Amount Of Plant Material Research Articles

DESIGN OF ANTI-EROSION HYDRAULIC AND FORESTRY MEASURES TO COMBAT RAVINES

The article discusses the design of anti-erosion hydraulic and forestry measures to combat ravines in the territory of Kopenkuvate village, Golovanivsky district, Kirovohrad region. It has been established that climatic conditions (especially the distribution of precipitation, both in terms of the growing season and the intensity of precipitation) in some periods contribute to the acceleration of water erosion, especially gully erosion. According to hydrological calculations, the volume of runoff from the 10% snowmelt was 6792 m3, and the total solid runoff was 476 m3. An earthen rampart 186 m long and 2.0 m high was designed to capture the runoff. Reducing erosion intensity to acceptable levels is the main criterion for selecting erosion control measures. In addition to the agrotechnical erosion control measures used, the farm selected and designed their combination with hydrotechnical and forest reclamation measures that meet this condition. Simultaneously with the installation of the water retention berm, we create a forest strip along the ravine, which is placed on both sides of the ravine along its sides at a distance of 5 meters from them. The strip is 0.526 km long, 12.5 m wide, and has 5 rows. Amount of planting material: common oak - 1.052 thousand pieces; pear - 0.375 thousand pieces; white acacia - 0.375 thousand pieces; common dog rose - 0.750 thousand pieces; sea buckthorn - 1.052 thousand pieces. To strengthen the bottom of the ravine, we plan to install 15 wicker dams. The bottom of the ravine will be used for continuous forestation. To reforest the ravine bottom with an area of 0.52 hectares, we need 1,733 thousand poplar plants and 0.867 thousand rose hips. The complex of anti-erosion measures will reduce the intensity of erosion processes and the formation of ravines on agricultural lands.

Structural reorganization in beech forests in central Germany as response to drought-induced mortality in the overstory

Prolonged periods of extreme heat and drought are changing forest health and forest structure. Loss of vitality, signs of dieback, and mortality of trees are reported in many regions of the world. There is a need for information on the quantification of drought effects on forest structure in order to detect drought stress at an early stage. Furthermore, forest structure is linked to growth processes that result from single tree and stand dynamics. In this study, we used mobile laser scanning to objectively assess forest structure in European beech (Fagus sylvatica L.) stands differently affected by drought. We used the box-dimension as a holistic measure of structural complexity that jointly describes both distribution and amount of plant material in a forest, and canopy cover as a measure for overall defoliation. We observed large canopy gaps in heavily damaged stands, but the overall structural complexity of the forest stands remained unchanged. However, when dividing the 3D forest stand model from mobile laser scanning into 5 m height layers, we noticed a “structural flip” with reduced structural complexity in the upper parts of the forests and increased structural complexity in the lower strata. This indicates that the forests are responding to drought with increased mortality of mature trees, resulting in increased light availability in the understory and consequently increased growth of the understory trees. The rather indifferent overall structural complexity of the investigated forests was due to the reorganization of structures by shifting the major foliage layer from top-to-bottom and is interpreted as a successful ecosystem-scale response to the drought events. However, it is unclear how these forests will respond to repeated droughts. The “structural flip” should therefore be regarded as an early warning signal pointing to increased ecosystem stress, though the stress level has not yet exceeded the adaptive capacity in the investigated forest.

The Development of Dry Extract Technology from Callus Culture Biomass of <i>Scutellaria baicalensis</i>

