Biologia plantarum, 2016 (vol. 60), issue 4

Original Papers

Age-associated alterations in DNA methylation and expression of methyltransferase and demethylase genes in Arabidopsis thaliana

Z. V. Ogneva, A. S. Dubrovina, K. V. Kiselev

Biologia plantarum 60:628-634, 2016 | DOI: 10.1007/s10535-016-0638-y

Little is known about the contributions of DNA methylation/demethylation to plant aging and senescence. We used Arabidopsis thaliana to study how increasing age of an annual plant species influences DNA methylation. Based on methylation-sensitive DNA fragmentation assay, it could be concluded that aging A. thaliana was accompanied by DNA demethylation. Bisulfite sequencing reveals that cytosine methylation within the Actin2 3' untranslated region and internal transcribed spacer with 5.8S rRNA (ITS1-5.8SrRNA-ITS2) DNA regions decreased with A. thaliana growth and aging. We show that transcription of methyltransferase...

Protein as a sole source of nitrogen for in vitro grown tobacco plantlets

H. Synková, V. Hýsková, K. Garčeková, S. Křížová, H. Ryšlavá

Biologia plantarum 60:635-644, 2016 | DOI: 10.1007/s10535-016-0639-x

We tested the capability of plants to utilize protein as the exclusive source of nitrogen. The aim of this study was to find out how such a nutrition affected plantlet growth, photosynthetic performance, and N assimilation metabolism in tobacco (Nicotiana tabacum L., cv. Petit Havana SR1) grown in vitro. Plantlets grown in a casein-supplemented (CA) medium were compared to plantlets grown in a complete Murashige-Skoog (MS) medium, plantlets grown in an ammonium-deficient medium (N1), or plantlets grown in a nitrate-reduced medium (N2). In addition, the plantlets were grown in the presence or absence of 1.5 % (m/v) saccharose as an additional...

Selection of reference genes for real-time quantitative PCR analysis of gene expression in Glycyrrhiza glabra under drought stress

A. Maroufi

Biologia plantarum 60:645-654, 2016 | DOI: 10.1007/s10535-016-0601-y

Licorice (Glycyrrhiza glabra L.) is an important medicinal plant accumulating high-value secondary metabolites. Real-time reverse transcription quantitative PCR (RT-qPCR) has become a common method for studying gene expression, and the availability of stable reference genes is a prerequisite to obtain accurate quantification of transcript abundance. Therefore, an experiment was designed to determine appropriate reference genes for gene expression studies in licorice. Based on reports in the literature and the availability of genomic sequences, eight putative reference genes were chosen. Further, the expression stabilities of these genes were...

Cloning PIP genes in drought-tolerant vetiver grass and responses of transgenic VzPIP2;1 soybean plants to water stress

S. B. Hu, Q. Zhou, J. An, B. J. Yu

Biologia plantarum 60:655-666, 2016 | DOI: 10.1007/s10535-016-0631-5

Vetiver grass [Vetiveria zizanioides (L.) Nash] displays comprehensive abiotic stress tolerance closely related to fine maintenance of plant water relation mediated by plasma membrane intrinsic proteins (PIPs). Two open reading frame sequences of PIPs (867 and 873 bp) were cloned from vetiver grass and named as VzPIP1;1 and VzPIP2;1, respectively. Expression of green fluorescent protein revealed only subcellular localization of VzPIP2;1 in the plasma membrane. Agrobacterium tumefaciens mediated transgenic (VzPIP2;1) soybean plants had a higher water content in above-ground parts under sufficient water...

