Biologia plantarum, 2013 (vol. 57), issue 2

Original Papers

In vitro regeneration of Aristolochia tagala and production of artificial seeds

M. Remya, V. Narmatha Bai, V. N. Mutharaian

Biologia plantarum 57:210-218, 2013 | DOI: 10.1007/s10535-012-0280-2

Protocols for in vitro plant multiplication from somatic tissues and production of artificial seeds through encapsulation of nodes were developed for Aristolochia tagala Cham., a rare and valuable medicinal plant, as a measure of conservation and as a prerequisite for genetic transformation procedure. A maximum number of adventitious shoots were regenerated from leaf-derived callus on Murashige and Skoog (MS) medium containing 6-benzylaminopurine (BAP; 2 μM), α-naphthaleneacetic acid (NAA; 0.5 μM), and phloroglucinol (PG; 10μM). Nodes collected from in vitro established shoot cultures were encapsulated in 3 % (m/v)...

Methylglyoxal with glycine or succinate enhances differentiation and shoot morphogenesis in Nicotiana tabacum callus

A. Ray, S. Ray, S. Mukhopadhyay, M. Ray

Biologia plantarum 57:219-223, 2013 | DOI: 10.1007/s10535-012-0272-2

The aim of this study was to evaluate the influence of methylglyoxal (MG) on organogenesis and regeneration of tobacco (Nicotiana tabacum L.) plants from callus in media containing glycine or succinate. The best improvement in shoot proliferation and shoot length was obtained in the medium supplemented with 0.1 mM MG and 0.5 mM glycine or 0.25 mM succinate. The histological studies showed vigorous development of corm like structures and shoot organogenesis from callus tissues cultured in MG supplemented media. Biochemical studies also revealed higher content of δ-aminolaevulinic acid (a precursor of chlorophyll) and of chlorophyll.

Transport of mRNA molecules coding NAC domain protein in grafted pear and transgenic tobacco

W. N. Zhang, X. W. Duan, C. Ma, T. Harada, T. Z. Li

Biologia plantarum 57:224-230, 2013 | DOI: 10.1007/s10535-012-0293-x

Grafting is an important cultivation method and recent research on the mechanism of interactions between rootstock nad scion is focused on the long-distance transport of mRNA and small RNAs in the phloem. Among these transportable molecules, NACP gene coding NAM, ATAF1/2, CUC2 (NAC) domain protein might be involved in apical meristem development. Here, we report the transport of NACP mRNA between Chinese pear (Pyrus bretschneideri) cv. Yali (scion) and the wild Pyrus betulaefolia Bunge (rootstock). Our results indicated that NACP mRNA can be transported in both directions from the 3^rd to 10^th...

Characterization of the GLP13 gene promoter in Arabidopsis thaliana

L. Yang, T. Li, S. C. Zhang, G. L. Gao, C. W. Yang

Biologia plantarum 57:231-237, 2013 | DOI: 10.1007/s10535-012-0273-1

In transgenic plants, for many applications it is important that the inserted genes are expressed in a tissue-specific manner. This in turn could help better understanding their roles in plant development. Germin-like proteins (GLPs) play diverse roles in plant development and defense responses. In order to understand the functions and regulation of the GLP13 gene, its promoter (762 bp) was cloned and fused with a β-glucuronidase (GUS) reporter gene for transient expression in Arabidopsis thaliana and tobacco (Nicotiana tabacum cv. K326). Histochemical analysis of the transgenic plants showed that GUS was specifically...

Molecular cloning and expression analysis of four actin genes (MiACT) from mango

C. Luo, X. -H. He, H. Chen, Y. Hu, S. -J. Ou

Biologia plantarum 57:238-244, 2013 | DOI: 10.1007/s10535-012-0278-9

Actin is the most abundant protein in eukaryotic cells and is a key cytoskeletal component controlling cell morphology and motility. In this study, four MiACT genes were isolated from mango by homological cloning and designated as MiACT1, MiACT4, MiACT7, and MiACT9. Sequence alignments and phylogenetic analysis demonstrated that the four MiACT genes of mango were highly similar to each other at the nucleotide and amino acid levels. All of four MiACT proteins showed high similarity to the known actin proteins from other species. Reverse transcription polymerase chain reaction revealed that the four MiACT...

