Overview of the Success of In Vitro Culture for Ex Situ Conservation and Sustainable Utilization of Endemic and Subendemic Native Plants of Romania
Abstract
:1. Introduction
2. Biotechnological Approaches for Ex Situ Conservation of Red-Listed Endemic and Subendemic Romanian Plants
2.1. Short-Term In Vitro Cultures
2.1.1. Initiation of In Vitro Culture
2.1.2. In Vitro Multiplication Stage
Taxon | Type of Explant | Disinfection of Explants | Initiation Media (Basic + PGRs) | Culture Initiation Success (%) | Multiplication Media (Basic + PGRs) | Time Interval after Inoculation | Multiplication Rate (No. of Shoots/Explant or %) | Shoot Length (cm) | Reference |
---|---|---|---|---|---|---|---|---|---|
Andryala laevitomentosa | petiole | - | MS + 1 mg/L 2,4-D + 1 mg/L Kin | - | MS, Free | - | - | - | [71] |
Andryala laevitomentosa | leaves | 3% NaOCl for 10–15 min | MS + 2 mg/L BAP + 0.2 mg/L IAA | - | MS, Free | - | - | - | [72] |
Astragalus peterfii | seeds | 0.1% HgCl2, NaOCl, Domestos, 98° C2H6O | MS ½, Free | - | MS + 0.2 mg/L NAA + 0.5 mg/L Kin + 40 mg/L AdSO4 | 1 month | 4–5 | 5.4 | [67] |
Astragalus pseudopurpureus | leaves, petiole | 70° C2H6O for 1 min, 0.1% HgCl2 for 6–7 min | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | - | MS + Vit B5 + 2 mg/L BAP + 2 mg/L Kin + 0.3 mg/L NAA | - | 7–10 | - | [45] |
Astragalus pseudopurpureus | leaves | 0.1% HgCl2 for 5–10 min | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | - | MS + Vit B5 + 1 mg/L 2,4-D + 0.5 mg/L Kin | - | - | - | [65] |
Campanula romanica | nodal segments | 70% C2H6O for 20 s, 0.5% NaDCC for 5 min | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | 80 | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | 40 days | 12 | - | [53] |
Centaurea reichenbachii | seeds | 96% C2H6O for 40 s, 10% H2O2 for 15 min | MS ½ + Vit B5, Free | 18 | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | - | 10–15 | - | [44] |
Cerastium transsilvanicum | single node stem fragments | 70° C2H6O for 30 s, HgCl2 for 5–6 min | Macro MS ½ + Micro MS + Vit B5 + 0.1 mg/L BAP + 0.01 mg/L NAA | - | Macro MS ½ + Micro MS + Vit B5 + 0.1 mg/L BAP + 0.01 mg/L NAA | - | 15–20 | - | [47] |
Cerastium transsilvanicum | young shoots | 0.1% HgCl2 for 5–10 min | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | - | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | - | - | - | [65] |
Dianthus callizonus | seeds | 0.1% HgCl2 for 10 min | MS, Free | 80 | - | - | - | - | [64] |
Dianthus callizonus | single node stem fragments | 70° C2H6O for 30 s, 0.1% HgCl2 for 5–6 min | - | - | MS + Vit B5 + 1 mg/L BAP + 1.5 mg/L Kin + 0.1 mg/L NAA + 0.1 mg/L 2,4-D + 0.5 mg/L GA3 | 2 months | 43 | - | [48] |
Dianthus callizonus | seeds | 0.1% HgCl2 for 20 min | MS, Free | - | MS + 1 mg/L BAP + 0.1 mg/L NAA | 10 days | 6 | - | [74] |
Dianthus giganteus subsp. banaticus | vegetative shoots (stolons) with 2–3 compact internodes | 10% Domestos for 5 min, 0.2% HgCl2 for 5 min | MS + 1 mg/L BAP + 1 mg/L NAA | 60–92 | MS + 1 mg/L BAP + 0.1 mg/L NAA | 30 days | 6.75–18.2 | - | [75] |
Dianthus giganteus subsp. banaticus | nodal segments | 5% Domestos, 80% C2H6O | MS + 1 mg/L BAP + 1 mg/L NAA | - | MS + 10 mg/L 2iP + 0.1 mg/L NAA | 110 days | 19 | - | [76] |
Dianthus giganteus subsp. banaticus | seeds | 2.4% Domestos for 10 min | MS + 1 mg/L BAP + 0.5 mg/L IBA | 83.33 | MS + 1 mg/L BAP + 0.1 mg/L NAA | 30 days | 12 | - | [50] |
