Somatic embryogenesis of pepper in bioreactors: A study of bioreactor type and oxygen uptake rates

Somatic embryogenesis of pepper, Capsicum annuum var. Ace, was performed in an airlift bioreactor and a magnetically stirred hanging-stirrer-bar bioreactor, each with 1.8 l working volume. All stages of embryogenesis, from growth of embryogenic suspension cultures to embryo maturation, were performed in the bioreactor as a series of drain-and-fill batches, keeping the cells and embryos in the bioreactor all the time. When two bioreactors were compared in terms of percentage embryogenesis and visually observed quality of mixing, under different rates of aeration and stirring, the performance of the magnetically stirred bioreactor was better. The effects of inoculum type and inoculum level on the percentage embryogenesis were also investigated. Under the optimum conditions, embryogenesis was 98%, with 57 embryos/ml. Oxygen-uptake rates of cultures in different stages of embryogenesis were different, the highest being in the embryogenic suspension culture and the lowest during embryo maturation

Artificial seeds of pepper somatic embryos

1st International Symposium on Solanacea for Fresh Market -- MAR 28-31, 1995 -- MALAGA, SPAINWOS: 000071774700011Artificial seeds are a potential method for delivering somatic embryos to the field. Mature somatic embryos were encapsulated in calcium alginate gel using different concentrations of sodium alginate (2-6%) and calcium chloride (25-100 mM) in order to determine the effect of rigidity of bead on the conversion frequency of somatic embryos, MS medium supplemented with 3% sucrose was used far germination. Variability was obtained between sodium alginate and calcium cloride concentration with respect to % germination. Plant regeneration frequency decreased with an increase in sodium alginate concentration above 3%. 3% sodium alginate and 75 mM calcium chloride gave the best conversion and optimum bead rigidity and shape. In vitro and in vivo conversion of naked and encapsulated embryos were compared.Int Soc Hort Sci, Spanish Soc Hort Sci, Consejo Super Invest Cient, Junta Andalucia, Consejeria Agr & Pesca, Plan Nacl Invest Cientif & Desarrollo Tecnol, Excma, Diputac Malaga, British Council, Cajas Rurales Granada & Malaga, Unicaja, MSD Agvet, Haifa Chem, Ferquis

Somatic embryogenesis and plant regeneration of pepper in liquid media

A protocol was developed for regeneration of pepper (Capsicum annuum var. Ace) through somatic embryogenesis in liquid media. For embryogenic callus formation, mature zygotic embryo explants were used on basal Murashige and Skoog medium with 9.05 µM 2,4-dichlorophenoxyacetic acid and 3% sucrose. Embryogenic callus was transferred to liquid basal Murashige and Skoog medium with 4.52 µM 2,4-dichlorophenoxyacetic acid and 3% sucrose in order to increase the mass of the embryogenic culture. After pretreatment with potassium citrate, cells were placed into embryo initiation medium with 6 g l-1 L-proline and a decreased (10 mM) ammonium concentration. Embryos were matured in 1.89 µM abscisic acid containing half-strength Murashige and Skoog medium and converted into plants both in vivo and in vitro at up to a 97% efficiency

Large scale production of pepper somatic embryos using bioreactors

1st International Symposium on Solanacea for Fresh Market -- MAR 28-31, 1995 -- MALAGA, SPAINWOS: 000071774700004Bioreactors can he used for large scale production of somatic embryos effectively as labour saving system. Two different types of bioreactor, an airlift and a magnetically stirred "hanging stirrer bar" bioreactor, each with 1.8 l working volume, and different mixing/aeration rates were examined to determine a system capable of in vitro pepper somatic embryo production. Production of pepper somatic embryos in a bioreactor was successful when a hanging stirrer bar bioreactor was used, Mixing, aeration, inoculum type and level affect directly the somatic embryogenesis in a bioreactor. The best results were obtained from hanging stirrer bar bioreactor at 0.45 vvm aeration and 80 rpm mixing rate in terms of percent embryogenesis. These results confirm that bioreactor culture offers promising aspects for large scale production.Int Soc Hort Sci, Spanish Soc Hort Sci, Consejo Super Invest Cient, Junta Andalucia, Consejeria Agr & Pesca, Plan Nacl Invest Cientif & Desarrollo Tecnol, Excma, Diputac Malaga, British Council, Cajas Rurales Granada & Malaga, Unicaja, MSD Agvet, Haifa Chem, Ferquis

ARTIFICIAL SEEDS OF PEPPER SOMATIC EMBRYOS

1st International Symposium on Solanacea for Fresh Market -- MAR 28-31, 1995 -- MALAGA, SPAINWOS: 000071774700011Artificial seeds are a potential method for delivering somatic embryos to the field. Mature somatic embryos were encapsulated in calcium alginate gel using different concentrations of sodium alginate (2-6%) and calcium chloride (25-100 mM) in order to determine the effect of rigidity of bead on the conversion frequency of somatic embryos, MS medium supplemented with 3% sucrose was used far germination. Variability was obtained between sodium alginate and calcium cloride concentration with respect to % germination. Plant regeneration frequency decreased with an increase in sodium alginate concentration above 3%. 3% sodium alginate and 75 mM calcium chloride gave the best conversion and optimum bead rigidity and shape. In vitro and in vivo conversion of naked and encapsulated embryos were compared.Int Soc Hort Sci, Spanish Soc Hort Sci, Consejo Super Invest Cient, Junta Andalucia, Consejeria Agr & Pesca, Plan Nacl Invest Cientif & Desarrollo Tecnol, Excma, Diputac Malaga, British Council, Cajas Rurales Granada & Malaga, Unicaja, MSD Agvet, Haifa Chem, Ferquis

