Biologia plantarum, 2011 (vol. 55), issue 3

ArticleReviewsOriginal PapersBrief Communications

Reviews

Beyond osmolytes and transcription factors: drought tolerance in plants via protective proteins and aquaporins

S. S. Hussain, M. T. Iqbal, M. A. Arif, M. Amjad

Biologia plantarum 55:401-413, 2011 | DOI: 10.1007/s10535-011-0104-9

Mechanisms of drought tolerance have been studied by numerous groups, and a broad range of molecules have been identified to play important roles. A noteworthy response of stressed plants is the accumulation of novel protective proteins, including heat-shock proteins (HSPs) and late embryogenesis abundant (LEA) proteins. Identification of gene regulatory networks of these protective proteins in plants will allow a wide application of biotechnology for enhancement of drought tolerance and adaptation. Similarly, aquaporins are involved in the regulation of water transport, particularly under abiotic stresses. The molecular and functional characterization...

Original Papers

Variability in shoot cultures regenerated from hairy roots of Gentiana punctata

B. Vinterhalter, S. Zdravković-Korać, S. Ninković, N. Mitić, T. Janković, J. Miljuš-Djukić, D. Vinterhalter

Biologia plantarum 55:414-422, 2011 | DOI: 10.1007/s10535-011-0105-8

Differences among three clones of Gentiana punctata L. hairy root shoot regenerants were investigated in relation to their growth patterns, production of secondary metabolites and 2D protein profiles. Prominent differences in growth parameters were stable thus qualifying regenerant clones as true somaclones. Marked differences in protein spots were registered among the regenerant clones but not in comparison with the non-transformed control. Southern blot hybridization of regenerants showed the absence of rolA, B and C genes, initially present in the main hairy root lines. Orf13 and rolD were present and orf8...

Effects of NaCl on the response of Mesembryanthemum crystallinum callus to Botrytis cinerea infection

E. Kuźniak, B. Gabara, M. Skłodowska, M. Libik-Konieczny, Z. Miszalski

Biologia plantarum 55:423-430, 2011 | DOI: 10.1007/s10535-011-0106-7

Callus of the halophyte Mesembryanthemum crystallinum was used to study the effect of NaCl on the response to Botrytis cinerea infection. The fungus easily colonized the callus surface and the intercellular spaces. However, in the NaCl-adapted tissues the incidence of penetration was 67 % lower than in the inoculated control tissue. The modification of the infection pattern found in the salt-adapted callus could be related to metabolic adaptations to salinity. This was manifested by the enhanced antioxidant potential of ascorbate, the up-regulated activities of ascorbate peroxidase, as well as guaiacol and syringaldazine peroxidases together...

Construction and characterization of a cDNA library from floral organs and fruitlets of Citrus reticulata

T. G. Zheng, W. M. Qiu, G. E. Fan, B. B. Zheng, W. W. Guo

Biologia plantarum 55:431-436, 2011 | DOI: 10.1007/s10535-011-0107-6

To explore and isolate genes related to flowering and fruit development, we constructed a cDNA library from floral organs and fruitlets of Ponkan mandarin (Citrus reticulata Blanco). A total of 661 high-quality expressed sequence tags (ESTs) were generated and submitted to GenBank with the accession numbers from GO343532 to GO344192. All these ESTs were assembled into 43 contigs and 296 singletons (totally 339 unigenes). The BLAST2GO software was employed to annotate the unigenes, among which 77 ones had no significant homology with the sequences in NCBI non-redundant proteins database by BLASTX analysis. Additionally, gene ontology...

cDNA-AFLP profiling in the embryo axes during common bean germination

B. Teulat-Merah, M. -C. Morere-Le Paven, C. Ricoult, C. Aubry, D. Peltier

Biologia plantarum 55:437-447, 2011 | DOI: 10.1007/s10535-011-0108-5

cDNA-AFLP fingerprinting was used to identify genes with modulated expression during germination in common bean (Phaseolus vulgaris L.). The analysis was performed on the embryo axes. Nine time points covering the whole germination were considered and 800 transcript-derived fragments (PvTDFs) were scored. Among them, 80 % showed no changes during germination. The 97 PvTDFs showing differential expressions during germination were sequenced along with 14 constant transcripts that were randomly chosen. The expression of seven variable PvTDFs was confirmed by real-time RT-PCR. We observed that 92 % of the transcript changes, including 35 % of appearing...