Differential expression of VRN1 and other MADS-box genes in Festuca pratensis selections with different vernalization requirements

Å. Ergon, H. Hamland, O. A. Rognli

Biologia plantarum 57:245-254, 2013 | DOI: 10.1007/s10535-012-0283-z

Most perennial and winter annual temperate grasses have a vernalization requirement (VR) for flowering, that is, they require a cold period before they can flower in response to long days. From a F1 mapping population of the outbreeding perennial forage grass Festuca pratensis Huds. (meadow fescue) previously used to map several quantitative trait loci (QTLs) for VR, we produced two F2 populations divergently selected for high or low VR. The two populations were characterised for flowering behaviour and gene expression of VRN1 as well as other MADS-box genes with a putative function in the induction of flowering. Expression of FpVRN1...

Microarray-based screening of the microRNAs associated with caryopsis development in Oryza sativa

H. Liu, D. Shen, S. Jia, W. Li, J. Li, J. Liu, S. Han, Y. Wang

Biologia plantarum 57:255-261, 2013 | DOI: 10.1007/s10535-012-0270-4

Plant microRNAs modulate diverse developmental processes by regulating expression of their target genes. To explore potential miRNA-guided gene regulation in developing rice (Oryza sativa L.) caryopses, a miRNA microarray was used to identify miRNAs present at the different developmental stages. We found that 27 miRNAs, of which 16 were conserved miRNAs, were present in developing caryopses. High expression levels were detected for miR159, miR167, and miR530 at the morphogenesis stage and for miR169, miR435, and miR528 at the stage of accumulation of metabolites. Next, 26 target genes were predicted for seven of the detected miRNAs and the expression...

Over-expression of BcFLC1 from non-heading Chinese cabbage enhances cold tolerance in Arabidopsis

T. Liu, Y. Li, J. Ren, C. Zhang, M. Kong, X. Song, J. Zhou, X. Hou

Biologia plantarum 57:262-266, 2013 | DOI: 10.1007/s10535-012-0287-8

A gene (named BcFLC1) homologous to the AtFLC gene, which encodes a floral repressor, was isolated from the nonheading Chinese cabbage (Brassica campestris L. ssp. chinensis) cv. NJ074. The gene showed high similarity to AtFLC. For studying the gene function, we designed to introduce the BcFLC1 gene into Arabidopsis thaliana. The results showed that BcFLC1 had effects on flowering time similar to AtFLC. We also found that Arabidopsis cold-tolerance was enhanced by BcFLC1 overexpression. Under low temperature stress, the BcFLC1 transgenic plants exhibited stronger growth...

Isolation and characterization of purple acid phosphatase gene during seedling development in mungbean

A. Wongkaew, P. Srinives, S. Nakasathien

Biologia plantarum 57:267-273, 2013 | DOI: 10.1007/s10535-012-0292-y

Purple acid phosphatases (PAPs), which are normally found in plant tissues, can hydrolyze a broad spectrum of phosphate esters. In this study, a mungbean [Vigna radiata (L.) Wilczek cv. KPS1] acid phosphatase gene (VrPAP1) was isolated from seedling cotyledons. The full-length of VrPAP1 cDNA contained an open reading frame of 1 644 bp encoding 547 amino acid residues with a predicted molecular mass of 62.07 kDa. Sequence analysis showed that VrPAP1 is purple acid phosphatase. RNA blot analyses indicated that the VrPAP1 accumulated during the first hour in cotyledons of germinating seeds and reached a maximum expression after...

