Biologia plantarum, 2014 (vol. 58), issue 4

Original Papers

In vitro culture of sweet basil: gas exchanges, growth, and rosmarinic acid production

C. Kiferle, M. Lucchesini, R. Maggini, A. Pardossi, A. Mensuali-Sodi

Biologia plantarum 58:601-610, 2014 | DOI: 10.1007/s10535-014-0434-5

Five in vitro culture systems with different ventilation rates were used to investigate the influence of vessel environment on photosynthesis, dark respiration, ethylene evolution, and rosmarinic acid (RA) production in sweet basil (Ocimum basilicum L.) micropropagated shoots. The systems under comparison were two bioreactors with either temporary (RITA™) or stationary (Growtek™) immersion, and three types of vessels (Magenta™, Microbox ECO ₂ ™, and PCCV25™) that are largely used for plant micropropagation. Shoots of green-leaved cv. Genovese and purple-leaved...

Exogenous sucrose can enhance tolerance of Arabidopsis thaliana seedlings to salt stress

Z. B. Qiu, Y. F. Wang, A. J. Zhu, F. L. Peng, L. S. Wang

Biologia plantarum 58:611-617, 2014 | DOI: 10.1007/s10535-014-0444-3

To investigate the physiological mechanisms of salt stress mitigated by exogenous sucrose, Arabidopsis thaliana seedlings grown on Murashige and Skoog medium were treated with 3 % (m/v) sucrose combined with 75, 150, and 225 mM NaCl for 3 d. Our results show that increased salinity significantly decreased the survival rate, fresh mass, content of proteins, chlorophyll a (Chl a), and chlorophyll b (Chl b), and activities of antioxidant enzymes, whereas enhanced the content of malondialdehyde. However, the treatment with sucrose significantly enhanced salt stress tolerance in the Arabidopsis seedlings by decreasing...

Isolation and expression analysis of anthocyanin biosynthetic genes in Morus alba L.

J. Li, R. -H. Lü, A. -C. Zhao, X. -L. Wang, C. -Y. Liu, Q. -Y. Zhang, X. -H. Wang, D. Umuhoza, X. -Y. Jin, C. Lu, Z. -G. Li, M. -D. Yu

Biologia plantarum 58:618-626, 2014 | DOI: 10.1007/s10535-014-0450-5

Anthocyanins from mulberry fruits are used in medicine. However, little anthocyanin can be detected in other tissues and sometimes also mulberry fruits are colorless. The aim of this study was to investigate which gene or genes have the strongest correlation with the anthocyanin biosynthesis. The expression of several anthocyanin synthesis genes were determined in different tissues of two white and two purple fruit cultivars. Genes encoding dihydroflavonol reductase (MaDFR) and anthocyanidin synthase (MaANS) showed a high expression only in fruit tissue of purple-fruit cultivars. During the development of mulberry fruits, the anthocyanin...

Natural genetic variation in MIR172 isolated from Brassica species

S. M. Shivaraj, P. Dhakate, P. Mayee, M. S. Negi, A. Singh

Biologia plantarum 58:627-640, 2014 | DOI: 10.1007/s10535-014-0441-6

The present study reports a natural variation in microRNA172 (MIR172) family members isolated from six species of genus Brassica. The analysis of nucleotide polymorphism across 44 Brassica MIR172 homologs revealed a higher conservation in the predicted precursors relative to flanking regions. Single nucleotide polymorphisms (SNPs) were detected in miRNA and miRNA*. The 21-nt miRNA sequence was conserved in all MIR172 members except MIR172a. However, the miRNA* sequence was conserved only in MIR172a compared to A. thaliana. Non-canonical Brassica variants of precursor miR172a were detected wherein...

Molecular cloning and expression analyses of FaFT, FaTFL, and FaAP1 genes in cultivated strawberry: their correlation to flower bud formation

R. Nakajima, S. Otagaki, K. Yamada, K. Shiratake, S. Matsumoto

Biologia plantarum 58:641-648, 2014 | DOI: 10.1007/s10535-014-0452-3

In this study, we cloned flowering-related genes FLOWERING LOCUS T (FT) and TERMINAL FLOWER1 (TFL1) from domesticated octaploid strawberries (Fragaria × ananassa) and analyzed their expression patterns in cultivars Tochiotome and Akihime. The floral meristem generation was induced under the short day and low temperature (SDLT), but not under the long day and high temperature (LDHT). We found that FaFT1, which is an orthologue of the Arabidopsis floral activator FT, was highly expressed in leaves under LDHT but not expressed in leaves under SDLT. On the other hand, the expression of FaTFL2,...

