Unusual developmental morphology and anatomy of vegetative organs in Utricularia dichotoma-leaf, shoot and root dynamics.

The terrestrial carnivorous species Utricularia dichotoma is known for a great phenotypic plasticity and unusual vegetative organs. Our investigation on 22 sources/populations revealed that after initiation of a leaf and two bladders on a stolon, a bud was formed in the proximal axil of the leaf, developing into a rosette with up to seven organs. The first two primordia of the bud grew into almost every possible combination of organs, but often into two anchor stolons. The patterns were generally not population specific. The interchangeability of organs increased with increasing rank in the succession of organs on stolon nodes. A high potential of switching developmental programs may be successful in a fluctuating environment. In this respect, we were able to show that bladders developed from anchor stolons experimentally when raising the water table. Anatomical structures were simple, lacunate and largely homogenous throughout all organs. They showed similarities with many hydrophytes, reflecting the plant's adaptation to (temporarily) submerged conditions. The principal component analysis was used in the context of dynamic morphology to illustrate correlations between organ types in the morphospace of U. dichotoma, revealing an organ specific patchwork of developmental processes for typical leaves and shoots, and less pronounced for a typical root. The concept and methods we applied may prove beneficial for future studies on the evolution of Lentibulariaceae, and on developmental morphology and genetics of unusual structures in plants.

Machine Learning Approaches to Improve Three Basic Plant Phenotyping Tasks Using Three-Dimensional Point Clouds.

Developing automated methods to efficiently process large volumes of point cloud data remains a challenge for three-dimensional (3D) plant phenotyping applications. Here, we describe the development of machine learning methods to tackle three primary challenges in plant phenotyping: lamina/stem classification, lamina counting, and stem skeletonization. For classification, we assessed and validated the accuracy of our methods on a dataset of 54 3D shoot architectures, representing multiple growth conditions and developmental time points for two Solanaceous species, tomato (Solanum lycopersicum cv 75 m82D) and Nicotiana benthamiana Using deep learning, we classified lamina versus stems with 97.8% accuracy. Critically, we also demonstrated the robustness of our method to growth conditions and species that have not been trained on, which is important in practical applications but is often untested. For lamina counting, we developed an enhanced region-growing algorithm to reduce oversegmentation; this method achieved 86.6% accuracy, outperforming prior methods developed for this problem. Finally, for stem skeletonization, we developed an enhanced tip detection technique, which ran an order of magnitude faster and generated more precise skeleton architectures than prior methods. Overall, our improvements enable higher throughput and accurate extraction of phenotypic properties from 3D point cloud data.

GABA operates upstream of H+-ATPase and improves salinity tolerance in Arabidopsis by enabling cytosolic K+ retention and Na+ exclusion.

The non-protein amino acid γ-aminobutyric acid (GABA) rapidly accumulates in plant tissues in response to salinity. However, the physiological rationale for this elevation remains elusive. This study compared electrophysiological and whole-plant responses of salt-treated Arabidopsis mutants pop2-5 and gad1,2, which have different abilities to accumulate GABA. The pop2-5 mutant, which was able to overaccumulate GABA in its roots, showed a salt-tolerant phenotype. On the contrary, the gad1,2 mutant, lacking the ability to convert glutamate to GABA, showed oversensitivity to salinity. The greater salinity tolerance of the pop2-5 line was explained by: (i) the role of GABA in stress-induced activation of H+-ATPase, thus leading to better membrane potential maintenance and reduced stress-induced K+ leak from roots; (ii) reduced rates of net Na+ uptake; (iii) higher expression of SOS1 and NHX1 genes in the leaves, which contributed to reducing Na+ concentration in the cytoplasm by excluding Na+ to apoplast and sequestering Na+ in the vacuoles; (iv) a lower rate of H2O2 production and reduced reactive oxygen species-inducible K+ efflux from root epidermis; and (v) better K+ retention in the shoot associated with the lower expression level of GORK channels in plant leaves.

Can Aluminum Tolerant Wheat Cultivar Perform Better under Phosphate Deficient Conditions?