Allelic differentiations and effects of the Rf3 and Rf4 genes on fertility restoration in rice with wild abortive cytoplasmic male sterility

J. Cai, Q. P. Liao, Z. J. Dai, H. T. Zhu, R. Z. Zeng, Z. M. Zhang, G. -Q. Zhang

Biologia plantarum 57:274-280, 2013 | DOI: 10.1007/s10535-012-0294-9

To reveal the allelic differentiations at the two genes for fertility restoration (Rf) on chromosomes 1 (Rf3) and 10 (Rf4), 15 chromosome single segment substitution lines (SSSLs) with the Rf3 locus and 18 SSSLs with the Rf4 locus were crossed with Bobai A (BbA), a cytoplasmic male sterility line with wild abortive type of cytoplasm (WA-CMS), respectively. Based on the pollen and seed fertility of the F₁ hybrids, the Rf3 and Rf4 genes were each classified into four alleles, namely Rf3-1, Rf3-2, Rf3-3, and Rf3-4 for Rf3, and Rf4-1, Rf4-2, Rf4-3,...

Wheat truncated hemoglobin interacts with photosystem I PSK-I subunit and photosystem II subunit PsbS1

D. Y. Kim, M. J. Hong, Y. J. Lee, M. B. Lee, Y. W. Seo

Biologia plantarum 57:281-290, 2013 | DOI: 10.1007/s10535-012-0268-y

Recently, the truncated hemoglobin gene (trHb) was discovered in plant species, however, its role has not yet been determined. In this study, the gene expression of wheat trHb (TatrHb) was analyzed under various biotic and abiotic stresses. TatrHb transcript levels increased in NaCl-treated leaves and gibberellic acid (GA₃)-treated roots. In addition, sodium nitroprusside (SNP), a nitric oxide donor, induced an increase in TatrHb transcript levels in roots and leaves. A yeast two-hybrid assay (YIIH) was used to screen a hypoxia-treated wheat seedling library with the goal of determining the putative function...

The role of spherosome-like vesicles in formation of cytomictic channels between tobacco microsporocytes

S. Mursalimov, Y. Sidorchuk, E. Deineko

Biologia plantarum 57:291-297, 2013 | DOI: 10.1007/s10535-012-0276-y

The formation of cytomictic channels (CCs) during the tobacco microsporogenesis has been analyzed by microscopy and cytochemical methods. Starting from the pachytene stage, CCs were formed between microsporocytes with involvement of specific organelles, the so-called spherosome-like vesicles. The presence of the enzyme callase, able to degrade callose and form CCs in the cell wall of microsporocytes, has been demonstrated for the first time in the spherosome-like vesicles. An active form of callase was detectable in the spherosome-like vesicles and cell wall but not in the endoplasmic reticulum and Golgi apparatus. The release of callase from spherosome-like...

Multilamellar bodies linked to two active plasmalemma regions in the pollen grains of Sarcocapnos pulcherrima

M. C. Fernández, M. A. Pérez-Gutierrez, V. N. Suarez-Santiago, M. J. Salinas-Bonillo, A. T. Romero-García

Biologia plantarum 57:298-304, 2013 | DOI: 10.1007/s10535-012-0295-8

The presence of visible multilamellar bodies in the cytoplasm of pollen grains of at least seven species of the family Papaveraceae has led us to study the behaviour of these bodies during pollen-grain ontogeny and in growing pollen tubes of Sarcocapnos pulcherrima C. Morales & R. Garcia germinated in vitro. Our transmission-electron-microscope (TEM) studies in pollen grains show that the multilamellar bodies may be classified as: 1) small, isolated and placed in the region of apertures in the cytoplasm; and 2) large, in clusters and in contact with the active plasmalemma apertures only when tubules are being...

Patterns of protein expression in water-stressed wheat chloroplasts

A. H. M. Kamal, K. Cho, J. -S. Choi, Y. Jin, C. -S. Park, J. S. Lee, S. H. Woo

Biologia plantarum 57:305-312, 2013 | DOI: 10.1007/s10535-012-0290-0

The performance of control and water-stressed 10-d-old wheat seedlings was compared. During short-term water stress (irrigation was withheld for 9 d), rates of photosynthesis and transpiration, stomatal conductance, and relative water content decreased whereas the proline content increased. Chloroplast proteins were extracted from the leaves, separated by iso-electric focusing through two-dimensional electrophoresis, and stained with CBB R-250. Differentially expressed proteins were detected and analyzed with MALDI-TOF/TOF mass spectrometry. Under water stress, 9 proteins were up-regulated whereas 11 proteins were not affected. The ribulose-1,5-bisphospate...

