状提供指导。     

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.     

Transgressive segregation for reduced glucosinolate content in Brassica carinata A. Braun

Successful commercial utilization of the meal by‐product of Brassica oilseed crops requires the cultivation of cultivars with low glucosinolate (GSL) content in the seeds; however, such cultivars are not yet available in Brassica carinata. The objective of the present research was to search for transgressive segregants with further‐reduced GSL content in the progeny of crosses involving four B. carinata lines with reduced GSL content (90 compared with 120 μmol/g seed in standard germplasm). The four lines were crossed following a diallel design and F₂ phenotypes (F₃ seed bulked) were analysed for GSL content. F₂ phenotypes with a transgressive GSL content lower than the parents were identified in all crosses involving the line S2–1241, suggesting that this line carries alleles for reduced GSL content not present in the other lines. F_{3 : 4} lines from transgressive F₂ phenotypes were evaluated for 2 years, which resulted in the selection of an F_{3 : 4} line with an average GSL content of 58 and 46 μmol/g seed, respectively compared with 84 and 62 μmol/g seed, respectively in S2–1241.     

Doubled haploid lines of Brassica carinata with modified erucic acid content through mutagenesis by EMS treatment of isolated microspores

Brassica carinata is a potential oilseed crop for the Mediterranean area. Chemical mutagenesis has been applied to microspores of B. carinata with the purpose of identifying lines with altered erucic acid content. From a population of nearly 400 doubled haploid plants recovered, nine lines have been identified that exhibit promising useful changes in erucic acid concentration in the seed oil. Three lines showed erucic acid contents below 25%, with a minimum of 17.1%, and in six lines the level of this fatty acid was greater than 52%. Changes in other fatty acids are also described and discussed.     

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 .     

甘蓝型油菜与埃塞俄比亚芥种间可交配性的胚胎学研究

在甘蓝型油菜×埃塞俄比亚芥中,杂种胚乳不甚发育并提前形成细胞,杂种胚发育缓慢,消耗早育的胚乳组织后便停滞在心形期或鱼雷形期.反交时胚乳早期发育旺盛,但在发育中期不能形成细胞并逐渐退化解体;杂种胚从原胚阶段即有退化现象发生,并可持续到鱼雷形期,胚退化时常伴有胚柄细胞的解体.甘蓝型油菜母本基因型对杂交的生殖过程有显著影响.对受精的影响表现在其柱头与异种花粉的亲和性不同,以Rucabo作母本时表现与反交相似的高亲和性和高受精率.以84014A作母本时近半的胚珠中发生体质不育现象.而华油8号作母本时产生的杂种胚和胚乳在后期的发育相对正常,可形成少量有生命力的杂种种子.文中提出通过杂交重组创造出种间受精率高,又具结实潜力的“桥梁品系”,以经济有效地克服甘蓝型油菜与埃塞俄比亚芥杂交的不可交配性.     

Development of Polima temperature-sensitive cytoplasmic male sterile lines of Brassica napus through isolated microspore culture

A novel Pol CMS named Polima temperature‐sensitive cytoplasmic male sterile (Pol TCMS) is sterile at high temperatures and partially fertile at low temperatures. Its temperature sensitivity is controlled by several minor genes, and probably influenced by the genetic background, making it quite difficult to breed a stable Pol TCMS line through the conventional breeding methods. The present study aimed to obtain stable homozygous Pol TCMS lines by microspore culture. The highest frequency of embryogenesis was induced from cultures of buds sampled at 4–7 days before first blossom. The second method of microspore cultures mentioned in this study could significantly increase the production of embryos. Analysis of fatty acid composition, glucosinolate content and genetic diversity by simple sequence repeat (SSR) markers indicated that microspore culture was effective for selecting Pol TCMS lines.     

Hydrotime Analysis of Ethiopian Mustard (Brassica carinata A. Braun) Seed Germination Under Different Temperatures

Ethiopian mustard (Brassica carinata A. Braun) is an oil‐seed species that has recently become an object of great interest as a promising crop for energy purposes (biodiesel and biolubricant production). In semi‐arid regions of South Italy, soil moisture at sowing time is often inadequate, delaying and reducing seed germination. In this study, the effect of reduced water potential on seed germination in three cultivars of B. carinata (ISCI 7, CT 180 and Sincron) was investigated in the laboratory. Germination behaviour at constant temperatures under low water availability was also analysed through the hydrotime model. Six water potentials (ψ_s) in PEG solution (0, ?0.2, ?0.4, ?0.6, ?0.8 and ?1.0 MPa) and three temperatures (10, 20 and 30 °C) were used for the germination tests. A thermo‐inhibiting effect was observed at the highest levels of water stress. The hydrotime analysis revealed that the increase of temperature to 30 °C reduced predicted hydrotime (θ_H) by hastening the rate of germination, but shifted median base water potential (ψ_b(50)) to higher values (less negative), whereas the lowering of temperature increased θ_H. These observations may have a great agronomic impact because although fewer seeds germinated at 30 °C at the highest levels of water stress, they may germinate faster in rapidly drying soil. However, genetic differences were observed among cultivars in terms of estimated θ_H and ψ_b(50). The hydrotime modelling approach offered in this study may help predict B. carinata seed germination under soil water deficit conditions occurring under early or late autumnal sowing.     

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.     

Variation and inheritance of erucic acid content in Brassica carinata germplasm collections from Ethiopia

Ethiopian mustard possesses a number of agronomic advantages over other oilseed crops with similar ecological adaptation in Ethiopia. However, its high erucic acid content is undesirable for a vegetable oil. Although efforts have been made to improve its quality, much has to be done to use natural variations that might exist within the species for fatty acid contents. This project was undertaken to study the variability of fatty acid contents, primarily erucic acid, in germplasm collections of Ethiopian origin, with an attempt to develop low (zero) erucic acid genotypes. The study used inbred lines as well as F₂ populations of 10 crosses between six parental lines. A wide variation in fatty acids was found. Oleic acid content varied from 5 to 34% and erucic acid content from 6 to 51%. Linoleic and linolenic acid contents were less variable. The high‐oleic genotypes exhibited not only low erucic but also higher linoleic (25%) and considerably lower linolenic acid (8%) contents. It was possible to classify the F₂ populations with the lowest erucic acid into three distinct classes. While the first class had an erucic acid content of 6–12%, the second and third classes had contents of 18–32% and 36–42%, respectively. The existence of a multiple allelic series of erucic acid in Ethiopian mustard would enable its fixation at zero levels without necessarily going into interspecific crossing.