Gynogenesis of Chinese long cucumber (Cucumis sativus L.) was obtained from unpollinated ovules cultured on cucumber basal medium (CBM) supplemented with thidiazuron (TDZ) and in some experiments AgNO₃. High induction frequencies (7.85-12.14 %) were induced from unpollinated ovules at the time of anthesis at 0.03-0.07 mg dm^-3 TDZ. Histological analysis indicated that embryo sacs developed completely at the time of anthesis. Further, the highest plant regeneration rate was achieved at CBM supplemented with 0.05 mg dm^-3 a-naphthaleneacetic acid, 0.2 mg dm^-3 6-benzyladenine and 5-10 mg dm^-3 AgNO₃. Flow cytometry analysis showed that 80 % of the regenerated plants were haploid. Histological micrographs and ploidy level analyses clearly revealed initiation, development, and germination of embryos from the unpollinated ovules.

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 profiles of these miRNAs and their corresponding target genes were examined in developing caryopses. Our results suggest that the miRNAs and their target genes examined at the two distinct stages could contribute to the developmental progress of rice caryopses in concert with phytohormone signalling.

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 g_s compared with controls; therefore the reduction in photosynthesis was caused by non-stomatal limitation. Trehalose treatment resulted in significantly higher effective photochemical efficiency of PS II (Φ_PSII) and did not affect maximum photochemical efficiency of PS II (F_v/F_m). At 24, 48, and 72 h, trehalose decreased non-photochemical quenching (NPQ) and increased photochemical quenching (qP). Our results suggest that trehalose did not damage photosynthetic reaction centers.

Temperature and mineral nutrition are major environmental factors regulating plant growth and development. Yet, cold impact on mineral contents and the ability of the plants to perform changes in specific elements as a part of the acclimation process received little attention. Using five Coffea genotypes previously characterized concerning their cold sensitivity, a mineral analysis was performed considering macro (N, P, K, Ca, Mg, and S) and micro (Na, Fe, Mn, Zn, Cu, and B) nutrients in order to predict their importance in cold tolerance. The results showed a cold-induced dynamics of mineral nutrients in recently mature leaves. The less cold sensitive Icatu, and partially Catuaí, accumulated N, Ca, Mn, Cu, and Zn with potential implications in the maintenance of photosynthetic performance, the reinforcement of the antioxidative defense system, lipid metabolism, and the expression of cold regulated genes, thus constituting interesting traits to evaluate the cold acclimation ability. After a principal component analysis (PCA), N, Fe, Mn, and Cu were further confirmed as strong candidates for an early cold tolerance evaluation due to their dynamics and to specific roles in the activities of Cu/Zn-SOD (Cu), APX (Fe), and PSII (Mn).

In vitro morphogenetic ability of plant cells has been demonstrated in diverse species of angiosperms and gymnosperms but no such report is available in the genus Selaginella till date. We have established an in vitro morphogenic root culture where indole butyric acid (IBA) induced profuse branched and unbranched roots in Selaginella microphylla. We observed inter-convertibility of root apical meristem (RAM) to shoot apical meristem (SAM) in presence of IBA and showed that intact roots are also capable of transformation. Friable callus was obtained from roots on prolonged (∼50 weeks) root cultures. By isolating total RNA from each of the developmental stages, we performed cDNA synthesis followed by random amplification, sequencing, and BLAST analysis of differentially expressed transcripts to correlate morphological events with the changes on molecular level. The results revealed sequence matches to genes involved in diverse cellular processes such as transcription, translation, photosynthesis, replication, secondary metabolism, stress response, and plant defense suggesting ancient origins of such proteins and the evolutionary conservation of biological function.

The effects of plant growth regulators (PRGs) on the induction of flowering and sex expression in micropropagated cucumbers are presented. The highest number of male flowers (6.0 ± 0.7 per plant) was produced by cv. Kmicic F1 on the Murashige and Skoog (MS) medium supplemented with 4.0 μM kinetin. The highest number of female flowers (3.1 ± 0.3) was also observed in cv. Kmicic F1 on either control (PRG-free) medium or medium supplemented with 6.4 μM indole-3-acetic acid (IAA). The MS medium supplemented with 4.4 μM benzyladenine inhibited flower formation. The highest percentage of flowering plantlets (67.5 ± 7.5) was observed on the control MS medium after 16 weeks of culture. Female-to-male flower ratio was influenced by the culture media and changed during cultivation. The highest pollen viability (60-70 %) was observed in anthers of plants cultured on the control medium and the medium with IAA.