Gene mutations in rye causing embryo lethality in hybrids with wheat: allelism and chromosomal localisation

N. Tikhenko, N. Tsvetkova, S. Priyatkina, A. Voylokov, A. Börner

Biologia plantarum 55:448-452, 2011 | DOI: 10.1007/s10535-011-0109-4

In crosses between hexaploid wheat and inbred lines of rye, a small number of rye genotypes produce seeds carrying undifferentiated, non-viable embryos. Hybrids between such lines and those not showing this phenotype were used as pollen donors in crosses with bread wheat in order to determine the genetic basis of disturbed embryo development. A single gene, designated Eml-R1b, is causing this character. Molecular markers associated with F₂ genotypes derived from a contrasting rye inbred progeny were used for a linkage study. Recombinant inbred lines of an F₅ population served as testers. Eml-R1b maps to chromosome...

Engineering tocopherol biosynthetic pathway in lettuce

Y. Li, G. Wang, R. Hou, Y. Zhou, R. Gong, X. Sun, K. Tang

Biologia plantarum 55:453-460, 2011 | DOI: 10.1007/s10535-011-0110-y

In order to increase tocopherol content, genes encoding Arabidopsis homogentisate phytyltransferase (HPT) and γ-tocopherol methyltransferase (γ-TMT) were constitutively over-expressed in lettuce (Lactuca sativa L. var. logifolia), alone or in combination. Over-expression of hpt could increase total tocopherol content, while over-expression of γ-tmt could shift tocopherol composition in favor of α-tocopherol. Transgenic lettuce lines expressing both hpt and γ-tmt produced significantly higher amount of tocopherol and elevated α-/γ-tocopherol ratio compared with...

NaCl tolerance in Lycopersicon pennellii introgression lines: QTL related to physiological responses

A. Frary, D. Keleş, H. Pinar, D. Göl, S. Doğanlar

Biologia plantarum 55:461-468, 2011 | DOI: 10.1007/s10535-011-0111-x

The growth and ion content of salt sensitive Lycopersicon esculentum Mill. cv. M82 and salt tolerant L. pennellii Correll accession LA716 were examined under both control and stress conditions (150 mM NaCl). L. esculentum grew more vigorously than L. pennellii under optimal conditions, however, L. pennellii was able to maintain its growth better than cultivated tomato when the plants were exposed to salinity. Sodium content of both L. esculentum and L. pennellii increased as a result of NaCl stress. In addition, both species showed reduced potassium and calcium content due to salinity. The physiological...

Microsporogenesis and pollen formation in cassava

C. Wang, Z. Lentini, E. Tabares, M. Quintero, H. Ceballos, B. Dedicova, C. Sautter, C. Olaya, P. Zhang

Biologia plantarum 55:469-478, 2011 | DOI: 10.1007/s10535-011-0112-9

This article describes the complete microsporogenesis and pollen formation in cassava (Manihot esculenta Crantz) at the various developmental stages (pollen mother cell, meiosis, tetrads, early free spore, mid uninucleate, late uninucleate, binucleate and mature pollen grain). Light microscopy, transmission electron microscopy and confocal laser scanning microscopy were used for the study. Floral bud size and other inflorescence characteristics were correlated with specific stages of the microspore development. This association allows a rapid selection of floral buds with similar microspore developmental stages, useful when a large number of...

Histomorphological changes in shoot apices of Lactuca sativa treated with gibberellic acid

O. Lee, N. Sugiyama

Biologia plantarum 55:479-484, 2011 | DOI: 10.1007/s10535-011-0113-8

Lettuce plants were treated with gibberellic acid (GA₃) and uniconazole (UZ; a gibberellin synthesis inhibitor) to investigate the influence of GA₃ on cell division frequency in the shoot apical meristem (SAM) during stem elongation and flower initiation in lettuce (Lactuca sativa) grown in a greenhouse. GA₃ (0.1 mM) was sprayed on the surface of outer leaves and uniconazole solution (0.86 mM) was applied to the soil. GA₃ increased cell division frequency in the peripheral zone and the rib meristem of shoot apices, and this was associated with the stimulation of stem elongation. UZ treatment decreased...