Over-expression of ApKUP3 enhances potassium nutrition and drought tolerance in transgenic rice

Z.-Z. Song, S.-Y. Yang, J. Zuo, Y.-H. Su

Biologia plantarum 58:649-658, 2014 | DOI: 10.1007/s10535-014-0454-1

ApKUPs are typical high-affinity potassium (K⁺) transporters of Alternanthera philoxeroides which are involved in its response to K⁺ starvation and abiotic stresses. In this study, the overexpression of ApKUP3 gene in rice resulted in enhanced K⁺ nutrition and drought tolerance of transgenic plants. Compared with wild-type (WT) plants, the transgenic plants showed a better growth performance and a strengthened K⁺ accumulation under different K⁺ supplies. The ApKUP3 overexpression in the rice plants also enhanced tolerance to a drought stress, as evidenced by a reduced leaf...

Cold-induced starch degradation in potato leaves - intercultivar differences in the gene expression and activity of key enzymes

D. Sitnicka, S. Orzechowski

Biologia plantarum 58:659-666, 2014 | DOI: 10.1007/s10535-014-0453-2

Exposure to low temperatures is one of the most important factors that generate abiotic stress in plants, and the rapid accumulation of soluble sugars belongs to significant metabolic responses to cold stress. The accumulation of soluble sugars may be at least partially triggered by an increased rate of starch degradation. The analysis of transcript profiles and starch degrading enzyme activities in leaves of two potato cultivars was performed during a 12-h exposure to 2 °C. An induction of β-amylase expression and activity as well as an accumulation of reducing sugars were observed in cv. Desiree. No accumulation of reducing sugars and no significant...

COR15B expression is affected by chloroplast functionality and its role in response to salt stress in Arabidopsis thaliana

D. Liu, L. Hou, W. C. Li, J. F. Cheng, Y. Q. Fu

Biologia plantarum 58:667-675, 2014 | DOI: 10.1007/s10535-014-0451-4

Cold-responsive (COR) genes participate in the response of plants to low-temperature stress. In this study, we isolated and characterized a cold-responsive and light-inducible gene COR15B from Arabidopsis thaliana. Chloroplast damage caused by mutations (albino mutants seca1, secy1, and tic20) or by a norflurazon (NF) treatment resulted in a reduction of COR15B transcription. A semi-quantitative RT-PCR analysis shows that COR15B was induced by the salt stress in an abscisic acid-dependent manner. An over-expression of COR15B in Arabidopsis resulted in transgenic lines more sensitive to...

Stomatal closure in sweet potato leaves induced by sulfur dioxide involves H₂S and NO signaling pathways

K. -D. Hu, J. Tang, D. -L. Zhao, L. -Y. Hu, Y. -H. Li, Y. -S. Liu, R. Jones, H. Zhang

Biologia plantarum 58:676-680, 2014 | DOI: 10.1007/s10535-014-0440-7

Sulfur dioxide (SO₂) is a well-known and widespread air pollutant but it also acts as signaling molecule in various processes in animals. However, there is limited information on the role of SO₂ in plants except of its toxicity. Here we studied the role of SO₂ on stomatal movements in sweet potato (Ipomoea batatas) leaves. SO₂, generated by Na₂SO₃/NaHSO₃ solutions, was applied on epidermal strips. We found that the SO₂ donor induced stomatal closure in a dose-dependent manner. Rapid increases in endogenous hydrogen sulfide and nitric oxide content levels...

The genetic basis of durum wheat germination and seedling growth under osmotic stress

M. Nagel, S. Navakode, V. Scheibal, M. Baum, M. Nachit, M. S. Röder, A. Börner

Biologia plantarum 58:681-688, 2014 | DOI: 10.1007/s10535-014-0436-3

Durum wheat (Triticum turgidum L. var. durum) is mainly produced under rainfed but often sub-optimal moisture conditions in the Mediterranean basin. A set of 114 durum wheat recombinant inbred lines (RILs) developed from the cross of cultivars Omrabi5 × Belikh2 were tested for the ability to tolerate moisture deficiency at the germination and early seedling growth stage. The stress was imposed by exposing the germinating grain to 12 % polyethylene glycol. It induced a measurable reduction in root length, shoot length, and the percentage of normal seedlings. The germination and seedling growth of Belikh2 were more strongly inhibited than...

