The enzyme 4-coumarate:coenzyme A ligase (4CL) plays an important role in phenylpropanoid metabolism. The 5'-upstream regions of two Sm4CL genes were isolated from danshen (Salvia miltiorrhiza Bunge) and their functions were characterized by promoter-directed GUS gene expression assay in transgenic Arabidopsis. Seedlings containing pSm4CL1 promoter:GUS fusions showed apparent GUS staining in hypocotyl and those harboring pSm4CL2 promoter:GUS fusions were clearly stained in cotyledon vasculars and roots. Mature Arabidopsis transformed with pSm4CL1 promoter:GUS exhibited GUS expression which was weak in the shoots and scarcely in roots and those modified with pSm4CL2 promoter:GUS displayed obvious GUS staining in roots, stigmatic papillae, stamens and sepal veins. Semi-quantitative RT-PCR revealed that Sm4CL2 was transcribed at the highest level in roots which was also shown to be the major accumulation site of salvianolic acid B. The results suggested that Sm4CL2 rather than Sm4CL1 might be responsible for the biosynthesis of salvianolic acid B in danshen roots.

Historically, sugar beets were selected from fodder beets. We used mitochondrial minisatellite loci to analyze cytoplasmic genetic diversity in fodder beet and sugar beet. Among the 8 sugar beet accessions examined we identified 3 multi-locus haplotypes. These 3 haplotypes were a subset of 5 haplotypes identified among the 29 fodder beet accessions examined. All but one haplotype in fodder beet comprised, in turn, a subset of 12 haplotypes identified previously in leaf beets. Such apparent decreases in cytoplasmic genetic diversity must result from genetic bottlenecks associated with domestication and the ensuing breeding processes. We also detected the haplotype associated with the male-sterile Owen cytoplasm of sugar beet in the fodder beet gene pool. Furthermore, the presence of a 39 kDa protein associated with the Owen cytoplasm was confirmed in two fodder beet plants by Western blot analysis. These results lead us to speculate that the Owen cytoplasm may have originated in fodder beet, from which sugar beet was derived.

C-repeat/dehydration-responsive element binding factor (CBF) plays important roles in cold response network in plants. Here, one member of CBF coding gene family in trifoliate orange (Poncirus trifoliata), designated as PtCBF, was isolated. Semi-quantitative reverse transcription-polymerase chain reactions showed up-regulation of PtCBF not only under low temperature but also induced by abscisic acid. Additionally, the CBF genomic fragments in four citrus species including trifoliate orange, sweet orange (Citrus sinensis), pummel (Citrus grandis) and rough lemon (Citrus jambhiri) were isolated with complete open reading frames. According to the results of alignment analysis between full length cDNA and genomic DNA sequences in trifoliate orange, there were no introns in PtCBF. Moreover, the results of multiple sequence alignment analysis and phylogenetic analysis on putative protein sequences suggested that the AP2 DNA binding domains and CBF signature sequences were highly conserved in four citrus CBF proteins. Finally, the CBF promoters in above citrus species were isolated, which provides some information concerning promoter function.

The polyamine (PA) content of in vitro-grown explants of Thymus moroderi Pau ex Martínez has been evaluated during minimal growth storage. The growth restriction was imposed by the combined action of osmotically-active compounds (15 g dm^-3 sucrose and 15 g dm^-3 mannitol) present in the Murashige and Skoog medium and the modification of the physical environment of the culture (4 °C and darkness). In these conditions, cultures were maintained up to 29 weeks without subculture. During this storage period, we analyzed contents of free, perchloric acid (PCA)-soluble and PCA-insoluble conjugated PA. Minimal growth storage brought about an increase in free putrescine (Put) coinciding with a reduction in PCA-soluble conjugated Put occurring during the first weeks of storage. PCA-insoluble conjugated spermidine (Spd) also accumulated in response to storage.

NADP-dependent isocitrate dehydrogenase (NADP-ICDH) catalyses the production of NADPH, which is an essential component in the cellular homeostasis. In Arabidopsis, the kinetic parameters (K _m and V _max) of cytosolic NADP-ICDH were different in leaves and roots. In vitro applied H₂O₂ did not affect the NADP-ICDH activity in either organ, however, the reduced glutathione inhibited the activity in leaves but not in roots. On the other hand, S-nitrosoglutathione (a NO donor) and peroxynitrite depressed NADP-ICDH activity in leaves and roots.