Introduction. Plant biologically active substances (BAS) are valuable for drug development in view of their pharmacological effects and metabolic proximity to the human body. As an example of a useful herbal raw material, the roots of Scutellaria baicalensis, which contain species-specific flavonoids, baicalin and scutellarin, possess the potency of creating antiviral drugs and functional nutrition systems with antioxidant and adaptogenic properties. In order to levelling the existing limitations of using intact plant in pharmaceutical practice, we proposed a nature-saving and ergonomic approach of cultivation of S. baicalensis cells under in vitro conditions. This technology allows to extract active substances of S. baicalensis without causing damage to the environment and contributes to the reduction of the time required to produce the necessary amount of plant material with an increase in the efficiency of production processes.Aim. The development of dry extract technology from S. baicalensis callus biomass.Materials and methods. The object of the study – biomass of S. baicalensis cells obtained from callus culture. Functional characteristics (friability, Hausner’s coefficient, Carr’s index, fractional composition, as well as porosity and hygroscopicity) of S. baicalensis biomass and dry extract on its basis were determined according to the methods of the EAEU Pharmacopoeia. Extraction of plant material was carried out by maceration method with heating in a water bath with reflux condenser. Qualitative and quantitative analysis of BAS in the dry extract was carried out by spectrophotometry and high-performance liquid chromatography. Quality indicators and functional characteristics of the finished product were evaluated in accordance with pharmacopeial requirements.Results and discussion. The functional characteristics of the S. baicalensis biomass were evaluated and the possibility of obtaining a dry extract on its basis was confirmed. We selected parameters of BAS extraction process, established the presence of baicalin in the biomass extract and carried out a comparative evaluation of aglycone profile in the extract based on callus culture and roots of intact plant. The properties of the finished product were evaluated for compliance with regulatory requirements, as well as technological characteristics.Conclusion. The quality indicators of the dry extract of the S. baicalensis biomass satisfy the normative conditions. The technological characteristics of dry extract are suitable for the utilisation as a phytosubstance for the development of medicines of combined type and dietary supplements.

PM 7/153 (1) Mechanical inoculation of test plants

PM 7/153 (1) Mechanical inoculation of test plants

One prep to catch them all: “2 in 1”, an efficient method for the simultaneous extraction of DNA and RNA from Grapevine tissues

Recent advances in our understanding of plant physiology and adaptation to the environment are tightly related to the development of ‘omics’ technologies such as metabolomics, transcriptomics, genomics and epigenomics that allow a more comprehensive view of the plant functioning. In this context, the ability to extract DNA and RNA from small amounts of plant material can be a limiting factor, worse in the case of non-model plants for which efficient nucleic extraction procedures are lacking. In the case of grapevine, extraction of high-quality DNA is typically limited by the high polyphenolic and polysaccharide contents of the different tissues. Here, we propose an adaptation of the method of Reid et al. (2006) that allows the simultaneous and efficient extraction of DNA and RNA from grapevine vegetative and berry tissues from in vitro grown grapevine plants and cells and from other plants. The protocol allows the extraction of high-quality RNA and DNA for standard molecular biology methods as well as for Next Generation Sequencing (NGS). It also works with a limited amount of plant material, such as young developing buds, and provides the means to analyse “omics” data from a single plant sample.

Combined effects of vermicompost and vermicompost leachate on the early growth of Meloidogyne incognitastressed Withania somnifera (L.) Dunal.

Roots of Withania somnifera, an important medicinal herb, are prone to the infection of Meloidogyne incognita (a root parasitic nematode). The infection lowers the quality and quantity of plant material and poses a challenge in crop cultivation and obtaining desirable yield. In the present study, in vitro inhibitory activity of vermicompost leachate (Vcom-L) was assessed against % hatch and survival of M. incognita in a 96h assay. Then, Vcom-L was used as soil supplement in combination with vermicompost (Vcom) to evaluate their nematode inhibitory and stress alleviating effect in W. somnifera, in a pot experiment. Root galling intensity and growth performance of nematode-stressed W. somnifera raised from seeds pre-soaked in distilled water (DW), Vcom-L, vermicompost tea (Vcom-T) and different dilutions of Vcom-L were assessed. We observed 79% suppression of egg hatching and 89% juvenile (J2) mortality after 96h compared to control, at 100% concentration of Vcom-L. Significant reduction in gall formation with increase in growth parameters of seedlings was observed after combined application of Vcom (60% or 100%) + Vcom-L and was evident as enhancement in seedling biomass and contents of chlorophyll and protein. However, proline, total phenolics and malondialdehyde (MDA) content declined significantly in these combinations compared to the control (0% Vcom). Activity of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidise (APX), guaiacol peroxidise (GPX) and glutathione reductase (GR) declined with Vcom as well as Vcom + Vcom-L and corresponded with decline in the accumulation of reactive oxygen species in leaves. Further, 1:5 and 1:10 dilutions of Vcom-L in combination with Vcom (60%) proved beneficial in mitigating the nematode-induced stress in W. somnifera. Present results showed the potential of Vcom and Vcom-L in standardised combination as an effective strategy in controlling the pathogenicity of M. incognita in medicinal plants such as W. somnifera.

