ABSTRACT
Double haploid technique is not routinely used in legume breeding programs, though recent publications report haploid plants via anther culture in chickpea (Cicer arietinum L.). The focus of this study was to develop an efficient and reproducible protocol for the production of double haploids with the application of multiple stress pre-treatments such as centrifugation and osmotic shock for genotypes of interest in chickpea for their direct use in breeding programs. Four genotypes, ICC 4958, WR315, ICCV 95423 and Arearti were tested for anther culture experiments. The yield was shown to be consistent with 3-5 nucleate microspores and 2-7 celled structures with no further growth. To gain a further insight into the molecular mechanism underlying the switch from microsporogenesis to androgenesis, bioinformatics tools were employed. The challenges on the roles of such genes were reviewed while an attempt was made to find putative candidates for androgenesis using Expressed Sequenced Tags (EST) and interolog based protein interaction analyses.
Subject(s)
Breeding , Cicer/genetics , Computational Biology , Expressed Sequence Tags , Fabaceae/genetics , Genotype , Haploidy , Plant Roots/genetics , Plant Roots/metabolism , Protein Interaction Maps/genetics , Stress, PhysiologicalABSTRACT
Background The quality of wheat grain depends on several characteristics, among which the composition of high molecular weight glutenin subunits, encoded by Glu-1 loci, are the most important. Application of biotechnological tools to accelerate the attainment of homozygous lines may influence the proportion of segregated genotypes. The objective was to determine, whether the selection pressure generated by the methods based on in vitro cultures, may cause a loss of genotypes with desirable Glu-1 alleles. Results Homozygous lines were derived from six winter wheat crosses by pollination with maize (DH-MP), anther culture (DH-AC) and single seed descent (SSD) technique. Androgenetically-derived plants that originated from the same callus were examined before chromosome doubling using allele-specific and microsatellite markers. It was found that segregation distortion in SSD and DH-MP populations occurred only in one case, whereas in anther-derived lines they were observed in five out of six analyzed combinations. Conclusions Segregation distortion in DH-AC populations was caused by the development of more than one plant of the same genotype from one callus. This distortion was minimized if only one plant per callus was included in the population. Selection of haploid wheat plants before chromosome doubling based on allele-specific markers allows us to choose genotypes that possess desirable Glu-1 alleles and to reduce the number of plants in the next steps of DH production. The SSD technique appeared to be the most advantageous in terms of Mendelian segregation, thus the occurrence of residual heterozygosity can be minimized by continuous selfing beyond the F6 generation.
Subject(s)
Triticum/genetics , Chromosome Segregation , Seeds/genetics , In Vitro Techniques , Microsatellite Repeats , Zea mays , Alleles , Genotype , Glutens/analysis , HomozygoteABSTRACT
The use of maize in anther culture has been limited because only few genotypes presented a high androgenetic potential. Obtaining the proper stage of microspore development at culture initiation is one of the most crucial factors for success in the androgenesis. For Brazilian maize genotypes there are no studies reporting a correlation between cytological features and morphological parameters. In this study, morphological parameters were recorded and associated with cytological specific stages of the the microsporogenesis in two Brazilian maize genotypes that were sowed in different places (field and growing chamber). For both genotypes, the plants of the growing chamber presented a delay in development. Spikelets length and anther length/spikelet length ratio are not good parameters since they can be greatly influenced by the environment. The anther length was the more reliable parameter to correlate with a specific developmental stage. Nevertheless, variations between genotypes and environment were detected.
A utilização do milho (Zea mays) na cultura de anteras é limitada devido ao baixo número de genótipos com alto potencial androgenético. A obtenção de micrósporos no estádio de desenvolvimento apropriado no início da cultura é um dos fatores cruciais para o sucesso do processo androgenético. Em genótipos brasileiros de milho não existem estudos relatando a correlação entre características citológicas e parâmetros morfológicos. Neste estudo, parâmetros morfológicos foram avaliados e associados com estádios específicos da microsporogênese em dois genótipos brasileiros de milho os quais foram semeados em diferentes locais (campo e câmara de crescimento). Para ambos os genótipos, as plantas crescidas na câmara de crescimento apresentaram atraso no desenvolvimento. O comprimento da espigueta e a razão comprimento da antera/comprimento da espigueta não são bons parâmetros uma vez que podem ser muito influenciados pelo ambiente. O comprimento da antera foi o melhor parâmetro para indicar o estádio de desenvolvimento do micrósporo. Todavia, variações entre genótipos e ambiente foram detectadas.
ABSTRACT
The development of in vitro haploid plants followed by spontaneous or induced genome duplication allows to achieve, in one generation, the recovery of total homozygosis. The efficiency of the haplodiploidization process through in vitro anther culture of barley is variable among genotypes. This study was aimed at determining the androgenetic response of nine barley genotypes from the breeding program of Embrapa Trigo, analysing proembryoid development and green plantlets regeneration in anthers cultivated in vitro. Cultivar 'BR2' presented the highest average of proembryoids (104/anther) and 'MN698' presented the highest average of green plantlets (0,41/anther). There was a significant variation among the average values of barley genotypes for embryo formation and green plantlets regeneration, making possible the selection to combine androgenetic capacity and good agronomic traits.
A obtenção, na cevada, de plantas haplóides in vitro e a posterior duplicação natural ou artificial do genoma permitem alcançar a homozigose completa, em uma geração. A eficiência da haplodiploidização pela cultura de anteras é variável entre os genótipos. Foi avaliada a resposta androgenética através da formação de pró-embrióides e da regeneração de plântulas verdes em nove cultivares do programa de melhoramento de cevada da Embrapa Trigo, em anteras cultivadas in vitro. A cultivar "BR2" apresentou maior média de pró-embrióides (104/antera), enquanto "MN698" mostrou a maior média de plântulas verdes (0,41/antera). Houve variação significativa entre os valores médios dos genótipos em relação à formação de pró-embrióides e à regeneração de plântulas verdes, indicando a possibilidade de seleção para combinar a capacidade androgenética com boas características agronômicas.