Papers by Angela Carra, Maurizio Sajeva, Loredana Abbate, Mirco Siragusa, Ranjith Pathirana, Francesco Carimi
Plants, 2021
This study is the first approach to in vitro asymbiotic germination of two species of Sicilian th... more This study is the first approach to in vitro asymbiotic germination of two species of Sicilian threatened terrestrial orchids, Anacamptis longicornu and Ophrys panormitana. Seeds were collected in the wild and cultured in two different media—Orchimax medium (OM) and Murashige and Skoog (MS)—and exposed to different photoperiods and temperatures to evaluate the best conditions for the specific stages of development. The germination of A. longicornu was very high on OM (95.5%) and lower on MS medium (21.4%), whereas O. panormitana germinated only on OM medium, with significantly lower percentages (12.0%), compared with A. longicornu. This difference is caused by variation in quality and quantity of nutrients used, primarily by nitrogen source. The results show that temperature and photoperiod widely affect seed germination and development. Although further investigations on asymbiotic and symbiotic germination are needed for the improvement of conservation of Mediterranean terrestrial...
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Synthetic Seeds, 2019
Although grapevine (Vitis spp.) is one of the most ancient and important fruit crops, there is no... more Although grapevine (Vitis spp.) is one of the most ancient and important fruit crops, there is no concerted international effort to conserve its genetic resources, which are estimated to consist of 10–14,000 cultivars. Synthetic seed technology offers opportunities to conserve clonal genetic resources either in the form of quiescent somatic embryos or as encapsulated regenerable somatic tissue. Since the first report of somatic embryogenesis in grapevine in 1976, much research has been conducted into synchronising the process, maturation, dehydration, encapsulation and testing longevity under cold storage. Since the development of vitrification-based cryopreservation methods, both somatic embryos and other somatic tissue with meristematic regions have been used in cryopreservation experiments, and methods have been optimised to reach post-thaw regeneration percentages that satisfy gene bank standards for implementing cryopreservation. Nevertheless, improved protocols for ‘difficult’ genotypes are still needed for induction of somatic embryos and synchronising their formation, maturation and germination, as well as cryopreservation. As a result of these difficulties, conservation by cryopreservation has progressed using encapsulated shoot tips or axillary buds of tissue culture plants. Some vitrification-based methods use a droplet of vitrification solution to protect the shoot tips on an aluminium strip allowing faster freezing of tissue, an important factor for post-cryo-survival. The novel V cryo-plate method combines the advantages of both encapsulating the shoot tips in alginate beads that then adhere to the aluminium of the V cryo-plate, meaning manipulations can be performed easily, and the high thermal conductivity of aluminium speeding processed of freezing and thawing. Cryopreservation of somatic embryos has been suggested as a way to conserve the diversity of wild V. vinifera ssp. sylvestris, and limited results obtained to date are promising.
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Cells, 2021
Polyploidy plays an important role in plant adaptation to biotic and abiotic stresses. Alteration... more Polyploidy plays an important role in plant adaptation to biotic and abiotic stresses. Alterations of the ploidy in grapevine plants regenerated via somatic embryogenesis (SE) may provide a source of genetic variability useful for the improvement of agronomic characteristics of crops. In the grapevine, the SE induction process may cause ploidy changes without alterations in DNA profile. In the present research, tetraploid plants were observed for 9.3% of ‘Frappato’ grapevine somatic embryos regenerated in medium supplemented with the growth regulators β-naphthoxyacetic acid (10 µM) and N6-benzylaminopurine (4.4 µM). Autotetraploid plants regenerated via SE without detectable changes in the DNA profiles were transferred in field conditions to analyze the effect of polyploidization. Different ploidy levels induced several anatomical and morphological changes of the shoots and mature leaves. Alterations have been also observed in stomata. The length and width of stomata of tetraploid l...