120 days | 46.3 | 8.35 | |||||||
Dianthus glacialis subsp. gelidus | seeds | 90–100% Domestos for 15 min or 4% H2O2 for 12 h, 99% C2H6O for 1 min, 10% H2O2 for 18 min | MS + Vit B5 | 100 | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | 56 days | 55 | - | [54] |
Dianthus glacialis subsp. gelidus | uninodal fragments | 70° C2H6O for few seconds, 0.1% HgCl2 for 5–6 min | MS + Vit B5 + 1 mg/L BAP + 1.5 mg/L Kin + 0.25 mg/L NAA + 0.5 mg/L GA3 | - | MS + Vit B5 + 1 mg/L BAP + 1.5 mg/L Kin + 0.25 mg/L NAA + 0.5 mg/L GA3 | - | - | - | [42] |
Dianthus glacialis subsp. gelidus | single node stem fragments | 70° C2H6O for 30 s, 0.1% HgCl2 for 5–6 min | - | - | MS + Vit B5 + 1 mg/L BAP + 1.5 mg/L Kin + 0.1 mg/L NAA + 0.1 mg/L 2,4-D + 0.5 mg/L GA3 | 2 months | 43.3 | - | [48] |
Dianthus henteri | seeds | 4% H2O2 for 13 h, 96% C2H6O for 2 min, 10% H2O2 for 19 min | MS + Vit B5, Free | 75 | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | 44 days | 6.9–7.7 | 2.9 | [56] |
83 days | 16 | 3.1 | |||||||
Dianthus henteri | seeds | 2.4% Domestos for 10 min | MS + 1 mg/L BAP + 0.5 mg/L IBA | 52.5 | MS + 1 mg/L BAP + 0.1 mg/L NAA | 30 days | 6.7 | - | [50] |
120 days | 28.4 | 5.88 | |||||||
Dianthus nardiformis | seeds | 70° C2H6O for 30 s, 2.4% Domestos for 10 min or 70° C2H6O for 30 s, 2.5% H2O2 for 16 h, 10% H2O2 for 15 min | MS + Vit B5 + 1 mg/L BAP + 1 mg/L Kin + 0.2 mg/L 2,4-D | - | MS + Vit B5 + 1 mg/L BAP + 1 mg/L Kin + 0.2 mg/L 2,4-D | 2 months | 40–50 | - | [55] |
Dianthus nardiformis | shoots fragments | 70° C2H6O for 30 s, 0.1% HgCl2 for 10 min | MS + Vit B5 + 1 mg/L BAP + 1 mg/L Kin + 0.2 mg/L 2,4-D | - | MS + Vit B5 + 1 mg/L BAP + 1 mg/L Kin + 0.2 mg/L 2,4-D | 40 days | 8.3 | - | [77] |
seeds | 70° C2H6O for 30 s, 2.4% Domestos for 10 min or 70° C2H6O for 30 s, 2.5% H2O2 for 16 h, 10% H2O2 for 15 min | ||||||||
Dianthus spiculifolius | seeds | - | MS | - | MS + 1 mg/L 2iP + 0.1 mg/L NAA | 4 weeks | 10.2 | - | [78] |
Dianthus spiculifolius | apical, uninodal fragments | 100% Domestos for 15 min | MS + Vit B5 + 1 mg/L Kin + 1 mg/L NAA | 30 | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | 28 days | 9.3 | - | [79] |
45 days | 31.8 | - | |||||||
Dianthus spiculifolius | apices with 2–3 nodes from young shoots | 0.2% HgCl2 for 10 min | MS + Vit B5 + 4.44 μM BAP + 5.37 μM NAA | 100 | MS + Vit B5 + 4.44 μM BAP + 0.54 μM NAA | 60 days | 17.6–199.2 | - | [80] |
Dianthus spiculifolius | shoot tips | 20% Domestos for 10 min | MS + Vit B5 + 4.44 µM BAP + 5.37 µM NAA | 99.3–100 | MS + Vit B5 + 4.44 μM BAP + 0.54 μM NAA | 60 days | 23.6–49.9 | - | [81] |
Dianthus spiculifolius | flower buds, single node stem fragments | 70° C2H6O for 30 s, HgCl2 for 5–6 min | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | - | Macro MS ½ + Micro MS + Vit B5 + 1 mg/L + BAP + 0.1 mg/L NAA | - | 30 | - | [47] |
Macro MS ½ + Micro MS + Vit B5 + 0.1 mg/L BAP + 0.01 mg/L NAA | - | 30 | - | ||||||
Dianthus spiculifolius | single node stem fragments | 70° C2H6O for 30 s, 0.1% HgCl2 for 5–6 min | - | - | MS + Vit B5 + 1 mg/L BAP + 1.5 mg/L Kin + 0.1 mg/L NAA + 0.1 mg/L 2,4-D + 0.5 mg/L GA3 | 2 months | 83.3 | - | [48] |
Dianthus spiculifolius | knot, apex | 3–10% NaOCl for 8–30 min | MS + 2 mg/L IBA + 2 mg/L BAP + 40 mg/L AdSO4 | 80 | MS + 2 mg/L IBA + 2 mg/L BAP + 40 mg/L AdSO4 | 2 months | 90 | - | [26] |