Dynamic QTL analysis of the Na⁺ content, K⁺ content, and Na⁺/K⁺ ratio in rice roots during the field growth under salt stress

J. Sun, D. T. Zou, F. S. Luan, H. W. Zhao, J. G. Wang, H. L. Liu, D. W. Xie, D. Q. Su, J. Ma, Z. L. Liu

Biologia plantarum 58:689-696, 2014 | DOI: 10.1007/s10535-014-0445-2

Rice (Oryza sativa L.) is seriously impacted by global soil salinization. To determine the quantitative trait loci (QTLs) related to salt tolerance in rice roots, F_2:3 and BC₁F_2:3 populations derived from a cross between the cv. Dongnong 425 of high quality and yield and the salt-tolerant cv. Changbai 10, were studied at different development stages. Two genetic linkage maps of F_2:3 and BC1F_2:3 populations were constructed. A 66 mM NaCl solution was used to irrigate the field and to analyze the dynamic QTL of some rice root traits. Using unconditional and conditional QTL mapping methods,...

Alleviation of osmotic stress in Brassica napus, B. campestris, and B. juncea by ascorbic acid application

M. M. Alam, K. Nahar, M. Hasanuzzaman, M. Fujita

Biologia plantarum 58:697-708, 2014 | DOI: 10.1007/s10535-014-0447-0

The roles of ascorbic acid (AsA, 1 mM) under an osmotic stress [induced by 15 % (m/v) polyethylene glycol, PEG-6000] were investigated by examining morphological and physiological attributes in Brassica species. The osmotic stress reduced the fresh and dry masses, leaf relative water content (RWC), and chlorophyll (Chl) content, whereas increased the proline (Pro), malondialdehyde (MDA), and H₂O₂ content, and lipoxygenase (LOX) activity. The ascorbate content in B. napus, B. campestris, and B. juncea decreased, increased, and remained unaltered, respectively. The dehydroascorbate (DHA) content increased only...

Calcium and L-histidine effects on ascorbate-glutathione cycle components under nickel-induced oxidative stress in tomato plants

Z. Asrar, H. Mozafari, F. Rezanejad, S. Pourseyedi, M. M. Yaghoobi

Biologia plantarum 58:709-716, 2014 | DOI: 10.1007/s10535-014-0443-4

The effects of NiSO₄, calcium, and L-histidine (His) on the components of ascorbate-glutathione cycle, antioxidant enzymes and lipid peroxidation in a tomato cultivar Early Urbana Y was investigated. The activities of enzymes including catalase (CAT), guaiacol peroxidase (GPX), ascorbate peroxidase (APX), superoxide dismutase (SOD), glutathione reductase (GR), lipoxygenase (LOX), and phenylalanine ammonia lyase (PAL) were measured. In addition, the content of H₂O₂, ascorbate (ASC), dehydroascorbate (DHA), reduced glutathione (GSH), chlorophyll (Chl) a+b, carotenoids, proteins, malondialdehyde (MDA),...

Aluminum alleviates boron-deficiency induced growth impairment in tea plants

R. Hajiboland, S. Bahrami-Rad, S. Bastani

Biologia plantarum 58:717-724, 2014 | DOI: 10.1007/s10535-014-0425-6

Interaction between aluminum (Al) and boron (B) in Al accumulator species has not been characterized so far. In this work, tea [Camellia sinensis (L.) O. Kuntze] plants were cultivated hydroponically and treated with adequate (control) or low B supply (-B) without or with 300 μM Al (-B+Al) for 14 weeks. Growth of B-deficient plants was completely resumed by Al supplementation or even surpassed control plants regarding shoot biomass. Net photosynthetic rate was negatively influenced by the low B supply, and the Al treatment increased it up to the level of the control plants that was reflected in the higher content of saccharides. The activity...

Aluminum tolerance in Arabidopsis thaliana as affected by endogenous salicylic acid

D. Y. Guo, S. Y. Zhao, L. L. Huang, C. Y. Ma, L. Hao

Biologia plantarum 58:725-732, 2014 | DOI: 10.1007/s10535-014-0439-0

Endogenous salicylic acid (SA) functions in plant response to an aluminum stress were assessed. We used different Arabidopsis thaliana genotypes including snc1 with a constitutively high content of SA, sid2 and nahG (transgenic lines) both with a low content of SA, SA insensitive mutant npr1-1, and snc1/nahG (i.e., the nahG expression in the snc1 background) with a similar SA content as in wild type (WT) plants. Results show that the snc1 plants displayed obvious growth retardation of roots and shoots under the Al³⁺ stress, whereas the sid2, nahG, and npr1-1...