The oxidative processes and antioxidative system in cucumber (Cucumis sativus L.) fruit were determined during development and senescence. Four distinct developmental stages could be delineated during fruit maturation: immature (3-8 d after anthesis, DAA), mature (9-16 DAA), breaker (17-22 DAA), and yellow (35-40 DAA). The electrolyte leakage, malondialdehyde content, superoxide anion production rate, and hydrogen peroxide content increased continuously during fruit development and senescence. Superoxide dismutase and peroxidase activities consistently increased during fruit maturation, and the catalase activity displayed a single peak at the mature stage. Ascorbate peroxidase and glutathione reductase activities declined during fruit development, but both were activated in yellow fruit. Monodehydroascorbate reductase activity declined and dehydroascorbate reductase (DHAR) activity increased during fruit growth. DHAR was repressed in yellow fruit. Ascorbate dramatically accumulated and its redox state increased, whereas glutathione was degraded and its redox state declined, with fruit maturation.

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 (GO) analysis revealed an overview of sequences distribution, which implied some specially expressed genes related to flower and fruit development. Furthermore, some abundantly expressed unigenes involved in several crucial metabolic pathways related to fruit quality were highlighted and three types of homologues of miraculin-like protein2 were analyzed by both semiquantitative RT-PCR and real-time PCR. The results showed different expression profiles of these genes, which meant that they contribute distinctly to fruit development.

Ethylene response factors (ERFs) are involved in many plant development events and stress defenses. In this study, an ERF gene, VvERF3b, was cloned from the leaves of Vitis vinifera. VvERF3b belongs to ERF group VIIIa. Expression of the gene was induced by abscisic acid, ethephon, and salicylic acid, but not by NaCl. Promoter sequence analysis of the VvERF3b gene revealed that there are several potential cis-acting elements that may be potentially recognized and bound by the transcription factors related to hormones and stress responses. Deletion analysis showed that the 5'-flanking sequence of -1047 to -585 from the transcriptional start site is essential to the high expression of the VvERF3b gene, whereas the sequence fragment of -1324 to -1047 revealed suppression effect. The result indicated that the region appears to contain cis-acting elements that can be bound by the proteins in a transcription complex to induce the inhibition of gene expression.

Nucleoside diphosphate kinase (NDPK) operates in the homeostasis of cellular nucleoside triphosphate (NTP) pools and the cytosolic NDPK1 is the main NDPK isoform in plants, accounting for more than 70 % of total NDPK activity in plant. A full length cDNA (697 bp), designated as CsNDPK1 was cloned from Camellia sinensis (L.) O. Kuntze leaves. Sequence analysis of CsNDPK1 shows several motifs, binding and catalytic sites which are highly conserved among other NDPKs. Southern blot analysis revealed that tea genome has two copies of CsNDPK1. Transcription pattern analysis indicated that CsNDPK1 is expressed in all tissues examined, but expressed more in buds than in other organs.

Elicitation of cell cultures of Silybum marianum with methyl jasmonate (MeJA) increases the production and release of the secondary metabolite silymarin into the culture medium and this process seems to be dependent on phospholipase D activity and its product phosphatidic acid (PA). However, MeJA did not alter total membrane lipid content or overall fatty acid composition. A progressive increase in some galactolipids was observed with elicitation time. Phospholipids were mainly represented by phosphatidylcholine (PC) followed by phosphatidylethanolamine (PE) and phosphatidylinositol (PI). MeJA caused losses of PC species that contain two unsaturated acyl species, 36:5 and 36:6 and an increase in 36:2 species. A drop in the ratio of compounds with 18:3 in PI and PE was also observed. The presence of the lysophospholipids (LP) LPC (16:0, 18:3, 18:2, 18:1) and LPE (16:0, 18:3, 18:2, 18:1) and the high contents of PA, represented by the molecular species 34:3, 34:2 and 36:5 and 36:4, indicates high basal level of phospholipase activity in cultures and a high phospholipid turnover. MeJA treatment did not quantitatively alter these lipid classes.