Dianthus spiculifolius | seeds | 2.4% Domestos for 10 min | MS + 1 mg/L BAP + 0.5 mg/L IBA | 70.83 | MS + 1 mg/L BAP + 0.1 mg/L NAA | 30 days | 7.8 | - | [50] |
120 days | 32.6 | 4.98 | |||||||
Dianthus trifasciculatus subsp. parviflorus | single node stem fragments | 70° C2H6O for 30 s, 0.1% HgCl2 for 10 min | MS + Vit B5 + 1 mg/L BAP + 1 mg/L Kin + 0.2 mg/L NAA | - | MS ½ + Vit B5 + 0.01 mg/L NAA | 30 days | 13.33 | - | [51] |
Doronicum carpaticum | leaves, petiole | 700 C2H6O for few seconds, 0.1% HgCl2 for 4–5 min | MS + Vit B5 + 1 mg/L BAP + 0.25 mg/L NAA + 1.5 mg/L Kin + 0.5 mg/L GA3 | - | MS + Vit B5 + 1 mg/L BAP + 0.2 mg/L NAA + 1 mg/L Kin + 0.08 g/L Ad | - | 2–5 | - | [42] |
MS + Vit B5 + 1 mg/L BAP + 0.2 mg/L NAA + 1 mg/L Kin + 0.08 g/L Ad | - | ||||||||
Gypsophila petraea | cotyledon, hypocotyl, root | - | MS + 1 mg/L Kin + 2 mg/L Ad + 0.1, 0.25, 0.5 or 1 mg/L 2,4-D | - | MS + 1 mg/L Kin + 2 mg/L Ad + 0.1 mg/L 2,4-D | - | - | - | [82] |
Gypsophila petraea | seeds | 700 C2H6O for 30 s, 0.1% HgCl2 for 10 min | MS + Vit B5 + 1 mg/L BAP + 0.5 mg/L NAA + 1 mg/L Kin | 100 | MS + Vit B5 + 1 mg/L BAP + 0.5 mg/L NAA + 1 mg/L Kin | - | 5 | - | [49] |
Hieracium pojoritense | leaves, petiole, floral bud | 700 C2H6O for 1 min, 0.1% HgCl2 for 5–7 min | MS + Vit B5 + 15 mg/L ascorbic acid + 1 mg/L BAP + 0.1 mg/L NAA | - | MS + Vit B5 + 15 mg/L ascorbic acid + 1 mg/L BAP + 0.1 mg/L NAA | 21 days | - | - | [46] |
Hieracium pojoritense | foliar cuttings | 0.1% HgCl2 for 5–10 min | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | - | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | 25 days | 20–40 | - | [65] |
Klasea bulgarica | seeds | 70% C2H6O for 40 s, 0.5% NaDCC for 5 min | MS | 87 | MS + 2.5 mg/L 2,4 D + 0.5 mg/L Kin | 30 days | - | - | [52] |
MS + 1 mg/L NAA + 0.1 mg/L BAP | 40 days | - | - | ||||||
MS + 0.1 mg/L NAA + 1 mg/L BAP + 2 mg/L GA3 | 50 days | - | - | ||||||
Moehringia jankae | seeds | 70% C2H6O for 30 s, 2.7% NaDCC for 10 min + 2–3 drops of Tween 20 | MS + 0.5 g/L active charcoal + 5 mg/L GA3 | 22 | MS + Vit B5 + 0.22 µM TDZ + 0.49 µM IBA | 2 months | 13 | 4.65 | [43] |
MS + Vit B5 + 4.4 µM BAP + 0.49 µM IBA | 2 months | 14 | 4.95 | ||||||
MS + Vit B5 + 4.5 µM ZEA + 0.49 µM IBA | 2 months | 13 | 3.93 | ||||||
Papaver alpinum subsp. corona-sancti-stephani | seeds | 0.1% HgCl2 for 10 min | MS | 10 | - | - | - | - | [64] |
Papaver alpinum subsp. corona-sancti-stephani | seeds | 70% C2H6O for 30 s, 0.01% HgCl2 for 10 min | MS + Vit B5 | - | MS + Vit B5 + 1 mg/L BAP + 1 mg/L Kin + 0.2 mg/L NAA | 3 months | 35 | - | [73] |
Saussurea porcii | seeds | 96% C2H6O + Tween-80 (2 ± 0.1 min) + NaOCl enriched with active chlorine, “Bilyzna” (TUU6-05743160.001-93) in the ratio 1 × 4 (15 ± 0.1 min) | MS + Fe2+ in chelated form + 60 mg/L cysteine + 0.1 mg/L IAA + 1 mg/L BAP | - | MS + Fe2+ in chelated form + 60 mg/L cysteine + 0.1 mg/L IAA + 1 mg/L BAP (at the first stage of in vitro culture) | - | - | - | [62] |
MS + Fe2+ in chelated form + 60 mg/L cysteine + 0.05 mg/L IAA + 0.5 mg/L BAP (after the fifth subculture passage) | 24–28 days | - | - | ||||||
Silene nivalis | seeds | HgCl2, H2O2, NaOCl | - | 60 | 0.5 or 1 mg/L BAP | 62 days | 30 | - | [68] |
0.5 or 1 mg/L TDZ | 62 days | 28 | - | ||||||
