Low expression of osmotically responsive genes 2 (LOS2) encodes an enolase (2-phospho-D-glycerate hydrolase, EC 4.2.1.11) that converts 2-phospho-D-glycerate (PGA) to phosphoenolpyruvate (PEP) in the glycolytic pathway in Arabidopsis. Meanwhile, it is a transcriptional activator of cold-responsive gene, negatively controlling the expression of STZ/ZAT10, a zinc finger transcriptional repressor of cold-responsive gene from Arabidopsis. A novel LOS2 gene, designated PtrLOS2 (GenBank accession number GQ144341), was isolated from trifoliate orange [Poncirus trifoliata (L.) Raf.]. The PtrLOS2 cDNA is 1 662 bp in length with a 1 338 bp open reading frame (ORF), encoding a deduced 445 amino acid residue protein with a predicted molecular mass of 47.79 kDa and an isoelectric point of 5.54. The deduced protein of the PtrLOS2 gene shares high identity (over 86 %) with other plant species enolase, which suggests that the PtrLOS2 probably encodes an enolase. Sequence alignment showed that PtrLOS2 protein has a conserved DNA-binding and a repression domain. Moreover, a conserved start site of alternative translation for the c-myc promoter binding protein (MBP-1) was also found in PtrLOS2 protein. PtrLOS2 was constitutively expressed in leaves, stems and roots. PtrLOS2 expression in roots and stems was much higher than that in leaves under normal conditions, however, the expression of PtrLOS2 was up-regulated in leaves, but down-regulated in roots after cold treatments. The PtrLOS2 expression in stems was firstly up-regulated and then down-regulated after cold treatments. Meanwhile, after ABA treatment, the expression of PtrLOS2 was up-regulated in leaves but in stems and roots firstly down-regulated followed with up-regulation.

Arabidopsis thaliana plants (wild type accessions Col and N1438) were grown in nutrient solution for 34 d with or without 50 mM NaCl. Salt stress inhibited plant growth rate more in Col than in N1438 and a decrease in K⁺, Ca²⁺ and nitrogen contents was observed in both accessions. NaCl diminished accumulation of malate, fumarate and citrate only in Col accession. To measure the effect of NaCl on transcript level of superoxide dismutase (SOD) isoforms and copper chaperone for SOD genes, a semi-quantitative polymerase chain reaction (RT-PCR) method was developed using cDNA normalized against the EF1a gene in parallel with quantitative real time RT-PCR (Q-PCR) technique. Both methods gave the same results. The abundance of transcripts of the three genes coding for Cu/Zn-SOD responded similarly to NaCl in both accessions: CSD1 gene was overexpressed, and CSD2 and CSD3 genes were repressed. However, the genes coding for Fe-SOD (FSD1), Mn-SOD (MSD1) and Cu-chaperone for SOD (CCS) responded to NaCl differently in Col and N1438: the former gene was overexpressed in Col and repressed in N1438, and the opposite behaviour was observed for the latter two genes.

The effects of two-day dark treatment, applied to whole plants or to individual organs, on the photosynthetic apparatus in cotyledons and first rosette leaves of young Arabidopsis thaliana plants were studied. Darkness affected the individually darkened pair of cotyledons as well as the cotyledons of whole darkened plants (DP) in a similar manner as revealed by the significant decrease in the actual yield of photosystem 2 electron transport and the down-regulation of the psaB and rbcL transcript levels. However, cotyledons and rosette leaves responded differently to darkness with respect to the non-photochemical quenching (NPQ) and the non-regulated energy dissipation (Φ_NO), indicating different capacity for photoprotection depending on the type of the applied dark treatment. Besides, the expression of the genes for the two plastid proteases FtsH5 and Deg1 involved in D₁ protein degradation was inhibited in both leaf organs, suggesting that these proteases function mainly under irradiance. Upon re-irradiation, dark-treated cotyledons recovered from the applied stress and during further senescence the changes in the photosynthetic parameters and the mRNA levels of psaB, rbcL and SAG12 were similar as in the control plants. However, in the course of recovery typical chloroplast senescence symptoms were observed only in individually darkened leaves while re-irradiated DP leaves maintained high photosynthetic capacity.