Functional analysis of the Malus domestica MdHMGR2 gene promoter in transgenic Arabidopsis thaliana

D. M. Lv, T. T. Zhang, S. Deng, Y. H. Zhang

Biologia plantarum 60:667-676, 2016 | DOI: 10.1007/s10535-016-0637-z

The first rate-limiting enzyme of the mevalonate pathway during isoprenoid biosynthesis is 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR). In this study, the expression pattern of the MdHMGR2 gene in Malus domestica suggests that MdHMGR2 was expressed in a tissue-specific manner and was significantly induced by ethephon (ETH), indoleacetic acid (IAA), methyl jasmonate (MeJA), and salicylic acid (SA). The MdHMGR2 promoter was isolated, sequenced, and analyzed through bioinformatics tools, and the results suggest the presence of various putative cis-acting elements responsive to different hormones. Activity of β-glucuronidase...

Suppression of tomato SlGGP aggravates methyl viologen-mediated oxidative stress

D.-Y. Yang, N.-N. Ma, Z.-M. Liu, X.-C. Ma, S.-J. Zhao, Q.-W. Meng

Biologia plantarum 60:677-685, 2016 | DOI: 10.1007/s10535-016-0628-0

Ascorbate (AsA) is an important antioxidant that can scavenge reactive oxygen species to protect plant cells against oxidative stress. Guanosine 5'-diphosphate (GDP)-L-galactose phosphorylase (GGP) is a key enzyme in the AsA biosynthetic pathway. To investigate the functions of GGP in AsA synthesis and oxidative stress tolerance in tomato, antisense lines with a reduced expression of SlGGP were obtained. Photobleaching after treatment of leaf disks with methyl viologen was more severe in transgenic lines compared to wild type (WT) plants. Moreover, compared with the WT plants, the transgenic plants showed a higher content of hydrogen peroxide,...

Salicylic acid and nitric oxide increase photosynthesis and antioxidant defense in wheat under UV-B stress

F. Yan, Y. Liu, H. Sheng, Y. Wang, H. Kang, J. Zeng

Biologia plantarum 60:686-694, 2016 | DOI: 10.1007/s10535-016-0622-6

The effects of exogenous salicylic acid (SA), sodium nitropusside (SNP, a nitric oxide donor), or their combination on dwarf polish wheat (Triticum polonicum L.) seedlings under UV-B stress were studied. The UV-B stress significantly decreased plant height, shoot dry mass, pigment content, net photosynthetic rate, intercellular CO₂ concentration, stomatal conductance, transpiration rate, and variable to maximum chlorophyll fluorescence ratio (F_v/F_m) in all plants, but less in the presence of SA, SNP, and their combination. On the other hand, there were considerable increases in malondialdehyde (MDA), proline,...

Tracing root permeability: comparison of tracer methods

E. Pecková, E. Tylová, A. Soukup

Biologia plantarum 60:695-705, 2016 | DOI: 10.1007/s10535-016-0634-2

Root epidermis and apoplastic barriers (endodermis and exodermis) are the critical root structures involved in setting up plant-soil interface by regulating free apoplastic movement of solutes within root tissues. Probing root apoplast permeability with "apoplastic tracers" presents one of scarce tools available for detection of "apoplastic leakage" sites and evaluation of their role in overall root uptake of water, nutrients, or pollutants. Although the tracers are used for many decades, there is still not an ideal apoplastic tracer and flawless procedure with straightforward interpretation. In this article, we present our experience with the most...

Impact of exogenous caffeine on morphological, biochemical, and ultrastructural characteristics of Nicotiana tabacum

R. Alkhatib, B. Alkhatib, N. Al-Quraan, L. Al-Eitan, N. Abdo, R. Muhaidat

Biologia plantarum 60:706-714, 2016 | DOI: 10.1007/s10535-016-0600-z

Caffeine (1,3,7-trimethylxanthine) is purine alkaloid, and it is secondary metabolite produced naturally in plants. It plays a crucial role in defense and stress tolerance. A hydroponic experiment was carried out to study the morphological, biochemical, and ultrastructural effects of caffeine treatment on seedlings of tobacco (Nicotiana tabacum L. cv. Turkish). The plants were grown in a growth chamber for 14 d in Hoagland's nutrient solution supplemented with 0 (control), 25, 50, 100, 1000, and 5000 μM caffeine. Shoot heights as well as root lengths significantly decreased in the plants treated with 1000 and 5000 μM caffeine. Total protein...