Low availability of inorganic phosphate (Pi), together with aluminum (Al), is a major constraint for plant growth and development in acidic soils. To investigate whether or not Al-resistant cultivars can perform better under Pi deficiency, we chose two wheat cultivars with different Al-responses-Atlas 66, being Al-tolerant, and Scout 66, which is Al-sensitive-and analyzed their responses to Pi deficiency. Results showed that, unexpectedly, the Al-sensitive cultivar Scout 66 contained comparatively higher amount of soluble phosphate (Pi) and total phosphorus (P) both in the roots and in the shoots than Atlas 66 under P deficiency. In addition, Scout 66 exhibited higher root biomass, root volume, and root tip numbers, compared with Atlas 66. The expression of Pi-responsive marker genes, TaIPS1, TaSPX3, and TaSQD2 was strongly induced in both cultivars, but the extents of induction were higher in Scout 66 than in Atlas 66 under long-term Pi starvation. Taken together, our results suggest that the Al-sensitive cultivar Scout 66 performed much better under sole Pi starvation, which sets the following experimental stage to uncover the underlying mechanisms of why Scout 66 can display better under Pi deficiency. Our study also raises an open question whether Al-resistant plants are more sensitive to Pi deficiency.

Bio-priming mitigates detrimental effects of salinity on maize improving antioxidant defense and preserving photosynthetic efficiency.

Salinity is an abiotic stress which seriously affects crop production over the world, particularly in arid and semi-arid regions, with harmful effects on germination, growth and yield. Maize (Zea mays L.), cultivated in a wide spectrum of soil and climatic conditions, is the third most important cereal crop after rice and wheat, moderately sensitive to salt stress. A saline level more than 250 mM NaCl damages maize plants, causing severe wilting. In this study, the effects of hydro-priming (distilled water) and bio-priming (Rosmarinus officinalis L. and Artemisia L. leaf extracts) on seed germination and seedling growth of maize, under 100 mM NaCl salinity were investigated. The factorial experiments were carried out in greenhouse under controlled condition (25 °C in 12/12 h day/night) based on a completely randomized design with three replicates. Results showed that both hydro- and bio-priming increased germination percentage and germination indexes in maize seeds. Rosmarinus extract was the most effective in inducing salt resistance in 30 days old seedlings, with beneficial effects in the strengthening of the antioxidant system and in the maintenance of a higher photosynthetic efficiency under salt stress condition.

Characterization of New Polyphenolic Glycosidic Constituents and Evaluation of Cytotoxicity on a Macrophage Cell Line and Allelopathic Activities of Oryza sativa.

Four new constituents, as 5, 7-dihydroxy-4'-methoxyflavonol-3-O-ß-d-arabinopyranosyl-(2''→1''')-O-ß-d-arabinopyrnosyl-2'''-O-3'''', 7''''-dimethylnonan-1''''-oate (1), 5-hydroxy-7, 4'-dimethoxyflavone-5-O-α-d-arabinopyranosyl-(2"→1''')-O-α-d-arabinopyranosyl-2'''-O-3'''', 7''''-dimethylnonan-1''''-oate (2), 5-hydroxy-7, 4'-dimethoxyflavone-5-O-ß-d-arabinofuranosyl-(2"→1''')-O-ß-d-arabinopyranosyl-2'''-O-lanost-5-ene (3) and 4',4''-diferuloxy feruloyl-O-α-d-arabinopyranosyl-(2a→1b)-O-α-d-arabinopyranosyl-(2b→1c)-O-α-d-arabinopyranosyl-(2c→1d)-O-α-d-arabinopyranosyl-(2d→1e)-O-α-d-arabinopyranosyl-2e-3''', 7'''-dimethylnonan-1'''-oate (4), along with three known compounds (5⁻7) were isolated from Oryza sativa leaves and straw. The structures of new and known compounds were elucidated by 1D (¹H and 13C NMR) and 2D NMR spectral methods, viz: COSY, HMBC, and HSQC aided by mass techniques and IR spectroscopy. The cytotoxicity of these constituents was assessed by using (RAW 264.7) mouse macrophage cell line, and allelopathic effects of compounds (1⁻7) on the germination and seedling growth characteristics such as seedling length and root length of barnyardgrass (Echinochloa oryzicola) were evaluated. Significant inhibitory activity was exhibited by compounds comprising flavone derivatives such as (1⁻3) on all of seed germination characteristics. The allelopathic effect of flavone derivatives were more pronounced on seedling length and root length than the germination characteristics. The higher concentration of flavone derivatives showed stronger inhibitory effects, whereas the lower concentrations showed stimulatory effects in some cases.