Syringa josikaea | apical meristems | - | WPM + 0.25 mg/L IAA + 0.1 mg/L NAA | 80 | WPM + 0.1 mg/L NAA + 7.5 mg/L BAP | - | - | 2 | [61] |
Syringa josikaea | - | - | MS + 1 mg/L BAP + 0.1 mg/L NAA | - | MS + 1 mg/L BAP + 0.1 mg/L NAA or MS + 0.1 mg/L 2iP + 0.1 mg/L IBA | 8 weeks | 1.66 | 4.46 / 7.4 | [83] |
MS + 0.1 mg/L 2iP + 0.1 mg/L IBA | - | ||||||||
MS + 2 mg/L 2iP + 1 mg/L IBA | - | ||||||||
Syringa josikaea | knot, apex | NaOCl | MS + 1 mg/L IBA + 0.1 mg/L BAP | 70 | MS + 1 mg/L IBA + 0.1 mg/L BAP | 2 months | 3 | - | [26] |
Syringa josikaea | seeds | - | MS ½ + 3 g/L Cv | - | MS ½ + 3 g/L Cv | - | 54% | - | [27] |
knot, apex | - | SH + 170 mg/L KH2PO4 + 0.1 mg/L IBA + 0.1 mg/L NAA | - | SH + 170 mg/L KH2PO4 + 0.1 mg/L IBA + 0.1 mg/L NAA | - | 54% | - |
2.1.3. In Vitro Rooting Stage
2.1.4. Acclimatization Stage
2.2. Medium-Term In Vitro Cultures
2.3. Cryopreservation
Taxon | Explants Used for Cryostorage | Osmoprotection Treatment | Dehydration Treatment | Cooling Treatment | Regrowth Treatment | Shoot Survival Following Cryostorage (%) | Shoot Regrowth Following Cryostorage (%) | No. of Shoots/Explant Following Cryostorage | Reference |
---|---|---|---|---|---|---|---|---|---|
Dianthus callizonus | axillary buds from micropropagated plants for two years | MS medium + 0.25 M sucrose, 24 h at 23 ± 1 °C | PVS2, 30 min at 23 ± 1 °C | liquid nitrogen (−196 °C), 24 h | MS medium + 30 g/L sucrose, 23 ± 1 °C | 71.6 | 65 | 4.6 | [116] |
Dianthus glacialis subsp. gelidus | shoot tips from micropropagated plants for two years | MS medium + 0.25 M sucrose, 24 h at 23 ± 1 °C | PVS2, 30 min at 23 ± 1 °C | liquid nitrogen (−196 °C), 24 h | MS medium + 30 g/L sucrose, 23 ± 1 °C | 70 | 63.3 | 3 | [116] |
Dianthus henteri | shoot tips from micropropagated plants for two years | MS medium + 0.25 M sucrose, 24 h at 23 ± 1 °C | PVS2, 30 min at 23 ± 1 °C | liquid nitrogen (−196 °C), 24 h | MS medium + 30 g/L sucrose, 23 ± 1 °C | 83.3 | 71.6 | 3 | [116] |
Dianthus nardiformis | shoot tips from micropropagated plants for two years | MS medium + 0.25 M sucrose, 24 h at 23 ± 1 °C | PVS2, 30 min at 23 ± 1 °C | liquid nitrogen (−196 °C), 24 h | MS medium + 30 g/L sucrose, 23 ± 1 °C | 73.3 | 73.3 | 5.6 | [116] |
Dianthus spiculifolius | shoot tips from micropropagated plants for two years | MS medium + 0.25 M sucrose, 24 h at 23 ± 1 °C | PVS2, 30 min at 23 ± 1 °C | liquid nitrogen (−196 °C), 24 h | MS medium + 30 g/L sucrose, 23 ± 1 °C | 68.3 | 66.6 | 3.6 | [116] |
Dianthus trifasciculatus subsp. parviflorus | shoot tips from micropropagated plants for two years | MS medium + 0.25 M sucrose, 24 h at 23 ± 1 °C | PVS2, 30 min at 23 ± 1 °C | liquid nitrogen (−196 °C), 24 h | MS medium + 30 g/L sucrose, 23 ± 1 °C | 71.6 | 70 | 5 | [116] |
3. Conclusions and Perspectives
- -
- Improvement of short-term in vitro propagation protocols, respective of each stage of micropropagation (initiation, multiplication, rooting, acclimatization), medium- and long-term conservation protocols developed so far. For conservation purposes, it is important to have optimized micropropagation protocols that result in a satisfactory regeneration rate, vigorous and rooted plants, and high viability of acclimatized plants, particularly when dealing with threatened endemic plants whose germplasm is very limited.