Role of peroxisomes in the oxidative injury induced by 2,4-dichlorophenoxyacetic acid in leaves of pea plants

I. McCarthy-Suárez, M. Gómez, L. A. Del Río, J. M. Palma

Biologia plantarum 55:485-492, 2011 | DOI: 10.1007/s10535-011-0114-7

The role of peroxisomes in the oxidative injury induced by the auxin herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) in leaves of pea (Pisum sativum L.) plants was studied. Applications of (2,4-D) on leaves or to root substrate increased the superoxide radical production in leaf peroxisomes. Foliar application also increased H₂O₂ contents in leaf peroxisomes. Reactive oxygen species (ROS) overproduction was accompanied by oxidative stress, as shown by the changes in lipid peroxidation, protein carbonyls, total and protein thiols, and by the up-regulation of the activities of superoxide dismutase, ascorbate peroxidase,...

Ultrastructure changes of seedlings of Kosteletzkya virginica under waterlogging conditions

J. Zhou, S. W. Wan, G. Li, P. Qin

Biologia plantarum 55:493-498, 2011 | DOI: 10.1007/s10535-011-0115-6

The changes in ultrastructure of leaf cell in waterlogged Kosteletzkya virginica seedlings were observed by transmission electron microscope. During waterlogging, the chloroplasts turned into a round shape and their volume was gradually reduced. Furthermore, the lamellae of thylakoids swelled, and the inclusions in chloroplast decreased. The shape of starch grains slightly changed, but their number and volume decreased, and they even disappeared in the end. On the other hand, plastoglobules enlarged and their amount enhanced gradually. The membrane and inner cristea of mitochondria gradually became unclear, while the mitochondria firstly enlarged...

Impact of homogenization and protein extraction conditions on the obtained tobacco pollen proteomic patterns

J. Fíla, V. Čapková, J. Feciková, D. Honys

Biologia plantarum 55:499-506, 2011 | DOI: 10.1007/s10535-011-0116-5

Mature pollen grain represents a highly desiccated structure with an extremely tough cell wall. Thanks to it, it resists common proteomic protocols. Instead, a robust homogenization has to be performed since proteins are needed to burst out of the cell to be included in the extracted proteome fraction. Here, a novel way of pollen homogenization employing Roche MagNA Lyser Instrument is presented, sparing time and laborious work. However, plant proteomics does not rely solely on perfect homogenization; also the choice of the extraction protocol is of key importance. The composition of the extraction buffer has a decisive influence on which proteome...

Arabidopsis Ca²⁺-dependent protein kinase CPK3 mediates relationship of putative inositol triphosphate receptor with slow-type anion channel

A. Cousson

Biologia plantarum 55:507-521, 2011 | DOI: 10.1007/s10535-011-0117-4

It has been suggested in Arabidopsis thaliana (L.) Heynh. cv. Columbia that, contrary to 30 μM abscisic acid (ABA), 20 μM ABA induces guard cell Ca²⁺ mobilization through activating phosphoinositide-specific phospholipase C (PI-PLC)-dependent inositol 1,4,5-triphosphate (IP₃) production. Here, it was investigated whether Ca²⁺-dependent protein kinase, CPK3 or CPK6 would mediate ABA-induced stomatal closure downstream of IP₃ production. In the knockout cpk3-1 mutant, the PLC inhibitor (U73122) adjusted 20 μM ABA-induced stomatal closure to the extent observed in the knockout cpk6-1...

Thiamine seed treatment enhances LOX expression, promotes growth and induces downy mildew disease resistance in pearl millet

H. G. Pushpalatha, J. Sudisha, N. P. Geetha, K. N. Amruthesh, H. Shekar Shetty

Biologia plantarum 55:522-527, 2011 | DOI: 10.1007/s10535-011-0118-3

Seeds of pearl millet [Pennisetum glaucum (L). R.Br.] susceptible cv. 7042S were treated with thiamine at 5, 10, 15, 20 and 25 mM concentrations and growth promotion and downy mildew resistance were tested. Seed treatment with 20 mM thiamine resulted in 72 and 70 % disease protection under greenhouse and field conditions, respectively, and enhanced vegetative and reproductive growth parameters. Analysis of lipoxygenase (LOX) activity in inoculated pearl millet seedlings at different time intervals indicated that increased LOX activity was initiated at 3 h after inoculation (hai) and maximum activity was observed at 24 hai. Northern analysis...

