Abstract More efficient haploidy protocols have the power to speed up breeding studies. We aimed ... more Abstract More efficient haploidy protocols have the power to speed up breeding studies. We aimed to increase microspore-derived embryo and in vitro plantlet yields in eggplant anther cultures. To achieve this, a comprehensive optimization study was carried out on the commonly used two-step DDV (Dumas de Vaulx et al., 1982) eggplant anther culture system. For optimizations of both induction and differentiation culture media, the effects of carbohydrate sources (maltose and sucrose) were investigated alone or in combination with two kinds of additives consisting of silver nitrate (AgNO3) and activated charcoal (AC). They were done in the presence of macro-micro salts, vitamins and PGRs of original DDV protocol. During the optimizations, we also compared the effects of genotype, the growth season of donor plants, anther culture type, and culture media, on embryo and in vitro plantlet yields. Regarding culture media, single effect of maltose use was considerable only at the highest concentration. The effects of maltose + AgNO3 combinations were not noteworthy while the effect of maltose + AC combination was negative. However, triple use of maltose (90 g/L) + AgNO3 (10 mg/L) + AC (1 g/L) had a remarkably positive synergistic effect depending on the genotype, season, and concentrations of optimization chemicals. We observed a triple synergistic effect of this aSC24-encoded modified induction medium on longer viability of anthers, embryo quality, direct embryogenesis, faster regeneration and increased embryo and in vitro plantlet yields. Through this extremely efficient modified induction medium, we achieved 3.9 times more embryo yields than the original DDV medium in two genotypes. The highest embryo and in vitro plantlet yields (320 embryos and 200 plantlets/100 anthers) were obtained from ‘A117’ eggplant anthers cultured in solid aSC24 + DDV-R combined cultures in the autumn season. In addition, as far as we know, embryos (up to 42 embryos/100 anthers) were produced for the first time in a liquid anther culture in eggplant.
ABSTRACT In the present study, anther culture method was used in Anemone coronaria, known as Mani... more ABSTRACT In the present study, anther culture method was used in Anemone coronaria, known as Manisa Tulip in Turkiye, in order to obtain haploid plants. The anthers with uninucleate microspores of the flower buds, collected from an A. coronaria var. coccinea population, were used as explant. In the treatments, two basic tissue culture media (NN and B5) were tested with 4 different auxin concentrations and also 4 different auxin:cytokinin ratios. Anthers were subjected to a chilling pre-treatment for 7 days at +7°C in the dark and then incubated under continuous dark conditions at 25 +1°C. The use of cytokinin together with auxin increased callus induction frequency. It was also found that the treatments with the combinations consisting of both kind of growth regulators in B5 medium showed more successful performance.
One of the most remarkable natural plants is Vaccaria hispanica (Mill.) Rauschert, which has a hi... more One of the most remarkable natural plants is Vaccaria hispanica (Mill.) Rauschert, which has a high economic, medicinal and industrial potential due to its valuable compounds. However, it is mostly an underused plant worldwide. To implement doubled haploid technology in plant breeding programs and exploit its potential, first knowing the particulars of the species is necessary. This study was aimed to explore the androgenic potential of different wild Turkish V. hispanica genotypes collected from different Turkish regions, as a starting point to identify bottlenecks to solve in future studies. As to induction of microspore embryogenesis, nearly all of them responded, with efficiencies reaching 300 embryos/100 buds in some cases. However, we also found three main bottlenecks. First, the presence of foam-producing saponins in flowers prevented an efficient isolation of microspores. Second, embryos showed a reduced ability to germinate. Third, a dense network of hairy roots prevented the formation of a true, fully functional root system. Together, these drawbacks led to the production of very few DH plants. The presence of rhizogenic endophytes may be the cause of most of these problems, which opens the door for possible solutions. Vaccaria hispanica is a highly responsive species to microspore embryogenesis. However, the two main bottlenecks, possibly caused by endophytes, were the formation of dense abnormal embryos and short hairy roots.