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Plant Cell, Tissue and Organ Culture (PCTOC), 2019
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The plant pathology journal, 2017
Prospecting of local grapevine (Vitis vinifera L.) germplasm revealed that Tunisia possesses a ri... more Prospecting of local grapevine (Vitis vinifera L.) germplasm revealed that Tunisia possesses a rich patrimony which presents diversified organoleptic characteristics. However, viral diseases seriously affect all local grapevine cultivars which risk a complete extinction. Sanitation programs need to be established to preserve and exploit, as a gene pool, the Tunisian vineyards areas. The presence of the Grapevine leafroll associated virus-3 (GLRaV-3), Grapevine stem pitting associated virus (GRSPaV) and Grapevine virus A (GVA), were confirmed in a Tunisian grapevine cultivar using serological and molecular analyses. The association between GRSPaV and GVA viruses induces more rugose wood symptoms and damages. For this reason the cleansing of the infected cultivar is highly advisable. Direct and recurrent somatic embryos of cv. 'Hencha' were successfully induced from filament, when cultured on Chée and Pool (1987). based-medium, enriched with 2 mg 1-1 of 2,4-dichlorophenoxyacet...
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Protoplasma, Jan 17, 2017
A new protocol for in vitro regeneration through direct somatic embryogenesis for two muskmelon c... more A new protocol for in vitro regeneration through direct somatic embryogenesis for two muskmelon cultivars (Cucumis melo L., "Mashhadi" and "Eivanaki") is reported. Somatic embryos were obtained culturing 4- and 8-day-old cotyledons, seeds, and hypocotyls on Murashige and Skoog medium supplemented with three different hormonal combinations never tested so far for melon (naphthoxyacetic acid (NOA) + thidiazuron (TDZ), NOA + 6-banzylaminopurine (BAP), and 2,4-dichlorophenoxyacetic acid (2,4-D) + N-(2-chloro-4-pyridyl)-N'-phenylurea (4-CPPU)). Results were compared with those obtained when explants were cultivated in the presence of 2,4-D + BAP, previously used on melon. Embryogenesis occurred more successfully in 4-day-old cotyledons and seeds than hypocotyls and 8-day-old cotyledons. The best result was achieved with NOA + BAP. Genotypes significantly affected embryogenesis. The number of embryos in "Eivanaki" was significantly higher than that in &qu...
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Plant Biosystems - An International Journal Dealing with all Aspects of Plant Biology, 2017
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European Journal of Plant Pathology, 2016
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Acta Horticulturae, 2016
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Scientia Horticulturae, 2016
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L’obiettivo è quello di identificare più tipologie di pectina aventi caratteristiche chimico-fisi... more L’obiettivo è quello di identificare più tipologie di pectina aventi caratteristiche chimico-fisiche, di purezza e trasparenza tali da poter essere applicate, da sole o in miscela con altre sostanze, in forma di gel. Tale studio permetterà di identificare quelle miscele che meglio si prestano ad essere utilizzate come film edibili da applicare sui prodotti ittici, con lo scopo di garantirne il mantenimento della shelf life.
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Economic Botany, 1992
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Plant Growth Regulation, 2012
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Proc. 20th Biennial Meeting of the International Association for Plant Biotechnology, Waiheke Island, New Zealand, Feb 25, 2013
Cryotherapy of grapevine (Vitis spp.) to remove leafroll viruses from infected plants
Ranjith Pa... more Cryotherapy of grapevine (Vitis spp.) to remove leafroll viruses from infected plants
Ranjith Pathirana1, Angela Carra2, Andrew McLachlan1, Duncan Hedderley1, Bart Panis3, Francesco Carimi2
1 The New Zealand Institute for Plant & Food Research Ltd, Private Bag 11600, Palmerston North 4442, New Zealand
2 Consiglio Nazionale delle Ricerche, Istituto di Genetica Vegetale, U.O.S. di Palermo, Corso Calatafimi 414, I-90129 Palermo, Italia
3Laboratory of Tropical Crop Improvement, Department of Biosciences, Katholieke Universiteit Leuven (K. U. Leuven), 3001 Leuven, Belgium.