MICROPROPAGATION AND ORGANOGENESIS OF ASTROPHYTUM ASTERIAS (ZUCC.) LEM. (CACTACEAE JUSS.), BLOSSFELDIA LILIPUTIANA WERDERM. AND STROMBOCACTUS DISCIFORMIS (DC.) BRITTON & ROSE

The research reveals results of the creation of sterile cultures and micropropagation of the species of three unique genera of Cactaceae: Astrophytum asterias (Zucc.) Lem. (І CITES); Blossfeldia liliputiana Werderm. (ІІ CITES); Strombocactus disciformis (DC.) Britton & Rose. (І CITES). The solutions for sterilization were selected, the time of sterilization, and the sterilization procedure of seeds was approbated. The expedience of usage of stems of juvenile plants grown in vitro for propagation of these species of cacti was proved. The possibility of an obtaining of the necessary quantity of plant material by the way of indirect organogenesis is shown. The proper content of nutrient media and favourable conditions of the cultivation for all stages of the micropropagation were approved.

Development of the first axillary in vitro shoot multiplication protocol for coconut palms

The coconut palm or “tree of life” is one of nature’s most useful plants and the demand for its fruit is increasing. However, coconut production is threatened by ageing plantations, pests and diseases. Currently, the palm is exclusively propagated via seeds, limiting the amount of planting material. A novel micropropagation method is presented, based on axillary shoot formation. Apical meristems of in vitro coconut seedlings are cultured onto Y3 medium containing 1 µM TDZ. This induces the apical meristem to proliferate through axillary shoots in ~ 27% of the initiated explants. These axillary shoots are seen as white clumps of proliferating tissue and can be multiplied at a large scale or regenerated into rooted in vitro plantlets. This innovative micropropagation method will enable the production of disease-free, high quality in vitro plantlets, which will solve the worldwide scarcity of coconut planting material.

Features of the Organisation of Rooting Cuttings of Decorative Woody Plants Under Artificial Lighting with Led Equipment

Obtaining a technology for rooting cuttings and standard seedlings of decorative woody plants under artificial lighting conditions is relevant in connection with the need to increase the amount of planting material required for green building. Currently, cuttings are carried out mainly in greenhouses during the growing season. The development of technology for rooting cuttings using lamps with different lighting parameters opens up the possibility of creating installations that are effective in various conditions. In the course of the study, organisational and technological measures were taken to modernise the greenhouse and install equipment for growing planting material of decorative plants. In accordance with the plan to study the characteristics of various crops using the technology of dynamic regulation of the spectral composition and temporal characteristics of LED lighting in a greenhouse, an assortment of decorative tree species has been developed. Heated greenhouses are equipped with automatic irrigation hotbeds. Cutting of coniferous, deciduous, fast- and slow-growing species of decorative woody plants (7 species and 3 cultivars in total) was carried out. Rooting was carried out under illumination with batch-type LED installations. The success of rooting is studied when using LEDs of various production, with different spectral composition and varying illumination intensity during year-round cultivation. The findings are necessary not only for the development of a regional technology for the year-round production of rooted cuttings of decorative woody plants, but also for the development of the engineering specifics of lighting equipment.