Short-term responses of Sedum alfredii roots to Cd exposure was compared in Cd hyperaccumulator (HE) and nonhyperaccumulating ecotype (NHE). Cadmium exposure significantly inhibited root elongation and induced loss of plasma membrane integrity and lipid peroxidation of roots tips in the NHE, whereas these effects were much less pronounced in the HE plants. A strong accumulation of reactive oxygen species with increasing Cd concentration was noted in the NHE root tips, but not in HE. After Cd exposure, a dose-dependent decrease in oxidized glutathione and marked increase in reduced glutathione and non-protein thiols were observed in root tips of HE, but were not seen in the NHE plants. These results suggest that the HE tolerates high Cd in the environment through the differential adaptations against Cd-induced oxidative stress.

Cinnamate 4-hydroxylase (C4H) is the second enzyme in the phenylpropanoid pathway which participates in the synthesis of numerous phenylpropanoid compounds such as flavonoids, lignins, suberins and others. We identified a gene putatively coding for Class I C4H in apricot and plum and we analyzed the expression pattern of this gene under different apricot/plum graft combinations with different degree of compatibility. The full-length cDNA is 1 739 bp with a 1 515 bp open reading frame encoding a protein of 504 amino acids. Like other C4Hs, the predicted C4H polypeptides included conserved domains of cytochrome P₄₅₀. The genomic sequence of the apricot C4H gene was interrupted by two introns 335 bp and 904 bp long. Several regulatory motifs including P-, A-, L- and H-boxes, which were conserved across phenylpropanoid metabolism-related genes in higher plants, were found in a 1 300 bp upstream promoter region of the apricot C4H gene. A phylogenetic analysis showed that all Prunus sequences clustered together and were closely related to Malus and Rubus C4H genes. The transcription of Class I PruC4H was detected in all the examined graft combinations, which suggested its rather constitutive character.

The potential of 5-aminolevulenic acid (ALA) to enhance the salt tolerance of cucumber (Cucumis sativus L.) seedlings was investigated. ALA was applied at various concentrations (0, 1, 10, 25, 50, and 100 mg dm^-3) as foliar spray or root watering. Then the seedlings were exposed to 0 or 75 mM NaCl for 5 d. NaCl stress reduced the root and leaf dry masses, leaf area, and the leaf net CO₂ assimilation rate. These reductions were counteracted by exogenous ALA, and the most efficient was 50 mg dm^-3 concentration via foliar spray. ALA decreased the H₂O₂ contents and increased the activities of ascorbate peroxidase (APX) and glutathione reductase (GR) in NaCl-treated cucumber roots and leaves and the activity of catalase (CAT) in leaves. The ALA application also up-regulated the expressions of CAT and cAPX genes in roots and leaves and the expression of GR gene in roots of the NaCl treated cucumber plants.

The ABC1 protein family (ABC1P), a new family of putative kinases, widely existed in procaryote and eucaryote. A comprehensive genome-wide analysis was carried out in this study to find all ABC1Ps in rice (Oryza sativa subsp. japonica). We identified 15 ABC1P genes in rice. All the ABC1Ps contained an ABC1 domain of about 120 conserved amino acid residues and conserved kinase motifs-VAIK (VAVK, VAMK) and DFG (DEG). The phylogenetic analysis showed that all the ABC1Ps were grouped into three subgroups, and formed a total of 12 sister pairs. Conserved motifs analysed by MEME program indicated that almost all ABC1Ps contains motifs 1, 3, 7, 8 and 9. Predictably, the ABC1Ps were localized in mitochondria or chloroplasts, which implied that the ABC1Ps might be involved in energy metabolism in plants. RT-PCR assays demonstrated that all 15 ABC1P genes were active, and some of them were affected by abiotic stresses (NaCl, high temperature, methyl viologen, abscisic acid and cadmium).

We compared stomatal ammonia compensation point (χ_s) and its metabolic regulation in tobacco (Nicotiana tabacum) leaves of a quick-leaf-senescence phenotype ZY90 and a slow-leaf-senescence phenotype NC89. Compared with NC89, ZY90 had significantly higher χ_s values between 40 and 60 d after leaf sprouting in spite of its lower nitrogen content. During the same time, a steeper decline in glutamine synthetase activity was detected in ZY90 leaves, simultaneously with a steep increase in χ_s. These results suggested that the quick leaf senescence phenotype exhibited high NH₃ emission potential due to efficient nitrogen recycling and remobilization, and glutamine synthetase played a key role in regulating χ_s in ZY90.