- -
- Development of a national database with in vitro protocols elaborated so far (published and unpublished) that can be easily accessed by scientists. For some endemic species, such as Silene zawadski and Silene dinarica, the in vitro protocols established within some research projects have not yet been published, access to these data being difficult.
- -
- Extension of research on conservation by biotechnological methods to other taxa with conservation priority that have not yet been studied. Among the 1453 threatened Romanian taxa, of which 95 are endemic and 90 subendemic, only 64 are conserved by in vitro cultures nationwide and cryopreservation protocols have been developed for only 8 taxa.
- -
- Medium- and long-term conservation of endemic species within the national germplasm bank. Currently, a small number of species are conserved only within some research institutes.
- -
- Organization of a researchers’ network from different fields: botany, genetics, ecology, micropropagation, conservation, etc., for the effective dissemination of information regarding in vitro conservation protocols. In this way, sustainable conservation can be achieved, especially for species that are threatened with extinction.
- -
- Allocation of funds for the development of a national database, gene banks, research programs, etc.
- -
- Elaboration of cryopreservation protocols for all endemic and subendemic plant species. Presently, there are known protocols only for some endemic species of the Dianthus genus [116].
- -
- Genetic diversity evaluation of the endemic and subendemic species using techniques such as AFLP, RAPD, RFLP, ISSR, etc. for the efficiency of cryopreservation. Until now, there are known studies concerning the genetic diversity of Dianthus callizonus, D. giganteus ssp. banaticus, D. glacialis ssp. gelidus, D. henteri, D. nardiformis, and D. spiculifolius using AFLP and RAPD technique [118,119].
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
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Family | Taxon | (sub)End. Ro [16] | Threat Category | |||||||
---|---|---|---|---|---|---|---|---|---|---|
Boșcaiu et al. [28] | Dihoru and Dihoru [29] | Oltean et al. [30] | Dihoru and Negrean [31] | Witkowski et al. [32] | Bilz et al. [33] | Turis et al. [34] | IUCN Red List [3] | |||
Asteraceae | Andryala laevitomentosa (Nyár.) Greuter | End. | EN | - | EN | CR | EN | DD | CR | DD |
Fabaceae | Astragalus peterfii Jav. | End. | R | EN | EN | CR | EN | DD | - | DD |
Fabaceae | Astragalus pseudopurpureus Gusul. | End. | R | VU | VU | EN | VU | DD | EN | DD |
Campanulaceae | Campanula romanica Savul. | End. | - | VU | VU | EN | - | DD | - | DD |
Asteraceae | Centaurea reichenbachii DC. | subEnd. | - | R | R | - | - | - | - | - |
Caryophyllaceae | Cerastium transsilvanicum Schur | End. | - | R | R | - | - | - | - | - |
Caryophyllaceae | Dianthus callizonus Schott & Kotschy | End. | R | R | R | LC | - | - | - | - |
Caryophyllaceae | Dianthus giganteus d’Urv. subsp. banaticus (Heuff.) Tutin | End. | - | VU | R | - | - | - | - | - |
Caryophyllaceae | Dianthus glacialis Haenke subsp. gelidus (Schott, Nyman & Kotschy) Tutin | subEnd. | - | R | R | - | - | - | - | - |