To determine the effects of leaf colour on gas exchange and chlorophyll fluorescence, two genotypes of Begonia semperflorens with green leaves or red leaves were compared. The red leaves showed a high accumulation of anthocyanins and high absorbance at 282 and 537 nm while the green leaves exhibited a higher net photosynthetic rate and lower thermal dissipation of light energy. It seems likely that anthocyanins in the vacuoles restricted the absorption of green light to the chloroplasts, leading to a decrease in the efficiency of excitation capture by open PS 2 centres, photochemical quenching and CO₂ assimilation.

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 traits were also measured in a population of 52 L. pennellii introgression lines grown under both normal and stress conditions. A total of 311 quantitative trait loci (QTL) were identified for the studied traits: plant height, stem diameter, leaf number, leaf and root fresh and dry mass, and sodium, potassium and calcium contents. Some of the loci (124) were identified under both control and stress conditions while 86 QTL were identified only under non-stress conditions and 101 loci were identified only under NaCl stress.

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 whorls and fruit but not in root, leaf, sepal, and petal, and that LbSEP3 constitutively expressed in root, leaf, fruit, and all the four floral whorls.

Sour pummelo (Citrus grandis) seedlings were irrigated with nutrient solution containing four boron concentrations (i.e., 2.5, 10, 25 and 50 μM H₃BO₃) and two aluminum concentrations [i.e., 0 (-Al) and 1.2 mM AlCl₃ . 6 H₂O (+Al)]. It was found that B did not affect, but Al increased, the Al content in the roots. The Al and citrate contents in the -Al leaves either did not change or slightly increased with increasing B concentration. On the other hand, the Al and citrate contents in the +Al leaves rapidly decreased as B concentration increased from 2.5 to 50 μM, then decreased at the highest B concentration. The Al and citrate contents were higher in the +Al than in the -Al leaves, except for at 25 μM B when they were similar. The leaf malate content did not change in response to B or Al, except for an increase in the +Al leaves and a decrease in the -Al leaves at 2.5 μM B. Similarly, the root malate and citrate contents did not change in response to B with or without Al, except for a decrease in the malate and citrate contents in the +Al roots at 50 μM B and an increase in the citrate content in the -Al roots at 50 μM B. The activities of acid-metabolizing enzymes were less affected by B-Al interactions in the roots than in the leaves.

Changes in pigments contents, leaf area, leaf dry mass per unit area (LMA), photosynthetic rate and chlorophyll a fluorescence were investigated in developing leaves of Fagopyrum dibotrys Hara. mutant. Anthocyanins transiently accumulate below the upper epidermis during leaf ontogeny of this mutant. Red leaves possessed lower Chl content, LMA, photosynthetic rate and apparent carboxylation efficiency than green leaves. However, content of anthocyanins declined and above mentioned parameters increased during further leaf development. In both red and green leaves, chronic photoinhibition did not take place according to variable to maximum chlorophyll fluorescence ratio (F_v/F_m). Red leaves had higher non-photochemical quenching (NPQ) and higher PS 2 efficiency.

Metallothioneins are involved in detoxification of heavy metals. A cDNA encoding type 3 metallothionein (PcMT3) was isolated from the salt stressed leaf cDNA library of Porteresia coarctata (Roxb.) Tateoka (wild rice) that grows well in the heavy metal laden estuarine soils. The PcMT3 cDNA (581 bp) encodes a protein of 64 amino acids. PcMT3 is highly homologous (82 %) to OsMT-I-3a of rice, but is unique from other type 3 plant MTs due to the presence of an additional glycine residue in the C-terminal domain. Analysis of the 5' upstream region of PcMT3 showed the presence of cis-acting elements like the CG box and STRE previously reported to be involved in gene expression under heavy metal stress. Southern analysis suggested the presence of more than one copy of PcMT3-like sequences in the P. coarctata genome. Analysis of genomic clone of PcMT3 revealed the presence of two introns. A comparison of the genomic sequence of PcMT3 with closely similar type 3 MTs from rice and mangrove species revealed conservation in the number and position of introns. Transcript profiling for PcMT3 in P. coarctata leaves in the presence of Cd, Cu and Zn showed an increase in transcript accumulation.