We found that cadmium promoted flowering in Arabidopsis and suppressed nitric oxide accumulation in leaves. Supplementation with NO donor SNP delayed flowering, whereas application of NO scavenger cPTIO further promoted the transition from vegetative to reproductive stage under Cd stress. Semi-quantitative RT-PCR showed that Cd treatment up-regulated the expression of CONSTANS and FLOWERING LOCUS T, whereas down-regulated the expression of FLOWERING LOCUS C.

Brassinosteroids (BRs) have been proposed to increase the resistance of plants to drought stress. The effect of foliar application of 0.1 μM 24-epibrassinolide (EBR) on chlorophyll (Chl) content, photosystem 2 (PS 2) photochemistry, membrane permeability, lipid peroxidation, relative water content (RWC), proline content, and the antioxidant system in drought-stressed Chorispora bungeana plants was investigated. The results showed that polyethylene glycol (PEG) induced water stress decreased RWC, Chl content and variable to maximum Chl fluorescence ratio (F_v/F_m) less in plants pretreated with EBR than in non-pretreated plants. In addition, lipid peroxidation, measured in terms of malondialdehyde content, membrane permeability and proline content in drought-stressed plants were less increased in EBR pretreated plants, while antioxidative enzyme activities and reduced ascorbate and glutathione contents were more increased in EBR pretreated than in non-pretreated plants. These results suggested that EBR could improve plant growth under drought stress

The differentiation of metaphloem sieve element (MSEs) in the developing caryopsis of wheat (Triticum aestivum L.) was a programmed cell semi-death process. We studied the changes of microtubules and polysaccharide contents during MSEs development. Some significant features are presented in MSEs, such as cell wall non-uniform thickening, chromatin condensation and so on. During the period of MSEs differentiation, numerous microtubules are distributed in the vicinity of the cell wall, but finally they vanished in mature MSEs. Large glycoconjugates in the cell wall and small glycoconjugates in the Golgi apparatus were observed in the developing MSEs. Programmed cell death (PCD) ceased in the mature MSEs after 6 d after flowering and higher aggregation of glycoconjugates appeared in the cytoplasm. All of these processes were in tight contact with the cell wall non-uniform thickening during PCD.

Somatic embryogenesis (SE) in Arabidopsis was induced using various systems, including auxin treatment of in vitro cultured explants (immature zygotic embryos, IZEs) and transgenic plants overexpressing embryogenesis-related transcription factors, e.g. LEC2 together with the GUS reporter gene under control of the auxin-induced DR5 promoter. The study indicated that the SE-systems used gave different embryogenic capacities for the production of true embryos. The highest ratio of true embryos (75 %) was found among embryo-like structures in transgenic seedlings overexpressing LEC2. Analysis of in vitro induced SE systems indicated that in somatic embryos produced in response to exogenous auxin treatment the formation of root poles is frequently disturbed. A lack of a properly formed root meristem was observed in 35-80 % of in vitro induced somatic embryos, in dependence on auxin concentration and duration of treatment.

The objective of this study was to examine the role of nitrate reductase, nitric oxide and non-symbiotic hemoglobin in imparting waterlogging tolerance in mung bean genotypes. Experiment was conducted with two cultivated mung bean [Vigna radiata (L.) Wilczek] genotypes T 44 (tolerant) and Pusa Baisakhi (susceptible) and a highly tolerant wild species Vigna luteola (Jacq.) Benth. The content of nitric oxide increased up to 6 d of waterlogging in Vigna luteola and T 44, and up to 4 d of treatment in Pusa Baisakhi. Increase in nitrate reductase (NR) activity was observed only up to 4 d of waterlogging in Vigna luteola and T 44, and up to 2 d of treatment in Pusa Baisakhi, and thereafter the activity decreased in all the genotypes. The increase in NO content and NR activity was greater in Vigna luteola and T 44 than in Pusa Baisakhi. Non-symbiotic hemoglobin (NSHb) and cNR mRNA expressions were observed only in waterlogging treated roots of Vigna luteola and T 44, while very little expression was observed in control plants of Vigna luteola and T 44, and in control and waterlogged plants of Pusa Baisakhi. PCR bands of Hb and cNR were cloned, and nucleotide and deduced amino acid sequences were obtained and conserved regions and domains were identified using database.