Special origin of stem sequence influence the resistance of hairpin expressing plants against PVY

F. Jiang, B. Wu, C. Zhang, Y. Song, H. An, C. Zhu, F. Wen

Biologia plantarum 55:528-535, 2011 | DOI: 10.1007/s10535-011-0119-2

In this study, 16 hairpin RNA (hpRNA) vectors were constructed, each harboring 50 bp viral RNA sequence as the stem. They all targeted the coat protein (CP) gene of Potato virus Y (PVY). Virus resistance assay revealed that hairpin constructs targeting the anterior 200 bp regions of the CP gene were unable to induce virus resistance, while the 12 hpRNA constructs targeting posterior 600 bp regions induced high virus resistance up to 77.78 %. Northern blot analysis revealed that 50 bp-length hpRNA constructs could be transcribed efficiently and processed into siRNAs; however, no correlation between siRNA accumulation and degree...

Brief Communications

Introduction of OsglyII gene into Oryza sativa for increasing salinity tolerance

S. H. Wani, S. S. Gosal

Biologia plantarum 55:536-540, 2011 | DOI: 10.1007/s10535-011-0120-9

Mature seed-derived embryogenic calli of indica rice (Oryza sativa L. cv. PAU201) were induced on semisolid Murashige and Skoog medium supplemented with 2.5 mg dm^-3 2,4-dichlorophenoxyacetic acid + 0.5 mg dm^-3 kinetin + 560 mg dm^-3 proline + 30 g dm^-3 sucrose + 8 g dm^-3 agar. Using OsglyII gene, out of 3180 calli bombarded, 32 plants were regenerated on medium containing hygromycin (30 mg dm^-3). Histochemical GUS assay of the hygromycin selected calli revealed GUS expression in 50 % calli. Among the regenerants, 46.87 % were GUS positive. PCR analysis confirmed the...

Evaluation of the effect of in vitro stress and competition on tissue culture response of flax

M. Yildiz

Biologia plantarum 55:541-544, 2011 | DOI: 10.1007/s10535-011-0121-8

This study was carried out to investigate the in vitro competition in tissue culture of three flax (Linum usitatissimum L.) cultivars using different distances among hypocotyl explants cultured. Hypocotyl fresh and dry masses, shoot regeneration percentage, shoot number per hypocotyl, regenerated shoot length and total chlorophyll content were examined during shoot regeneration, while plantlet height, number of roots and length of roots were recorded during rooting. With decreasing distance among explants we observed increased shoot regeneration and rooting till a certain point from where stress initiated and significant decreases in...

Involvement of ABA in reduced photosynthesis and stomatal conductance in Cuscuta campestris - Mikania micrantha association

H. Chen, H. Shen, W. Ye, H. Cao, Z. Wang

Biologia plantarum 55:545-548, 2011 | DOI: 10.1007/s10535-011-0122-7

We investigated whether changes in abscisic acid (ABA) content in leaves of Mikania micrantha infected by the holoparasite Cuscuta campestris at five growth stages, influenced the host stomatal conductance (g_s), transpiration rate (E) and net photosynthetic rate (P_N). C. campestris infection caused a negative effect on g_s, E and P_N of the host plants. ABA content in host leaves infected by C. campestris was significantly lower at 6 d after parasitization (DAP) and significantly higher at 13 and 33 DAP, relative to uninfected controls. In the parasite, ABA content was lowest at...

Extracellular matrix as the early structural marker for Centella asiatica embryogenic tissues

K. S. Lai, K. Yusoff, M. Maziah

Biologia plantarum 55:549-553, 2011 | DOI: 10.1007/s10535-011-0123-6

Embryogenic and non-embryogenic calli were induced from the Centella asiatica leaf explants on Murashige and Skoog medium supplemented with kinetin and 2,4-dichlorophenophenoxyacetic acid. The extracellular matrix (ECM) layer was seen on the surface of embryogenic cells but not on the non-embryogenic cells. The ECM formed bridges with net-like material between the embryogenic cells. This network like structure was believed to play an important role in plant morphogenesis and can serve as an early structural marker of embryogenic competence in Centella asiatica calli culture.