A stable isotope dilution method for a highly accurate analysis of karrikins

BackgroundKarrikins (KARs) are recently described group of plant growth regulators with stimulatory effects on seed germination, seedling growth and crop productivity. So far, an analytical method for the simultaneous targeted profiling of KARs in plant tissues has not been reported.ResultsWe present a sensitive method for the determination of two highly biologically active karrikins (KAR1 and KAR2) in minute amounts of plant material (< 20 mg fresh weight). The developed protocol combines the optimized extraction and efficient single-step sample purification with ultra-high performance liquid chromatography-tandem mass spectrometry. Newly synthesized deuterium labelled KAR1 was employed as an internal standard for the validation of KAR quantification using a stable isotope dilution method. The application of the matrix-matched calibration series in combination with the internal standard method yields a high level of accuracy and precision in triplicate, on average bias 3.3% and 2.9% RSD, respectively. The applicability of this analytical approach was confirmed by the successful analysis of karrikins in Arabidopsis seedlings grown on media supplemented with different concentrations of KAR1 and KAR2 (0.1, 1.0 and 10.0 µmol/l).ConclusionsOur results demonstrate the usage of methodology for routine analyses and for monitoring KARs in complex biological matrices. The proposed method will lead to better understanding of the roles of KARs in plant growth and development.

Biofertlizer Lumbrical improves the growth and ex vitro acclimatization of micropropagated pear plants

In vitro micropropagation of plants is highly useful for obtaining large quantities of planting material with valuable economic qualities. However, plantlets grow in vitro in a specific environment and the adaptation after the transfer to ex vitro conditions is difficult. Therefore, the acclimatization is a key step, which mostly determines the success of micropropagation. The aim of this investigation was to study the effect of the biofertlizer Lumbrical on ex vitro acclimatization of micropropagated pear rootstock OHF 333 (Pyrus communis L.). Micropropagated and rooted plantlets were potted in peat and perlite (2:1) mixture with or without Lumbrical. They were grown in a growth chamber at a temperature of 22&amp;plusmn;2 &amp;deg;C and photoperiod of 16/8 hours supplied by cool-white fluorescent lamps (150 &amp;micro;mol m-2 s-1 Photosynthetic Photon Flux Density, PPFD). The plants were covered with transparent foil to maintain the high humidity, and ten days later, the humidity was gradually decreased. Biometric parameters, anatomic-morphological analyses, net photosynthetic rate and chlorophyll a fluorescence (JIP test) were measured 21 days after transplanting the plants to ex vitro conditions. The obtained results showed that the plants, acclimatized ex vitro in the substrate with Lumbrical, presented better growth (stem length, number of leaves, leaf area and fresh mass) and photosynthetic characteristics as compared to the control plants. This biostimulator could also be used to improve acclimatization in other woody species

Determination of AB-FUBINACA and 5F-NPB-22 in rats exposed to "Bonsai" via inhalation and analysis of seized product.

Synthetic cannabinoids (SCs) are the most rapidly growing class of recreational designer drugs. Illicit drug manufacturers began to produce herbal smoking materials under a variety of brands names, e.g. "Spice, K2, Bonsai, Yucatan Fire". They were appeared on the European market in 2008. In this study, types of SCs in the herbal product sold as "Bonsai" in Turkey were determined and the identification of these substances in biological samples collected from rats depending on the inhalation of different amounts of plant material were aimed. To determine the SC species in the content of the plant product, analysis was performed via gas chromatography-mass spectrometry. Liquid-liquid extraction methods were utilized for blood and organ samples, while solid-phase extraction with β-glucuronidase enzyme treatment was applied for urine sample preparation. The relationship between the amount of burned plant and the amount of SCs accumulated in the blood, urine and organ samples of rats exposed to the plant product by inhalation was examined by liquid chromatography-tandem mass spectrometry. AB-FUBINACA and 5F-NPB-22 were detected in the herbal product. A significant correlation was found between the amount of herbal product inhaled and the prevalence of SCs, especially in lung tissues while no SCs were detected in the blood and urine samples of rats. There is currently no study on biological samples of individuals exposed to herbal products containing SCs by inhalation. Regarding the findings obtained in this study, the overall increase in the amounts of herbal product inhaled was demonstrated to pose a potential risk to humans.