Harpin, an elicitor molecule of bacterial origin induces hypersensitive response (HR) in non-host plants. In an attempt to induce male sterility, harpin was tagged with a signal peptide and expressed downstream to tapetum-specific TA29 promoter resulting in extracellular secretion, subsequent degeneration of tapetum and development of male sterility in tobacco. Putative transgenics were analyzed by PCR amplification of transgene, semiquantitative RT-PCR analysis from total RNA extracts from anther tissue with transgene specific probe, Western blotting using polyclonal antibody raised against harpin, by transmission and scanning electron microscopy, and by confocal microscopy of anthers and pollen at various stages of development. Varying degrees of male sterility (30-100 %) was observed with plants showing complete and partial male sterility as well as several morphological variations were seen especially in leaves and flowers. Further, some of the transgenics showed un-induced of HR-like local lesions in the vegetative tissues. Harpin_Pss got deposited on the pollen grains upon tapetal degeneration resulting in significant alterations in the morphology of pollen cell wall. However, megagametogenesis was not affected in complete and partial male sterile plants and female gametes were completely fertile. The complete male sterility was attributed to premature tapetal cell death due to sufficient extracellular harpin_Pss accumulation whereas insufficient protein content might be the reason for partial male sterility. These findings indicate the possible use of cytotoxic harpin_Pss for the development of male sterile plants.

To induce virus resistance in tobacco and rice we constructed hairpin RNA expression system harbouring inverted repeat fragments of coat protein cDNA of Potato virus Y (PVY) or Rice stripe virus (RSV). These structures were driven by three promoters [cauliflower mosaic virus 35S (CaMV 35S), polyubiqutin gene of maize (Ubi), and Pharbitis nil leucine zipper gene (PNZIP)] which have different tissue-specific activity. PVY resistance ratios were 65.18, 24.33 and 83.54 % in transgenic tobacco plants harboring p35S-PVY, pUbi-PVY and pPNZIP-PVY. RSV resistance was 16.21, 28.61 and 29.33 % in transgenic rice plants harboring p35S-RSV, pUbi-RSV and pPNZIP-RSV. Northern blotting and GUS assay demonstrated that virus resistance levels were related to promoter activity. Therefore, choice of the more effective and tissue-specific promoter to reinforce transcription of hpRNAs will favour the cultivation of highly virusresistant transgenic plants.

The aim of this study was to evaluate the growth and development of Aechmea blanchetiana Baker L.B. Sm. in vitro on medium with 0.0, 0.145, 1.45 and 14.5 μM Cu and 0.0, 2.75, 27.5 and 275 μM Zn. Significant accumulation of Cu and Zn occurred at 14.5 μM Cu and 27.5 and 275 μM Zn, respectively, and there were no significant changes in contents of the other macro- and micronutrients. Superoxide dismutase (SOD) activity significantly changed in the presence of both metals. Spermine content increased as Zn concentration increased and decreased with increasing concentrations of Cu. There was an accumulation of H₂O₂ in the leaf tissue of plants grown in 1.45 and 14.5 μM Cu and 27.5 and 275 μM Zn. A. blanchetiana was found tolerant to the Cu and Zn in concentrations used in this study and displays the capacity to accumulate these metals.

The production of reactive oxygen species (ROS) and the ways by which ROS are generated are very important facts related to heavy metal toxicity in plants. In this work, superoxide anion (O₂ ^.-) generation diminished in cadmium treated sunflower (Helianthus annuus L.) leaf discs, and this reduction was time and Cd-concentration dependent. In line with these findings, we observed that NADPH-dependent oxidase activity was significantly inhibited by 0.1 and 0.5 mM Cd²⁺ treatments and the expression of the NADPH oxidase putative gene related to O₂ ^.- synthesis in sunflower leaves was 83 % inhibited by 0.1 mM CdCl₂ and almost completely depleted by 0.5 mM CdCl₂.