The effect of polyploidization on tree hydraulic functioning.

PREMISE OF THE STUDY: Recent research has highlighted the importance of living tissue in wood. Polyploidization can impact amounts and arrangements of living cells in wood, potentially leading to increased drought tolerance. Tetraploid variants were created from the apple cultivar Malus ×domestica 'Gala' (Gala-4x), and their vulnerability to drought-induced cavitation and their hydraulic capacitance were compared to those of their diploid predecessors (Gala-2x). Assuming a positive correlation between polyploidy and drought tolerance, we hypothesized lower vulnerability and higher capacitance for the tetraploid. METHODS: Vulnerability to drought-induced cavitation and the hydraulic capacitance were quantified through acoustic emission and continuous weighing of shoots during a bench-top dehydration experiment. To underpin the hydraulic trait results, anatomical variables such as vessel area, conduit diameter, cell wall reinforcement, and ray and vessel-associated parenchyma were measured. KEY RESULTS: Vulnerability to drought-induced cavitation was intrinsically equal for both ploidy variants, but Gala-4x proved to be more vulnerable than Gala-2x during the early phase of desiccation as was indicated by its significantly lower air entry value. Higher change in water content of the leafy shoot, higher amount of parenchyma, and larger vessel area and size resulted in a significantly higher hydraulic capacitance and efficiency for Gala-4x compared to Gala-2x. CONCLUSIONS: Both ploidy variants were typified as highly sensitive to drought-induced cavitation, with no significant difference in their overall drought vulnerability. But, when water deficit is short and moderate, Gala-4x may delay a drought-induced decrease in performance by trading hydraulic safety for increased release of capacitively stored water from living tissue.

The origin and early evolution of vascular plant shoots and leaves.

The morphology of plant fossils from the Rhynie chert has generated longstanding questions about vascular plant shoot and leaf evolution, for instance, which morphologies were ancestral within land plants, when did vascular plants first arise and did leaves have multiple evolutionary origins? Recent advances combining insights from molecular phylogeny, palaeobotany and evo-devo research address these questions and suggest the sequence of morphological innovation during vascular plant shoot and leaf evolution. The evidence pinpoints testable developmental and genetic hypotheses relating to the origin of branching and indeterminate shoot architectures prior to the evolution of leaves, and demonstrates underestimation of polyphyly in the evolution of leaves from branching forms in 'telome theory' hypotheses of leaf evolution. This review discusses fossil, developmental and genetic evidence relating to the evolution of vascular plant shoots and leaves in a phylogenetic framework.This article is part of a discussion meeting issue 'The Rhynie cherts: our earliest terrestrial ecosystem revisited'.

Evaluation of heme oxygenase 1 (HO 1) in Cd and Ni induced cytotoxicity and crosstalk with ROS quenching enzymes in two to four leaf stage seedlings of Vigna radiata.

Research on heme oxygenase in plants has received consideration in recent years due to its several roles in development, defense, and metabolism during various environmental stresses. In the current investigation, the role of heme oxygenase (HO) 1 was evaluated in reducing heavy metal (Cd and Ni) uptake and alleviating Cd and Ni toxicity effects in the hydroponically grown seedlings of Vigna radiata var. PDM 54. Seedlings were subjected to Cd- and Ni-induced oxidative stress independently at different concentrations ranging from 10 to 100 µM. After 96 h (fourth day) of treatment, the stressed plants were harvested to study the cellular homeostasis and detoxification mechanism by examining the growth, stress parameters (LPX, H2O2 content), and non-enzymatic and enzymatic parameters (ascorbate peroxidase (APX), guaicol peroxidase (GPX), and catalase (CAT)) including HO 1. At 50 µM CdCl2 and 60 µM NiSO4, HO 1 activity was found to be highest in leaves which were 1.39 and 1.16-fold, respectively. The greatest HO 1 activity was reflected from the reduction of H2O2 content at these metal concentrations (50 µM CdCl2 and 60 µM NiSO4) which is correlated with the increasing activity of other antioxidant enzymes (CAT, APX). Thus, HO 1 works within a group that generates the defense machinery for the plant's survival by scavenging ROS which is confirmed by a time-dependent study. Hence, it is concluded that seedlings of V. radiata were more tolerant towards metal-induced oxidative stress in which HO 1 is localized in its residential area (plastids).

