Genome‑wide identification and analysis of the trihelix transcription factors in sunflowerOriginal Papers

J. SONG, W.Y. SHEN, S. SHAHEEN, Y.Y. LI, Z.R. LIU, Z. WANG, H.B. PANG, Z. AHMED

Biologia plantarum 65:80-87, 2021 | DOI: 10.32615/bp.2021.006  

The trihelix genes encode plant-specific transcription factors, which play a vital role in plant morphological and developmental processes. However, information about the presence of trihelix genes in sunflower (Helianthus annuus L.) is scarce. Sunflower belongs to composite family and possesses strong drought and salt-alkali tolerance. In this study based on H. annuus genome data, we have identified and analyzed the trihelix genes with a complete description of their physical and chemical properties, phylogenetic relationships, motif composition, chromosome distribution, exon-intron structure, cis-acting elements,...

HSP70 plays an ambiguous role during viral infections in plantsReviews

V. HÝSKOVÁ, K. BĚLONOŽNÍKOVÁ, N. ČEŘOVSKÁ, H. RYŠLAVÁ

Biologia plantarum 65:68-79, 2021 | DOI: 10.32615/bp.2021.001  

Heat shock and almost all types of stresses associated with oxidative stress are accompanied by heat shock protein (HSP) expression. HSPs are involved in refolding denatured proteins and directing unrepairable proteins for degradation. Thus, under stress conditions, HSPs help to restore cellular balance. However, in virus-infected plants, HSP70 can have both positive and negative effects because viruses usually recruit HSP70. HSP70 can promote the replication and translation of the viral genome, the formation of viral replication complexes, and the propagation of viral particles from cell to cell and throughout the plant. HSP gene silencing...

Cloning and functional analysis of the promoter of the sesquiterpene synthase gene ASS1 in Aquilaria sinensis Original Papers

Y.H. XU, F.F. LV, P.W. SUN, M.H. TIAN, J.H. WEI

Biologia plantarum 65:60-67, 2021 | DOI: 10.32615/bp.2020.141  

Agarwood, the resin part of Aquilaria spp., is valued in medicine, perfumes, and incense. The most important components of agarwood are sesquiterpenes, which are produced only when a healthy tree is wounded. Agarwood sesquiterpene synthase 1 (ASS1) is one of key enzymes responsible for the biosynthesis of sesquiterpenes in Aquilaria sinensis (Lour.) Gilg, and it is a typical wound-inducible synthase. To elucidate its regulatory mechanism at the transcriptional level, a 978-bp sequence upstream of the translation initiation codon ATG of the promoter for ASS1 was cloned. Computational analysis revealed that this promoter contained...

Molecular cloning and functional analysis of the thioredoxin gene SikTrxh from Saussurea involucrataOriginal Papers

L. ZHANG, C. LIU, F.F. CHENG, X.Y. GUO, Y.X. LI, A.Y. WANG, J.B. ZHU

Biologia plantarum 65:47-59, 2021 | DOI: 10.32615/bp.2020.155  

Thioredoxins are oxidoreductases that help to maintain redox homeostasis in plants under abiotic stress. In this study, a new thioredoxin gene, SikTrxh, was cloned from Saussurea involucrata (Kar. & Kir.), a perennial herb that grows in the high alpine mountains of Central Asia. Bioinformatics analysis shows that the full-length cDNA of SikTrxh consisted of 565 bp with a 354-bp open reading frame and encoded a 117 amino acid protein. Using quantitative reverse transcription (RT) PCR, we found that the expression of the SikTrxh gene was induced by salt, cold, and drought stresses, suggesting that this protein played a...

Growth response of Oryza sativa seedlings to graphene oxide and its variability among genotypesOriginal Papers

Y. HE, H.M. WEI, S.J. LIU, Y.C. XU, Z.Y. ZHU, H. YAN, J.X. LI, Z.H. TIAN

Biologia plantarum 65:39-46, 2021 | DOI: 10.32615/bp.2020.124  

With the extensive utilization of graphene nanomaterials, they inevitably enter our environment. The potential phytotoxicity and environmental impact of graphene oxide (GO) have recently attracted much attention. We designed the experiment based on seed germination, seedling morphology, physio-biochemical properties, and antioxidant enzyme activities of five rice genotypes (9311, MH63, R527, K866, and Nipponbare) under six concentrations of GO (0, 5, 10, 50, 100, and 150 mg dm^-3). We studied the effects of different concentrations of GO on germination index (GI), shoot length (SL) and root length (RL), adventitious root number, shoot and...