Caryophyllaceae | Dianthus henteri Heuff. ex Griseb. & Schenk | End. | - | - | LC | - | - | - | VU | - |
Caryophyllaceae | Dianthus nardiformis Janka | subEnd. | - | VU | VU | VU | - | - | - | - |
Caryophyllaceae | Dianthus spiculifolius Schur | subEnd. | - | VU | R | - | - | - | - | - |
Caryophyllaceae | Dianthus trifasciculatus Kit. subsp. parviflorus Stoj. & Aht. | subEnd. | - | R | R | CR | - | - | - | - |
Asteraceae | Doronicum carpaticum (Griseb. & Schenk) Nym. | subEnd. | - | - | R | - | - | - | - | - |
Caryophyllaceae | Gypsophila petraea (Baumg.) Rchb. | subEnd. | - | - | R | - | - | - | - | - |
Asteraceae | Hieracium pojoritense Wol. | End. | - | R | R | - | - | - | - | - |
Asteraceae | Klasea bulgarica (Acht. & Stoj.) J. Holub [syn. Serratula bulgarica Achtaroff et Stoj] | subEnd. | - | VU | VU | VU | - | - | - | - |
Caryophyllaceae | Moehringia jankae Griseb. ex Janka | subEnd. | R | VU | R | VU | - | DD | - | DD |
Papaveraceae | Papaver alpinum L. subsp. corona-sancti-stephani (Zapal.) Borza | End. | VU | R | R | - | - | - | - | - |
Asteraceae | Saussurea porcii Degen | subEnd. | EX | EX | EX | EX | CR | - | CR | - |
Caryophyllaceae | Silene nivalis (Kit.) Rohrb. [syn. Lychnis nivalis Kit.] | End. | R | VU | VU | VU | VU | - | VU | - |
Oleaceae | Syringa josikaea J. Jacq. ex Rchb. f. | subEnd. | EN | R | VU | LC | EN | DD | NT | EN |
Taxon | Rooting Media (Basic + PGRs) | Rooting Rate (%) | No. Roots/Shoot | Acclimatization Rate (%) | Reference |
---|---|---|---|---|---|
Andryala laevitomentosa | - | - | - | - | [71] |
Andryala laevitomentosa | - | - | - | - | [72] |
Astragalus peterfii | MS + 0.2 mg/L NAA + 0.5 mg/L Kin + 40 mg/L AdSO4 | 36.6 | - | - | [67] |
Astragalus pseudopurpureus | Macro MS ½ + Micro MS + Vit B5 + 0.25 mg/L Kin + 1 mg/L IBA + 1 mg/L 2,4-D | - | - | - | [45] |
Astragalus pseudopurpureus | MS + Vit B5 + 1 mg/L 2,4-D + 0.5 mg/L Kin | - | 6–12 | - | [65] |
Campanula romanica | - | - | - | - | [53] |
Centaurea reichenbachii | MS + Vit B5 + 3 g/L active coal | - | - | - | [44] |
Cerastium transsilvanicum | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | - | - | - | [47] |
Cerastium transsilvanicum | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | - | - | - | [65] |
Dianthus callizonus | - | - | - | - | [64] |
Dianthus callizonus | - | - | - | - | [48] |
Dianthus callizonus | MS + 1 mg/L BAP + 0.1 mg/L NAA | - | - | - | [74] |
Dianthus giganteus subsp. banaticus | - | - | - | - | [75] |
Dianthus giganteus subsp. banaticus | MS + 0.5 mg/L Kin + 0.1 mg/L NAA | - | 23.27 | - | [76] |
Dianthus giganteus subsp. banaticus | MS + 1 mg/L BAP + 0.5 mg/L NAA | 40 | - | - | [50] |
Dianthus glacialis subsp. gelidus | MS + Vit B5 + 1 mg/L Kin + 1 mg/L NAA | - | - | - | [54] |
Dianthus glacialis subsp. gelidus | - | - | - | - | [42] |
Dianthus glacialis subsp. gelidus | - | - | - | - | [48] |
Dianthus henteri | MS + Vit B5 + 0.1 mg/L BAP + 1 mg/L NAA | - | 8.6 | - | [56] |
Dianthus henteri | MS + 1 mg/L BAP + 0.5 mg/L NAA | 20 | - | - | [50] |
Dianthus nardiformis | - | - | - | - | [55] |
Dianthus nardiformis | MS ½ + 0.01 mg/L NAA | - | 7.17 | - | [77] |
Dianthus spiculifolius | MS + 1 mg/L 2iP + 1 mg/L NAA | - | 9.2 | 50–60 | [78] |
