Biologia plantarum, 2016 (vol. 60), issue 1

Original Papers

Effect of asparagine, cysteine, citrulline, and glutamine on in vitro rooting and biochemical constituents in cherry rootstocks

V. Sarropoulou, C. Chatzissavvidis, K. Dimassi-Theriou, I. Therios

Biologia plantarum 60:1-12, 2016 | DOI: 10.1007/s10535-015-0562-6

Effects of four amino acids, L-asparagine, L-cysteine, L-citrulline, and L-glutamine in different concentrations (0, 0.5, 1, and 2 mg dm-3) combined with 2 mg dm-3 indole-3-butyric acid, on in vitro rooting and biochemical constituents of cherry rootstocks CAB-6P (Prunus cerasus L.) and Gisela 6 (P. canescens × P. cerasus) were investigated. In CAB-6P, root number and root fresh mass (FM) were maximum at 0.5 mg dm-3 cysteine. All amino acids reduced root length in CAB-6P and root number as well as root FM in Gisela 6. In Gisela 6, 0.5 mg dm-3 asparagine or 2 mg dm-3...

Practical guidelines for the characterization of tobacco BY-2 cell lines

M. Srba, A. Černíková, Z. Opatrný, L. Fischer

Biologia plantarum 60:13-24, 2016 | DOI: 10.1007/s10535-015-0573-3

Plant cell lines represent useful models in plant cell biology. They allow simple analysis of the effects of various factors including modulated gene expression at cellular and subcellular levels. The tobacco BY-2 cell line is a favoured model due to its high proliferation rate, capability of effective synchronization, and accessibility to transformation. A relatively high uniformity of BY-2 cultures allows morphological phenotyping and assessment of growth parameters like mitotic index, viability, or cell density. Here we review already published and newly introduced optimized guidelines to carry out reliable, reproducible and efficient characterization...

Precise karyotyping of carrot mitotic chromosomes using multicolour-FISH with repetitive DNA

A. Nowicka, E. Grzebelus, D. Grzebelus

Biologia plantarum 60:25-36, 2016 | DOI: 10.1007/s10535-015-0558-2

Carrot (Daucus carota L.) chromosomes are small and uniform in shape and length. Here, mitotic chromosomes were subjected to multicolour fluorescence in situ hybridization (mFISH) with probes derived from conserved plant repetitive DNA (18-25S and 5S rDNA, telomeres), a carrot-specific centromeric repeat (Cent-Dc), carrot-specific repetitive elements (DCREs), and miniature inverted-repeat transposable elements (MITEs). A set of major chromosomal landmarks comprising rDNA and telomeric and centromeric sequences in combination with chromosomal measurements enabled discrimination of carrot chromosomes. In addition, reproducible and unique...

Functional analysis of CAX2-like transporters isolated from two ecotypes of Sedum alfredii

M. Zhang, J. Zhang, L. L. Lu, Z. Q. Zhu, X. E. Yang

Biologia plantarum 60:37-47, 2016 | DOI: 10.1007/s10535-015-0557-3

Two genes encoding CAX2-like proteins were isolated from a Zn/Cd hyperaccumulating ecotype (HE) and nonhyperaccumulating ecotype (NHE) of Sedum alfredii Hance, and they were named SaCAX2h and SaCAX2n, respectively. Both SaCAX2h:eGFP and SaCAX2n:eGFP proteins were localized to the vacuolar membrane of tobacco epidermal cells and yeast mutants. Heterologous expression of SaCAX2h or SaCAX2n in the Δzrc1 yeast mutant increased Cd content in yeast cells. Yeast complementary assay also revealed that both the transporters could suppress Ca and Mn hypersensitivity and enhance Ca and Mn accumulation in a K667 yeast mutant. The expression...

Generation and characterization of expressed sequence tags in young roots of tea (Camellia assamica)

A. Das, M. Mukhopadhyay, T. K. Mondal

Biologia plantarum 60:48-54, 2016 | DOI: 10.1007/s10535-015-0565-3

Assam tea (Camellia assamica) is perennial crop susceptible to moisture stress. We used its tender roots to construct a cDNA library for the identification, functional annotation, and analysis of transcripts. A total of 811 full-length expressed sequence tags were generated. After processing and assembly, 207 unigenes comprising 58 contigs and 149 singletons were registered. Finally, 35.75 % of the unigenes could be assigned to functional categories based on the Arabidopsis proteome. There was 43 % of a coding GC content and 1 272 coding DNA sequences found in the unigenes. Codon usage analysis shows leucine as the highest (9.92 %) and...