Overexpression of the Panax ginseng MYB4 gene enhances stress tolerance in transgenic Arabidopsis thalianaOriginal Papers

W.H. LIAN, T.X. SUN, X.Y. MENG, R. SUN, F. HUI, Y.N. JIANG, Y. ZHAO

Biologia plantarum 65:27-38, 2021 | DOI: 10.32615/bp.2020.164  

The myeloblastosis (MYB) transcription factors are essential for plant stress responses. They can enhance plant tolerance to abiotic stresses (e.g., drought, salinity, and cold) via improved physiological and biochemical responses including the accumulation of metabolites. In this study, we constructed a Panax ginseng MYB4 (PgMYB4) gene expression vector and established the stable transgenic Arabidopsis thaliana lines to study the effects of this gene on plant stress tolerance. The germination rate and seedling taproot length were greater for the PgMYB4-overexpressing plants than for the wild-type plants. Accordingly,...

Ectopic expression of a wheat superoxide dismutase gene TaSOD5 enhances salt and oxidative stress tolerance in ArabidopsisOriginal Papers

Y. WANG, Q. LIU, Y. LIU, G. LI, G. XIA, M. WANG

Biologia plantarum 65:19-26, 2021 | DOI: 10.32615/bp.2020.147  

Superoxide dismutase (SOD) is a crucial reactive oxygen species (ROS) scavenger, which converts superoxide radical to H₂O₂, so it is thought to enhance abiotic stress tolerance by reducing ROS and thus avoiding oxidative damage. In this study, we isolated a salt- and oxidative stress-responsive copper-zinc (Cu/Zn) SOD encoding gene TaSOD5 from wheat. The ectopic overexpression of TaSOD5 in Arabidopsis thaliana increased total SOD and Cu/Zn SOD activities and enhanced tolerance to salt stress. Arabidopsis ectopically expressing TaSOD5 possessed a superior resistance to oxidative stress stimulated...

Genome-wide association study of low nitrogen tolerance traits at the seedling stage of rapeseedOriginal Papers

C.L. ZENG, H.P. WAN, X.M. WU, X.G. DAI, J.D. CHEN, Q.Q. JI, F. QIAN

Biologia plantarum 65:10-18, 2021 | DOI: 10.32615/bp.2020.144  

The large application of nitrogen fertilizer will cause soil deterioration and pollute the environment. Reduction of nitrogen inputs and maintaining high yields are therefore essential to ensure a more sustainable agriculture. However, little information is available about rapeseed (Brassica napus L.) low nitrogen tolerance. We evaluated low nitrogen tolerance of 304 rapeseed accessions at seedling stage and performed a genome-wide association study to detect low nitrogen tolerance-related quantitative trait loci. A natural population comprising 304 B. napus inbred lines was genotyped with a Brassica 60K Illumina Infinium...

EditorialEditorial

H. Plchová

Biologia plantarum 65:2021  

Molecular cloning and characterization of a novel gene MsKMS1 in Medicago sativaOriginal Papers

B. HAN, P. ZHANG, Z.-Q. ZHANG, Y.-F. WANG, T.-M. HU, P.-Z. YANG

Biologia plantarum 65:1-9, 2021 | DOI: 10.32615/bp.2020.059  

Vacuole membrane proteins play a critical role in the regulation of plant physiological processes including normal growth and development, and responses to stresses. The killing me slowly 1 (KMS1) gene that encodes a soluble N-ethylmaleimide-sensitive fusion attachment receptor (SNARE) domain-containing vacuole membrane protein was first reported in Arabidopsis. Currently, the function of KMS1 in other plants under stress is poorly understood. In this study, we report cloning, expression, and characterization of a novel KMS1 gene in alfalfa (Medicago sativa L.), designated MsKMS1 (GenBank accession No....