Innate immune system is employed by plants to defend against phytopathogenic microbes through specific perception of non-self molecules and subsequent initiation of resistance responses. Current researches elucidate that plants mostly rely on cell surface-located pattern recognition receptors (PRRs) and intracellular nucleotide-binding leucine-rich repeat proteins (NB-LRRs) to recognize pathogen-associated molecular patterns (PAMPs) and effector proteins from microbial pathogens, initiating PAMP- and effector-triggered immunity (PTI and ETI), respectively. Some pathogenic bacterial effector proteins are usually secreted into plant cells and play a virulence function by suppressing plant PTI, implying an evolutionary process of plant immunity from PTI to ETI. In the past several years, a great progress has been achieved to reveal fascinating molecular mechanisms underlying the pathogenic recognition, resistance signaling transduction, and plant immunity evolution. Here, we summarized the latest breakthroughs about these topics, and offered an integral understanding of plant molecular immunity.

Gentiana straminea is native to the Qinghai-Tibet plateau, where it is exposed to extremes of cold and strong UV-B radiation. Here we showed that low temperature, but not UV-B radiation, affected the accumulation of the triterpenoid oleanolic acid. Neither of these stresses altered the expression of known triterpenoid synthesis genes. However, the application of methyl jasmonate (MeJA), but not salicylic acid (SA), significantly enhanced the accumulation of oleanolic acid, and up-regulated the triterpenoid synthesis genes, especially the expression of βAS, the gene encoding β-amyrin.

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.

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 that all transgenic plants harboured single and complete T-DNA copies. Homozygous transgenic lines were first identified in the T₁ generation by Southern blot analysis and were subsequently confirmed by segregation analysis of T₂ plants. Accumulation of transcripts encoded by the transgenes was confirmed in T0 plants and homozygous T₂ plants by Northern blot analysis. The homozygous T2 plants harbouring tlp-D34 + chi11 genes showed 2.8- to 4.2-fold higher chitinase activity. Western blot analysis revealed the accumulation of thaumatin-like protein and chitinase in the respective transgenic plants. Upon infection with Rhizoctonia solani, the disease index reduced from 100 % in control plants to 65 % in a T₃ homozygous transgenic line T4 expressing the tlp-D34 gene alone. In a T2 homozygous transgenic line CT22 co-expressing tlp-D34 and chi11 genes, the disease index reduced to 39 %.

In order to produce human lactoferrin (Lf) in alfalfa (Medicago sativa L.), a construct containing human Lf cDNA under the control of cauliflower mosaic virus 35S promoter was engineered. As selectable marker bar gene, whose expression in plant cells confers tolerance to L-phosphinothricin (ppt) was used. Plants from a highly embryogenic alfalfa clone from the Bulgarian cultivar Obnova 10 were transformed using Agrobacterium tumefaciens mediated leaf disc method. Transgenic alfalfa plants were established from ppt-resistant calli via indirect somatic embryogenesis. The presence of human Lf cDNA in the genome of the selected regenerants was confirmed by polymerase chain reaction (PCR). Reverse transcriptase (RT)-PCR and Western blot showed expression of human Lf in leaf tissue. Studies on antibacterial effect of the recombinant glycoprotein were conducted and resistance of the transgenic alfalfa plants to two phytopathogens, Pseudomonas syringae pv. syringae and Clavibacter michiganensis, was demonstrated. The obtained results suggest that the expression of human Lf in alfalfa could be beneficial not only for producing recombinant protein for clinical application but also for crop quality improvement.