Repetitive sequences in Eragrostis curvula cDNA EST libraries obtained from genotypes with different ploidy

J. Romero, J. P. Selva, S. Pessino, V. Echenique, I. Garbus

Biologia plantarum 60:55-67, 2016 | DOI: 10.1007/s10535-015-0569-z

Eragrostis curvula (Schrad) Nees (weeping lovegrass) represents important cultivated forage in semiarid regions, and the most useful cultivars are tetraploid and reproduce by pseudogamous diplosporous apomixis. We previously produced a series of genetically related E. curvula lines that provide a suitable system for the identification of gene(s) involved in diplosporous apomixis and ploidy, including a natural apomictic tetraploid (T), a diploid sexual line (D), and a tetraploid sexual plant (C). A collection of expressed sequence tags (ESTs) was generated from cDNA libraries obtained from panicles of the D, T, and C, and leaves of the...

The mitochondrial DNA markers for distinguishing Phalaenopsis species and revealing maternal phylogeny

B. -Y. Lin, C. -D. Chang, L. L. H. Huang, Y. -C. Liu, Y. -Y. Su, T. -C. Chen, R. -H. Lee, H. -J. Huang, W. -L. Wu, C. -C. Chang

Biologia plantarum 60:68-78, 2016 | DOI: 10.1007/s10535-015-0566-2

Moth orchids (Phalaenopsis) are among the top-traded blooming potted plants in the world. To explore mitochondrial DNA (mtDNA) markers for species identification, we located simple sequence repeats in the mtDNA of Phalaenopsis aphrodite subsp. formosana and then pre-screened them for polymorphic markers by their comparison with corresponding mtDNA regions of P. equestris. The combination of 13 selected markers located in intergenic spacers could unambiguously distinguish 15 endemic moth orchids. Five most variable markers with polymorphic information content (PIC) ≥ 0.7 could be combined to classify 18 of 19 endemic moth...

Differential expression of iron-sulfur cluster biosynthesis genes during peach flowering

Z.-Z. Song, B.-B. Zhang, C.-H. Zhang, R.-J. Ma, M.-L. Yu

Biologia plantarum 60:79-85, 2016 | DOI: 10.1007/s10535-015-0577-z

Iron is required for the Fe-S cluster assembly which occurs in chloroplasts, mitochondria, and cytosol and here we characterized 44 Fe-S cluster biosynthesis genes and investigated their expression profiles during different peach flowering stages. Quantitative real-time PCR analysis shows that the highest expression of most peach Fe-S cluster biosynthesis genes appeared in the full bloom stage. Also, the highest Fe accumulation occurred in the full bloom stage followed by beginning bloom, petal fall, and bud swell stages. Activities of nitrite reductase (NiR) and succinate dehydrogenase (SDH) were closely correlated to the flower Fe content, whereas...

A rice mutant defective in antioxidant-defense system and sodium homeostasis possesses increased sensitivity to salt stress

K. -C. Lin, W. -S. Jwo, N. N. P. Chandrika, T. -M. Wu, M. -H. Lai, C. -S. Wang, C. -Y. Hong

Biologia plantarum 60:86-94, 2016 | DOI: 10.1007/s10535-015-0561-7

Screening salt-sensitive mutants is a powerful method to identify genes associated with salt tolerance. We used forward genetic screening with sodium azide-mutated rice (Oryza sativa L. cv. Tainung 67) to identify mutants showing hypersensitivity to salt stress. A new mutant line, named salt hypersensitive 1 (shs1) and exhibiting a severe salt-sensitivity when grown under a high NaCl concentration, was identified; the salt hypersensitivity was caused by duplicate recessive epistasis with mutations likely in two different loci. The shs1 salt sensitive phenotypes included a decreased seed germination rate, reduced shoot height...

BvPRR7 is a cold responsive gene with a clock function in beet

O. Omolade, A. E. Müller, C. Jung, S. Melzer

Biologia plantarum 60:95-104, 2016 | DOI: 10.1007/s10535-015-0568-0

The life cycle of flowering plants is partially defined by environmental cues like day length and temperature. In the model plant Arabidopsis thaliana and temperate cereals, such as barley (Hordeum vulgare) and wheat (Triticum spp.), differences in life cycle control have been associated with a natural variation in FLOWERING LOCUS C (FLC) and VERNALIZATION 1-3 (VRN1-3). In sugar beet (Beta vulgaris L.), variation in vernalization requirement and life cycle is determined by a major gene at the B locus. This gene has recently been identified as a pseudo-response regulator (PRR) gene BOLTING TIME...

Identification of a drought responsive gene encoding a nuclear protein involved in drought and freezing stress tolerance in Arabidopsis

H. -D. Moon, M. -S. Lee, S. -H. Kim, W. -J. Jeong, D. -W. Choi

Biologia plantarum 60:105-112, 2016 | DOI: 10.1007/s10535-015-0567-1

Plants have developed adaptive strategies to survive under different abiotic stressors. To identify new components involved in abiotic stress tolerance, we screened unannotated expressed sequence tags (ESTs) and evaluated their cold or drought response in Arabidopsis. We identified a drought response gene (DRG) encoding a 39.5-kDa polypeptide. This protein was expressed specifically in siliques and was induced by drought stress in most tissues. When a DRG-GFP construct was introduced into Arabidopsis protoplasts, GFP signals were detected only in the nucleus. The drg mutant plant was more sensitive to mannitol-induced...