Cloning, characterization, and subcellular localization of a novel JAZ repressor from Eleusine coracana

S. Sen, S. K. Dutta

Biologia plantarum 60:715-723, 2016 | DOI: 10.1007/s10535-016-0652-0

Jasmonate ZIM domain (JAZ) proteins are key regulators of the jasmonic acid (JA) signaling pathway. Repressors of JAZ remain bound to the myelocytomatosis 2 (MYC2) or MYC3/MYC4 transcription factors in the absence of JA and negatively regulate transcription of the JA responsive genes. In the presence of JA, JAZ proteins interact with coronative insensitive 1 (COI1), the recognition molecule of E3 ubiquitin ligase SCF^COI1 (COI1 stabilized by Skp, cullin, F-box containing complex), get ubiquitinated, and subsequently degraded by the 26S proteasome. However, there is a dearth of knowledge about this gene family in monocot cereals, specifically...

Cytochrome P₄₅₀, CYP93A1, as defense marker in soybean

A. J. Kinzler, Z. A. Prokopiak, M. M. Vaughan, P. W. Erhardt, J. G. Sarver, J. A. Trendel, Z. Zhang, N. J. Dafoe

Biologia plantarum 60:724-730, 2016 | DOI: 10.1007/s10535-016-0629-z

Cytochrome P₄₅₀, CYP93A1, is involved in the synthesis of the phytoalexin glyceollin in soybean (Glycine max L. Merr). The gene encoding CYP93A1 has been used as defense marker in soybean cell cultures, however, little is known regarding how this gene is expressed in the intact plant. To further understand the tissue-specific role of CYP93A1 in soybean defense, we analyzed the expression of this gene in mechanically damaged leaves and stems. In leaves, CYP93A1 was constitutively expressed; its expression did not change in response to mechanical damage. In stems, however, expression of CYP93A1 was induced as quickly as...

Salt-induced NO₃^- uptake inhibition in cowpea roots is dependent on the ionic composition of the salt and its osmotic effect

R. M. Aragão, E. N. Silva, P. C. C. Silva, J. A. G. Silveira

Biologia plantarum 60:731-740, 2016 | DOI: 10.1007/s10535-016-0604-8

Salinity remarkably inhibits NO₃ ^- uptake but the mechanisms are not well understood. This study was addressed to elucidate the role of ionic and osmotic components of salinity on NO₃ ^- influx and efflux employing classic kinetics involving a low affinity transport system (LATS) and a high affinity transport system (HATS). In the presence of KCl, NaCl, and Na₂SO₄ at 100 mM concentrations, in both LATS and HATS, Michaelis constant (K_m) was similar for the three salts and maximum rate (V_max) decreased as follows: KCl > NaCl > Na₂SO₄, compared...

Sucrose-metabolizing enzymes and their genes in the arils of two Dimocarpus longan cultivars

L. Shuai, J. Li, J. J. Niu, P. H. Qian, W. H. Liu, X. Q. Xue, D. M. Han, Z. X. Wu

Biologia plantarum 60:741-748, 2016 | DOI: 10.1007/s10535-016-0602-x

This study aimed to investigate sucrose-metabolizing enzymes and their genes in fruits of two longan (Dimocarpus longan Lour) cultivars Cihezhong (CHZ) and Lidongben (LDB). Content of sucrose, glucose, and fructose were measured by high-performance liquid chromatography. The genes of sucrose-metabolizing enzymes were cloned by combining reverse transcription polymerase chain reaction and rapid amplification of cDNA ends, and enzyme activities were analyzed at various points in the fruiting cycle. The total soluble solid (TSS) content of longan arils rose and was positively correlated with sucrose content during maturation and then declined as...