Universal (consensus) primers are those primers that have the ability to amplify the targeted region of DNA across a broad range of individuals in a certain group of organisms. In plants, such universal primers have been designed to target regions in the nuclear, mitochondrial or chloroplast genome. Among these three genomes, the chloroplast genome is the most suited for the design of consensus primers due to the lower rate of evolution and hence conservation of gene order and sequence of the genome among the different plant species compared to the other two genomes. Several molecular studies in plants have developed and used chloroplast-specific universal primers. In this review, I present some examples of the nuclear DNA-specific universal primers and discuss the features of the chloroplast DNA that make it the most suited for the design of such primers. I then refer to all chloroplast-specific primers developed so far and provide some examples of molecular studies and applications that made use of them.

Grape (Vitis vinifera L.) seedlings grown in vitro were treated with either 200 mM NaCl or 350 mM mannitol for 7 d. Both salinity and osmotic stress caused significant increase in electrolyte leakage. From the three commonly occurring free polyamines (PA), only conspicuous accumulation of putrescine was found in the NaCl-treated seedlings. Four PA biosynthetic genes encoding arginine decarboxylase (pVvADC), S-adenosylmethionine decarboxylase (pVvSAMDC), spermidine synthase (pVvSPDS) and spermine synthase (pVvSPMS) were successfully isolated. While induction of pVvADC was observed from the 1^st day of salt treatment, pVvSAMDC and pVvSPMS were induced only at late stage of stress. As for expression levels of genes in the mannitol-treated seedling, either temporary (pVvADC at day 1) or late (pVvSPMS at days 5 and 7) induction was observed.

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 with the increased detoxification capacity of glutathione transferase in the NaCl-adapted callus. The post-inoculation changes in NaCl-adapted and non-adapted calli were roughly similar and supported the prooxidative nature of B. cinerea infection.

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 13 DAP and then sharply increased to the maximum at 26 DAP. Moreover, the ABA content in the parasite was always lower than in the infected host leaves. The results suggest that an increase in host ABA concentration contributes to reduced host g_s, E and P_N in the holoparasitic C. campestris - M. micrantha association.

LAF1 (Long after far-red radiation 1) is a R2R3 Myb transcription factor and a signal transducer of far-red radiation. To investigate the role of LAF1 in leaf and root development, the leaf growth and vein patterning in laf1 mutants under short day conditions were examined. The length of rosette leaves was reduced and the width of the midvein was increased in laf1 mutants compared to their wild-type (WT) counterparts. In addition, cell size and cell number were both decreased in the laf1 mutant in comparison to the WT plant. A comparative analysis of gene expression showed that the transcript levels of PIN and IAA genes, encoding auxin carrier and response proteins, were decreased in laf1 mutants. LAF1 expression was also shown to be induced by 1-naphthaleneacetic acid. These results suggest that both auxin transport and auxin responses are impaired in laf1 mutants, and that LAF1 is involved in the regulation of leaf and root development mediated by auxin signaling under short day conditions.

The objective of this study was to examine the role of NADPH oxidase on superoxide radical production under waterlogging in mung bean (Vigna radiata) cvs. T 44 (tolerant) and Pusa Baisakhi (PB) (susceptible), and wild species Vigna luteola. Two days of waterlogging caused decline in superoxide radical (O₂ ^.-) contents in all the genotypes, however, further waterlogging up to 8 d caused significant increase in O₂ ^.- contents. In control and revived plants O₂ ^.- contents were higher in PB, while under waterlogging stress T 44 and V. luteola showed greater increases in the O₂ ^.- contents. During waterlogging the increase in O₂ ^.- content was found to be due to the diphenylene iodonium chloridesensitive NADPH oxidase (NOX). This was further confirmed by the waterlogging induced increase in NOX activity, which was higher in tolerant genotypes T 44 and V. luteola compared with PB. Gene expression studies showed enhanced expression of NOX in the roots of waterlogged V. luteola and T 44, while little expression was observed in control or treated plants of PB. PCR band products were cloned and sequenced, and partial cDNAs of NOX was obtained. Results suggest that increase in O₂ ^.- content during waterlogging could be due to the induction of membrane linked NOX.

