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EN
Genetic stability of plants during in vitro propagation and conservation is one of the important aspects of plant biotechnology. In the present study, micropropagated P. lentiscus L. shoot cultures, which are cultivated for the mastic resin, have been cold stored up to 12 months at 4°C in the dark for different durations (2, 4, 6, 8, 10 and 12 months) and genetic alterations in cold storage conditions were evaluated. Growth parameters such as proliferation rate, shoot numbers per explant, shoot lengths and shoot forming capacity were also calculated. Since the highest proliferation rate (100 %) was obtained in 6 month-stored shoot cultures without any severe influence of cold stress on proliferation ability, amplified fragment length polymorphism (AFLP) and inter-retrotransposon amplified polymorphism (IRAP) marker systems were used to determine genetic stability in the plantlets after this storage period. Totally, 702 scorable bands were produced by 10 AFLP primer pairs. Genetic similarity value of the non-stored (control) plant and coldstored clones ranged from 0.66 to 0.84 with a mean of 0.74. In the case of IRAP, 159 bands were produced by 8 IRAP primers. Genetic similarity value of the non-stored plant and cold-stored clones varied from 0.65 to 0.83 and the average genetic similarity value was determined as 0.72. The genetic similarity indices revealed that genetic variability was similar in both techniques. Our results showed that tissue culture and especially cold storage of P. lentiscus L. may result transposons activation, thus could cause genetic instability.
EN
This study was aimed at developing an efficient protocol for regeneration of Pseudostellaria heterophylla plantlets and induction of polyploidy. Calli of P. heterophylla (Miq) from stems, leaves and buds as explants could not differentiate into plantlets. However, young embryo segments used as primary explants produced embryonic calli on MS medium containing 5.0 mg/L 2,4-D and 0.5 mg/L KT. After the embryonic calli with granular protuberances were transferred to MS medium containing 0.5 mg/L BA, they developed shoots and then rooted to form plantlets. Polyploidy was induced when embryonic calli were placed in liquid MS medium containing 0.5% colchicine for 4 days, followed by culturing in solid medium to induce differentiation. Polyploidy was identified by the number of chromosomes and the size of plantlet stomata. The tetraploid plantlets produced larger root tubers than the diploid plantlets.
EN
In vitro plantlets and callus of M. longifolia were established and their volatile constituents characterized by GC-MS analysis of their headspaces (HSs) and essential oils (EOs). Significant quali-quantitative differences were found in the aromatic fingerprints in comparison with the M. longifolia parent plants. In fact, limonene and carvone were the main constituents in the EOs of the mother plants, while the aroma of the in vitro plant material were especially enriched in oxygenated terpenes. In particular, huge amounts of piperitenone and piperitenone oxide (75 %) were found for in vitro plantlets, while trans-carvone oxide (19 %) and trans-piperitone epoxide (9 %) were found in callus EO. However, the established in vitro plant material showed lack of pulegone and menthofurane, thus preserving an important feature observed in the volatile fingerprint of the parent plants. In fact, because of their well-known toxicity significant amounts of pulegone and menthofurane may compromise the safety using of mint essential oil. Therefore the in vitro M. longifolia plantlets and callus may be regarded as a potential source of a safe flavouring agent.
EN
 The essential oils (EOs) and static headspaces (HSs) of in vitro plantlets and callus of Mentha x piperita were characterized by GC-MS analysis. Leaves were used as explants to induce in vitro plant material. The EO yields of the in vitro biomass were much lower (0.1% v/w) than those of the parent plants (2% v/w). Many typical mint volatiles were emitted by the in vitro production, but the callus and in vitro plantelet EOs were characterized by the lack of both pulegone and menthofuran. This was an important difference between in vitro and in vivo plant material as huge amounts of pulegone and menthofuran may jeopardise the safety of mint essential oil. Regarding the other characteristic volatiles, menthone was present in reduced amounts (2%) in the in vitro plantlets and was not detected in the callus, even if it represented the main constituent of the stem and leaf EOs obtained from the cultivated mint (26% leaves; 33% stems). The M. piperita callus was characterized by menthol (9%) and menthone (2%), while the in vitro plantlet EO showed lower amounts of both these compounds in favour of piperitenone oxide (45%). Therefore, the established callus and in vitro plantlets showed peculiar aromatic profiles characterized by the lack of pulegone and menthofuran which have to be monitored in the mint oil for their toxicity.
EN
The aim of the present work was to determine the carbohydrate content in plantlets of Clematis pitcheri cultured in vitro depending on the temperature (15°C, 20°C and 25°C) and sucrose (10 and 30 g·dm⁻³) and nitrogen (100% and 50% of normal MS strength) level in the medium. 100% N meant 1.90 g KNO₃ and 1.65 g NH₄NO₃, and 50% N meant 0.950 g KNO₃ and 0.825 g NH₄NO₃ in 1 dm⁻³ of MS medium. The highest accumulation of starch in shoots took place at 30 g·dm⁻³ of sucrose and lower level of nitrogen compounds in the medium. Also, at 30 g·dm⁻³ of sucrose in the medium the content of fructose in shoots was higher and independent from the level of nitrogen. The temperature of 25°C stimulated glucose accumulation in shoots at the highest degree at 30 g·dm⁻³ of sucrose and lower level of nitrogen. Regardless of the treatment, no sucrose was found in plantlets. Generally, the higher concentration of sucrose in the medium the higher content of sum of carbohydrates in plantlets. A lower nitrogen compound level (50% N) led to increase of glucose and starch content in shoots, and had a smaller effect on the level of fructose. Shoots cultured at the temperature of 20°C and 25°C accumulated more carbohydrates than at 15°C.
PL
W pracy oznaczono zawartość węglowodanów w pędach Clematis pitcheri rosnących in vitro w zależności od temperatury (15°C, 20°C i 25°C) oraz stężenia sacharozy (10 i 30 g·dm⁻³) i związków azotowych (100% i 50% standardowego poziomu) w pożywce MS. Stosowano 1,90 g KNO₃ i 1,65 g NH₄NO₃ (100% N) oraz 0,950 g KNO₃ i 0,825 g NH₄NO₃ (50% N) w 1 dm⁻³ pożywki. Pędy akumulowały najwięcej skrobi na pożywce z 30 g·dm⁻³ sacharozy i niskim poziomem związków azotowych. Zawartość fruktozy w pędach była wyższa na pożywce z 30 g·dm⁻³ sacharozy niezależnie od poziomu azotu. Temperatura 25°C stymulowała akumulację glukozy w pędach, przy czym zawartość glukozy była najwyższa w pędach rosnących na pożywce z 30 g·dm⁻³ sacharozy i niskim poziomem związków azotowych. Niezależnie od traktowań nie stwierdzono obecności sacharozy w pędach. Ogólnie można stwierdzić, że im wyższe było stężenie sacharozy w pożywce, tym wyższa była zawartość sumy węglowodanów w pędach. Obniżenie poziomu azotu w pożywce (do 50%) zwiększało zawartość glukozy i skrobi w pędach i w mniejszym stopniu wpływało na zawartość fruktozy. Pędy rosnące w 20°C i 25°C akumulowały więcej węglowodanów niż w 15°C.
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