Dianthus spiculifolius | MS + Vit B5 + 1 mg/L Kin + 1 mg/L NAA | - | - | 60–80 | [79] |
Dianthus spiculifolius | MS + Vit B5 | - | 5.3–10.1 | 50–70 | [80] |
Dianthus spiculifolius | MS + Vit B5 | 61.1–86.1 | 5.1–11.1 | - | [81] |
Dianthus spiculifolius | Macro MS ½ + Micro MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L NAA | - | - | - | [47] |
Macro MS ½ + Micro MS + Vit B5 + 0.1 mg/L BAP + 0.01 mg/L NAA | - | - | - | ||
Dianthus spiculifolius | - | - | - | - | [48] |
Dianthus spiculifolius | MS + 2 mg/L IBA + 2 mg/L BAP + 40 mg/L AdSO4 | - | 40 | 80 | [26] |
Dianthus spiculifolius | MS + 1 mg/L BAP + 0.5 mg/L NAA | 30 | - | - | [50] |
Dianthus trifasciculatus subsp. parviflorus | MS ½ + Vit B5 + 0.01 mg/L NAA | - | 9.33 | 80 | [51] |
Doronicum carpaticum | - | - | - | - | [42] |
Gypsophila petraea | - | - | - | - | [82] |
Gypsophila petraea | MS + Vit B5 | - | - | - | [49] |
Hieracium pojoritense | MS + 0.1 mg/L NAA + 1 mg/L GA3 | - | - | - | [46] |
Hieracium pojoritense | - | - | - | - | [65] |
Klasea bulgarica | MS + 0.1 mg/L IAA | - | - | - | [52] |
Moehringia jankae | MS + Vit B5 + 0.22 µM TDZ + 0.49 µM IBA | 100 | - | 50 | [43] |
MS + Vit B5 + 4.4 µM BAP + 0.49 µM IBA | 100 | - | 50 | ||
MS + Vit B5 + 4.5 µM ZEA + 0.49 µM IBA | 100 | - | 50 | ||
Papaver alpinum subsp. corona-sancti-stephani | - | - | - | [64] | |
Papaver alpinum subsp. corona-sancti-stephani | MS + Vit B5 + 1 mg/L BAP + 0.1 mg/L Kin + 0.2 mg/L NAA | - | - | - | [73] |
Saussurea porcii | Macro MS ½ + Micro MS ½ + Vit MS + 0.1 mg/L IAA + 0.01 mg/L BAP | - | - | 80 | [62] |
Silene nivalis | - | - | - | - | [68] |
Syringa josikaea | WPM + NAA | 48.4 | - | - | [61] |
Syringa josikaea | MS + 2 mg/L 2iP + 0.1 mg/L IBA | - | 1–2 | - | [83] |
Syringa josikaea | MS + 1 mg/L IBA + 0.1 mg/L BAP | - | 3–4 | 50 | [26] |
Syringa josikaea | MS ½ + 3 g/L Cv | - | - | 50 | [27] |
SH + 170 mg/L KH2PO4 + 0.1 mg/L IBA + 0.1 mg/L NAA | - | - | 50 |
Taxon | Basic Media + PGRs | Growth Inhibitory Factors | Time Intervals | Regeneration Rate (No. of Shoots/Explant) | Shoot Length (cm) | Viable Shoots (%) | Rooting Rate (%) | Survival Rate (%) | Reference |
---|---|---|---|---|---|---|---|---|---|
Dianthus callizonus | MS + Vit B5, free | 3% mannitol | 3 months | 26.60 | - | - | - | - | [48] |
Dianthus callizonus | MS1/10 + Vit B5, free | reduced mineral concentration | 12 months | - | - | 100 | 100 | - | [63] |
MS + Vit B5, free | reduced temperature (10 °C) | 12 months | - | - | 100 | - | - | ||
Dianthus glacialis subsp. gelidus | MS + Vit B5, free | 3% mannitol | 3 months | 14.30 | - | - | - | - | [48] |
Dianthus glacialis subsp. gelidus | MS + Vit B5, free | 0.32 M mannitol | 1 months | 2–5 | 0.5 | - | - | - | [92] |
2 months | 5–10 | 0.5 | - | - | - | ||||
0.49 M mannitol | 1 months | 2–4 | < 0.5 | - | - | - | |||
2 months | 10–15 | <0.5 | - | - | - | ||||
Dianthus glacialis subsp. gelidus | MS + Vit B5, free | 0.16 M mannitol | 3 months | 15–20 | 1 | - | - | - | [94] |
6 months | 20 | 1 | - | - | - | ||||
Dianthus nardiformis | MS + Vit B5, free | 329 mM mannitol | 40 days | 2.60 | 1.80 | - | - | - | [96] |
80 days | 4.86 | 2.80 | - | - | - | ||||
120 days | 22.26 | - | - | - | - | ||||