Photosynthetic rate (P_N) and chlorophyll (Chl) fluorescence induction of source leaves in response to a low sink demand created by girdling the branch (GB) between the root-tuber-system and the leaves were studied in Dahlia pinnata L. cv. Rigolet during the stage of rapid tuber growth in the greenhouse. GB resulted in significantly lower values of P_N, stomatal conductance (g_s), and transpiration rate (E), but in higher leaf temperature (T_l) compared with those of controls. With exception of maximum quantum yield of photosystem 2 (PS 2) photochemistry (F_v/F_m) and maximum ratio of quantum yields of photochemical and concurrent non-photochemical processes in PS 2 (F_v/F₀), no significant differences were observed in Chl fluorescence parameters between girdled and control leaves on days 1 and 2 after GB, indicating no apparent damage in the photosynthetic apparatus. However, longer girdling duration resulted in higher non-photochemical Chl fluorescence quenching (NPQ), but lower F_v/F₀, actual efficiency of energy conversion in PS 2 under steady-state conditions (Φ_PS2), and photochemical quenching coefficient (qP) in comparison with controls from 10:00 to 16:00 or 15:00 on days 4 and 5, respectively, indicating reversible injury in the photosynthetic apparatus.

The effect of increased Cd²⁺ concentrations in the watering solution on citrus physiology was studied by using two citrus genotypes, Cleopatra mandarin and Carrizo citrange. Cadmium content in roots and leaves was tested together with measurements of leaf damage, gas exchange parameters, and hormonal contents. Citrus roots efficiently retained Cd²⁺ avoiding its translocation to the shoots and Cleopatra mandarin translocated less Cd²⁺ than Carrizo. With increasing Cd²⁺ concentration all gas exchange parameters were decreased more in Carrizo than in Cleopatra mandarin. Cd-induced increases in abscisic acid and salicylic acid contents were observed in leaves but not in roots of both genotypes.

Pseudomonas fluorescens was used as a biological control agent against bacterial wilt disease caused by Ralstonia solanacearum. Activities of phenylalanine ammonia lyase (PAL), guaiacol peroxidase (POX), polyphenol oxidase (PPO) and lipoxygenase (LOX) increased in P. fluorescens pretreated tomato seedlings, which were later inoculated with R. solanacearum and activities of PAL, POX, PPO and LOX reached maximum at 12, 9, 15 and 9 h, respectively, after inoculation. Reverse transcription - polymerase chain reaction (RT-PCR) confirmed the maximum induction of all these enzymes in P. fluorescens pretreated seedlings.

Traditionally, tetraploid Paspalum notatum was considered an obligate or a facultative apomict according to cytoembryological analyses. The degree of facultativeness was usually determined by the relative amount of mature ovules bearing aposporous or meiotic (sexual) embryo sacs, or both together. We established, through progeny tests conducted with the aid of AFLP markers, the degree of residual sexuality expressed in four selected biotypes. The results showed it to be substantially and significantly lower than predicted by previous embryological analyses for the same biotypes. Moreover, the lowest expression of residual sexuality was coincident with maximum flowering period. Seed development in facultative apomictic P. notatum shows a definite bias against meiotic embryo sacs.

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 and cpk3-1cpk6-1 mutants and the wild type, whereas, in the wild type, the inhibitor of IP₃-induced Ca²⁺ mobilization, xestospongin C (XeC), adjusted this closure to the extent observed in the cpk3-1 mutant. The Ca²⁺ buffer, EGTA and XeC positively interacted with the slow anion channel blocker, anthracene-9-carboxylic acid (9-AC) to inhibit 20 μM ABA-induced stomatal closure, which was suppressed in the dexamethasone-inducible AtPLC1 antisense transgene or the knockout cpk3-1, cpk6-1, cpk3-1cpk6-1 and NADPH oxidase atrbohD/F mutants. Discrete concentrations of 9-AC or another slow anion channel blocker (probenecid) negatively interacted with the Ca²⁺ buffer, BAPTA or the inhibitor of cyclic ADP-ribose-induced Ca²⁺ mobilization, ruthenium red, to inhibit 30 μM ABAinduced stomatal closure in the wild type but not in the cpk6-1, cpk3-1cpk6-1 and atrbohD/F mutants. Based on so far revealed features of the tested compounds and plant materials, interpretation of the results confirmed that guard cell ABA concentration discriminates between two Ca²⁺ mediations and outlined that one of them sequentially implicates CPK6, PLC1, a putative IP₃ receptor homologue, CPK3, and the slow anion channel, whereas the other one excludes AtPLC1-dependent IP₃ production and CPK3.