Metabolomic profiling delineate taste qualities of tea leaf pubescence.

The amount of pubescence on leaf epidermis is an important morphological marker for the quality of green tea, and the tea with plenty of pubescence is generally recognized as having a better taste. However, there is no systematic study on chemical compositions of tea leaf pubescence. The contributions of pubescence to taste properties are far from clear. In this research, 114 components were identified from the tea leaf pubescence of yunkang 10, a broad-leaf tea cultivar with plenty leaf pubescence, for the first time with a non-targeted metabolomics approach using ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOF-MS). Compared to the tea leaf with the pubescence removed (non-pubescent leaf), the pubescence obtained from the same shoots had relatively higher contents of amino acids and lower contents of polyphenols. It was also found that the umami of pubescence was elevated, while the bitterness and astringency were significantly declined. Partial least-squares (PLS) analysis suggested that the polyphenols and amino acids accounted for the taste quality. To the best of our knowledge, this is the first time that the metabolites in tea leaf pubescence were profiled. The results offer the direct concrete evidence on the contributions of pubescence to the tea taste properties.

Sulphur alters chromium (VI) toxicity in Solanum melongena seedlings: Role of sulphur assimilation and sulphur-containing antioxidants.

The present study investigates modulation in hexavalent chromium [Cr(VI) 25 µM] toxicity by sulphur (S; 0.5, 1.0 and 1.5 mM S as low (LS), medium (MS) and high sulphur (HS), respectively) in Solanum melongena (eggplant) seedlings. Biomass accumulation (fresh and dry weights), photosynthetic pigments, photosynthetic oxygen evolution and S content were declined by Cr(VI) toxicity. Furthermore, fluorescence characteristics (JIP-test) were also affected by Cr(VI), but Cr(VI) toxicity on photosystem II photochemistry was ameliorated by HS treatment via reducing damaging effect on PS II reaction centre and its reduction side. Enhanced respiration, Cr content and oxidative biomarkers: superoxide radical, hydrogen peroxide, lipid peroxidation and membrane damage were observed under Cr(VI) stress. Though Cr(VI) enhanced adenosine triphasphate sulfurylase (ATPS) and o-acetylserine(thiol)lyase (OASTL), glutathione-S-transferase (GST), glutathione reductase (GR) and ascorbate peroxidase (APX) activity, and content of total glutathione, cysteine and NP-SH, however, their levels/activity were further enhanced by S being maximum with HS treatment. The results show that Cr(VI) toxicity does increase under LS treatment while HS protected Cr(VI)-induced damaging effects in brinjal seedlings. Under HS treatment, in mitigating Cr(VI) toxicity, S assimilation and its associated metabolites such as cysteine, glutathione and NP-SH play crucial role.

Tetraploidy enhances the ability to exclude chloride from leaves in carrizo citrange seedlings.