Differential ozone sensitivity interferes with cadmium stress in poplar clones

A. Castagna, D. Di Baccio, R. Tognetti, A. Ranieri, L. Sebastiani

Biologia plantarum 57:313-324, 2013 | DOI: 10.1007/s10535-012-0274-0

Information on plant responses to combined ozone and cadmium stresses are scarce and limited to herbaceous species. In this research, two poplar clones (I-214 and Eridano), differently sensitive to O₃, were grown for 5 weeks in pots supplied with 0, 53.5, and 160.5 mg(Cd) kg^-1 (soil d.m.) and then exposed to 15-d O₃ fumigation (0.06 mm³ dm^-3, 5 h a day). The effects of the two stressors, alone or in combination, on Cd, Ca, Fe, and Zn accumulation in above-nad below-ground organs, photosynthesis, leaf pigments, and accumulation of H₂O₂ and NO were investigated. Cadmium induced...

Metabolism and aluminum accumulation in Plantago almogravensis and P. algarbiensis in response to low pH and aluminum stress

N. Martins, S. Gonçalves, A. Romano

Biologia plantarum 57:325-331, 2013 | DOI: 10.1007/s10535-012-0271-3

We investigated the impact of low pH and aluminum on the metabolism and capacity for Al accumulation in shoots of the plantain species Plantago algarbiensis and P. almogravensis. We found that increasing the concentration of Al in the medium increased accumulation of it in the shoots of both plants (although more in P. almogravensis than in P. algarbiensis). The presence of Al in the medium induced proline and saccharide synthesis in P. almogravensis without affecting lipid peroxidation, but increased proline synthesis and lipid peroxidation in P. algarbiensis without affecting the saccharide content. Lipid...

Lithium alters elicitor-induced H₂O₂ production in cultured plant cells

N. Orbán, K. Bóka

Biologia plantarum 57:332-340, 2013 | DOI: 10.1007/s10535-012-0279-8

Lithium pollution may seriously influence the metabolic and signalling processes of plants. In the present paper, we investigate the effect of lithium chloride on fungal elicitor-triggered H₂O₂ generation in Rubia tinctorum L. cell cultures. Our results show that Li⁺ strongly influences elicitor-induced H₂O₂ formation and time-course in the cells nad culture medium. Neomycin, a phospholipase C inhibitor, and 2-APB, an inositol-1,4,5-triphosphate (IP₃) receptormediated Ca²⁺ release blocker, strongly affected the elicitor-induced H₂O₂ production...

Identification of promoter regions in the Arabidopsis thaliana atExt1 extensin gene controlling late responses to wounding and pathogen attack

K. Roberts, G. Merkouropoulos, A. H. Shirsat

Biologia plantarum 57:341-350, 2013 | DOI: 10.1007/s10535-012-0281-1

The Arabidopsis thaliana (L.) Heynh. atExt1 extensin gene is expressed in a cell and tissue-specific manner, in response to developmental cues, and is inducible by a wide range of biotic and abiotic stresses. Over-expression of this gene has been shown to alter stem morphology and to limit the invasiveness of virulent bacterial pathogens, indicating that this cell wall protein gene plays an important role in plant development and defense. A detailed sequence analysis of 3.2 kb of the atExt1 gene promoter region has identified a large number of putative 5'cis-acting elements. Based on the location of clusters of putative...

Transgenic rice lines constitutively co-expressing tlp-D34 and chi11 display enhancement of sheath blight resistance

J. M. Shah, R. Singh, K. Veluthambi

Biologia plantarum 57:351-358, 2013 | DOI: 10.1007/s10535-012-0291-z

Transgenic rice (Oryza sativa L. subsp. indica cv. White Ponni) constitutively expressing the rice thaumatin-like protein gene (tlp-D34, PR-5) individually or in combination with the rice chitinase gene (chi11, PR-3) was generated using an Agrobacterium vir helper strain with multiple copies of pTiBo542 virB and virG. Transformation with the tlp-D34 gene alone and tlp-D34 + chi11 genes yielded five and seven single-copy transgenic lines, respectively. Southern blot analysis with two probes, one flanking the right T-DNA border and the second flanking the left T-DNA border, confirmed...