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 (sect. Ellobium) produced at the University of Auckland, New Zealand, showed both large and normal pollen grains in the same anther indicating the presence of unreduced gametes. Cytological investigation carried out on the hybrid progeny of F. fulgens (diploid, 2n=22, sect. Ellobium) × F. magellanica (tetraploid, 2n=44, sect. Quelusia) and F. triphylla (diploid, sect. Fuchsia) × F. arborescens (diploid, sect. Schufia) revealed unexpected chromosome numbers of 2n=44 and 2n=33, respectively. In general, the hybrids showed low fertility caused by genetically unbalanced gametes resulted from random disjunction of chromosomes at anaphase I. Studies on meiosis together with the presence of different shapes and sizes of pollen grains in Fuchsia proved indirectly that unreduced gametes are the products of first division meiotic nuclear restitution. These unreduced gametes were viable irrespective of pollen shape, their predominance in the hybrids, nuclear DNA amount and species phylogenetic position.

This study investigated guard cells on the adaxial and the abaxial epidermes during Erythrina corallodendron sepal development. On the adaxial epidermis, the morphology of guard cells was highly variable and changes in aperture induced by abscisic acid (ABA) were observed in 9.1 % stomata, while on the abaxial epidermis 86.7 % stomata responded to ABA. On the adaxial epidermis, stomata did not close even when guard cell pressure potential was reduced to zero by plasmolysis, even if fluorescein diacetate revealed that guard cells were alive. It was concluded that guard cells on the adaxial and the abaxial epidermes of sepals sensed environmental conditions differently, maybe due to different guard cell wall elasticity.

Strawberry (Fragaria ananassa Duch.) seedlings were pretreated with hexanoic acid 2-(diethylamino)ethyl ester (DA-6) in concentrations of 0, 10, 20 and 40 mg dm^-3 and then subjected to chilling and rewarming. The effects of applied DA-6 on the generation of reactive oxygen species (O₂ ^-, H₂O₂), lipid peroxidation, proline accumulation and photosynthesis were evaluated. Pretreatment with DA-6 alleviated the inhibition of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) activities caused by chilling stress thus reducing O₂ ^- and H₂O₂ production and lipid peroxidation in pretreated plants. DA-6 pretreatment also accelerated accumulation of proline and reduce the decrease in proline content after rewarming. DA-6 pretreatment increases maximum quantum yield of photosystem 2 (F_v/F_m), actual photochemical efficiency of photosystem 2 (Φ_PS2), photochemical quenching coefficient (qP) and net photosynthetic rate (P_N) and decreases non-photochemical quenching coefficient (qNP) of the seedlings under chilling stress. DA-6 pretreatment also increased the recovery rate of photosynthesis after rewarming.

To identify genes that are up and down-regulated by water deficit in sugarcane we used the macroarray methodology and the expression level of 3 575 independent sugarcane cDNAs was measured by hybridization with RNA extracted from plants submitted to mild, moderate and severe water deficit. We identified approximately 1 670 differentially expressed genes from which 62 % were up-regulated by different stress-conditions, whereas many repressed genes were exclusive for each time-point. Analysis of similarity showed that approximately 24 % of the differentially expressed genes shared homology with proteins involved in different processes such as signal transduction, hormone metabolism, photosynthesis, transcription and stress response. Transcripts with no known function accounted for approximately 39 % and those without similarity represented 36 % of the sequences. Five genes analyzed by RT-PCR confirmed the macroarray results.

In vitro cultures of Azadirachta indica A. Juss. were raised by first culturing the root segments on modified Murashige and Skoog (MS) medium supplemented with 8.88 μM 6-benzylaminopurine (BAP), 9.84 μM N⁶-(2-isopentenyl) adenine (2iP), 5.71 μM indole-3-acetic acid (IAA), 81.43 μM adenine hemisulphate and 2.27 μM putrescine for 2 d followed by their transfer to the same medium except containing one-tenth of the initially used concentrations of BAP, 2iP and IAA. The regenerated shoots sustained proliferation in the basal medium supplemented with 1.11 μM BAP, 1.43 μM IAA and 135.72 μM adenine hemisulphate. The isolated shoots were rooted to produce plantlets in the presence of 2.46 μM indole-3-butyric acid (IBA). The plantlets showed uniform luxuriant growth under field conditions. True-to-type nature of the field-grown root-regenerated plants was ascertained by random amplified polymorphic DNA (RAPD) analysis.