Infections by viruses and other pathogens are a threat to the grapevine industry. A robust method for removing all microorganisms from infected tissue is important for cultivar imports, germplasm maintenance and to produce grafted material for the industry. The demonstration of virus and phytoplasma eradication by methods used in cryopreservation of plants has led to the establishment of cryotherapy: a new method for cleaning infected plant material of vegetatively propagated species. Vitrification-based cryopreservation techniques have been shown to be the most adaptable across species. In droplet vitrification, plant tissue pre-treated with vitrification solution is placed on aluminium foil in a droplet of vitrification solution and directly immersed in liquid nitrogen. Only highly cytoplasmic, non-vacuolar meristematic cells survive the freezing process because only these cells can tolerate the dehydration caused by the vitrification solution; therefore, cryotherapy can be considered a precise method of meristem culture. Using in vitro-sourced apical and axillary buds as explants, we developed a droplet vitrification protocol and plants were regenerated from cryopreserved tissues of the 13 Vitis genotypes used, although regeneration ability was influenced by genotype. Growing plantlets in salicylic acid-supplemented media followed by pre-treatment of explants sourced from those in high sucrose solutions increased regeneration rates after cryopreservation, including that of the previously recalcitrant 41B rootstock. Regeneration from virus-infected plant material was generally poorer than for “clean” plants. The regeneration rates achieved are such that for most varieties cryopreservation of 6 – 15 explants will be sufficient to regenerate at least one plant at 95% probability, providing a cost-effective way of maintaining clonal plant material that avoids the threats associated with field-based collections. To test the suitability of cryotherapy for virus eradication, we used Lakemont Seedless and Chardonnay infected with Grapevine leafroll associated-virus-3 (GLRaV-3, an Ampelovirus), Sauvignon blanc and Pinot gris infected with Grapevine leafroll associated virus-2 (a Closterovirus), and another clone of Sauvignon blanc infected with both Grapevine leafroll associated virus -1 (an Ampelovirus) and GLRaV-3. Plants regenerated after cryotherapy tested negative (DAS-ELISA) for all three viruses, whereas untreated control plants tested positive. Droplet vitrification has the potential to be a novel and precise tool for virus eradication and establishment of high-health grapevine germplasm collections.
Acknowledgement: This work was funded by New Zealand Winegrowers (NZW 10-107 – “Cryopreserved grapevine: a new way to maintain high-health germplasm and cultivar imports with less rigorous quarantine”) and was part of COST Action 871. Work in Italy was part of PO-FESR Linea di intervento 4.1.1.1: “Recupero e valorizzazione dei vitigni tradizionali Siciliani”.
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Plant Science, 2001
Vegetative propagation of cuttings is a widespread method to multiplicate plants. Adventitious ro... more Vegetative propagation of cuttings is a widespread method to multiplicate plants. Adventitious root formation is a key step in vegetative propagation and considerable progress has recently been made in understanding root formation. But, in spite of the ...
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Plant Growth Regulation, 2003
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Papers by Angela Carra, Maurizio Sajeva, Loredana Abbate, Mirco Siragusa, Ranjith Pathirana, Francesco Carimi
Ranjith Pathirana1, Angela Carra2, Andrew McLachlan1, Duncan Hedderley1, Bart Panis3, Francesco Carimi2
1 The New Zealand Institute for Plant & Food Research Ltd, Private Bag 11600, Palmerston North 4442, New Zealand
2 Consiglio Nazionale delle Ricerche, Istituto di Genetica Vegetale, U.O.S. di Palermo, Corso Calatafimi 414, I-90129 Palermo, Italia
3Laboratory of Tropical Crop Improvement, Department of Biosciences, Katholieke Universiteit Leuven (K. U. Leuven), 3001 Leuven, Belgium.
Infections by viruses and other pathogens are a threat to the grapevine industry. A robust method for removing all microorganisms from infected tissue is important for cultivar imports, germplasm maintenance and to produce grafted material for the industry. The demonstration of virus and phytoplasma eradication by methods used in cryopreservation of plants has led to the establishment of cryotherapy: a new method for cleaning infected plant material of vegetatively propagated species. Vitrification-based cryopreservation techniques have been shown to be the most adaptable across species. In droplet vitrification, plant tissue pre-treated with vitrification solution is placed on aluminium foil in a droplet of vitrification solution and directly immersed in liquid nitrogen. Only highly cytoplasmic, non-vacuolar meristematic cells survive the freezing process because only these cells can tolerate the dehydration caused by the vitrification solution; therefore, cryotherapy can be considered a precise method of meristem culture. Using in vitro-sourced apical and axillary buds as explants, we developed a droplet vitrification protocol and plants were regenerated from cryopreserved tissues of the 13 Vitis genotypes used, although regeneration ability was influenced by genotype. Growing plantlets in salicylic acid-supplemented media followed by pre-treatment of explants sourced from those in high sucrose solutions increased regeneration rates after cryopreservation, including that of the previously recalcitrant 41B rootstock. Regeneration from virus-infected plant material was generally poorer than for “clean” plants. The regeneration rates achieved are such that for most varieties cryopreservation of 6 – 15 explants will be sufficient to regenerate at least one plant at 95% probability, providing a cost-effective way of maintaining clonal plant material that avoids the threats associated with field-based collections. To test the suitability of cryotherapy for virus eradication, we used Lakemont Seedless and Chardonnay infected with Grapevine leafroll associated-virus-3 (GLRaV-3, an Ampelovirus), Sauvignon blanc and Pinot gris infected with Grapevine leafroll associated virus-2 (a Closterovirus), and another clone of Sauvignon blanc infected with both Grapevine leafroll associated virus -1 (an Ampelovirus) and GLRaV-3. Plants regenerated after cryotherapy tested negative (DAS-ELISA) for all three viruses, whereas untreated control plants tested positive. Droplet vitrification has the potential to be a novel and precise tool for virus eradication and establishment of high-health grapevine germplasm collections.