Future agroclimatic conditions and implications for European grasslandsSpecial Issue: Festulolium

M. TRNKA, J. BALEK, M.A. SEMENOV, D. SEMERÁDOVÁ, M. BĚLÍNOVÁ, P. HLAVINKA, J.E. OLESEN, J. EITZINGER, A. SCHAUMBERGER, P. ZAHRADNÍČEK, D. KOPECKÝ, Z. ŽALUD

Biologia plantarum 64:865-880, 2020 | DOI: 10.32615/bp.2021.005  

Grasslands play a significant role in livestock fodder production and thus, contribute to food security worldwide while providing numerous additional ecosystem services. However, how agroclimatic conditions and adverse weather events relevant for grasslands will change across the European grassland areas has not been examined to date. Using a single reference setup for soil and management over 476 European sites defined by climate stations, we show the probability of eight selected adverse weather events with the potential to significantly affect grassland productivity under climate change and how these events vary regionally across Europe. Changes...

Persistency, yield, and silage quality of Festulolium cultivars over a consecutive five-year period under a mild Atlantic climateSpecial Issue: Festulolium

N. CURRAN, D. GROGAN, D. MILBOURNE, S.L. BYRNE, E. O'RIORDAN, M. HANLEY, J. GRANT, T.R. HODKINSON, S. BARTH

Biologia plantarum 64:856-864, 2020 | DOI: 10.32615/bp.2020.157  

Festulolium are hybrids between species within the Lolium and Festuca genera. They are attractive candidates for hybridisation due to their complementary characteristics in the context of forage production and environmental challenges, with Lolium exhibiting greater digestibility and Festuca possessing superior resilience to abiotic stress. To establish the extent of the representation of these characteristics in different Festulolium hybrids, a five-year field experiment was undertaken in a mild Atlantic climate. Four alternative species combinations and six pure species (controls) were evaluated for dry matter yield...

Genome-wide identification and expression analysis of the potato ZIP gene family under Zn-deficiencyOriginal Papers

X. B. LI, H.C. SUO, J.T. LIU, L. WANG, C.C. LI, W. LIU

Biologia plantarum 64:845-855, 2020 | DOI: 10.32615/bp.2020.125  

Zinc deficiency is a worldwide problem for crops including potato (Solanum tuberosum L.), the fourth most important crop worldwide. The zinc/iron-regulated transporter-like protein (ZIP) transporter family is thought to play key roles in Zn uptake and transport. However, little is known about the potato ZIP family. In this study, 12 genes encoding members of the ZIP family were identified in the potato genome. The 12 StZIP genes were predicted to encode proteins of 220 - 407 amino acids harboring 5 - 9 putative transmembrane domains (TMDs), and 11 of these proteins had a variable region rich in histidine residues between TMDIII and TMDIV....

Reference gene selections for real time quantitative PCR analysis of gene expression in different oat tissues and under salt stressOriginal Papers

Z.L. DUAN, W.H. HAN, L. YAN, B. WU

Biologia plantarum 64:838-844, 2020 | DOI: 10.32615/bp.2020.153  

Appropriate choice of reference genes for data normalization is of critical importance for accurate real time reverse transcription quantitative PCR (RT-qPCR) analysis of gene expression. Oat is an agriculturally important crop cultivated widely around the world for grain or forage, and appropriate reference genes for reliable gene expression analysis remain to be identified. In this study, we selected nine candidate reference genes based on available oat RNA-seq data. We then conducted a systematic evaluation of the relative expression stability of these genes in different tissues and under salt stress using statistical algorithms, geNorm,...

Simultaneous induction of anthocyanin and peroxidase by sucrose in hypocotyls and roots of Chinese red radish seedlingsOriginal Papers

L. WANG, X.-X. JING, P.-P. ZHAO, L.-F. WANG, Y. YIN, Y.-F. LI

Biologia plantarum 64:828-837, 2020 | DOI: 10.32615/bp.2020.156  

Anthocyanin and peroxidase (POD) are important active compounds in Chinese red radish (Raphanus sativus L.). The effects of exogenous sucrose, mixture of fructose and glucose (F/G 1:1), and mannose on anthocyanin accumulation and POD activity were investigated in hypocotyls and roots of red radish seedlings. Sucrose was most effective in inducing anthocyanin accumulation and POD activity in both organs, followed by F/G; mannose caused only a little increase of anthocyanin content and POD activity in hypocotyls, but a decrease in roots. The distribution of anthocyanin accumulation was different between hypocotyls and roots in the presence or...