The protein elicitor cerato-platanin (CP) is known to induce defence-related responses in various plants. Some of these responses occur very quickly. In the present work, transcriptional changes caused by CP in leaves from Platanus × acerifolia (Aiton) Willd. were studied. With a cDNA microarray, 131 differentially regulated transcripts were identified as responsive to CP after 24 h of treatment. Eighty-six of these were cold-or ozone-modulated transcripts, thus revealing a significant overlap between genes responsive to CP and to cold/ozone stress. The transcriptional changes caused by CP were compared with the CP-orthologous protein Pop1 in a time-course analysis performed after 3, 6, 12, and 24 h of treatment by real-time RT-PCR on five defence-related genes. Despite some differences, CP and Pop1 were both able to induce early transcriptional changes (WRKY was overexpressed after only 3 h) confirming that pathogenassociated molecular patterns (PAMPs) act very quickly on gene transcription.

Programmed cell death (PCD) is a genetically controlled and conserved process in eukaryotes during development as well as in response to pathogens and other stresses. BAX inhibitor-1 (BI-1) has been implicated as an anti-PCD factor which is highly conserved in plants. Sequence of putative cucumber BI-1 protein exhibited 77.7 % identity and 91.2 % positive value with the homologue Blast BI-1 protein of Arabidopsis thaliana (AtBI-1). This highly homologous protein to the AtBI-1 protein was named CsBI-1. This protein contains an open reading frame (ORF) of 250 amino acids with a BAX inhibitor domain and five transmembrane regions conserved among members of the BI-1 family. Primers designed by the cDNA of CsBI-1gene were used for further sequencing. Cell death in cold-stored cucumber developed concomitantly with increased expression of the CsBI-1 gene and reached maximum at day 6. However, cell death accelerated significantly after 9 d when sharp decrease of the CsBI-1 expression occurred. After warming to 20 °C, expression of the CsBI-1 gene was the highest at day 3, decreased afterwards, and the lowest expression was detected at day 9 when PCD obviously appeared. The overall results indicate that CsBI-1 is cucumber homologue of Arabidopsis thaliana AtBI-1 gene. CsBI-1 is a conserved cell death suppressor induced by cold stress and a negative regulator of PCD.

The ultrastructure of leaf mesophyll cells of in vitro cultured Hypericum perforatum L. plants regenerated after cryopreservation was studied. Electron microscopy analysis revealed that the chloroplasts in plants pretreated with abscisic acid and regenerated after cryopreservation were round, with increased amount of starch, rather small volume of the thylakoid system, and destroyed envelope. Plants pretreated with 0.3 M mannitol and cooled at rates of 0.1 or 0.3 °C min^-1 possessed chloroplasts with high starch content that resulted in a reduction of a membrane system. However, the pretreatment with 0.3 M mannitol and cooling at a rate of 0.2 °C min^-1 was the best as chloroplast ultrastructure resembled the controls regenerated without cryopreservation.

Proteins WCS120 and DHN5 are known as the major cold-inducible dehydrins in wheat and barley plants, respectively. WCS120 and DHN5 relative accumulation increased exponentially along with a growth temperature decline in the range from optimum to cold temperatures. Even at optimum growth temperatures, the most frost-tolerant wheat and barley cultivars can be distinguished from the remaining ones according to dehydrin relative accumulation. The highly tolerant wheat and barley cultivars started accumulating dehydrins at higher growth temperatures and reached higher dehydrin amounts than the less tolerant ones. Statistically significant correlations between lethal temperature for 50 % of the samples (LT50) and dehydrin relative accumulation have been found at all growth temperatures (5, 10, 15 and 20 °C) for WCS120 in wheats and at 5 and 10 °C for DHN5 in barleys. Analogous relationships between dehydrin relative accumulation at different growth temperatures and plant acquired frost tolerance have been proved for wheat WCS120 and barley DHN5.

Aquaporins (AQPs) are proteinaceous channels known to regulate transmembrane water transport, and therefore may be important component of imbibition during osmopriming and germination. To explore the association between AQPs and osmopriming-led enhanced germination performance, we studied the expression patterns of four spinach (Spinacia oleracea) AQP coding genes (SoPIP1;1, SoPIP1;2, SoPIP2;1, and SoδTIP) during osmopriming and subsequent germination under optimal conditions, chilling and drought. All these genes were up-regulated within 2-4 d of priming (phase II-imbibition). We hypothesize such up-regulation to facilitate the pressure potential-driven cell expansion and increase germination potential of primed seeds. Our data during post-priming germination suggest that SoPIP1;1 and SoδTIP were more closely associated with enhanced germination performance. In general, all AQPs were downregulated under chilling and drought. However, under chilling, SoPIP2;1 was expressed at relatively higher level in primed seeds that also exhibited greater chilling tolerance, while SoPIP1;2 and SoδTIP exhibited opposite pattern. Similarly, SoPIP1;1, SoPIP2;1, and SoδTIP exhibited higher expression in primed seeds that also had greater drought tolerance.