Evaluación de la propagación asexual por esquejes en Sesuvium verrucosum Raf. (Aizoaceae)

La sequía y la salinidad son condiciones de algunos suelos que limitan la producción agrícola a nivel mundial. Para combatir estos efectos, se ha hecho énfasis en una técnica llamada fitodesalinización, la cual consiste en el uso de especies halófitas con la capacidad para extraer cantidades importantes de sodio (Na+) del medio afectado. Sin embargo, ésta es una práctica agrícola que demanda una gran cantidad de material vegetal por hectárea, lo cual dificulta su implementación en los sistemas productivos con problemas de salinidad. El objetivo de este trabajo fue determinar la capacidad de enraizamiento de los esquejes de la especie halófita S. verrucosum y analizar los cambios de magnitud en sus parámetros biométricos, a partir de diferentes concentraciones en el tratamiento hormonal, en un periodo de 90 días. Para el desarrollo del experimento se utilizó un diseño experimental completamente al azar y se consideraron dos factores: las concentraciones del ácido indol butírico (AIB) (0, 800, 1 200, 1 600, 2 000 y 4 000 mg L-1) y la posición de los esquejes (basal-intermedia e intermedia-apical). Los resultados obtenidos demostraron diferencias significativas entre los diferentes tratamientos. Los esquejes de la porción intermedia-apical fueron los que obtuvieron el mayor porcentaje de enraizamiento, número de raíces, peso fresco y seco, tanto de la raíz como de la parte área de la planta, diámetro del tallo y altura de la planta, principalmente a una concentración de 2 000 mg L-1. A través de este método, es posible implementar la fitodesalinización de manera extensiva.

Tagging and catching: rapid isolation and efficient labeling of organelles using the covalent Spy-System in planta

BackgroundUp-to-now, several biochemical methods have been developed to allow specific organelle isolation from plant tissues. These procedures are often time consuming, require substantial amounts of plant material, have low yield or do not result in pure organelle fractions. Moreover, barely a protocol allows rapid and flexible isolation of different subcellular compartments. The recently published SpySystem enables the in vitro and in vivo covalent linkage between proteins and protein complexes. Here we describe the use of this system to tag and purify plant organelles.ResultsWe developed a simple and specific method to in vivo tag and visualize, as well as isolate organelles of interest from crude plant extracts. This was achieved by expressing the covalent split-isopeptide interaction system, consisting of SpyTag and SpyCatcher, in Nicotiana benthamiana leaves. The functionality of the SpySystem in planta, combined with downstream applications, was proven. Using organelle-specific membrane anchor sequences to program the sub-cellular localization of the SpyTag peptide, we could tag the outer envelope of chloroplasts and mitochondria. By co-expression of a cytosolic, soluble eGFP-SpyCatcher fusion protein, we could demonstrate intermolecular isopeptide formation in planta and proper organelle targeting of the SpyTag peptides to the respective organelles. For one-step organelle purification, recombinantly expressed SpyCatcher protein was immobilized on magnetic microbeads via covalent thiol-etherification. To isolate tagged organelles, crude plant filtrates were mixed with SpyCatcher-coated beads which allowed isolation of SpyTag-labelled chloroplasts and mitochondria. The isolated organelles were intact, showed high yield and hardly contaminants and can be subsequently used for further molecular or biochemical analysis.ConclusionThe SpySystem can be used to in planta label subcellular structures, which enables the one-step purification of organelles from crude plant extracts. The beauty of the system is that it works as a covalent toolbox. Labeling of different organelles with individual tags under control of cell-specific and/or inducible promoter sequences will allow the rapid organelle and cell-type specific purification. Simultaneous labeling of different organelles with specific Tag/Catcher combinations will enable simultaneous isolation of different organelles from one plant extract in future experiments.