Tetraploid citrus seedlings are more tolerant to salt stress than diploid genotypes. To provide insight into the causes of differences in salt tolerance due to ploidy and thus to better understand Cl- exclusion mechanisms in citrus, diploid and tetraploid seedlings of Carrizo citrange (CC) were grown at 0 (control) and 40mM NaCl (salt-treated) medium for 20 days. Chloride uptake and root-to-shoot translocation rates were on average 1.4-fold higher in diploid than in tetraploid salt-treated plants, which resulted in a greater (1.6-fold) Cl- build up in the leaves of the former. Root hydraulic conductance and leaf transpiration rate were 58% and 17% lower, respectively, in tetraploid than in diploid control plants. Differences remained after salt treatment which reduced these parameters by 30-40% in both genotypes. Morphology of the root system was significantly influenced by ploidy. Tetraploid roots were less branched and with lower number of root tips than those of diploid plants. The cross-section diameter and area were lower in the diploid, and consequently specific root length was higher (1.7-fold) than in tetraploid plants. The exodermis in sections close to the root apex was broader and with higher deposition of suberin in cell walls in the tetraploid than in the diploid genotype. Net CO2 assimilation rate in tetraploid salt-treated seedlings was 1.5-fold higher than in diploid salt-treated plants, likely due to the loss of photosynthetic capacity of diploid plants induced by Cl- toxicity. Leaf damage was much higher, in terms of burnt area and defoliation, in diploid than in tetraploid salt-treated plants (8- and 6-fold, respectively). Salt treatment significantly reduced (37%) the dry weight of the diploid plants, but did not affect the tetraploids. In conclusion, tetraploid CC plants appear more tolerant to salinization and this effect seems mainly due to differences in morphological and histological traits of roots affecting hydraulic conductance and transpiration rate. These results may suggest that tetraploid CC used as rootstock could improve salt tolerance in citrus trees.

Branching Shoots and Spikes from Lateral Meristems in Bread Wheat.

Wheat grain yield consists of three components: spikes per plant, grains per spike (i.e. head or ear), and grain weight; and the grains per spike can be dissected into two subcomponents: spikelets per spike and grains per spikelet. An increase in any of these components will directly contribute to grain yield. Wheat morphology biology tells that a wheat plant has no lateral meristem that forms any branching shoot or spike. In this study, we report two novel shoot and spike traits that were produced from lateral meristems in bread wheat. One is supernumerary shoot that was developed from an axillary bud at the axil of leaves on the elongated internodes of the main stem. The other is supernumerary spike that was generated from a spikelet meristem on a spike. In addition, supernumerary spikelets were generated on the same rachis node of the spike in the plant that had supernumerary shoot and spikes. All of these supernumerary shoots/spikes/spikelets found in the super wheat plants produced normal fertility and seeds, displaying huge yield potential in bread wheat.

[Morphological Features of the Transgenic Tobacco Plant Shoot Expressing the 3-Hydroxy-3-Methylglutagyl-CoA Reductase (HMG1) Gene in the Direct and Reverse Orientations Towards the Promoter].

3-Hydroxy-3-methylglutaryl-CoA reductase (HMG1) catalyzes the formation of mevalonic acid, the key intermediate of the cytosolic isoprenoid synthesis pathway. The parameters of stem and leaf growth were studied in the transgenic tobacco plants that express the HMG1 gene in both sense and antisense orientations towards the constitutive promoter. The transgenic plant height did not significantly differ from that of the control plants, though the plants carrying the sense copy of the HMG1 gene were considerably taller than plants that carried the antisense gene copy. Plants carrying an extra copy of the HMG1 gene were also characterized by increased leaf area. The number of mesophyll cells calculated per square unit of transgenic plants leaves was smaller than in the control plant leaves, though their volume was not considerably changed in any of the variants, suggesting changes in the cell packing density in leaves.

Heterologous expression of chloroplast-localized geranylgeranyl pyrophosphate synthase confers fast plant growth, early flowering and increased seed yield.

Geranylgeranyl pyrophosphate synthase (GGPS) is a key enzyme for a structurally diverse class of isoprenoid biosynthetic metabolites including gibberellins, carotenoids, chlorophylls and rubber. We expressed a chloroplast-targeted GGPS isolated from sunflower (Helianthus annuus) under control of the cauliflower mosaic virus 35S promoter in tobacco (Nicotiana tabacum). The resulting transgenic tobacco plants expressing heterologous GGPS showed remarkably enhanced growth (an increase in shoot and root biomass and height), early flowering, increased number of seed pods and greater seed yield compared with that of GUS-transgenic lines (control) or wild-type plants. The gibberellin levels in HaGGPS-transgenic plants were higher than those in control plants, indicating that the observed phenotype may result from increased gibberellin content. However, in HaGGPS-transformant tobacco plants, we did not observe the phenotypic defects such as reduced chlorophyll content and greater petiole and stalk length, which were previously reported for transgenic plants expressing gibberellin biosynthetic genes. Fast plant growth was also observed in HaGGPS-expressing Arabidopsis and dandelion plants. The results of this study suggest that GGPS expression in crop plants may yield desirable agronomic traits, including enhanced growth of shoots and roots, early flowering, greater numbers of seed pods and/or higher seed yield. This research has potential applications for fast production of plant biomass that provides commercially valuable biomaterials or bioenergy.