Stability of sheath blight resistance in transgenic ASD16 rice lines expressing a rice chi11 gene encoding chitinase

T. Rajesh, S. Maruthasalam, K. Kalpana, K. Poovannan, K. K. Kumar, E. Kokiladevi, D. Sudhakar, R. Samiyappan, P. Balasubramanian

Biologia plantarum 60:749-756, 2016 | DOI: 10.1007/s10535-016-0594-6

Development of transgenic plants by introducing defense genes is one of the strategies to engineer disease resistance. Transgenic ASD16 rice plants harbouring rice chitinase chi11 gene, belonging to a PR-3 group of defense gene conferring sheath blight (Rhizoctonia solani Kuhn) resistance, were used in this study. Three T₂ homozygous lines (ASD16-4-1-1, 5-1-1, and 6-1-1) were identified from seven putative (T₀) transgenic lines expressing chi11 using Western blotting analysis. The inheritance of sheath blight resistance in those lines was studied over generations. The stability of chi11 expression up...

Finding genomic regions and candidate genes governing water use efficiency in rice

V. Roja, S. Patil, D. A. Deborah, A. Srividhya, N. Ranjitkumar, G. Kadambari, P. V. Ramanarao, E. A. Siddiq, L. R. Vemireddy

Biologia plantarum 60:757-766, 2016 | DOI: 10.1007/s10535-016-0651-1

Water use efficiency (WUE) is an worth attempting trait to discover the genomic regions governing it, especially in view of the diminishing water resources for the crop plants in general and rice in particular. In order to address this, the present investigation was aimed at identification of genomic regions governing WUE employing a recombinant inbred line population derived from a cross between INRC10192, a high WUE landrace, and IR64, a high yielding cultivar. A total of 36 quantitative trait loci (QTLs) were detected under control as well as drought conditions on chromosomes 1, 2, 4, 8, 9, 10, and 11. Among all, the QTLs with the marker intervals...

A novel double T-DNA system for producing stack and marker-free transgenic plants

X. J. Wang, Y. Y. Su, Y. F. Dong, Q. L. Tang, Z. X. Wang

Biologia plantarum 60:767-773, 2016 | DOI: 10.1007/s10535-016-0653-z

This study aimed to develop a new vector system to remove selection genes and to introduce two or more genes of interest into plants in order to express them in a coordinated manner. A multigene expression vector was established based on pCamBIA2300 using a selectable marker gene (SMG)-free system based on the combination of the isocaudamer technique and double T-DNA. The vector DT7 containing seven target genes was constructed and introduced into tobacco using Agrobacterium-mediated transformation. Twenty-one of 27 positive transgenic plants contained both T-DNA regions. The co-transformation frequency was 77.8 %. The frequency of unlinked...

Effects of salt and alkali stress on growth, accumulation of oxalic acid, and activity of oxalic acid-metabolizing enzymes in Kochia sieversiana

Y. Ma, X. P. Wang, S. F. Zhang, D. C. Shi, L. X. Sheng

Biologia plantarum 60:774-782, 2016 | DOI: 10.1007/s10535-016-0650-2

Kochia sieversiana (Pall.) C.A. Mey. is a forage plant that can grow in extremely alkalinized grasslands at pH 10 or higher. Accumulation of a large amount of oxalic acid (OxA) is a primary characteristic of K. sieversiana. In our study, seedlings of K. sieversiana were exposed to the following conditions: non-stress, salinity (200 mM, a molar ratio of NaCl and Na₂SO₄ 1:1), and alkali stress (200 mM, a molar ratio of NaHCO₃ and Na₂CO₃ 1:1). Growth, water content, content of organic acids (including OxA), Na⁺, and K⁺, and activities of some OxA metabolism-related...