Some cytological observations on pollen grains of wild poppy anemone varieties from Turkey

There are four native varieties of poppy anemone, namely, Anemone coronaria var. coccinea (red flowered), Anemone coronaria var. alba (white flowered) Anemone coronaria var. rosea (pink flowered) and Anemone coronaria var. cyanea (purple flowered) in Turkey. In this study, flowers of the four varieties were used as plant material in order to determine viability, germination rate and production of the pollen grains. For the determination of pollen viability, TTC (Triphenyl tetrazolium chloride) and FDA (Fluorescein diacetate) were used. In vitro pollen germination was performed using an agar method and pollen production was counted with the hemacytometric lam method. The maximum pollen viability rate was obtained from the violet-flowered variety in TTC whilst the difference was not found to be statistically important among the varieties when FDA was used. However, viability values of FDA were higher than that of TTC. As far as the pollen germination was concerned, the maximum germination was observed from the red-flowered variety in a medium with 1% agar + 25% sucrose + 150 ppm H3BO3. Additionally, the average number of anthers, the pollen production per anther and per flower, morphological homogeneity level and dimensions of pollen were determined for each variety

Response of Turkish melon genotypes to Fusarium oxysporum f. sp melonis race 1 determined by inoculation tests and RAPD markers

WOS: 000251999200006The response to Fusarium oxysporum f. sp. melonis (F.o.m.) race 1 of 79 Cucumis melo L. genotypes - 56 accessions collected from different parts of Turkey, 5 local cultivars and 18 foreign genotypes - was determined by using pathogenicity tests and RAPD markers (E07 and G17). For pathogenicity tests, at least 15 individuals of each melon genotype were artificially inoculated by either conidia suspension culture or sand culture of F.o.m. race 1. Melon seedlings were evaluated on a 0-3 scale for the presence of vascular browning two weeks after inoculation with conidia suspension culture resp. four weeks after inoculation with sand culture. The disease incidence on the 79 melon genotypes ranged from 0 to 100%. Seven melon genotypes were resistant (CU129, CU258, T8, Y9, Y10, Y15, and Y63), three melon genotypes (CU280, CU309, and T1) were heterogeneous, and all the others were susceptible to F.o.m. race 1 based on disease incidence and vascular browning. The success of the E07 marker in detecting susceptible genotypes was higher than that of G17; the mistmatch ratios of E07 and G17 markers were 5.06 and 58.23%, respectively

Ampelographic and molecular diversity among grapevine (Vitis spp.) cultivars

This study presents the ampelographic and molecular characterization of 44 grapevine cultivars. Ampelographic data were obtained during two vegetation periods using the latest version of the descriptors. Based on the mean values transformed by the method indicated in IBPGR publications, a dendrogram was constructed. ISSR analysis was also employed to characterize the genotypes at the DNA level. Twenty primers, selected on the basis of their discriminating potential, generated a total of 157 bands, of which 140 were polymorphic. The dendrograms constructed by the two approaches were largely similar in both the clustering position and divergence of varietal groups. The least distance was observed between Yuvarlak Cekirdeksiz and Superior Seedless. The clustering position of cultivars throughout the dendrograms was basically related to the genetic distances and main uses, as well as to geographic origins

Callus initiation for indirect pollen embryogenesis in Anemone coronaria

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

Response of Turkish melon genotypes to Fusarium oxysporum f. sp. melonis race 1 determined by inoculation tests and RAPD markers

The response to Fusarium oxysporum f. sp. melonis (F.o.m.) race 1 of 79 Cucumis melo L. genotypes - 56 accessions collected from different parts of Turkey, 5 local cultivars and 18 foreign genotypes - was determined by using pathogenicity tests and RAPD markers (E07 and G17). For pathogenicity tests, at least 15 individuals of each melon genotype were artificially inoculated by either conidia suspension culture or sand culture of F.o.m. race 1. Melon seedlings were evaluated on a 0-3 scale for the presence of vascular browning two weeks after inoculation with conidia suspension culture resp. four weeks after inoculation with sand culture. The disease incidence on the 79 melon genotypes ranged from 0 to 100 %. Seven melon genotypes were resistant (CU129, CU258, T8, Y9, Y10, Y15, and Y63), three melon genotypes (CU280, CU309, and T1) were heterogeneous, and all the others were susceptible to F.o.m. race 1 based on disease incidence and vascular browning. The success of the E07 marker in detecting susceptible genotypes was higher than that of G17; the mistmatch ratios of E07 and G17 markers were 5.06 and 58.23 %, respectively. © Verlag Eugen Ulmer KG