Effect of crude oil contamination on the chlorophyll content and morpho-anatomy of Cyperus brevifolius (Rottb.) Hassk.

Chlorophyll plays a pivotal role in the plant physiology and its productivity. Cultivation of plants in crude oil contaminated soil has a great impact on the synthesis of chlorophyll pigment. Morpho-anatomy of the experimental plant also shows structural deformation in higher concentrations. Keeping this in mind, a laboratory investigation has been carried out to study the effect of crude oil on chlorophyll content and morpho-anatomy of Cyperus brevifolius plant. Fifteen-day-old seedling of the plant was planted in different concentrations of the crude oil mixed soil (i.e., 10,000, 20,000, 30,000, 40,000, and 50,000 ppm). A control setup was also maintained without adding crude oil. Results were recorded after 6 months of plantation. Investigation revealed that there is a great impact of crude oil contamination on chlorophyll content of the leaves of the experimental plant. It also showed that chlorophyll a, chlorophyll b, and total chlorophyll content of leaves grown in different concentrations of crude oil were found to be lower than those of the control plant. Further, results also demonstrated that chlorophyll content was lowest in the treatment that received maximum dose of crude oil. It also showed that chlorophyll content was decreased with increased concentration of crude oil. Results also demonstrated that there was a reduction in plant shoot and root biomass with the increase of crude oil concentration. Results also revealed that the shoot biomass is higher than root biomass. Morphology and anatomy of the experimental plant also show structural deformation in higher concentrations. Accumulation of crude oil on the cuticle of the transverse section of the leaves and shoot forms a thick dark layer. Estimation of the level of pollution in an environment due to oil spill is possible by the in-depth study of the harmful effects of oil on the morphology and anatomy and chlorophyll content of the plants grown in that particular environment.

Effect of shooting medium and source of material on grapevine (Vitis vinifera L.) shoot tip recovery after cryopreservation.

BACKGROUND: Selecting experimental material at the optimal physiological stage is of paramount importance for successful cryopreservation. OBJECTIVE: The study was to investigate the effect of the physiological state of grapevine buds on their regrowth after liquid nitrogen exposure. METHODS: In a first set of experiments, we tested the regrowth of cryopreserved buds sampled from microcuttings cultured on shooting medium containing benzylaminopurine or zeatin riboside for various durations. In a second set of experiments, we studied the regrowth after liquid nitrogen exposure of buds sampled from different positions on the stem of in vitro plantlets. RESULTS: Regrowth of cryopreserved buds sampled from microcuttings was higher (30%), compared to buds sampled directly from in vitro plantlets (23%), for all culture durations of microcuttings on shooting medium tested (2-6 weeks). Addition of cytokinin in the shooting medium improved regrowth of cryopreserved buds compared to buds sampled from microcuttings cultured on medium devoid of growth regulators; however similar results were obtained with the two cytokinins tested. Buds sampled on nodes 3-4 and 6-7 (from the top of the stem) displayed higher regrowth compared to shoot tips. No significant differences were noted in regrowth after cryopreservation between buds sampled from microcuttings produced from the terminal node, or nodes 3-4 and 6-7. CONCLUSION: The physiological state of the plant material is important for cryopreservation success. Actively growing buds sampled from microcuttings displayed higher regrowth compared to buds sampled directly on in vitro plantlets.

Cytogenetic and anatomic behavior of cytochimeras and total polyploids in cassava.

Cassava periclinal cytochimeras, cultivars, and interspecific hybrid and polyploid types were studied in relation to embryonic, cytogenetic, and anatomical behavior. Their apical shoots, pollen grains, male and female buds, roots, stomata, and flowering period were analyzed. Chimeras exhibited increased size of L1 and L2 cells. Polyploidy led to enlargement of stomata in chimeras whereas L2 gave tetraploid chromosome configurations, tetrad irregularity, decrease of pollen viability, and increase in frequency of polyembryo sacs. The chimeric composition of tetraploids L1 and L2 and diploid L3 expressed a notable epigenetic effect seen in a marked enlargement of edible roots compared to total diploid. One of the chimeric types was accompanied by complete flowering inhibition. Pollen viability and diameter appeared to be reliable markers to determine ploidy levels.

