Módosított életformájú és alkaloidspektrumú mák (Papaver somniferum L.) kemotípusok előállítása = Development of poppy (Papaver somniferum L.) chemotypes of modified life cycle and alkaloidspectrum

Célunk az egyik legfontosabb gyógy- és élelmiszernövényünk, a mák áttelelő genotípusainak tudományosan eddig nem ismert kémiai-genetikai-élettani tulajdonságainak feltárása volt. Tíz mákfajtát/törzset és ezek F1, F2 hibridgenerációit tanulmányoztuk in vitro és szabadföldi körülmények között. Kidolgoztunk egy reprodukálható hidegtűrési tesztprogramot az 'őszi' és 'tavaszi' jellegű genotípusok elkülönítésére. Megállapítottuk, hogy a fagytűrést a mákban valószínűleg dominancia hatások szabályozzák, ami F2-ben csökken. Az utódok fagytűrése a toleráns és a szenzitív szülőtől is függ. Igazoltuk, hogy hidegedzés a mákban a prolinszint növekedését indukálja, ami a fagytűrő genotípusokban erőteljesebb, de a fajták között eltérések vannak. A hidegedzés során az oldható cukrok koncentrációja is jelentősen megnő, a komponensek spektruma átalakul, de nincs szignifikáns eltérés a tavaszi illetve őszi ökotípusú fajták között. Több ezer egyedi analízis alapján összefüggéseket állapítottunk meg az alkaloidszint, ezen belül a morfin, kodein, tebain illetve narkotin öröklődésére és a vegyületek korrelációira vonatkozóan. A mák alkaloidfelhalmozódása és hidegtűrése között -klímakamrás és szabadföldi kísérletek alapján- nem igazolható határozott kapcsolat. Így célirányos nemesítéssel előállíthatók mind ipari, mind étkezési célra alkalmas, megnövekedett fagytoleranciájú genotípusok. Az előállított populációkból kiválasztottunk a fajtaelőállítás céljára perspektivikus illetve szokatlan spektrumú, tudományos szempontból érdekes vonalakat. | Aim of the recent investigation was to clear up the chemical-genetical and physiological characteristics of the less-known overwintering ecotype of poppy, one of the most improtant medicinal and culinary plant species. We examined 10 genotypes and their F1, F2 hybrid generations under in vitro and open field conditions. A reliable in vitro test method was established for checking the frost tolerance and distinguishing of poppy genotypes. We concluded that the regulation of winter hardiness in poppy might be under dominance genetic control which decreases in F2 generations. However, cold tolerance of poppy hybrids depends both on sensitive and tolerant parent partners. It was proved that proline level increases significantly during hardening and it is more empasized in tolerant genotypes. Nevertheless, there are differences among cultivars. The concentration and spectrum of soluble sugars also increased during hardening, but there is not a significant difference between tolerant and sensitive varieties. Based on several thousand analysis, we established conclusions on the inheritance of alkaloids and especially that of morphine, codein, thebaine and narcotine as well as their correlations. No interaction between the alkaloid accumulation and winterhardiness could be proved. Thus, both industrial nad culinary varieties of good frost tolerance can be obtained by well oriented breeding. We selected several accessions both for further breeding as well as for scientific studies

Changes in the volatile compounds of two wormwood (Artemisia absinthium L.) accessions under controlled weather conditions

Two wormwood accessions (originating from Spain and Hungary) were grown in climatic chambers in order to determine the effect of temperature and light intensity on the composition of their volatile compounds. Young plants were grown in a controlled environment termed as “warm” (increasing temperature from 18 °C/10 °C to 27 °C/19 °C and 16 klx light intensity with a 14 h/10 h light-dark rhythm, respectively) and “cold” ones (increasing temperature from 13 °C/8 °C to 18 °C/ 10 °C and 8 klx light intensity with a 14 h/10 h light-dark rhythm, respectively) for 14 weeks.                The EO yield of the investigated accessions varied from 0.188 mL/100 g (“Hungarian” accession grown in the “warm” chamber) to 1.092 mL/100 g (“Spanish” accession grown in the “cold” chamber) and the installed weather programs had no effect on the EO yield of any of the accessions. Evaluating the components higher than 1% of all detected GC areas, 33 compounds were identified with the total identified percentage varying from 88.8% (“cold” treatment of the “Hungarian” accession) to 92.5% (“cold” treatment of the “Spanish” accession). The well-established chemical differences between the two investigated accessions of wormwood, determined in our former study [1], have been confirmed by the present data. The major components of the oils were sabinene (0-10.8%), b-myrcene (1.7-16.5%), cis-epoxyocimene (1.2-57.7%), cis-chrysanthenyl acetate (0-13.8%), and (Z)-nuciferyl isobutyrate (1.7-10%). The “Spanish” accession represents a “cis-epoxyocimene” chemotype while the “Hungarian” accession exhibits a much more variable profile with sabinene and β-myrcene as the most characteristic components. The results showed that the accumulation of volatile compounds was not influenced by the weather under the investigated parameters. However, the different weather conditions induced quantitative changes in the EO profile of both chemotypes. The relative amount of cis-chrysanthenyl acetate increased from 8.0 (“cold” chamber) to 13.8% (“warm” chamber) in the oil of the “Spanish” plants while sabinene increased from 2.3 to 10.8% and b-myrcene rose from 8.0 to 16.5% in the “cold” and “warm” chambers, respectively, for the “Hungarian” oil samples

