Considerable progress has been accomplished in the cellular and molecular biology of Brassica species in the past few years. Plant regeneration has been increasingly optimized via organogenesis and somatic embryogenesis using various explants; with tissue culture improvements focusing on factors such as age of the explant, genotype, and media additives. The production of haploids and doubled haploids using microspores has accelerated the production of homozygous lines in the Brassica species. Somatic cell fusion has facilitated the development of interspecific and intergeneric hybrids in the sexually incompatible species of Brassica. Crop improvement using somaclonal variation has also been achieved. The use of molecular markers in marker-assisted selection and breeding, transformation technology for the introduction of desirable traits, and a comparative analysis of these as well as their future prospects are important parts of the current research that is reviewed.

Invited review: brassica biotechnology: progress in cellular and molecular biology

Related or distant species of cultivated cs are a large pool of many desirable genes. Gene transfer from these species through conventional breeding is difficult owing to post- and pre-zygotic sexual incompatibilities. Somatic hybridization via protoplast fusion is a possible alternative for gene transfer from these species to cultivated crops. Since the early days of somatic hybridization many intergeneric somatic hybrids have been developed through symmetric fusion, asymmetric fusion and microfusion. Somatic hybrids are mainly selected by using markers such as specific media or fusion parents with special features, biochemical mutants, antibiotic resistance and complementation strategy. The hybridity of the regenerants is determined based on morphological, cytological and molecular analysis. The inheritance patterns of nuclear and cytoplasmic genomes in the somatic hybrids are diverse. Nuclear DNA from both fusion parents co-exists congruously in some hybrids with translocation and rearrangement of chromosomes, but spontaneous elimination of chromosomes from either or both fusion parents has been observed very often. In asymmetric fusion, chromosome elimination is an important issue that is a complicated process influenced by many factors, such as irradiation dose, phylogenetic relatedness, ploidy level of fusion parent and regenerants. As for chloroplast genome, uniparental segregation is mainly detected, though co-existence is also reported in some cases. The mitochondrial genome, in contrast to chloroplast, undergoes recombination and very frequent rearrangements. Somatic cell fusion has potential applications for crop genetic improvement by overcoming sexual incompatibility or reproductive barriers, and by realizing novel combinations of nuclear and/or cytoplasmic genomes.

Intergeneric somatic hybridization and its application to crop genetic improvement

We compared the structural, biochemical, and physiological characteristics involved in photorespiration of intergeneric hybrids differing in genome constitution (DtDtR, DtDtRR, and DtRR) between the C(3)-C(4) intermediate species Diplotaxis tenuifolia (DtDt) and the C(3) species radish (Raphanus sativus; RR). The bundle sheath (BS) cells in D. tenuifolia included many centripetally located chloroplasts and mitochondria, but those of radish had only a few chloroplasts and mitochondria. In the hybrids, the numbers of chloroplasts and mitochondria, the ratio of centripetally located organelles to total organelles, and the mitochondrial size in the BS cells increased with an increase in the constitution ratio of the Dt:R genome. The P-protein of glycine decarboxylase (GDC) was confined to the BS mitochondria in D. tenuifolia, whereas in radish, it accumulated more densely in the mesophyll than in the BS mitochondria. In the hybrids, more intense accumulation of GDC in the BS relative to the mesophyll mitochondria occurred with an increase in the Dt:R ratio. These structural and biochemical features in the hybrids were reflected in the gas exchange characteristics of leaves, such as the CO(2) compensation point. Our data indicate that the leaf structure, the intercellular pattern of GDC expression, and the gas exchange characteristics of C(3)-C(4) intermediate photosynthesis are inherited in the hybrids depending on the constitution ratio of the parent genomes. Our findings also demonstrate that the apparent reduced photorespiration in C(3)-C(4) intermediate plants is mainly due to the structural differentiation of mitochondria and chloroplasts in the BS cells combined with the BS-dominant expression of GDC.

Structural and biochemical dissection of photorespiration in hybrids differing in genome constitution between Diplotaxis tenuifolia (C3-C4) and radish (C3).

The subjects of this article are protoplast isolations and protoplast fusions, in particular their history, a review of factors influencing the protoplasts isolation and fusion, selection of hybrid plants and utilization of somatic hybrids in plant breeding. Somatic hybridization through protoplast fusion can overcome sexual incompatibility among plant species or genera; transfer genes of resistance to diseases (viral, bacterial, fungal), pests, herbicides and others stress factors; obtain cybrid plants; transfer cytoplasmic male sterility or incease content of secondary metabolites in hybrid plants. The article is focussed mainly on the family Brassicaceae because among representatives are significant crops for the human population. Various successful combination of intraspecific, interspecific and intergeneric protoplast fusion were reported between representatives of the family Brassicaceae with the genus Brassica which belonged to the first agricultural crops used for the isolation of protoplast.

/pdf/protoplast-cultures-and-protoplast-fusion-focused-on-3oxben7ny7.pdf

Protoplast cultures and protoplast fusion focused on Brassicaceae: A review

Somatic Hybridization and Applications in Plant Breeding

Protoplast fusion between Brassica oleracea and Moricandia nitens, a C3–C4 intermediate wild species, was carried out. Four hundred and twenty five plants were regenerated from 1995 calli. More than 90% of the regenerated plants were verified as true intergeneric hybrids on the basis of morphological observation and molecular-marker analysis. The hybrids were morphologically intermediate between both fusion parents. Variations in flower color and petal number were also observed. The chromosome number and pollen fertility varied across the individual hybrids. Although after self-pollination pollen germinated on the stigma and pollen tubes were visible in the style, the pods did not develop properly without in vitro culture. Measurements of the CO2 compensation point revealed that six out of eight hybrid plants expressed a gas-exchange character that was intermediate between the C3–C4M. nitens and C3B. oleracea parents.

Production of somatic hybrids between Brassica oleracea and the C3–C4 intermediate species Moricandia nitens

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Production of somatic hybrids between Brassica oleracea and the C3–C4 intermediate species Moricandia nitens