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 formed in the apertural intine. Similar types of multilamellar bodies to those observed in the pollen apertures can be seen near the apex of the growing pollen tube (small and isolated) and in contact with the apex plasmalemma (large and clustered). Our results support the hypothesis that the multilamellar bodies are functionally linked to moments when the cytoplasmic membrane is very active. We have also linked the multilamellar bodies to Golgi vesicles as they both react positively to acid-phosphatase (AP) staining and also to the plasmalemma by the thiocarbohydrazide-silver proteinate-staining (TCH-Sp) electron-contrasting technique.

An efficient, rapid, and reproducible plant regeneration protocol was successfully developed for Abrus precatorius L. using mature nodal explants excised from a 5-year-old field grown plant. The highest shoot regeneration frequency (87 %) with maximum number of multiple shoots (15.0) and shoot length (4.8 cm) were recorded on Murashige and Skoog (MS) medium amended with 2.5 μM thidiazuron, 120 mg dm^-3 polyvinylpyrrolidone, and 0.5 μM α-naphthalene acetic acid. The best treatment for maximum root (4.0) induction was half strength MS medium supplemented with 1.5 μM indole-3-butyric acid. The in vitro plantlets with well-developed shoots and roots were successfully transferred into plastic cups with Soilrite and acclimatized in a culture room under photon flux density (PFD) of 150 μmol m^-2 s^-1, thereafter transferred to a greenhouse with PFD of 300 μmol m^-2 s^-1, and finally to a field with 70 % survival rate. During the acclimatization period (0-49 d), leaf chlorophyll and carotenoid content increased whereas malondialdehyde and H₂O₂ content decreased probably due to increasing activities of antioxidant enzymes (catalase, superoxide dismutase, glutathione reductase, and ascorbate peroxidase). Our work suggests that micropropagated plants developed an antioxidant enzymatic protective system to avoid oxidative stress during establishment under ex vitro environment.

Viral resistance can be effectively induced in transgenic plants through their silencing machinery. Thus, we designed nine short hairpin RNAs (shRNA) constructs to target nuclear inclusion protein b (NIb), helper component proteinase (HC-Pro), cylindrical inclusion protein (CI) and viral protein genome linked (VPg) genes of Potato virus Y (PVY^N) and Tobacco etch virus (TEV-SD1). The shRNAs were completely complementary to the genes of PVY^N, and contained 1-3 nt mismatches to the genes of TEV-SD1. To study the specificity of gene silencing in shRNA-mediated viral resistance, the constructs were introduced into tobacco plants. The results of viral resistance assay revealed that these nine kinds of transgenic tobacco plants can effectively induce viral resistance against both PVY^N and TEV-SD1, and the shRNA construct targeting the NIb gene showed higher silencing efficiency. Northern blot and short interfering RNA (siRNA) analyses demonstrated that the viral resistance can be attributed to the degradation of the target RNA through the RNA silencing system. Correlation analysis of siRNA sequence characteristics with its activity suggested that the secondary structure stability of the antisense strand did not influence siRNA activity; 1 to 3 nt 5' end of the sense strand caused a significant effect on siRNA activity where the first base such as U was favourable for silencing; the base mismatch between the siRNA and the target gene may be more tolerated in the 5' end.