Abstract More efficient haploidy protocols have the power to speed up breeding studies. We aimed ... more Abstract More efficient haploidy protocols have the power to speed up breeding studies. We aimed to increase microspore-derived embryo and in vitro plantlet yields in eggplant anther cultures. To achieve this, a comprehensive optimization study was carried out on the commonly used two-step DDV (Dumas de Vaulx et al., 1982) eggplant anther culture system. For optimizations of both induction and differentiation culture media, the effects of carbohydrate sources (maltose and sucrose) were investigated alone or in combination with two kinds of additives consisting of silver nitrate (AgNO3) and activated charcoal (AC). They were done in the presence of macro-micro salts, vitamins and PGRs of original DDV protocol. During the optimizations, we also compared the effects of genotype, the growth season of donor plants, anther culture type, and culture media, on embryo and in vitro plantlet yields. Regarding culture media, single effect of maltose use was considerable only at the highest concentration. The effects of maltose + AgNO3 combinations were not noteworthy while the effect of maltose + AC combination was negative. However, triple use of maltose (90 g/L) + AgNO3 (10 mg/L) + AC (1 g/L) had a remarkably positive synergistic effect depending on the genotype, season, and concentrations of optimization chemicals. We observed a triple synergistic effect of this aSC24-encoded modified induction medium on longer viability of anthers, embryo quality, direct embryogenesis, faster regeneration and increased embryo and in vitro plantlet yields. Through this extremely efficient modified induction medium, we achieved 3.9 times more embryo yields than the original DDV medium in two genotypes. The highest embryo and in vitro plantlet yields (320 embryos and 200 plantlets/100 anthers) were obtained from ‘A117’ eggplant anthers cultured in solid aSC24 + DDV-R combined cultures in the autumn season. In addition, as far as we know, embryos (up to 42 embryos/100 anthers) were produced for the first time in a liquid anther culture in eggplant.
ABSTRACT In the present study, anther culture method was used in Anemone coronaria, known as Mani... more ABSTRACT In the present study, anther culture method was used in Anemone coronaria, known as Manisa Tulip in Turkiye, in order to obtain haploid plants. The anthers with uninucleate microspores of the flower buds, collected from an A. coronaria var. coccinea population, were used as explant. In the treatments, two basic tissue culture media (NN and B5) were tested with 4 different auxin concentrations and also 4 different auxin:cytokinin ratios. Anthers were subjected to a chilling pre-treatment for 7 days at +7°C in the dark and then incubated under continuous dark conditions at 25 +1°C. The use of cytokinin together with auxin increased callus induction frequency. It was also found that the treatments with the combinations consisting of both kind of growth regulators in B5 medium showed more successful performance.
One of the most remarkable natural plants is Vaccaria hispanica (Mill.) Rauschert, which has a hi... more One of the most remarkable natural plants is Vaccaria hispanica (Mill.) Rauschert, which has a high economic, medicinal and industrial potential due to its valuable compounds. However, it is mostly an underused plant worldwide. To implement doubled haploid technology in plant breeding programs and exploit its potential, first knowing the particulars of the species is necessary. This study was aimed to explore the androgenic potential of different wild Turkish V. hispanica genotypes collected from different Turkish regions, as a starting point to identify bottlenecks to solve in future studies. As to induction of microspore embryogenesis, nearly all of them responded, with efficiencies reaching 300 embryos/100 buds in some cases. However, we also found three main bottlenecks. First, the presence of foam-producing saponins in flowers prevented an efficient isolation of microspores. Second, embryos showed a reduced ability to germinate. Third, a dense network of hairy roots prevented the formation of a true, fully functional root system. Together, these drawbacks led to the production of very few DH plants. The presence of rhizogenic endophytes may be the cause of most of these problems, which opens the door for possible solutions. Vaccaria hispanica is a highly responsive species to microspore embryogenesis. However, the two main bottlenecks, possibly caused by endophytes, were the formation of dense abnormal embryos and short hairy roots.
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