High frequency plant regeneration from the mature seeds of Garcinia indica

M. Baskaran, S. Krishnan

Biologia plantarum 55:554-558, 2011 | DOI: 10.1007/s10535-011-0124-5

A high frequency plant regeneration system was developed for the production of high yielding elite clones of Garcinia indica via direct organogenesis. A maximum number of 86.2 shoot buds per explant were induced from the mature seed segments cultured on Woody plant medium (WPM) supplemented with 2.0 mg dm^-3 N⁶-benzyladenine and 1 mg dm^-3 indole-3-acetic acid. Rooting was achieved on half-strength WPM medium supplemented with 3 mg dm^-3 indole-3-butyric acid. Rooted plantlets were acclimatized and transferred to greenhouse for further growth. The highest survival rate of 95 % was recorded using a mixture...

Improved growth and acclimatization of somatic embryo-derived Oplopanax elatus plantlets by ventilated photoautotrophic culture

S. Y. Park, H. K. Moon, H. N. Murthy, Y. W. Kim

Biologia plantarum 55:559-562, 2011 | DOI: 10.1007/s10535-011-0125-4

To improve large-scale in vitro production of Oplopanax elatus Nakai, we cultured somatic embryo-derived plantlets under a heterotrophic condition (semi-solid culture with sucrose), photoautotrophic condition (semi-solid culture without sucrose), or modified photoautotrophic condition (liquid culture with forced ventilation). The plantlets grown under the modified photoautotrophic condition had more leaves as well as higher chlorophyll content, and higher net photosynthetic rate than those grown under the conventional conditions. Further, the photoautotrophically grown plantlets acclimatized better and sooner upon ex vitro transplantation...

Changes of leaf antioxidant system, photosynthesis and ultrastructure in tea plant under the stress of fluorine

C. Li, Y. Zheng, J. Zhou, J. Xu, D. Ni

Biologia plantarum 55:563-566, 2011 | DOI: 10.1007/s10535-011-0126-3

Seedlings of Camellia sinensis were grown hydroponically for 30 d in order to study the effect of fluorine (F) on growth parameters, antioxidant defence system, photosynthesis and leaf ultrastructure. Fresh and dry mass, chlorophyll (Chl) content and net photosynthetic rate (P_N) decreased with increasing F concentration. Superoxide dismutase (SOD) activity decreased significantly, catalase (CAT) and guaiacol peroxidase (GPX) activities reached maximun under 0.21 and 0.32 mM F, respectively. Proline, malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) contents increased significantly. These results suggested, that...

Isolation and characterization of two MADS-box genes from Lycium barbarum

S. H. Zeng, Y. Q. Xu, Y. Wang

Biologia plantarum 55:567-571, 2011 | DOI: 10.1007/s10535-011-0127-2

To broaden our knowledge of flower development, two floral homeotic genes, LbAG and LbSEP3, were isolated from the flower of Lycium barbarum L. The open reading frame length of LbAG and LbSEP3 were 1090 and 992 bp encoding 249 and 242 amino acids, respectively. Sequence alignment and phylogenetic analysis indicated that LbAG belonged to C-type MADS-box gene and that LbSEP3 was E-type MADS-box gene. Compared with other floral homeotic proteins, LbAG held the conserved AG motif I, II and LbSEP3 conserved SEP3 motif I, II. Expression profile showed that LbAG transcripts were abundant in inner two...

Selection of co-transformed Dendrobium Sonia 17 using hygromycin and green fluorescent protein

C. S. Tee, M. Maziah, C. S. Tan, M. P. Abdullah

Biologia plantarum 55:572-576, 2011 | DOI: 10.1007/s10535-011-0128-1

Dendrobium Sonia 17 calluses were used for co-transformation study using particle bombardment. The bombarded transformed callus tissues were selected using half-strength MS medium containing 25 mg dm^-3 hygromycin. Expression of green fluorescent protein (GFP) was observed in the callus and protocorm-like body (PLBs) tissues survived on the selection medium. The presence of green fluorescence protein (sgfp), hygromycin-B-phosphotransferase (hptII) and β-glucuronidase (uidA) genes in the transformed tissues were verified using PCR, Southern blot and dot blot analyses. Based on the results from PCR and expression...