Variability of thujone content in essential oil due to plant development and organs from Artemisia absinthium L. and Salvia officinalis L.

The study compared changes in essential oil content and its thujone ratio in two popular herbs (Artemisia absinthium L. and Salvia officinalis L.), pertaining to plant development and plant organs. Both species were harvested in 2018 at the vegetative, floral budding, flowering and after flowering phases; flowers and leaves were sampled separately. The essential content is always higher in the flowers than in the leaves at the same phenophase in both species we examined. Decreased essential oil content in both organs during the developmental phases was also common to both species. In S. officinalis, both leaf and flower oils showed quantitatively different compositional profiles. During plant development, the main component α-thujone decreased significantly in leaf oils, while both thujone isomers demonstrated statistically stable values in flower oils. In A. absinthium, leaf and flower oils exhibited similar thujone ratios. During plant development, neither of the thujone isomers changed significantly in leaf oils but the ratio of α-thujone increased in the flowers. It was established, that only the distribution and dynamics of total volatiles showed common features in the two species, while the variability of thujone ratios represents differences specific to each target species. For the praxis, in S. officinalis timing of harvest seems to be more important while in A. absinthium the ratio of organs may play a more significant role in reaching lower thujone levels of the drug

Small changes in the distillation method result in variable quality of yarrow (Achillea collina) essential oil

Achillea collina (Becker ex Rchb.f.) Heimerl (Asteraceae) is one of the yarrow species providing the chamazulene containing, characteristic blue essential oil by water distillation. The chamazulene content varies on a large scale [1,2]. Besides several other factors–like the determination of the species and taxon, geographical and ecological characteristics of the habitat, sampling, and GC analytical methods–which have been studied and discussed more frequently, the method of distillation and oil recovery might have a large influence on the quality of the oil, too. Unfortunately, the applied solvents, evaporation/drying methods and/or further dilution is almost never described accurately in manuscripts. Therefore, a well-established comparison and evaluation of the results is at least questionable. In our recent experiment, this aspect was investigated in detail. As plant material, a high chamazulene-containing strain of A. collina was used, selected and maintained at our experimental station. Dried flowering shoots were distilled in a Clevenger type apparatus: a) as in Pharmacopoeia Hungarica VII, b) as in Ph.Hg.VIII (=Ph.Eur.). For the second treatment, different solvents were applied for washing the sesquiterpene-rich oil of large viscosity: a) n-hexane, b) n-pentane, c) xylol, d) ethanol (96%), e) acetone. The recovered oil was analyzed a) immediately after the washing down, b) after the evaporation of the solvent–as for measuring the yield–diluted by different amounts of hexane again (resulting in 0.3, 1.0, 5.0, 10% concentrations) for the injection into the GC apparatus. The GC-MS analysis was carried out as in [3]. The results show that based on the above circumstances and factors, the number of GC peaks varied between 4 and 68. Chamazulene content of the oil samples changed between 45 and 78% of the total GC area percentage and a higher dilution rate of the extracts increased the ratio of chamazulene. Evaporation of the solvents resulted in a severe loss of volatile monoterpenes. At the same time, the proportion of α-bisabolol remained more constant (18-25%) but its ratio increased with the b-type of apparatus. Although internal standards might help quantification, however, the studied factors may influence severely the analytical results and any comparison. A well-established, standardized method of distillation and subsequent oil recovery process would be of high importance in the case of sesquiterpene-rich species

Effect of harvest date on yield and secondary compounds of lemon balm (Melissa officinalis L.)