Spatial and Temporal Trends in Diet for Pinfish (Lagodon rhomboides) from Turtle Grass (Thalassia testudinum) Beds with Contrasting Environmental Regimes in the Lower Laguna Madre, Texas

Seagrass meadows in far South Texas are dominated by turtle grass (Thalassia testudinum), and juvenile pinfish (Lagodon rhomboides) are abundant grazers of seagrass epifauna. As they grow, pinfish exhibit an ontogenetic dietary shift and begin consuming increasing amounts of plant material. In this study, contrasting environmental regimes in two seagrass meadows were examined to identify how they might affect pinfish diets, diet shifts, and growth across the growing season. The eastern site was sheltered from prevailing southeasterly winds by a barrier island, while the western site was shallower and subjected to higher wind energy and turbidity. Gut contents from 290 pinfish were quantified from May through October 2016 and multivariate analyses showed strong but overlapping separation of diets and temporal change in feeding habits. Although there were spatial and temporal differences between the two sites, a site by time interaction indicated different temporal trends. Nonetheless, ooze (amorphous detritus) and aquatic invertebrates (primarily amphipods) were the primary diet items consumed at both localities across time. Pinfish at both sites consumed seagrass, which increased in importance as the growing season progressed, although volume and consumption rate did not differ between sites. Growth rates of pinfish were similar at both sites, but pinfish from the western site were larger and fewer in number across the study. Although pinfish diets differed under different environmental regimes, they were generalist consumers in South Texas seagrass meadows with high diet variability that trended towards herbivory in late summer and fall.

Утворення додаткових коренів у стеблових живців ялини Канадської (форма конічна) залежно від оброблення росторегулятивною речовиною

Наведено результати дослідження утворення та розвитку кореневої системи у стеблових живців ялини канадської (форма конічна) з однорічною деревиною та живців із відрізком дворічної деревини (п'яткою). Встановлено, що кількість коренів, які утворюються у стеблових живців ялини канадської (форма конічна) та їхня загальна довжина залежать від типу живця й оброблення розчинами росторегулятивної речовини. Більша кількість коренів утворюється у живців з відрізком дворічної деревини. Довжина коренів у живців цього типу також більша. Виявлено, що найефективнішим для стимулювання утворення додаткових коренів у живців ялини канадської з однорічною деревиною є розчин ІМК з концентрацією 50 мг/л, під впливом дії якої кількість коренів у них становить 17,8 шт. і достовірно переважає контрольний варіант на 2,7 шт. У живців з дворічною деревиною кількість адвентивних коренів становить 25,3 шт., перевага над контролем – 4,2 шт. Досліджено, що в усіх варіантах із застосуванням для оброблення живців розчинів ІМК одержано достовірне зростання довжини основних коренів порівняно з контрольним варіантом. Живці ялини канадської (форма конічна) з дворічною деревиною (п'яткою) формують найбільшу за довжиною кореневу систему, яка має 114,2 см, живці з однорічною деревиною – 96,0 см під впливом оброблення ІМК із концентрацією 50 мг/л.

Influence of Phenology and Post-Harvest Processing of Vegetal Material on the Allelopathy of Annoni Grass (Eragrostis plana) Extracts

ABSTRACT: The phenological stage, post-harvest processing and quantity of material can interfere in the allelopathic activity of extracts. The objective of this study is to test the allelopathic effect of aqueous extracts of the leaves of Annoni grass (Eragrostis plana), an invader of natural pastures. Twelve extracts were prepared by combining the phenological stage of the plants when harvesting the leaves (vegetative; flowering), post-harvest processing before extraction (fresh; dry at 40 oC), and the amount of plant material (5, 15, and 25 g 100 mL-1 distilled water). Two bioassays were conducted in a germination chamber, with evaluation of germination and growth of seedlings of lettuce (Lactuca sativa) and white clover (Trifolium repens). The extracts with higher allelopathic activity were further developed with 15 and 25 g of dried leaves and harvested in plants at the vegetative stage. Germination was attributed to being the greatest contribution to the divergence between the extracts. White clover was more sensitive to extracts, for which 50% to 67% of the extracts were effectively allelopathic (≥50% inhibition); in lettuce, between 8% and 58% of the extracts exhibited this potential. The phenological stage was the factor with a greater individual effect on the allelopathic activity on lettuce and white clover seedlings, and on lettuce germination. The post-harvest processing responded by the greater variation on germination and germination speed index of white clover. The sensitivity of the white clover to extracts of Annoni grass suggests a compromise of its establishment in pastures with the presence of the invader.

