Development of a Feasible and Efficient In Vitro Rescue Protocol for Immature <i>Prunus</i> spp. Embryos
The major factors affecting the in vitro immature embryo rescue efficiencies from <i>Prunus persica</i> or <i>P. armeniaca</i> accessions have been identified, along with improving the feasibility. Variations in the woody plant medium (WPM) were used depending on the embryo s...
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MDPI AG
2024-10-01
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| author | Maria Casanovas Elisabet Claveria Ramon Dolcet-Sanjuan |
| author_facet | Maria Casanovas Elisabet Claveria Ramon Dolcet-Sanjuan |
| author_sort | Maria Casanovas |
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| description | The major factors affecting the in vitro immature embryo rescue efficiencies from <i>Prunus persica</i> or <i>P. armeniaca</i> accessions have been identified, along with improving the feasibility. Variations in the woody plant medium (WPM) were used depending on the embryo size. Embryos less than 5 mm long were cultured in WPM supplemented with 1 μM BAP and 1 μM GA<sub>3</sub>, while embryos bigger than 5 mm long were cultured in hormone-free medium, with or without vermiculite. The environmental in vitro culture conditions consisted of three phases: a (I) stratification at 4 °C during a 3- to 5-month-long period in the dark, followed by (II) growth of germinated embryos at 14 °C for a 4-week-long period, with 12 h light a day, which favors plantlet development, and finally, (III) growth at 24 °C, with 16 h light a day, until the plantlets were acclimatized in the greenhouse. The germination of smaller embryos, at the end of phase I, ranged from 82.2% to 22.1% for apricots and flat peaches, respectively, whereas for bigger embryos, the germination varied from 97.3% to 53.2% for the same species. The embryo germination for peaches and nectarines ranged from 40.1% to 30.3% for smaller embryos, and from 91.9% to 63.0% for bigger embryos. Endo- and epiphytic contamination, affecting from 7.4% to 52.9% of cultured embryos, depending on the fruit type and conservation conditions, and the capacity to acclimate to soil conditions, ranging from 50.4% to 93.2%, were the two most important factors influencing the protocol’s efficiency and feasibility. Considering the overall efficiencies, expressed as hardened plants transferred to field plots over clean uncontaminated embryo, the values ranged from 55.8% for nectarines, 54.0% for peaches, 45.6% for apricots, and 23.3% for flat fruits. The addition of vermiculite to the culture medium significantly improved the plantlet development, avoiding subculture to fresh medium when an extension of phase III was required before acclimatization. Compared to laboratory glassware, the use of food glass containers with air-permeable sealing film, along with vermiculite-containing medium, significantly reduced the costs when handling the large number of embryos required for breeding programs. |
| format | Article |
| id | doaj-art-01ffac4e01e54c02b5f9979fae456a0c |
| institution | Kabale University |
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| spelling | doaj-art-01ffac4e01e54c02b5f9979fae456a0c2024-11-08T14:39:02ZengMDPI AGPlants2223-77472024-10-011321295310.3390/plants13212953Development of a Feasible and Efficient In Vitro Rescue Protocol for Immature <i>Prunus</i> spp. EmbryosMaria Casanovas0Elisabet Claveria1Ramon Dolcet-Sanjuan2IRTA, Fruitcentre, Plant In Vitro Culture Laboratory, Fruticulture Program, Parc AgroBiotech, 25003 Lleida, SpainIRTA, Torre Marimon, Serveis Corporatius de Proximitat, Caldes de Montbui, 08140 Barcelona, SpainIRTA, Fruitcentre, Plant In Vitro Culture Laboratory, Fruticulture Program, Parc AgroBiotech, 25003 Lleida, SpainThe major factors affecting the in vitro immature embryo rescue efficiencies from <i>Prunus persica</i> or <i>P. armeniaca</i> accessions have been identified, along with improving the feasibility. Variations in the woody plant medium (WPM) were used depending on the embryo size. Embryos less than 5 mm long were cultured in WPM supplemented with 1 μM BAP and 1 μM GA<sub>3</sub>, while embryos bigger than 5 mm long were cultured in hormone-free medium, with or without vermiculite. The environmental in vitro culture conditions consisted of three phases: a (I) stratification at 4 °C during a 3- to 5-month-long period in the dark, followed by (II) growth of germinated embryos at 14 °C for a 4-week-long period, with 12 h light a day, which favors plantlet development, and finally, (III) growth at 24 °C, with 16 h light a day, until the plantlets were acclimatized in the greenhouse. The germination of smaller embryos, at the end of phase I, ranged from 82.2% to 22.1% for apricots and flat peaches, respectively, whereas for bigger embryos, the germination varied from 97.3% to 53.2% for the same species. The embryo germination for peaches and nectarines ranged from 40.1% to 30.3% for smaller embryos, and from 91.9% to 63.0% for bigger embryos. Endo- and epiphytic contamination, affecting from 7.4% to 52.9% of cultured embryos, depending on the fruit type and conservation conditions, and the capacity to acclimate to soil conditions, ranging from 50.4% to 93.2%, were the two most important factors influencing the protocol’s efficiency and feasibility. Considering the overall efficiencies, expressed as hardened plants transferred to field plots over clean uncontaminated embryo, the values ranged from 55.8% for nectarines, 54.0% for peaches, 45.6% for apricots, and 23.3% for flat fruits. The addition of vermiculite to the culture medium significantly improved the plantlet development, avoiding subculture to fresh medium when an extension of phase III was required before acclimatization. Compared to laboratory glassware, the use of food glass containers with air-permeable sealing film, along with vermiculite-containing medium, significantly reduced the costs when handling the large number of embryos required for breeding programs.https://www.mdpi.com/2223-7747/13/21/2953acclimatizationapricotembryo rescueflat fruitsnectarinepeach |
| spellingShingle | Maria Casanovas Elisabet Claveria Ramon Dolcet-Sanjuan Development of a Feasible and Efficient In Vitro Rescue Protocol for Immature <i>Prunus</i> spp. Embryos Plants acclimatization apricot embryo rescue flat fruits nectarine peach |
| title | Development of a Feasible and Efficient In Vitro Rescue Protocol for Immature <i>Prunus</i> spp. Embryos |
| title_full | Development of a Feasible and Efficient In Vitro Rescue Protocol for Immature <i>Prunus</i> spp. Embryos |
| title_fullStr | Development of a Feasible and Efficient In Vitro Rescue Protocol for Immature <i>Prunus</i> spp. Embryos |
| title_full_unstemmed | Development of a Feasible and Efficient In Vitro Rescue Protocol for Immature <i>Prunus</i> spp. Embryos |
| title_short | Development of a Feasible and Efficient In Vitro Rescue Protocol for Immature <i>Prunus</i> spp. Embryos |
| title_sort | development of a feasible and efficient in vitro rescue protocol for immature i prunus i spp embryos |
| topic | acclimatization apricot embryo rescue flat fruits nectarine peach |
| url | https://www.mdpi.com/2223-7747/13/21/2953 |
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