The quality of the drug of lemon balm (Melissa officinalis L.) is influenced by several factors, among which the effect of ontogenesis has practically not been studied before. Five varieties (‘Lorelei’, ‘Lemona’, ‘Quedlinburger Niederliegende,’, ‘Gold Leaf’, ‘Soroksár’) were sampled at vegetative, budding, full flowering and after flowering phases at two locations (Budapest and Poznań) and their dried leaves analysed. The accumulation of volatile compounds showed maximum values (0.08-0.46 ml/100 g dry weight) in budding phase (Budapest) or during flowering (Poznań). The content of total phenolics was highest (226-431 mg gallic acid equivalent/g dry weight) in vegetative stage and in some cases similar values were measured until budding. After a sharp decrease at flowering time in several cases, a second peak was detected at the end of the vegetation period. Similarly to the total phenolics, also the total flavonoid content reached the highest level (0.239-1.152% dry weight) at the first half of the vegetation period however, with characteristic differences between habitats. In cultivation, the highest essential oil content may be reached later than highest polyphenol content, however harvesting at budding time may assure a good quality from both aspects with advantageous fresh and drug yields. The described dynamics of the accumulation of the investigated secondary metabolites was depending more on the habitat and less from the cultivar

Variability of essential oil composition of wormwood (Artemisia absinthium L.) affected byplant organ

The essential oil composition of leaves and roots of twenty individuals of wormwood (Artemisia absinthium L.) belonging to different chemotypes has been investigated. They were obtained from two accesions coming from Hungarian and Spanish wild populations. Essential oils were extracted by hydrodistillation and analyzed by GC/MS and GC/FID. Results showed a great individual variability of wormwood accesions. Based on leaves, three chemotypes (sabinene + myrcene, -thujone and new sesquiterpene type accounting up to 80.8% of sesquiterpenic fraction) were found in the Hungarian population and two chemotypes ((Z)--epoxyocimene and (Z)--epoxyocimene + (Z)-chrysanthemyl acetate types) were present in the Spanish one. The composition of EO of the roots from both locations was predominated by monoterpenic esters (14.5-80.2% and 59.9-90.3%, in Hungarian and Spanish samples respectively) but characteristic quantitative and qualitative differences were present. No relationship was found between the composition of EO of roots and leaves of the same plant.Llorens Molina, JA.; Vacas González, S.; Castell Zeising, V.; Németh Zámboriné, É. (2016). Variability of essential oil composition of wormwood (Artemisia absinthium L.) affected byplant organ. Journal of Essential Oil Research. 29(1):11-21. doi:10.1080/10412905.2016.1202152S112129

Az életforma és a hatóanyagprodukció összefüggései néhány termesztett gyógynövényfaj esetében = Relationship between life form and production of active materials in case of some cultivated medicinal plants

2002-2005 között szabadföldi és laboratóriumi munkával tisztáztuk a Salvia sclarea, a Verbascum phlomoides és a Carum carvi gyógynövényfajok ún. egy- és kétéves intraspecifikus biotípusainak eltéréseit, fenológiai, morfológiai sajátosságaik, drogprodukciójuk, néhány biokémiai jellemzőjük és hatóanyag felhalmozódásuk alapján. A három modellfaj virágzásindukciós szükséglete egymáshoz nagyon hasonló. Valamennyi kvalitatív (obligát) vernalizációs igényű növény, ahol a hideghatás fagypont feletti hőmérsékleteken is jelentkezik. Fajspecifikus azonban az indukcióra megfelelő fenofázis. Az egy- és kétévesnek biotípusok eltérése ebben nem éles, hanem mennyiségi, elsősorban a vernalizációs periódus hosszára vonatkozik. A kétféle ökotípus mindhárom faj esetében stabilan eltér fagytűrésében, több morfológiai és hatóanyag jellemzőjében is, amelyek alapján varietas szintű megkülönböztetésük indokolt. Párhuzamos termőhelyi vizsgálatokkal bizonyítottuk, hogy a termőhely hatással van az elvetett magvak kelésére, fejlődésére, áttelelésére valamint a hatóanyagok felhalmozódására. Az állományok virágzási hányadára azonban az adott évjárat jellege hat döntően. Termőhelyenként meghatároztuk az optimális vetésidőt, amivel mindhárom faj mindkét változatában maximális drog- és hatóanyagprodukció érhető el. | During 2002-2005 in open field and laboratory research we described the differences of intraspecific biotypes of Salvia sclarea, Verbascum phlomoides and Carum carvi based on phenological, morphological, production properties, some biochemical markers and active materials. Flowering induction requirements of the three species are similar. All of them have qualitative vernalisation requirements, temperatures above zero are also effective. The optimal phenophase for induction depends on the species. Difference of annual and biennial varieties is not rigid, but quantitative. The two ecotypes of all of the species are different also in frost tolerance, several morphological and chemical characteristics. Based on these stable traits, they are proposed to be varietas. We proved by parallell investigations in two areas, that the gowing site influences germination, development, overwintering and accumulation of active materials. The flowering proportion of the stands is influenced more strongly by the weather conditions of the given year. We decided the optimal sowing period for both growing sites, in order to get maximum yields in drug and active agents in all of the three species