Imaging fluorometry was applied to investigate the tolerance of Japanese honeysuckle (Lonicera japonica Thunb.) of different ploidy levels to heat stress. Seedlings of L. japonica, the diploid cv. Damaohua and the tetraploid cv. Jiufengyihao, were exposed to heat stress of 42 °C for 6 h and a recovery for 10 h. Heat stress significantly decreased maximum photochemical efficiency, electron transport rate, effective quantum yield of photosystem 2, and photochemical quenching of both cultivars, but it decreased the non-photochemical quenching (NPQ) only in the tetraploid. Heat stress increased the content of total soluble sugars, proline, and malondialdehyde in both cultivars while it increased NPQ only in the diploid. Our findings suggest that the tetraploid showed to be more resistance to heat stress than the diploid of L. japonica, which was indicated by different chlorophyll fluorescence imaging techniques and metabolic changes. Moreover, the degree of recovery in the tetraploid was higher than that of the diploid. The tetraploid also possessed thicker epidermis (both upper and lower) and palisade tissue as well as denser pubescence.

Anther-derived pre-embryogenic masses (PEMs) of callus, established via suspension cultures, were encapsulated to form synthetic seeds suitable for cryopreservation. The synchronised suspension culture proliferation necessitated the optimisation of plant growth regulators for different cultivars. The growth phase and density of the culture were also important as well as the exposure of cells to vitrification solution containing 0.75 M sucrose with 0.1 M CaCl₂ and 2.0 % sodium alginate (pH 5.7). Pre-treatment of the encapsulated cells for 2 d with Nitsch and Nitsch (NN) medium containing 0.75 M sucrose solution followed by dehydration for 4 h in a laminar flow box provided maximum cell viability, which varied from 0 to 40 %. The embryo proliferation from the cryopreserved beads involved warming them and then transfer to NN medium containing glutamine (50 mg dm^-3) and activated charcoal (2.5 %). The maximum number of embryos obtained was 31-53 per bead. Subculture into the same medium induced secondary embryogenesis, which was initiated from the meristematic region, radicle, and root cap. Proliferation and maturation of secondary embryos was faster than of primary embryos. No phenotypic variation or abnormal structures compared to the control were observed in the regenerated plantlets.

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 homogeneous cells are needed for analysis and for in vitro induction of androgenesis. This article also compares methods for digestion the exine wall in microspores.

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 of sgfp and uidA genes in the calluses and PLBs survived from hygromycin selection, we reported the co-transformation of sgfp, hptII and uidA genes into Dendrobium Sonia 17. GFP-expressing tissues were also observed in the regenerated transformed plantlets.

In this study, we selected two known pathogen-inducible cis-acting elements, F and E17, to construct synthetic pathogen-inducible promoters for analysis in transformed canola (Brassica napus L.). The synthetic promoter approach was used, which involved the insertion of dimers and combining two cis-acting elements (E17 and F) upstream of the minimal CaMV 35S promoter. Canola plants were transformed by three constructs, pGEE, pGFF, pGFFEE containing synthetic promoters (SP), SP-EE, SP-FF and SP-FFEE, respectively. Analyses of histochemical and fluorometric GUS expression indicated that synthetic promoters responded to fungal elicitors and phytohormone treatments. The SP-FF promoter showed high responses against methyl jasmonate and Sclerotinia sclerotiorum, while SP-EE demonstrated inducibility only in response to salicylic acid and Rhizoctonia solani. The SP-EE promoter similar to SP-FFEE, did not respond to S. sclerotiorum and methyl jasmonate. However, SP-FFEE was highly induced by R. solani elicitors and showed that the level of GUS expression was greater than that by either of E17 or F elements alone. These three synthetic promoters did not activate the expression of the reporter gene in response to cold, heat, UV and wounding.