A greener approach towards isolating clove oil from buds of Syzygium aromaticum using microwave radiation

The selection of suitable technique and apposite operating conditions are the key attributes for efficient extraction of phytochemicals from plant materials. Microwave assisted extraction (MAE) was incorporated to isolate essential oil from clove buds and the parameters were optimized using the grey based Taguchi method to maximize the yield of clove oil (Y, % w/w), eugenol content (y, % w/w) and bacterial inhibition. The results of single response analysis and multi-response analysis were compared. It was possible to achieve the best results (Y: 13.11%, w/w; y: 11.93%, w/w; zone of inhibition (ZOI): 24.80 mm) using the optimized conditions of multi-response study viz., amount of plant material − 30 g; volume of water − 200 mL; power − 600 W and extraction time − 30 min. The wastes generated during extraction were explored for their utilization as a by-product or raw material for other processes. Moreover, the performance of MAE was compared with that of hydrodistillation in terms of processing conditions, responses, energy and environmental impact. It was possible to reduce water usage, extraction time, energy requirement and CO2 emission while achieving the improved quantity and quality of essential oil along-with a higher ZOI using the green MAE technique.

A Simple and Rapid DNA Isolation Method from Dry Pea Seeds Suitable for PCR Analyses

Pea (Pisum sativum L.) is an important legume grown and consumed extensively worldwide. As a rich source of proteins, carbohydrates and vitamins, peas are important in human nutrition. In this study, we compared four different DNA isolation methods from four pea cultivars (F95-927, Specter, Windham and Nicoleta), with some modifications to the original extraction protocols: one SDS extraction buffer without PVP and one SDS extraction buffer with 1% PVP; one CTAB extraction buffer without PVP and one with 1% (1.5%) PVP. For all four extraction methods we used the same quantity of plant material (0.05 g) and an equal quantity of extraction buffer, so the comparison between methods to be very accurate. To establish which is the most efficient extraction method, after DNA isolation and purification, we submitted our samples to PCR analyzes with two markers: ISSR marker 17899A and SSR marker AA175. In my study, based on spectrophotometric measurements and PCR results, I concluded that both CTAB extraction methods were not suitable for DNA extraction from dry pea seeds because they did not show amplification products. The most appropriate DNA extraction method was SDS1 which provided a good quality DNA.

Micropropagation and in vitro elicitation of licorice (Glycyrrhiza spp.)

The Glycyrrhiza genus, in the Fabaceae family, includes numerous medicinal and edible herbs that contain a wide range of phytochemicals which are used pharmaceutically and commercially. Glycyrrhiza spp. are under the threat of overexploitation and depletion; therefore, there is an urgent need for conservation. It is advantageous to develop in vitro techniques not only for propagation, multiplication and preservation but also for elicitation of secondary metabolite production. This review provides information on the micropropagation of different Glycyrrhiza spp. related to the use of different explants, the combination of plant growth regulators with different sterilization strategies, the culture conditions and additional factors influencing in vitro propagation (such as light, temperature, humidity and pH). Enhanced production of bioactive secondary metabolites by different elicitors is discussed. Different strategies for conservation of germplasm from Glycyrrhiza spp., namely encapsulation, cryopreservation and slow-growth system, are deliberated. The storage and availability of germplasm for commercial cultivation over the long run along with their germplasm evaluation using random amplification of polymorphic DNA and inter-simple sequence repeat techniques have also been presented. Successful multiplication, elicitation and downstream processing will lead to the production of not only greater quantities of planting material with improved quality but also commercially desired metabolites. The production of clinically novel compounds through biotransformation by cultured Glycyrrhiza spp. cells which can be used in future studies on somaclonal variation, genetic transformation and drug discovery is also reviewed.