Evaluation of amino acid profile in contrasting arsenic accumulating rice genotypes under arsenic stress

A. Kumar, S. Dwivedi, R. P. Singh, D. Chakrabarty, S. Mallick, P. K. Trivedi, B. Adhikari, R. D. Tripathi

Biologia plantarum 58:733-742, 2014 | DOI: 10.1007/s10535-014-0435-4

Amino acids (AAs) play significant roles in metal binding, antioxidant defense, and signaling in plants during heavy metal stress. In the present study, the essential amino acids (EAAs), non-essential amino acids (NEAAs), as well as the enzymes of proline and cysteine biosynthetic pathways were studied in contrasting arsenic accumulating rice genotypes grown in hydroponic solutions with addition of arsenate (As^V) or arsenite (As^III). Under a mild As stress, the total AAs content significantly increased in both the rice genotypes with a greater increase in a low As accumulating rice genotype (LAARG; IET-19226) than in a high As...

Identification of a cold-inducible gene encoding calmodulin-binding protein from Eucalyptus dunnii through suppression subtractive hybridization

H. Z. Peng, H. B. Li, Q. Y. Jin, L. L. Wang, Y. Wang, N. Li, X. M. Zhang, Z. H. Gao

Biologia plantarum 58:743-750, 2014 | DOI: 10.1007/s10535-014-0457-y

Low temperature is the main limiting factor for cultivation expansion, fast growth, and high yield of Eucalyptus species. To investigate the mechanism of their cold tolerance, a cDNA subtraction library representing the cold-induced genes of Eucalyptus dunnii was constructed using suppression subtractive hybridization (SSH) technique. A gene encoding a calmodulin-binding protein (CaMBP) was identified from the SSH library, and the expression pattern of CaMBP under cold stress was further evaluated through reverse transcription - quantitative polymerase chain reaction (RT-qPCR). The expressions of EguCBF1a and EdCaMBP...

Zinc improves salt tolerance by increasing reactive oxygen species scavenging and reducing Na⁺ accumulation in wheat seedlings

L. H. Xu, W. Y. Wang, J. J. Guo, J. Qin, D. Q. Shi, Y. L. Li, J. Xu

Biologia plantarum 58:751-757, 2014 | DOI: 10.1007/s10535-014-0442-5

Salt decreases the uptake of Zn and other minerals and causes nutritional disorders in plants. Zn is an essential micronutrient for all organisms and it is reasonable to hypothesize that Zn status is essential for maintaining salt tolerance in plants. In this study, the physiological and molecular mechanisms of Zn-based alleviation of salt stress in wheat seedlings were investigated. Our results indicate that sufficient Zn nutrition maintained antioxidative enzyme activities and decreased a reactive oxygen species over-accumulation in wheat seedlings. Our data also reveal that sufficient Zn nutrition improved the expression of Na⁺/H⁺...

Expression of genes related to flavonoid and stilbene synthesis as affected by signaling chemicals and Botrytis cinerea in grapevines

S. Y. Ahn, S. A. Kim, K. S. Cho, H. K. Yun

Biologia plantarum 58:758-767, 2014 | DOI: 10.1007/s10535-014-0437-2

Recent studies have shown that the expression of genes related to the synthesis of flavonoids, such as the phenylalanineammonia lyase (PAL), chalcone synthase (CHS), chalcone isomerase (CHI), and stilbene synthase (STS) genes were induced in response to different signaling molecules and Botrytis cinerea inoculation in grapevine leaves. Therefore, in the present study, the nucleotide and deduced amino acid sequences of STS genes from cultivars Campbell Early and Kyoho were compared. The deduced amino acid sequences of VlKSTS12, VlKSTS11, VICESTS13, and VlKSTS1 showed 100 % homology to VlCESTS12, VlCESTS11, VICESTS24,...

Brief Communications

Cell membrane integrity, callose accumulation, and root growth in aluminum-stressed sorghum seedlings

E. J. Too, A. S. Carlsson, A. O. Onkware, B. A. Were, M. Geleta, T. Bryngelsson, S. Gudu

Biologia plantarum 58:768-772, 2014 | DOI: 10.1007/s10535-014-0455-0

Aluminum stress usually reduces plant root growth due to the accumulation of Al in specific zones of the root apex. The objectives of this study were to determine the localization of Al in the root apex of Sorghum bicolor (L.) Moech. and its effects on membrane integrity, callose accumulation, and root growth in selected cultivars. Seedlings were grown in a nutrient solution containing 0, 27, or 39 μM Al³⁺ for 24, 48, and 120 h. The Al stress significantly reduced root growth, especially after 48 and 120 h of exposure. A higher Al accumulation, determined by fluorescence microscopy after staining with a Morin dye, occurred in the...