Reviews

Recent advances in molecular events of fruit abscission

R. J. Xie, L. Deng, L. Jing, S. L. He, Y. T. Ma, S. L. Yi, Y. Q. Zheng, L. Zheng

Biologia plantarum 57:201-209, 2013 | DOI: 10.1007/s10535-012-0282-0

It is widely accepted that fruit abscission is a highly regulated developmental process that is both influenced and activated in response to changing environment and plays crucial roles in the health and reproductive success of plants. Recent evidences showed that numerous genes related to metabolic and signalling pathways were coordinately implicated in regulating fruit abscission. Cross talks within hormones, between saccharides and hormones, as well as between polyamines and ethylene result in synergetic or antagonistic interactions which together play an important role in adjusting fruit abscission. Although hormones are the most studied internal...

Brief Communications

Over-expression of PaSOD in transgenic potato enhances photosynthetic performance under drought

A. K. Pal, K. Acharya, S. K. Vats, S. Kumar, P. S. Ahuja

Biologia plantarum 57:359-364, 2013 | DOI: 10.1007/s10535-012-0277-x

Drought stress enhances the production of superoxide radical (O₂ ^._) and superoxide dismutase catalyses dismutation of it to H₂O₂ and O₂, and hence provides a first line of defense against oxidative stress. Over-expression of a cytosolic copper-zinc superoxide dismutase, cloned from Potentilla atrosanguinea (PaSOD), in potato (Solanum tuberosum ssp. tuberosum L. cv. Kufri Sutlej) resulted in enhanced net photosynthetic rates (P_N) and stomatal conductance (g_s) compared to that in the wild type (WT) plants under control (irrigated) as well...

Calcium-mediated enhancement of copper tolerance in Elodea canadensis

H. L. Min, S. J. Cai, Z. Rui, S. Sha, K. B. Xie, Q. S. Xu

Biologia plantarum 57:365-369, 2013 | DOI: 10.1007/s10535-012-0275-z

The alleviative effects of exogenous calcium on copper phytotoxicity were investigated in Elodea canadensis plants. There was a significant accumulation of Cu in the plants after their exposure to 0.01 mM Cu accompanied by many symptoms of toxicity. Increased uptake of Cu severely reduced content of photosynthetic pigments, soluble proteins, and free proline. The total antioxidant capacity (T-AOC), reduced glutathione (GSH), and non-protein thiol (NP-SH) were severely suppressed in Cu-stressed plants resulting in a rapid increase in content of superoxide anion (O₂ ^.-), hydrogen peroxide, lipid peroxidation, and cell death....

Drought induced programmed cell death and associated changes in antioxidants, proteases, and lipid peroxidation in wheat leaves

A. Hameed, M. Goher, N. Iqbal

Biologia plantarum 57:370-374, 2013 | DOI: 10.1007/s10535-012-0286-9

Two wheat (Triticum aestivum L.) genotypes with varying degree of drought tolerance were used to analyze programmed cell death (PCD) and related biochemical changes under drought stress. Drought induced PCD in leaves, as evident by internucleosomal nDNA fragmentation, was observed in sensitive genotype Nesser. Drought tolerant genotype (FD-83) showed higher peroxidase, superoxide dismutase, and catalase activities and ascorbate content under drought stress compared to sensitive genotype. Total phenolic content increased whereas lipid peroxidation remained un-changed under drought in FD-83. In contrast, drought enhanced the proteases and ascorbate...