Festulolium field performance under fluctuating growing conditions in LithuaniaSpecial Issue: Festulolium

V. KEMEŠYTĖ, K. JAŠKŪNĖ, G. STATKEVIČIŪTĖ

Biologia plantarum 64:821-827, 2020 | DOI: 10.32615/bp.2020.165  

Festulolium cultivars are widely utilized in Lithuania because they are persistent under abiotic stresses and are high yielding. However, changing climate challenges the existing Festulolium cultivars to adapt to new growing conditions and still maintain the yield. In this study, we aimed at evaluating the yield stability of two Festulolium cultivars in field trials under fluctuating Lithuanian conditions. The mean total dry matter yield (DMY) of both Festulolium cultivars fluctuated greatly between the years and ANOVA analysis showed a significant effect of environment on total DMY as well as DMY of each cut, but the genotype...

Morphological diversity of seeds of Polish Festulolium cultivars depending on weather conditionsSpecial Issue: Festulolium

B. BORAWSKA-JARMUŁOWICZ, G. MASTALERCZUK, E. MAŁUSZYŃSKA, W. BORUCKI

Biologia plantarum 64:814-820, 2020 | DOI: 10.32615/bp.2020.146  

Morphological traits of seeds of Festulolium braunii (K. Richt.) A. Camus cultivars were studied in relation to weather conditions. The obtained results show that morphological traits of seeds varied among cultivars and years of cultivation. The values of seed characteristics were higher in year 2016 due to more favorable weather conditions for plant growth. It was found that the husked seeds of Festulolium cultivars were characterized by a similar length and width regardless of the year, whereas naked seeds significantly differed in these characteristics. Mean values (from the research years) of seed parameters, such as thousand seed...

Virus induced PhFTRv gene silencing results in yellow-green leaves and reduced cold tolerance in petuniaOriginal Papers

L. SANG, L. PENG, Z. QIU, F. LUO, G. CHEN, L. GAO, Y. YU, J. LIU

Biologia plantarum 64:807-813, 2020 | DOI: 10.32615/bp.2020.151  

Ferredoxin-thioredoxin reductase (FTR) is an iron-sulfur protein that supplies electrons from photochemically reduced ferredoxin (Fd) to thioredoxin (Trx) in the ferredoxin/thioredoxin system in chloroplasts. The FTR is a heterodimer with a variable subunit (FTRv) and a catalytic subunit (FTRc). The function of FTRv is not well known. In petunia (Petunia hybrida), FTRv is a single-copy gene, which is named PhFTRv. In this study, the spatio-temporal expression of PhFTRv in petunia was analyzed, and PhFTRv transcription was found to be high in leaves and stems. A tobacco rattle virus gene silencing was used in...

Transcriptome-based screening and the optimal reference genes for real-time quantitative PCR in Rehmannia chingii and R. henryi Original Papers

X. ZUO, F.-Q. WANG, X.-R. LI, M.-M. LI

Biologia plantarum 64:798-806, 2020 | DOI: 10.32615/bp.2020.154  

Real time quantitative PCR (qPCR) is a powerful tool for studying the expression of specific genes. The accuracy and reliability of qPCR analysis data require the selection of reference genes with stable expression. However, the reference genes that can be used for qPCR of Rehmannia chingii and R. henryi have not yet been identified. In this study, based on the transcriptome data of R. chingii and R. henryi, we initially selected genes with relatively stable expression in different samples. We screened six candidate reference genes in R. chingii and R. henryi and calculated their expression abundance by real...

Breeding and genetics of two new amphiploid Festulolium synthetics with improved yield and digestibilitySpecial Issue: Festulolium

J. BAERT, K. VAN LAERE, C. VAN WAES, A. GHESQUIERE, J. APER

Biologia plantarum 64:789-797, 2020 | DOI: 10.32615/bp.2020.138  

In order to introduce drought tolerance and improved cell wall digestibility from fescue in fodder ryegrasses, we developed two amphiploid Festulolium synthetics. One is a synthetic composed of three selected drought tolerant F1 hybrid genotypes from a cross between tetraploid Lolium multiflorum and hexaploid Festuca arundinacea, further on called LMFA. The other is a synthetic composed of five selected genotypes with soft leaves from a cross between tetraploid Lolium perenne and tetraploid Festuca pratensis, further on called LPFP. We produced seeds in polycrosses of two generations of both amphiploids, i.e.,...