Long-distance transport of Gibberellic Acid Insensitive mRNA in Nicotiana benthamiana.

BACKGROUND: The Gibberellic Acid (GA) signal is governed by the GAI (Gibberellic Acid Insensitive) repressor, which is characterized by a highly conserved N-terminal DELLA domain. Deletion of the DELLA domain results in constitutive suppression of GA signaling. As the GAI transcript is transportable in phloem elements, a Δ-DELLA GAI (gai) transgenic stock plant can reduce the stature of a scion through transport of gai mRNA from the stock. However, little is known about the characteristics of a scion on a gai stock. RESULTS: Arabidopsis Δ-DELLA GAI (gai) was fused with a T7 epitope tag and expressed under the control of a companion cell-specific expression promoter, Commelina yellow mottle virus promoter (CoYMVp), to enhance transport in the phloem. The CoYMVp:Atgai-T7 (CgT) transgenic Nicotiana benthamiana exhibited a dwarf phenotype and lower sensitivity to GA enhancement of shoot stature. A wild-type (WT) scion on a CgT stock contained both Atgai-T7 mRNA and the translated product. Microarray analysis to clarify the effect of the CgT stock on the gene expression pattern in the scion clearly revealed that the WT scions on CgT stocks had fewer genes whose expression was altered in response to GA treatment. An apple rootstock variety, Malus prunifolia, integrating CoYMVp:Atgai moderately reduced the tree height of the apple cultivar scion. CONCLUSIONS: Our results demonstrate that Atgai mRNA can move from companion cells to sieve tubes and that the translated product remains at the sites to which it is transported, resulting in attenuation of GA responses by reducing the expression of many genes. The induction of semi-dwarfism in an apple cultivar on root stock harbouring Atgai suggests that long-distance transport of mRNA from grafts would be applicable to horticulture crops.

Genetic transformation of European chestnut somatic embryos with a native thaumatin-like protein (CsTL1) gene isolated from Castanea sativa seeds.

The availability of a system for direct transfer of antifungal candidate genes into European chestnut (Castanea sativa Mill.) would offer an alternative approach to conventional breeding for production of chestnut trees tolerant to ink disease caused by Phytophthora spp. For the first time, a chestnut thaumatin-like protein gene (CsTL1), isolated from chestnut cotyledons, has been overexpressed in three chestnut somatic embryogenic lines. Transformation experiments have been performed using an Agrobacterium tumefaciens Smith and Townsend vector harboring the neomycin phosphotransferase (NPTII) selectable and the green fluorescent protein (EGFP) reporter genes. The transformation efficiency, determined on the basis of the fluorescence of surviving explants, was clearly genotype dependent and ranged from 32.5% in the CI-9 line to 7.1% in the CI-3 line. A total of 126 independent transformed lines were obtained. The presence and integration of chestnut CsTL1 in genomic DNA was confirmed by polymerase chain reaction (PCR) and Southern blot analyses. Quantitative real-time PCR revealed that CsTL1 expression was up to 13.5-fold higher in a transgenic line compared with its corresponding untransformed line. In only one of the 11 transformed lines tested, expression of the CsTL1 was lower than the control. The remaining 115 transformed lines were successfully subjected to cryopreservation. Embryo proliferation was achieved in all of the transgenic lines regenerated and the transformed lines showed a higher mean number of cotyledonary stage embryos and total number of embryos per embryo clump than their corresponding untransformed lines. Transgenic plants were regenerated after maturation and germination of transformed somatic embryos. Furthermore, due to the low plantlet conversion achieved, axillary shoot proliferation cultures were established from partially germinated embryos (only shoot development), which were multiplied and rooted according to procedures already established. Transgenic plants were acclimatized and grown in a greenhouse. No phenotypic differences were found with control plants, suggesting no potential cytotoxic effects of the green fluorescent protein. The results reported in the present work could be considered as a first step toward the production of fungal-disease tolerant cisgenic chestnut plants.