Acknowledgement: This work was funded by New Zealand Winegrowers (NZW 10-107 – “Cryopreserved grapevine: a new way to maintain high-health germplasm and cultivar imports with less rigorous quarantine”) and was part of COST Action 871. Work in Italy was part of PO-FESR Linea di intervento 4.1.1.1: “Recupero e valorizzazione dei vitigni tradizionali Siciliani”.
Ranjith Pathirana1, Angela Carra2, Andrew McLachlan1, Duncan Hedderley1, Bart Panis3, Francesco Carimi2
1 The New Zealand Institute for Plant & Food Research Ltd, Private Bag 11600, Palmerston North 4442, New Zealand
2 Consiglio Nazionale delle Ricerche, Istituto di Genetica Vegetale, U.O.S. di Palermo, Corso Calatafimi 414, I-90129 Palermo, Italia
3Laboratory of Tropical Crop Improvement, Department of Biosciences, Katholieke Universiteit Leuven (K. U. Leuven), 3001 Leuven, Belgium.
Infections by viruses and other pathogens are a threat to the grapevine industry. A robust method for removing all microorganisms from infected tissue is important for cultivar imports, germplasm maintenance and to produce grafted material for the industry. The demonstration of virus and phytoplasma eradication by methods used in cryopreservation of plants has led to the establishment of cryotherapy: a new method for cleaning infected plant material of vegetatively propagated species. Vitrification-based cryopreservation techniques have been shown to be the most adaptable across species. In droplet vitrification, plant tissue pre-treated with vitrification solution is placed on aluminium foil in a droplet of vitrification solution and directly immersed in liquid nitrogen. Only highly cytoplasmic, non-vacuolar meristematic cells survive the freezing process because only these cells can tolerate the dehydration caused by the vitrification solution; therefore, cryotherapy can be considered a precise method of meristem culture. Using in vitro-sourced apical and axillary buds as explants, we developed a droplet vitrification protocol and plants were regenerated from cryopreserved tissues of the 13 Vitis genotypes used, although regeneration ability was influenced by genotype. Growing plantlets in salicylic acid-supplemented media followed by pre-treatment of explants sourced from those in high sucrose solutions increased regeneration rates after cryopreservation, including that of the previously recalcitrant 41B rootstock. Regeneration from virus-infected plant material was generally poorer than for “clean” plants. The regeneration rates achieved are such that for most varieties cryopreservation of 6 – 15 explants will be sufficient to regenerate at least one plant at 95% probability, providing a cost-effective way of maintaining clonal plant material that avoids the threats associated with field-based collections. To test the suitability of cryotherapy for virus eradication, we used Lakemont Seedless and Chardonnay infected with Grapevine leafroll associated-virus-3 (GLRaV-3, an Ampelovirus), Sauvignon blanc and Pinot gris infected with Grapevine leafroll associated virus-2 (a Closterovirus), and another clone of Sauvignon blanc infected with both Grapevine leafroll associated virus -1 (an Ampelovirus) and GLRaV-3. Plants regenerated after cryotherapy tested negative (DAS-ELISA) for all three viruses, whereas untreated control plants tested positive. Droplet vitrification has the potential to be a novel and precise tool for virus eradication and establishment of high-health grapevine germplasm collections.
Acknowledgement: This work was funded by New Zealand Winegrowers (NZW 10-107 – “Cryopreserved grapevine: a new way to maintain high-health germplasm and cultivar imports with less rigorous quarantine”) and was part of COST Action 871. Work in Italy was part of PO-FESR Linea di intervento 4.1.1.1: “Recupero e valorizzazione dei vitigni tradizionali Siciliani”.