Effects of cadmium and lead on the growth and the activity of peroxidase and superoxide dismutase of blueberry plantlets in vitro Brief Communications

J.Z. CHEN, L.P. ZHOU, C.Y. TIAN, F. YANG, G.Z. WANG

Biologia plantarum 64:784-788, 2020 | DOI: 10.32615/bp.2020.143  

To study the effects of cadmium and lead on the growth of blueberry (Vaccinium ashei Reade) plantlets in vitro and on the activity of peroxidase (POD) and superoxide dismutase (SOD) of blueberry shoots, Cd²⁺ and Pb²⁺ were added separately to a cultivation medium. The results show that 0.01 mM Cd²⁺ significantly inhibited the growth of blueberry shoots, and the height and fresh mass of the shoots were significantly lower than those of the control; 0.05 mM Cd²⁺ significantly inhibited the proliferation of the shoots; the inhibitory effect on the growth and proliferation of blueberry in vitro...

Ultraviolet-B radiation induces cell death in root tips and reprograms metabolism in ArabidopsisOriginal Papers

P. ZHANG, R. WANG, Y. WANG, J. XU

Biologia plantarum 64:764-772, 2020 | DOI: 10.32615/bp.2020.122  

Ultraviolet-B (UV-B) radiation inhibits root system growth, however, the influence of UV-B radiation on the regulation of root development remains unclear. Here, we investigated the effects of UV-B radiation on root growth and metabolism in Arabidopsis using physiological, pharmacological, and transcriptome approaches. Our previous study has demonstrated that UV-B radiation depresses auxin accumulation in roots by reducing auxin biosynthesis, transport, and response. In this study, we found that UV-B radiation inhibited primary root (PR) growth by inducing root tip cell death and thereby disrupting cell division and elongation in root tips....

Identifying citrus CBL and CIPK gene families and their expressions in response to drought and arbuscular mycorrhizal fungi colonizationOriginal Papers

B. SHU, D. CAI, F. ZHANG, D.J. ZHANG, C.Y. LIU, Q.S. WU, C. LUO

Biologia plantarum 64:773-783, 2020 | DOI: 10.32615/bp.2020.123  

The calcineurin B-like protein (CBL)-interacting protein kinase (CIPK) complex is an essential calcium sensor and contributes to biotic and abiotic stress responses. However, citrus CBL and CIPK gene family members and their underlying roles during drought and arbuscular mycorrhizal fungi (AMF) colonization remain relatively unknown. In the present study, CBLs and CIPKs were characterized in Citrus sinensis by analyzing the presence of specific domains such as the elongation factor (EF)-hand motif in CBLs, and a protein kinase and an Asn-Ala-Phe domain in CIPKs. After mining the C. sinensis genome, we identified...

Sex-specific responses of Populus deltoides to combined salinity and calcium under waterlogging conditionsOriginal Papers

L.-F. MIAO, D.-D. LI, F. YANG, Z.-H. TAN

Biologia plantarum 64:753-763, 2020 | DOI: 10.32615/bp.2020.116  

In this study, we investigated the sex-specific ion uptake and physiological and biochemical responses to combined salinity and calcium treatments of male and female Populus deltoides under waterlogging conditions. Results indicate that both P. deltoides clones were sensitive to salinity and calcium. Under well-watered conditions, salinity stress could especially increase the Na content in female P. deltoides clones, whereas salinity and calcium could increase the K and Ca content in male clones. Waterlogging could evidently stimulate Na content in leaves under salinity, especially in female P. deltoides clones. However,...

γ-Aminobutyric acid induces transcriptional changes contributing to salt tolerance in creeping bentgrassOriginal Papers

Z. LI, B.Z. CHENG, Y. PENG, Y. ZHANG

Biologia plantarum 64:744-752, 2020 | DOI: 10.32615/bp.2020.117  

γ-Aminobutyric acid (GABA) regulates plant tolerance to abiotic stresses; however, a transcriptomic change and key stress-related genes induced by GABA have not been investigated in plants during a prolonged period of salt stress. Roots of creeping bentgrass (Agrostis stolonifera) cv. Penncross were pretreated with or without 0.5 mM GABA solution for 2 days and then subjected to salt stress for 20 days (150 mM NaCl solution for 3 d, 200 mM NaCl for another 3 d, and 250 mM NaCl for 14 d) in controlled growth chambers. The application of GABA significantly increased GABA content in roots and alleviated a salt-stress induced decrease in GABA...

