The effect of sucrose concentration on the micropropagation of axillary shoots of willow was investigated. The following factors were examined: the culture system (semisolid medium in glass jars versus liquid medium in temporary immersion... more
The effect of sucrose concentration on the micropropagation of axillary shoots of willow was investigated. The following factors were examined: the culture system (semisolid medium in glass jars versus liquid medium in temporary immersion bioreactors), the type of explant (apical and basal sections), the frequency of immersion, and CO2 enrichment. Shoots and leaf growth were significantly higher in RITA® bioreactors than in the jars for all the sucrose treatments. Apical or basal sections of willow cultured in bioreactors under high light intensity (150 µmol m−2 s−1) and ventilated six times a day with CO2-enriched air were successfully proliferated without sucrose, whereas shoots cultured in jars did not proliferate well if sucrose concentration was 0.5% or lower. More roots were formed when sucrose was added to the medium. Shoots cultured in bioreactors were successfully acclimatized irrespective of the sucrose treatment and the root biomass when transferred to ex vitro conditions...
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The genus Castanea includes several tree species that are relevant because of their geographical extension and their multipurpose character, that includes nut and timber production. However, commercial exploitation of the trees is... more
The genus Castanea includes several tree species that are relevant because of their geographical extension and their multipurpose character, that includes nut and timber production. However, commercial exploitation of the trees is hindered by several factors, particularly by their limited regeneration ability. Regardless of recent advances, there exists a serious limitation for the propagation of elite genotypes of chestnut due to decline of rooting ability as the tree ages. In the present review, we summarize the research developed in this genus during the last three decades concerning the formation of adventitious roots (ARs). Focusing on cuttings and in vitro microshoots, we gather the information available on several species, particularly C. sativa, C. dentata and the hybrid C.sativa × C. crenata, and analyze the influence of several factors on the achievements of the applied protocols, including genotype, auxin treatment, light regime and rooting media. We also pay attention to...
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The present study investigated how auxin concentration and the method of application affected the formation of adventitious roots in microshoots of chestnut (Castanea sativa) and oak (Quercus robur). The activity of two urea derivatives... more
The present study investigated how auxin concentration and the method of application affected the formation of adventitious roots in microshoots of chestnut (Castanea sativa) and oak (Quercus robur). The activity of two urea derivatives (2, 3-MDPU and 3, 4-MDPU) was also evaluated. Microshoots were derived from basal sprouts of two mature chestnut trees (P1 and P2) and one adult oak genotype (Sainza). In chestnut, rooting percentage was positively affected by auxin in a dose-dependent manner, particularly in shoots treated with the hormone for 24 h. The effect of auxin on rooting also differed depending on the application method. In shoots treated for 24 h, the highest concentration of auxin produced the healthiest rooted plantlets, in terms of the root system and shoot quality. By contrast, in shoots treated by the basal quick-dip method, the shoot quality was best at the lowest auxin concentration. The effect of urea derivatives on the root system depended on the species as well a...
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Adventitious rooting of cuttings is a complex developmental process in forest species, with several exogenous and endogenous factors influencing the outcome of the process. In this study we applied an in vitro working system, comprising... more
Adventitious rooting of cuttings is a complex developmental process in forest species, with several exogenous and endogenous factors influencing the outcome of the process. In this study we applied an in vitro working system, comprising two lines of microshoots with the same genotype but at a different ontogenetic stages, in two different tree species (chestnut and oak). We analyzed the expression of a gene encoding an AP2/ERF transcription factor from group VII in the initial hours of the adventitious rooting induction, both in rooting competent and incompetent microshoots. The analysis revealed that expression of this gene is related to wounding, ontogenetic stage and auxin in a complex and species-specific manner. Putative induction of the gene by auxin was also analyzed in the presence of naphthyl-phthalamic acid (NPA), an auxin transport inhibitor. In situ expression analysis in chestnut relates the gene activity to cambial divisions and root primordia in rooting competent micr...