Cold hardening and sucrose treatment improve cryopreservation of date palm meristems

L. Fki, N. Bouaziz, O. Chkir, R. Benjemaa-Masmoudi, A. Rival, R. Swennen, N. Drira, B. Panis

Biologia plantarum 57:375-379, 2013 | DOI: 10.1007/s10535-012-0284-y

Date palm (Phoenix dactylifera L.) cv. Khenizi caulogenic meristems were initiated from achlorophyllous leaves excised from in vitro shoot cultures and then proliferated on a specific culture medium supplemented with 70 g dm^-3 sucrose. Regeneration rates obtained when using standard vitrification, droplet-vitrification, and encapsulation-vitrification protocols reached 26.7, 60.0, and 40.0 %, respectively. Only explants smaller than 3 mm in diameter were found to survive cryogenic treatments. Sucrose preculture, cold hardening and loading solution pretreatments showed significant effects on regeneration rates. Moreover, our...

Exogenous application of trehalose induced H₂O₂ production and stomatal closure in Vicia faba

J. Gao, N. Wang, S. -S. Xu, Y. Li, Y. Wang, G. -X. Wang

Biologia plantarum 57:380-384, 2013 | DOI: 10.1007/s10535-012-0285-x

Trehalose can reduce stomatal aperture by a hydrogen-peroxide-dependent pathway in Vicia faba L. (cv. Daqingpi) resulting in significantly lower values of net photosynthetic rate (P_N), stomatal conductance (g_s), and transpiration rate (E). At 8 and 24 h, the lower P_N in trehalose-treated plants was accompanied by significant decrease in intercellular CO₂ concentration (c_i) suggesting that the reduction of P_N was caused by stomatal limitation. At 48 and 72 h, trehalose decreased apparent carboxylation efficiency (P_N/c_i) and did not decrease c_i and...

The effect of arsenic on pigment composition and photosynthesis in Hydrilla verticillata

S. Srivastava, A. K. Srivastava, B. Singh, P. Suprasanna, S. F. D'souza

Biologia plantarum 57:385-389, 2013 | DOI: 10.1007/s10535-012-0288-7

The present study evaluated the effects of 100 and 500 μM arsenate (Na₂HAsO₄) on pigment composition and photosynthesis in Hydrilla verticillata (L.f.) Royle. Arsenic accumulation increased in concentration and duration dependent manner. The maximum accumulation [568 μg(As) g^-1(d.m.)] was observed at 500 μM concentration and 96-h exposure. This concentration led to a significant decline in chlorophyll a content and PS II efficiency during the whole experiment, and in chlorophyll b and carotenoids after 96 h, but no significant changes in photosynthetic pigments were noticed at 100 μM...

Glucose, sucrose, and steviol glycoside accumulation in Stevia rebaudiana grown under different photoperiods

S. Ceunen, J. M. C. Geuns

Biologia plantarum 57:390-394, 2013 | DOI: 10.1007/s10535-012-0289-6

The amount of free glucose and sucrose within the leaves of Stevia rebaudiana has been evaluated under different photoperiods and related to steviol glycoside (SVgly) accumulation. The diurnal dynamics of glucose and sucrose in leaves, internodes, apices, and roots revealed considerable fluctuations. Within leaves, glucose and sucrose content decreased up to three-fold during the night whereas SVgly amounts did not show any significant diurnal fluctuations. Ontogenetic variations in glucose content was markedly different under long-day (LD) and short-day (SD) regimes. Under LD, glucose content increased two-fold after the onset of flower bud...

Overexpression of homogentisate phytyltransferase (HPT) and tocopherol cyclase (TC) enhances α-tocopherol content in transgenic tobacco

M. C. Harish, P. Dachinamoorthy, S. Balamurugan, S. Bala Murugan, R. Sathishkumar

Biologia plantarum 57:395-400, 2013 | DOI: 10.1007/s10535-012-0298-5

Photosynthetic organisms synthesize the amphipathic antioxidants called tocopherols which are essential components of the human diet. To increase the α-tocopherol (vitamin E) content, Arabidopsis genes encoding homogentisate phytyltransferase (HPT) and tocopherol cyclase (TC) were constitutively expressed individually and in combination (HPT:TC) in tobacco plant by Agrobacterium mediated transformation. The transgene was confirmed by polymerase chain reaction (PCR), transgene expression was studied by reverse transcriptase (RT)-PCR, integration of the transgene in the plant genome was confirmed by Southern blot,...