Abstract
High efficiency of the cultivation of unfertilized sugar beet ovules and preparation of haploid regenerants (microclones) of pollinators – maintainers of О-type sterility and MS forms of the RMS 120 hybrid components has been shown. A technological method that accelerates the creation of new uniform starting material is proposed. It speeds up the breeding process two to threefold. The identification of haploid regenerants with sterile cytoplasm in initial populations is of great theoretical and practical importance for breeding, as it facilitates the production of homozygous lines with cytoplasmic male sterility and high-performance hybrids on sterile basis. As shown by molecular analysis, a single-nucleotide polymorphism never reported hitherto is present in the mitochondrial genome of the haploid plant regenerants. It allows identification of microclones as fertile and sterile forms. It has been found that DNA markers of the sugar beet mitochondrial genome belonging to the TR minisatellite family (TR1 and TR3) enable reliable enough identification of haploid microclonal plants as MS- or O-type forms. Fragments of 1000 bp in length have been detected in monogenic forms in the analysis of 11 sugar beet plants cultured in vitro by PCR with the OP-S4 random RAPD primer. Testing of the OP-S4 marker’s being in the same linkage group as the genes responsible for expression of the economically valuable trait monogermity demonstrates its relative reliability. By the proposed method, dihaploid lines (DH) of the male-sterile form and the О-type sterility maintainer of the RMS 120 sugar beet hybrid have been obtained in in vitro culture. These lines are highly uniform in biomorphological traits, as proven under field conditions.
Highlights
Sugar beet (Beta vulgaris L.) is an important source of sucrose, and sugar is one of the essential ingredients in the human diet and a source of readily available energy for the body
Approximately 98 percent of areas under sugar beet was planted in Russia with imported seeds of foreign breeding, which has a highly negative impact on the technological and economic sustainability of the whole sugar-beet industry in Russia
The combination of biotechnology and conventional breeding methods permits one to increase the productivity of sugar beet hybrids, and to improve the quality of seed material
Summary
Sugar beet (Beta vulgaris L.) is an important source of sucrose, and sugar is one of the essential ingredients in the human diet and a source of readily available energy for the body. The use of modern biotechnological and molecular techniques in breeding practice accelerates twofold the development of genetically uniform material, which ensures a high uniformity of root morphology parameters (size, weight, height of head protrusion, depth of the fibrous root system, etc.), as well as sustainable implementation of major commercially valuable traits (monogermity, crop capacity, sugar content, abiotic and biotic stress tolerance during the growing stage, prolonged viability during storage, etc.) during the reproduction. The combination of biotechnology and conventional breeding methods permits one to increase the productivity of sugar beet hybrids, and to improve the quality of seed material. The selected well-developed haploid regenerants are stabilized by in vitro micropropagation on agar medium. The step includes the diploidization of haploid material by colchicination, stabilization of colchicined regene rants, selection based on biochemical and molecular traits, and formation of in vitro DH lines
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