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This study reports on the cryostorage of embryogenic lines derived from selected mature Quercus robur trees, following application of the PVS2-vitrification based procedure. In seven oak genotypes, embryo recovery levels ranging from... more
This study reports on the cryostorage of embryogenic lines derived from selected mature Quercus robur trees, following application of the PVS2-vitrification based procedure. In seven oak genotypes, embryo recovery levels ranging from 57-92% were obtained when 4-6 mg embryo clumps were precultured for 3 days on 0.3 M sucrose basal medium, treated with PVS2 solution for 60 min at 24 degrees C, and then immersed in liquid nitrogen (LN). Embryos of six out of seven lines were cryostored for one week and one year and used to evaluate cryopreservation tolerance, germination ability and to assess genetic fidelity by random amplified polymorphic DNA (RAPD) markers. There were no significant differences between the recovery frequencies of samples retrieved from LN after 1 week and 1 year of cryostorage. In five out of six lines, RAPD profiles of cryopreserved somatic embryos and regenerated plantlets were identical to those of the controls. Although polymorphisms were detected in only one cr...
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This study reports on the cryostorage of embryogenic lines derived from selected mature Quercus robur trees, following application of the PVS2-vitrification based procedure. In seven oak genotypes, embryo recovery levels ranging from... more
This study reports on the cryostorage of embryogenic lines derived from selected mature Quercus robur trees, following application of the PVS2-vitrification based procedure. In seven oak genotypes, embryo recovery levels ranging from 57-92% were obtained when 4-6 mg embryo clumps were precultured for 3 days on 0.3 M sucrose basal medium, treated with PVS2 solution for 60 min at 24 degrees C, and then immersed in liquid nitrogen (LN). Embryos of six out of seven lines were cryostored for one week and one year and used to evaluate cryopreservation tolerance, germination ability and to assess genetic fidelity by random amplified polymorphic DNA (RAPD) markers. There were no significant differences between the recovery frequencies of samples retrieved from LN after 1 week and 1 year of cryostorage. In five out of six lines, RAPD profiles of cryopreserved somatic embryos and regenerated plantlets were identical to those of the controls. Although polymorphisms were detected in only one cr...
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European chestnut is currently being ravaged by ink and blight diseases. Cryopreservation can constitute a complementary approach to field collections for long-term conservation of germplasm. A two-step vitrification methodology (LS-PVS2)... more
European chestnut is currently being ravaged by ink and blight diseases. Cryopreservation can constitute a complementary approach to field collections for long-term conservation of germplasm. A two-step vitrification methodology (LS-PVS2) was successful in cryopreserving micropropagated Castanea sativa shoot apices so that they could be recovered for plant regeneration. In order to optimize shoot performance after cryopreservation different parameters, such as explant type, duration of PVS2 treatment, sucrose content in preculture medium, loading pre-treatment, and sequence of media used after thawing, were studied. INTRODUCTION European chestnut (Castanea sativa Mill), seriously damaged by ink disease (caused by the fungi Phytophtora cambivora and P. cinnamomi) and by blight (caused by Cryphonectria parasitica), survives by means of naturally resistant C. sativa trees as well as by hybrids of C. sativa with the Asian species C. mollissima Blume and C. crenata Sieb. and Zucc. (Vieit...
This study reports on the cryostorage of embryogenic lines derived from selected mature Quercus robur trees, following application of the PVS2-vitrification based procedure. In seven oak genotypes, embryo recovery levels ranging from... more
This study reports on the cryostorage of embryogenic lines derived from selected mature Quercus robur trees, following application of the PVS2-vitrification based procedure. In seven oak genotypes, embryo recovery levels ranging from 57-92% were obtained when 4-6 mg embryo clumps were precultured for 3 days on 0.3 M sucrose basal medium, treated with PVS2 solution for 60 min at 24 degrees C, and then immersed in liquid nitrogen (LN). Embryos of six out of seven lines were cryostored for one week and one year and used to evaluate cryopreservation tolerance, germination ability and to assess genetic fidelity by random amplified polymorphic DNA (RAPD) markers. There were no significant differences between the recovery frequencies of samples retrieved from LN after 1 week and 1 year of cryostorage. In five out of six lines, RAPD profiles of cryopreserved somatic embryos and regenerated plantlets were identical to those of the controls. Although polymorphisms were detected in only one cr...
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ABSTRACT
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ABSTRACT