Isoflavone synthase (IFS; CYP93C) plays a key role in the biosynthesis of phenolic secondary metabolites, isoflavonoids. These compounds, which are well-known for their benefits to human health and plant defence, are produced mostly in legumes. However, more than 200 of them have been described in 59 other plant families without any knowledge of their respective IFS orthologue genes (with the sole exception of sugar beet). In this study, we selected IFS from Pisum sativum L. (CYP93C18) for functional expression. CYP93C18 was isolated, cloned, and introduced into Arabidopsis thaliana. The presence of the gene was shown by Southern blot analysis and its expression in the transgenic Arabidopsis was proven by RT-PCR and Western blots. The functional activity of the heterologous IFS was verified by HPLC-MS analysis of the metabolite levels: the isoflavone genistein and its derivatives tectorigenin and biochanin A were detected in the overexpressing lines. In addition, 35S::CYP93C18::GFP fused proteins were transiently expressed in the leaves of Nicotiana benthamiana and the localization of the GFP signal was observed on the endoplasmic reticulum using confocal microscopy which is consistent with the data from the literature and with our in silico predictions. The putative mode of attachment of IFS to the endoplasmic reticulum membrane is suggested. The undemanding methodology presented in this paper is applicable to the functional analysis of newly-identified isoflavone synthase genes from various species.

This study aimed to investigate the effects of irradiance on plant growth and content of proline and phytohormones during ex vitro acclimatization of micropropagated Ulmus minor plants. In vitro rooted plants were acclimatized to ex vitro conditions in a climate chamber with two irradiances, 200 μmol m^-2 s^-1 (high irradiance, HI) and 100 μmol m^-2 s^-1 (low irradiance, LI) for 40 d. Immediately after the ex vitro transfer, the plants experienced a water deficit [wilting leaves with the reduced relative water content (RWC)], but following the experiment, the recovery of the RWC was more pronounced in the HI treatment. Also, the content of proline, ABA, and JA-Ile were higher in HI treatment. Growth analyses revealed that HI improved growth and biomass production.

Calcium is a critical component in a number of plant signal transduction pathways and the calcineurin B-like protein (CBL) family is a unique group of calcium sensors regulating a family of CBL-interacting protein kinases (CIPKs). In this study, two poplar CBL genes, PeCBL6 (GenBank acc. No. DQ907710) and PeCBL10 (GenBank acc. No. DQ899956), were characterized in transgenic Arabidopsis thaliana, particularly with regard to its role in abiotic stress resistance. Expression of the two CBL genes in poplar was induced by cold, drought, or high salinity, but not by abscisic acid (ABA) treatment. In Arabidopsis thaliana, PeCBL6 was found in the nucleus and PeCBL10 in the tonoplast. Transgenic Arabidopsis plants overexpressing PeCBL6 or PeCBL10 showed enhanced tolerance to high salinity, drought and low temperature. These results suggested that PeCBL6 and PeCBL10 may function as positive regulators of salt, drought and temperature responses.

Brassinosteroids are known to protect plants against various abiotic and biotic stresses, however, very limited information is available about the role of progesterone. Therefore the effects of Pseudomonas syringae pv. syringae (P.s.) wild type strain 61, its hrcC mutant, and the saprophytic P. fluorescens (P.f.) strain 55 were investigated in wild type Arabidopsis thaliana cv. Columbia and its rbohF knock-out mutant, with and without progesterone pre-treatment. The reactions of wild type and rbohF mutant Arabidopsis to bacterial inoculations were similar, although 2 h after injection of P.s. a larger increase of electrolyte leakage was measured in wild type than in rbohF knockout mutant leaves. The hrcC mutant caused weak necrotic symptoms and increased leakage in both types of Arabidopsis, although to a much lesser extent than P.s. The P.f. did not induce any visible symptom, but slightly increased the electrolyte leakage in both types of Arabidopsis. Inoculation by all Pseudomonas bacteria led to significant alterations in photosystem 2 efficiency as compared to control plants. Pre-treatment of leaves with progesterone diminished the necrotic symptoms, the electrolyte leakage and improve the efficiency of photosystem 2 caused by Pseudomonas bacteria.

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 arsenate. Net photosynthetic rate, electron transport rate, and water use efficiency declined whereas transpiration rate increased, and stomatal conductance and photochemical quenching did not show any effect or increased. The content of reactive oxygen species increased and content of reduced ascorbate declined at 500 μM arsenate in comparison to the control.