Effects of 5-aminolevulinic acid on osmotic adjustment and antioxidant system in mung bean under chilling stressOriginal Papers

J.J. ZHAO, N. ZHOU, N.J. FENG, D.F. ZHENG

Biologia plantarum 64:736-743, 2020 | DOI: 10.32615/bp.2020.101  

Temperature is an important factor for growth, development, productivity and geographical distribution of many plants (Nahar et al. 2015). Chilling stress is a major abiotic stress of crop production in Northeast China. Chilling stress exposure has been shown to enhance production of reactive oxygen species (ROS) and oxidative stress occurs (Nahar et al. 2015). The ROS, which include superoxide radical (O₂^*-), hydrogen peroxide (H₂O₂), hydroxyl radical (.OH), and singlet oxygen (¹O₂), cause damage to structural proteins, nucleic acids, enzymes, cell membranes, and...

Analysis of ABC1 protein family members in Lepidium apetalum seeds and the expression of LaAbc1 in seedlings in response to abiotic stressesOriginal Papers

Q.L. YANG, Z.Y. CHEN, H. LU, H.T. XIE, J.Y. LI, Y. DU, S.C. HAN, H.P. ZHAO, H.X. ZHAO

Biologia plantarum 64:725-735, 2020 | DOI: 10.32615/bp.2020.104  

To study the biological function of activity of bcl complex (ABC1) proteins in Lepidium apetalum Willd., genes encoding ABC1 family proteins were identified from the seed transcriptome. The sequence most closely related to germination at a low temperature was selected and gene expressions in response to low temperature stress further studied. The results show that 21 ABC1 genes were expressed in seeds germinating at the low temperature: 4 genes were upregulated, 6 were downregulated, and 11 were not significantly different from controls. The results of fluorescence quantification of the low-temperature stress on the seedlings of 7-d-old...

Transcriptome-sequencing analyses reveal flower color formation in Strelitzia reginae Original Papers

R.H. FAN, B. LIN, N.Y. FANG, X.X. YE, M.L. HUANG, H.Q. ZHONG

Biologia plantarum 64:717-724, 2020 | DOI: 10.32615/bp.2020.102  

Strelitzia reginae is a popular cut flower that has blue petals and orange sepals. Flower color is an important plant trait; however, little is known about its molecular mechanisms in S. reginae. In this study, cDNA libraries were constructed for blue petals and orange sepals of S. reginae. A total of 75 487 unigenes were obtained from transcriptome sequencing and de novo assembly, of which 41.86 % were annotated by public databases. Ultra-high performance liquid chromatography analysis revealed that anthocyanins were the main pigment in blue petals, and that carotenoids controled pigment formation in the orange sepals. Using a...

Rapid increases in β-1,3-glucanase and chitinase activities are markers of resistance to Microdochium nivale in grasses of the Lolium‑Festuca complexSpecial Issue: Festulolium

K. MARZEC-SCHMIDT, K. HURA, A. PŁAŻEK

Biologia plantarum 64:710-716, 2020 | DOI: 10.32615/bp.2020.088  

Microdochium nivale causes pink snow mould - a destructive disease of seedlings, stem bases, and ears of winter grasses and cereals. Glucanase and chitinase belong to pathogenesis-related proteins and exhibit a specific activity in response to pathogens. The aim of this study was to investigate changes in the activity of these enzymes in the leaves of chosen forage grasses during Microdochium nivale infection. Different cultivars of Festuca arudinacea, F. pratensis, Festulolium brauni, Lolium multiflorum, and L. perenne were prehardened at 12 °C and hardened at 2 °C and then inoculated with M. nivale...

Prolonged drought stress induced changes in yield and physiological processes of Trifolium repens and Festulolium brauniiSpecial Issue: Festulolium

M. STANIAK, J. BOJARSZCZUK, P. KRASKA, C. KWIATKOWSKI, E. HARASIM

Biologia plantarum 64:701-709, 2020 | DOI: 10.32615/bp.2020.114  

Water deficit is one of the main environmental factors, and the recognition of plant response to drought stress seems to be crucial for the yield improvement of the important crops. The aim of the study was to investigate the effect of prolonged drought stress on dry matter yield, gas exchange parameters, and relative chlorophyll content in the leaves of Festulolium braunii and Trifolium repens grown in pure stands and in mixture. In the pot experiment, different levels of soil moisture (well-watered conditions and drought stress) and of cultivation method (pure stand and mixture) have been applied. The study has shown that all measured...