Reviews

Some key physiological and molecular processes of cold acclimation

R. John, N. A. Anjum, S. K. Sopory, N. A. Akram, M. Ashraf

Biologia plantarum 60:603-618, 2016 | DOI: 10.1007/s10535-016-0648-9

Agricultural production worldwide has been severely impacted by cold and freezing stresses. Plant capacity to acclimate to environmental conditions in their immediate vicinity largely control their survival, growth, and productivity. Molecular as well as biochemical mechanisms underpinning plant cold acclimation are very complex and interwoven. The cold-impacted plants try to modulate expression of variety genes controlling cell membrane lipid composition, mitogen-activated protein kinase cascade, total soluble proteins, polyamines, glycinebetaine, proline, reactive oxygen species (ROS) scavengers, cryoprotectants, and a large number of cold responsive...

A plant biologists' guide to phylogenetic analysis of biological macromolecule sequences

F. Cvrčková

Biologia plantarum 60:619-627, 2016 | DOI: 10.1007/s10535-016-0649-8

Phylogenetic analysis has become a common step in characterization of gene and protein sequences. However, despite the availability of numerous affordable and more-or-less intuitive software tools, construction of biologically relevant, informative phylogenetic trees remains a process involving several critical steps that are inherently non-algorithmic, i.e., dependent on decisions made by the user. These steps involve, but are not limited to, setting the aims of the phylogenetic study, choosing sequences to be analyzed, and selecting methods employed in sequence alignment construction, as well as algorithms and parameters used to construct...

Brief Communications

Enhancement of antioxidant enzyme activities in rice callus by ascorbic acid under salinity stress

A. N. Alhasnawi, C. M. Z. Che Radziah, A. A. Kadhimi, A. Isahak, A. Mohamad, W. M. W. Yusoff

Biologia plantarum 60:783-787, 2016 | DOI: 10.1007/s10535-016-0603-9

Ascorbic acid (AsA) is naturally occurring compound with antioxidant activity and plays a pivotal role in plant cell adaptation to salinity stress. The objective of this work was to assess the influence of exogenous AsA on the embryogenic callus of indica rice (Oryza sativa L.) cv. MRQ74 cultivated under saline conditions. NaCl (200 mM) decreased callus fresh and dry masses, relative growth rate, and K⁺ and Ca⁺² content, and increased Na⁺ content and Na⁺/K⁺ ratio. Application of AsA (0.5 or 1 mM) alleviated these effects of salinity. Activities of peroxidase, catalase, superoxide dismutase,...

Activity of DNA polymerase λ in spikelets of rice and maize

S. Bakshi, S. Sihi, D. N. Sengupta

Biologia plantarum 60:788-792, 2016 | DOI: 10.1007/s10535-016-0605-7

Involvement of DNA polymerase (pol) enzymes in meiotic DNA repair has not been clearly understood in plants. DNA polymerase λ is involved in short patch DNA synthesis and base excision repair in both plants and animals. The presence and activity of the pol λ enzyme was studied in a protein isolated from spikelets during flower development stages of rice (Oryza sativa L.) and maize (Zea mays L.). Western blot analysis showed a 2- to 2.5-fold higher accumulation of pol λ in spikelets than in shoots. Assays of pol and in-gel activity showed the dideoxynucleotide triphosphate sensitive pol λ enzyme in spikelets...

Changes in free polyamines and expression of polyamine metabolic genes under drought and high-temperature in Citrus sinensis

X. Z. Fu, Y. Huang, F. Xing, C. P. Chun, L. L. Ling, L. Cao, L. Z. Peng

Biologia plantarum 60:793-798, 2016 | DOI: 10.1007/s10535-016-0636-0

Polyamines play important roles in plant responses to various environmental stresses. In this study, changes in free polyamine content and transcription of genes participating in polyamine metabolism were investigated in sweet orange (Citrus sinensis Osbeck cv. Jincheng) seedlings under drought (DR), high-temperature (HT), and combined DR and HT (DRHT) stresses. Content of putrescine (Put) and spermidine (Spd) was the highest and that of spermine (Spm) was the lowest under the DR stress, whereas Put and Spd content was the lowest, and Spm content was the highest under the HT stress. Content of the three polyamines under the DRHT stress ranged...