Isolation and characterization of a tonoplast Na+/H+ antiporter from the halophyte Nitraria sibirica

L. Wang, Y. K. Ma, N. N. Li, W. B. Zhang, H. P. Mao, X. F. Lin

Biologia plantarum 60:113-122, 2016 | DOI: 10.1007/s10535-015-0560-8

Na+/H+ exchanger (NHX)-mediated Na+ and H+ antiport is an important mechanism for salt tolerance in plants. In this study, an Na+/H+ antiporter gene, referred to as NsNHX1, was isolated from the halophyte Nitraria sibirica Pall. using degenerate polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE). The resulting 2 182 bp NsNHX1 cDNA contained a 1 635 bp open reading frame (ORF) that encoded 544 amino acids and showed striking sequence similarity to tonoplast-localized NHXs from other plants. Subcellular localization analysis confirmed NsNHX1...

Exogenous gamma-aminobutyric acid increases salt tolerance of wheat by improving photosynthesis and enhancing activities of antioxidant enzymes

M. F. Li, S. J. Guo, X. H. Yang, Q. W. Meng, X. J. Wei

Biologia plantarum 60:123-131, 2016 | DOI: 10.1007/s10535-015-0559-1

Gamma-aminobutyric acid (GABA) is a non-protein amino acid that accumulates in a number of plant species under various environmental stresses. In this paper, the ability of applied GABA for the alleviation of NaCl stress was investigated in view of growth parameters, gas exchange, photosynthetic pigments, chlorophyll fluorescence, activities of antioxidant enzymes, malondialdehyde (MDA) content, and electrolyte conductivity (REC) in wheat seedlings. Germination rate and shoot dry mass decreased with an increasing NaCl concentration and this decrease was less pronounced when 0.5 mM GABA was applied. In the NaCl-treated seedlings, exogenous GABA partially...

Characterization of the γ-aminobutyric acid shunt pathway and oxidative damage in Arabidopsis thaliana pop 2 mutants under various abiotic stresses

N. A. Al-Quraan, A. T. Al-Share

Biologia plantarum 60:132-138, 2016 | DOI: 10.1007/s10535-015-0563-5

In the present study, three Arabidopsis thaliana pop2 mutant lines with different T-DNA insertions in a gene coding γ-aminobutyric acid transaminase (GABA-TA) were screened for seed germination percentage, stress-induced oxidative damage, and GABA content and metabolism under various abiotic stresses including high temperature (42 °C), low temperature (4 °C), salinity (NaCl), and osmotic stress (mannitol). All mutant lines showed a decreased germination under all the stress treatments with a significant reduction in the pop2-1 and pop2-3 mutant lines. Content of GABA and MDA increased significantly in all pop2...

Effect of salicylic acid on the antioxidant system and photosystem II in wheat seedlings

Y. E. Chen, J. M. Cui, G. X. Li, M. Yuan, Z. W. Zhang, S. Yuan, H. Y. Zhang

Biologia plantarum 60:139-147, 2016 | DOI: 10.1007/s10535-015-0564-4

To study the effects of application of salicylic acid (SA) on the antioxidant system and photosystem II (PS II) in wheat seedlings we used two different experiments. The first method was carried out by immersing roots in Hoagland's nutrient solution containing 0, 0.25, or 2.5 mM SA, and the second method was performed by spraying two-week-old seedlings with the same SA concentrations. After 24 h, chlorophyll fluorescence, thylakoid membrane proteins, antioxidant enzyme activities, and reactive oxygen species were measured. The low concentration of SA caused a significant increase in the antioxidant enzyme activities. However, the treatment with 2.5...

Effects of chilling and high temperatures on photosynthesis and chlorophyll fluorescence in leaves of watermelon seedlings

W. Hou, A. H. Sun, H. L. Chen, F. S. Yang, J. L. Pan, M. Y. Guan

Biologia plantarum 60:148-154, 2016 | DOI: 10.1007/s10535-015-0575-1

The effects of chilling (CT, day/night temperatures of 12/10 °C, an irradiance of 250 μmol m-2 s-1), chilling combined with a low irradiance (CL, 12/10 °C, 80 μmol m-2 s-1), and a high temperature (HT, 42/40 °C, 250 μmol m-2 s-1) on chlorophyll content, chlorophyll fluorescence, and gas exchange were studied in two watermelon cultivars, ZJ8424 and YS01, differing in their resistance. The chlorophyll content, net photosynthetic rate (PN), stomatal conductance (gs), and transpiration rate (E) decreased substantially, whereas the intercellular CO2...