Comparison of genomic SSR and EST-SSR markers for estimating genetic diversity in cucumber

J. Hu, L. Wang, J. Li

Biologia plantarum 55:577-580, 2011 | DOI: 10.1007/s10535-011-0129-0

Thirteen genomic microsatellite (gSSR) and sixteen expressed sequence tag (EST)-SSR (eSSR) markers were compared to estimate genetic diversity among 29 cucumber (Cucumis sativus L.) accessions. gSSR markers detected mean 4.46 alleles with a mean polymorphic information content (PIC) of 0.664, against eSSR markers with mean 3.38 alleles and a mean PIC of 0.397. gSSRs amplified more null alleles than eSSRs. Genetic diversity within the accession set was estimated by construction of dendrograms using gSSR or eSSR data. There was a clear consistency between gSSR and eSSR trees in terms of positioning of most cucumber germplasms. gSSR markers could...

Fatty acid content during reconstitution of the photosynthetic apparatus in the air-dried leaves of Xerophyta scabrida after rehydration

K. Georgieva, A. Ivanova, S. Doncheva, S. Petkova, D. Stefanov, E. Péli, Z. Tuba

Biologia plantarum 55:581-585, 2011 | DOI: 10.1007/s10535-011-0130-7

Desiccation of Xerophyta scabrida caused considerable damage of chloroplast ultrastructure together with a complete loss of chlorophyll. Upon rehydration, the relative water content of the pale-green leaves almost reached that of the dark-green ones, however, the Chl content and photosynthetic activity remained lower. The process of reconstitution of the photosynthetic apparatus in the re-greening leaves was accompanied by changes in fatty acid (FA) content. The amount of the FA methyl esters was more than 2-fold higher in the green leaves as compared to the dry ones and slightly increased after rehydration in the pale-green leaves. Among the...

Differential proteomic analysis of cadmium-responsive proteins in wheat leaves

Y. Wang, H. Hu, Y. Xu, X. X. Li, H. J. Zhang

Biologia plantarum 55:586-590, 2011 | DOI: 10.1007/s10535-011-0131-6

To gain a comprehensive understanding of plant response to Cd, physiological and proteomic changes in wheat (Triticum aestivum L.) leaves exposed to a range of Cd concentrations (10, 100 and 200 μM) were investigated. Leaf elongation was decreased, whereas the H₂O₂ and malondialdehyde content increased significantly at higher Cd concentrations. Changes in protein profiles were analyzed by two-dimensional electrophoresis. Twenty-one proteins which showed 1.5-fold change in protein abundance in response to Cd were identified. These proteins can be functionally grouped into three groups: 1) oxidative stress response,...

Mannitol, polyethylene glycol and NaCl induced polypeptide changes during in vitro culture of three tomato cultivars

R. Roy, V. Agrawal, S. C. Gupta

Biologia plantarum 55:591-595, 2011 | DOI: 10.1007/s10535-011-0132-5

Three cultivars of tomato Pusa Ruby, Arka Vikas and Pusa Early Dwarf were subjected to osmotic stresses induced by mannitol, polyethylene glycol and NaCl in vitro. Polypeptide patterns were analyzed during each stress treatment to differentiate between tolerant and sensitive cultivars. The stresses induced more stress proteins in cv. Pusa Ruby compared to other two cultivars indicating it to be relatively osmotic stress tolerant.

Gametes with somatic chromosome number and their significance in interspecific hybridization in Fuchsia

R. S. Talluri

Biologia plantarum 55:596-600, 2011 | DOI: 10.1007/s10535-011-0133-4

Sexual polyploidization has both a theoretical as well as an applied significance. Morphological screening for large pollen grains and shape of pollen produced by the individual, cytological investigation of hybrid progeny, and unbalanced separation of chromosomes at anaphase I in pollen mother cells were used to detect the gametes with somatic chromosome number in Fuchsia. The interspecific hybrids of F. fulgens (sect. Ellobium) × F. magellanica (sect. Quelusia), F. fulgens (sect. Ellobium) × F. splendens (sect. Ellobium), and F. triphylla (sect. Fuchsia) × F. splendens...