Dianthus spiculifolius | MS + Vit B5, free | 3% mannitol | 3 months | 25.00 | - | - | - | - | [48] |
Dianthus spiculifolius | MS + Vit B5, free | 0.32 M mannitol | 1 months | 3–5 | 0.5 | - | - | - | [92] |
2 months | 5–10 | 0.5–1 | - | - | - | ||||
0.49 M mannitol | 1 months | 1–2 | <0.5 | - | - | - | |||
2 months | 7–10 | <0.5 | - | - | - | ||||
Dianthus spiculifolius | MS + Vit B5, free | 0.16 M mannitol | 3 months | 20–30 | 1–1.5 | - | - | - | [94] |
6 months | - | 1.8–2 | - | - | - | ||||
Dianthus spiculifolius | MS + Vit B5, free | 0.16 M mannitol | 6 months | - | - | - | - | - | [93] |
Dianthus trifasciculatus subsp. parviflorus | MS + Vit B5, free | 6% PEG 4000 | 40 days | 2.87 | 3.40 | - | - | - | [95] |
80 days | 13.40 | 3.93 | - | - | - | ||||
120 days | 35.13 | 6.00 | - | - | - | ||||
3% mannitol | 40 days | 5.67 | 3.60 | - | - | - | |||
80 days | 9.93 | 3.87 | - | - | - | ||||
120 days | 47.13 | 4.67 | - | - | - | ||||
Gypsophila petraea | MS1/4 + Vit B5, free | reduced mineral concentration | 12 months | - | - | 100 | 100 | - | [63] |
MS + Vit B5, free | reduced temperature (10 °C) | 12 months | - | - | 100 | 45 | - | ||
Moehringia jankae | MS, free | 32 μM flurprymidol | 1 month | 1.36 | 0.56 | - | 0 | - | [97] |
2 months | 2.52 | 0.75 | - | 36 | - | ||||
3 months | 4.24 | 0.98 | - | 76 | - | ||||
6 months | 4.60 | 0.55 | - | - | 88 | ||||
48 μM flurprymidol | 1 month | 1.24 | 0.36 | - | 0 | - | |||
2 months | 1.60 | 0.65 | - | 0 | - | ||||
3 months | 2.40 | 0.81 | - | 0 | - | ||||
6 months | 7.08 | 0.32 | - | - | 88 | ||||
0.16 M mannitol | 1 month | 1.80 | 0.42 | - | 32 | - | |||
2 months | 3.88 | 1.06 | - | 69 | - | ||||
3 months | 5.68 | 1.11 | - | 81 | - | ||||
6 months | 5.64 | 1.17 | - | - | 88 | ||||
2.5 mM clormerquat | 1 month | 1.72 | 1.62 | - | 0 | - | |||
2 months | 3.84 | 2.37 | - | 28 | - | ||||
3 months | 5.68 | 2.33 | - | 72 | - | ||||
6 months | 6.68 | 2.06 | - | - | 96 | ||||
Papaver alpinum subsp. corona-sancti-stephani | MS + 3% sucrose + 3% mannitol, free | reduced temperature (4 °C and −20 °C) | 3 weeks | - | - | - | - | > 65 | [98] |
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Radomir, A.-M.; Stan, R.; Florea, A.; Ciobotea, C.-M.; Bănuță, F.M.; Negru, M.; Neblea, M.A.; Sumedrea, D.I. Overview of the Success of In Vitro Culture for Ex Situ Conservation and Sustainable Utilization of Endemic and Subendemic Native Plants of Romania. Sustainability 2023, 15, 2581. https://doi.org/10.3390/su15032581
Radomir A-M, Stan R, Florea A, Ciobotea C-M, Bănuță FM, Negru M, Neblea MA, Sumedrea DI. Overview of the Success of In Vitro Culture for Ex Situ Conservation and Sustainable Utilization of Endemic and Subendemic Native Plants of Romania. Sustainability. 2023; 15(3):2581. https://doi.org/10.3390/su15032581
Chicago/Turabian StyleRadomir, Ana-Maria, Ramona Stan, Alina Florea, Cristina-Magdalena Ciobotea, Florina Mădălina Bănuță, Magdalena Negru, Monica Angela Neblea, and Dorin Ioan Sumedrea. 2023. "Overview of the Success of In Vitro Culture for Ex Situ Conservation and Sustainable Utilization of Endemic and Subendemic Native Plants of Romania" Sustainability 15, no. 3: 2581. https://doi.org/10.3390/su15032581