Malate as substrate for catabolism and gluconeogenesis during ripening in the pericarp of different grape cultivars

F. Famiani, D. Farinelli, T. Frioni, A. Palliotti, A. Battistelli, S. Moscatello, R. P. Walker

Biologia plantarum 60:155-162, 2016 | DOI: 10.1007/s10535-015-0574-2

Malate is accumulated in grape pericarp until the start of ripening and then it is dissimilated. One aim of this study was to determine if the potential contribution of stored malate to the substrate requirements of metabolism in ripening grape pericarp is dependent on the cultivar. Two Vitis vinifera L. cultivars which accumulated different amounts of malate and had ripening periods of a different length were compared. The potential contribution of stored malate over the whole period of ripening was around 20 % in the cv. Sagrantino and 29 % in the cv. Pinot Noir. The contribution was higher in Pinot Noir because it contained more malate and...

Glutathione metabolism in Urtica dioica in response to cadmium based oxidative stress

L. Tarhan, B. Kavakcioglu

Biologia plantarum 60:163-172, 2016 | DOI: 10.1007/s10535-015-0570-6

To investigate the antioxidative response of glutathione metabolism in Urtica dioica L. to a cadmium induced oxidative stress, activities of glutathione reductase (GR), glutathione-S-transferase (GST), and glutathione peroxidase (GSH-Px), content of reduced (GSH) and oxidized (GSSG) glutathione, lipid peroxidation (LPO), and also accumulation of Fe, Zn, Mn, Cu besides Cd were determined in the roots, stems, and leaves of plants exposed to 0 (control), 0.045, and 0.09 mM CdCl2 for 58 h. Whereas the Cd content continuously increased in all organs, the Fe, Zn, Mn, and Cu content decreased in dependence on the applied Cd concentration...

Arsenic toxicity: cell signalling and the attenuating effect of nitric oxide in Eichhornia crassipes

H. M. Andrade, J. A. Oliveira, F. S. Farnese, C. Ribeiro, A. A. Silva, F. V. Campos, J. L. Neto

Biologia plantarum 60:173-180, 2016 | DOI: 10.1007/s10535-015-0572-4

Nitric oxide (NO) is an important molecule involved in the perception of stress induced by toxic compounds such as arsenic (As). The present study investigated the role of NO applied as sodium nitroprusside (SNP) in cell signalling and the ability of NO to attenuate the toxic effects of As (in the form of sodium arsenate) in water hyacinth (Eichhornia crassipes). Water hyacinth plants were collected and assigned to one of the following treatments: control; 100 μM SNP; 20 μM As; or 20 μM As + 100 μM SNP. The plants remained under these conditions for 0, 4, 12, and 24 h. After each time interval, the plants were collected and As absorption,...

Overexpression of a harpin-encoding gene popW in tobacco enhances resistance against Ralstonia solanacearum

Chao Wang, Cui Wang, H. -W. Li, T. Wei, Y. -P. Wang, H. -X. Liu

Biologia plantarum 60:181-189, 2016 | DOI: 10.1007/s10535-015-0571-5

PopW, a harpin protein identified from Ralstonia solanacearum, has multiple beneficial effects in plants, promoting plant growth and development, increasing crop yield, and inducing resistance to pathogens. Tobacco plants transformed with popW, the PopW-encoding gene, exhibited a promoted growth rate and enhanced resistance to Tobacco mosaic virus (TMV). Here, it is documented that the transgenic tobacco plants overexpressing popW exhibited a higher resistance to R. solanacearum YN10 infection compared with that of the wild-type plants. In the popW-expressing tobacco lines, an enhanced H2O2 accumulation and...

Brief Communications

Metabolic regulation of ammonia emission in different senescence phenotypes of Nicotiana tabacum

Y. J. Wu, T. Z. Yang, Y. Y. Song, X. Q. Zhang, S. X. Xu, G. Xue, X. X. Xing

Biologia plantarum 60:190-194, 2016 | DOI: 10.1007/s10535-015-0556-4

In order to reveal the character of ammonia emission in senescent tobacco (Nicotiana tabacum), the content of NH4 +, total nitrogen, and soluble protein, and the activities of nitrogen metabolism-related enzymes were measured in leaves of a quick-leaf-senescence phenotype ZY90 and a slow-leaf-senescence phenotype NC89. Compared with NC89, ZY90 had a higher NH4 + accumulation, a lower glutamine synthetase activity, and a significantly higher stomatal ammonia compensation point, and ammonia emission during 40 to 60 d after leaf emergence. During senescence, the quick-leafsenescence phenotype was characterized...