This study was carried out to better understand the role of salicylic acid (SA) applied before cold stress in the cold tolerance mechanism. Two barley (Hordeum vulgare) cultivars, cold-sensitive (Akhisar) and cold-tolerant (Tokak), were used and 0.1 mM SA was applied to 7-d-old barley seedlings growing under control conditions (20/18 °C). The seedlings were transferred to cold chamber (7/5 °C) at the age 14, 21, and 28 d. After three days, the leaves were harvested to determine the activities of apoplastic antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT), and peroxidase (POX) and ice nucleation activity and electrophoretic patterns of apoplastic proteins. Cold treatment decreased the activities of all enzymes in cold-sensitive cultivar, however, it increased CAT and POX activities in cold-tolerant cultivar. Exogenous SA increased enzyme activities in both cultivars. Ice nucleation activity increased by cold treatment, especially in 17-d-old seedlings in both cultivars. In addition, SA treatment increased ice nucleation activity in all examined samplings in both cultivars. SA treatment caused accumulation or de novo synthesis of some apoplastic proteins. The results of the present study show that exogenous SA can improve cold tolerance by regulating the activities of apoplastic antioxidative enzymes, ice nucleation activity, and the patterns of apoplastic proteins.

Sex-related differences in the responses of plants to CO₂ enrichment are still rarely studied. In this study, we examined the effects of elevated atmospheric CO₂ (720 μmol mol^-1) on the activities of polyphenoloxidases (PPOs) and guaiacol peroxidases (PODs) in male and female plants of two ecologically divergent willow species Salix repens and S. phylicifolia. We detected that females invested more in PPO-based defence than did males, whose PPO activity decreased as a result of CO₂ enrichment. Moreover, we found that the inherently slow-growing S. repens had markedly higher POD activity than did the more rapid-growing S. phylicifolia. The PODs of these two species also differed in their biochemical properties.

A new ethylene response factor (ERF), GmERF6, was isolated from soybean. Protein sequence alignment of GmERF6 revealed an AP2/ERF domain, two putative nuclear localization signals (NLSs) and an ERF-associated amphiphilic repression (EAR) motif. Real-time quantitative PCR analysis revealed that the expression of GmERF6 was differentially induced in soybean seedlings by drought, salt, cold, salicylic acid, ethylene, abscisic acid and methyl jasmonate. Transient expression experiments demonstrated that GmERF6 functions as a transcriptional repressor to downregulate the transcriptional levels of the reporter gene and repress the activated ability of other transcriptional activator. Transgenic Arabidopsis lines constitutively expressing GmERF6 showed an increased tolerance to drought compared to wild-type plants.

Wood ontogeny patterns were determined during the ex vitro acclimatization period in micropropagated plantlets of hybrid poplar clones T-14 [Populus tremula × (Populus × canescens)] and T-50 [(Populus × canescens) × Populus tremula]. The temporal course of developmental changes in the woody tissue was characterized on a weekly basis starting from the day of transfer to the ex vitro environment until full acclimatization was achieved on day 28. In vitro rooted plantlets had already initiated lignification of secondary xylem cells. The greatest increase in the amount of woody tissue was observed on days 21 and 28. At the end of the acclimatization period, T-14 plantlets contained on average 41.4 % of secondary xylem tissue compared to 30.3 % found in T-50 plantlets. During the course of acclimatization, both clones displayed identical patterns of vessel lumen size distribution from small vessel lumen area to large vessel lumen area. This pattern differs from the characteristic diffuse-porous pattern of approximately evensized vessel lumen area distribution typical of mature wood. At the end of acclimatization, the differences in vessel lumen area and relative conductivity between the clones were negligible. Development of secondary xylem tissue during ex vitro acclimatization promotes the establishment of vigorous regenerants with stems that show increased bending strength and stiffness.

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 day after micro-grafting. It can also be transported to the shoot apex 30 to 70 cm away from graft-union in 2-year-old grafted trees. For further investigation, transgenic tobaccos with 35S: P. betulaefolia-NACP construct were grafted on wild-type tobaccos (Nicotiana tabacum L. cv. Samsun). The sustainable transport of Pyrus-NACP mRNA through the graft-union occurred from the 15^th day after grafting.