DNA methylation is known to play a crucial role in regulating plant development and organ or tissue differentiation. Here, we focused on the DNA methylation dynamics during the germination of wheat seeds using the adapted AFLP technique so called methylation-sensitive amplified polymorphism (MSAP). The MSAP profiles of genomic DNA in embryo and endosperm tissues of germinating seeds, as well as dry seeds were characterized and notable changes of cytosine methylation were detected. Comparisons of MSAP profiles in different tissues tested showed that the methylation level in dry seeds is the highest. The alteration analysis of cytosine methylation displayed that the number of demethylation events were three times higher than that of de novo methylation, which indicated that the demethylation was predominant in germinating wheat seeds, though the methylation events occurred as well. Sixteen differentially displayed DNA fragments in MSAP profiles were cloned and the sequencing analysis confirmed that nine of them contained CCGG sites. The further BLAST search showed that four of the cloned sequences were located in coding regions. Interestingly, three of the sixteen candidates were homologous to retrotransposons, which indicated that switches between DNA methylation and demethylation occurred in retrotransposon elements along with the germination of wheat seeds.

Acyl carrier protein (ACP), as an essential protein cofactor, plays an important role in de novo synthesis of fatty acids in plastids. In this study, the expression profile of peanut (Arachis hypogaea) AhACP1-1 and AhACP1-2 was analyzed in different tissues. The expression level of AhACP1-1 was highest in the seed, whereas expression was barely detected in the shoot, and AhACP1-2 was expressed in every tissue analyzed with the highest expression level detected in the leaf and seed. Overexpression (OE) and antisense-inhibition (AT) of AhACP1 in transgenic tobacco modified the transcript level of endogenous NtACPs, and the content of total lipids and composition of fatty acid in leaves were altered compared with the wild-type control. Transgenic OE-AhACP1 or AT-AhACP1 tobacco exhibited a significant increase or decrease in polyunsaturated C18:2 and C18:3 fatty acid content, and were more tolerant or sensitive to cold stress, respectively. It is suggested that AhACP1 bound with C18:1 might be the specific substrate of oleoyl-ACP thioesterase or glycerol-3-phosphate acyltransferase, and participates in membrane lipid synthesis.

This article describes the usage of non-commercial environmental scanning electron microscope (ESEM) for the visualization of plant extracellular matrix in Abies alba and Abies numidica. Non sputter-coated samples free of using the common fixation technique observed at the relatively low humidity of the air environment with the pressure 550 Pa and the low temperature of the sample from -18 to -22°C give surprisingly very good results that show the natural structure of the tissues. This seems to be generally applicable. Moreover, a specially designed ionization detector of secondary electrons and a YAG:Ce³⁺ detector of backscattered electrons were used for better comparison.

The effects of lead toxicity on leaf gas exchange, chlorophyll content, chlorophyll fluorescence, chloroplast ultrastructure, and opening of stomata were examined in tobacco (Nicotiana tabacum L.) plants. Plants were grown in a growth chamber for 7 d in Hoagland nutrient solution supplemented with 0.0 (control), 5, 10, 25, 50, 100, 300 and 500 μM Pb(NO₃)₂. Plants treated with 5, 10, and 25 μM Pb(NO₃)₂ exhibited no abnormalities. Root and shoot growth, net photosynthetic rate and stomatal conductance were significantly reduced in plants treated with 100, 300 and 500 μM Pb(NO₃)₂. In plants treated with 500 μM Pb(NO₃)₂, the majority of stomata were closed. The effect of Pb(NO₃)₂ on chlorophyll content and chlorophyll fluorescence parameters was negligible. However, in plants exposed to 100, 300, and 500 μM Pb(NO₃)₂, the mesophyll cells showed altered chloroplasts with disrupted thylakoid membranes.

Biotechnological improvement of monocots is often hampered by the lack of efficient regeneration systems, requisite wound responses and low cell competence. Despite these limitations, the biolistic and Agrobacterium methods have been successfully used to produce several transgenic monocots by adjusting the parameters that govern efficient delivery and integration of transgene(s) into plant genome. It is now possible to transform even difficult monocots using tailor-made gene constructs and promoters, suitable A. tumefaciens strains and a proper understanding of the entire process. This success has been reviewed in the present article and a special emphasis was laid on the measures that were taken in overcoming the difficulties that arise due to the differential responses of monocots and dicots. This information is necessary for biotechnological improvement of still newer monocotyledonous plants that have been hitherto difficult to transform.