Effect of cadmium on antioxidative enzymes, glutathione content, and glutathionylation in tall fescue

D. Del Buono, T. Mimmo, R. Terzano, N. Tomasi, S. Cesco

Biologia plantarum 58:773-777, 2014 | DOI: 10.1007/s10535-014-0412-y

The aim of this work was to assess the effect of different Cd2+concentrations on some antioxidant enzymes in Festuca arundinacea. Increased activities of ascorbate peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase, glutathione S-transferase, and glutathione reductase were ascertained in response to low Cd²⁺ concentrations (0-20 μM), whereas the enzyme activities were less increased or decreased at a higher Cd²⁺ dosage (50 μM) and a longer exposure. The content of reduced glutathione (GSH) decreased significantly with increasing Cd²⁺ concentrations, whereas the content of oxidized glutathione...

The localization of auxin transporters PIN3 and LAX3 during lateral root development in Arabidopsis thaliana

F. M. Perrine-Walker, E. Jublanc

Biologia plantarum 58:778-782, 2014 | DOI: 10.1007/s10535-014-0446-1

Fluorophore tagged proteins are used in Arabidopsis thaliana to understand their functional role in plant development. This requires the analysis of their spatial localization in planta. However, the localization analysis is often perturbed by a significant overlap of the fluorophores used to label proteins of interest and the optical filtering methods available on the confocal microscope. This problem can be addressed by the use of spectral imaging with linear unmixing the image data. We applied this method to help us identify double transgenic A. thaliana lines which expressed two fluorescently tagged auxin transporter proteins:...

Potato virus X induces DNA damage in leaf nuclei of the host plant Nicotiana tabacum L. var. xanthi

N. Cerovska, H. Plchova, P. Vaculik, T. Moravec, T. Gichner

Biologia plantarum 58:783-787, 2014 | DOI: 10.1007/s10535-014-0448-z

We employed the comet assay (single cell gel electrophoresis) to evaluate induced DNA damage in nuclei isolated from tobacco leaves (Nicotiana tabacum var. xanthi) inoculated with Potato virus X (PVX). The highest DNA damage, expressed by the tail moment value, was observed in the inoculated leaves and decreased in the 1^st to 4^th systemic leaves. DNA damage increased with the time after the inoculation (from day 3 to day 21) and was higher in nuclei isolated from a part of the leaf at the petiole compared to nuclei isolated from the leaf tip. A Pearson moment correlation (r = 0.94) between the induced...

The occurrence of dicotyledonar embryos in Agave tequilana

C. Ayala-González, A. Gutiérrez-Mora, B. Rodríguez-Garay

Biologia plantarum 58:788-791, 2014 | DOI: 10.1007/s10535-014-0456-z

Agave tequilana Weber is a monocot plant species member of the Asparagaceae family. One of the characteristics of monocot species is that their embryos show only one cotyledon. In this work, the occurrence of embryos with two cotyledons and fused cotyledons in A. tequilana is reported for the first time. The occurrence of dicotyledonar embryos in a species that by definition should have only one cotyledon could bring an opportunity to elucidate the mechanisms that have given the origin to the only one cotyledon present in monocots. Syncotyly is considered in this work as the possible mechanism that gave rise to the only cotyledon...

Marker assisted gene pyramiding for enhanced Tomato leaf curl virus disease resistance in tomato cultivars

A. Kumar, K. L. Tiwari, D. Datta, M. Singh

Biologia plantarum 58:792-797, 2014 | DOI: 10.1007/s10535-014-0449-y

The present research is aimed towards molecular marker assisted pyramiding Tomato leaf curl virus (ToLCV) disease resistance genes into two ToLCV susceptible tomato (Solanum lycopersicum L.) cvs. Pbc and H-86 (resistance genes recipient parents). Resistance gene donors were EC-538408 (Solanum chilense) and EC-520061 (S. peruvianum) in the case of cv. Pbc, and EC-520061 (S. peruvianum) and H-24 (S. lycopersicum) in the case of cv. H-86. A ToLCV resistance gene associated co-dominant simple sequence repeat (SSR) marker SSR-218 was used to discriminate between homozygotes and heterozygotes at the seedling stage...