Overexpression of potato ORANGE (StOR) and StOR mutant in Arabidopsis confers increased carotenoid accumulation and tolerance to abiotic stress.

ORANGE (OR) plays essential roles in regulating carotenoid homeostasis and enhancing the ability of plants to adapt to environmental stress. However, OR proteins have been functionally characterized in only a few plant species, and little is known about the role of potato OR (StOR). In this study, we characterized the StOR gene in potato (Solanum tuberosum L. cv. Atlantic). StOR is predominantly localized to the chloroplast, and its transcripts are tissue-specifically expressed and significantly induced in response to abiotic stress. Compared with wild type, overexpression of StOR increased ß-carotene levels up to 4.8-fold, whereas overexpression of StORHis with a conserved arginine to histidine substitution promoted ß-carotene accumulation up to 17.6-fold in Arabidopsis thaliana calli. Neither StOR nor StORHis overexpression dramatically affected the transcript levels of carotenoid biosynthetic genes. Furthermore, overexpression of either StOR or StORHis increased abiotic stress tolerance in Arabidopsis, which was associated with higher photosynthetic capacity and antioxidative activity. Taken together, these results indicate that StOR could be exploited as a potential new genetic tool for the improvement of crop nutritional quality and environmental stress tolerance.

Meta-Analysis of the Effect of Nitric Oxide Application on Heavy Metal Stress Tolerance in Plants.

Substantial single-species studies have reported the facility of nitric oxide (NO) in alleviating heavy metal-induced stress in plants. Understanding the mechanisms of NO-involved stress alleviation is progressing; however, a quantitative description of the alleviative capacity of NO against heavy metal stress is still lacking. We combined the results of 86 studies using meta-analysis to statistically assess the responses of heavy metal-stressed plants to NO supply across several metal stresses and plant families. The results showed that plant biomass was consistently improved following NO supply to metal-stressed plants. NO played an important role in mitigating oxidative damage caused by heavy metal stress by significantly stimulating the activities of antioxidant enzymes. Moreover, NO supply consistently increased the Ca, Fe, and Mg contents in both leaves and roots. Plant tissues accumulated less heavy metals when exposed to heavy metal stress after NO addition. Additionally, the best concentration of SNP (an NO donor) for hydroponic culture is in the range of 75-150 µM. We further confirmed that NO application can generally alleviate plant heavy metal stress and its action pathway. The results presented here can help guide future applications of NO as a plant growth regulator in agriculture and breeding plants for heavy metal stress tolerance.

Melatonin Alleviates Chromium Toxicity in Maize by Modulation of Cell Wall Polysaccharides Biosynthesis, Glutathione Metabolism, and Antioxidant Capacity.

Melatonin, a pleiotropic regulatory molecule, is involved in the defense against heavy metal stress. Here, we used a combined transcriptomic and physiological approach to investigate the underlying mechanism of melatonin in mitigating chromium (Cr) toxicity in Zea mays L. Maize plants were treated with either melatonin (10, 25, 50 and 100 µM) or water and exposed to 100 µM K2Cr2O7 for seven days. We showed that melatonin treatment significantly decreased the Cr content in leaves. However, the Cr content in the roots was not affected by melatonin. Analyses of RNA sequencing, enzyme activities, and metabolite contents showed that melatonin affected cell wall polysaccharide biosynthesis, glutathione (GSH) metabolism, and redox homeostasis. During Cr stress, melatonin treatment increased cell wall polysaccharide contents, thereby retaining more Cr in the cell wall. Meanwhile, melatonin improved the GSH and phytochelatin contents to chelate Cr, and the chelated complexes were then transported to the vacuoles for sequestration. Furthermore, melatonin mitigated Cr-induced oxidative stress by enhancing the capacity of enzymatic and non-enzymatic antioxidants. Moreover, melatonin biosynthesis-defective mutants exhibited decreased Cr stress resistance, which was related to lower pectin, hemicellulose 1, and hemicellulose 2 than wild-type plants. These results suggest that melatonin alleviates Cr toxicity in maize by promoting Cr sequestration, re-establishing redox homeostasis, and inhibiting Cr transport from the root to the shoot.

Physiological and Differential Proteomic Analyses of Imitation Drought Stress Response in Sorghum bicolor Root at the Seedling Stage.

Drought is one of the most important constraints on the growth and productivity of many crops, including sorghum. However, as a primary sensing organ, the plant root response to drought has not been well documented at the proteomic level. In the present study, we compared physiological alteration and differential accumulation of proteins in the roots of sorghum (Sorghum bicolor) inbred line BT×623 response to Polyethylene Glycol (PEG)-induced drought stress at the seedling stage. Drought stress (up to 24 h after PEG treatment) resulted in increased accumulation of reactive oxygen species (ROS) and subsequent lipid peroxidation. The proline content was increased in drought-stressed plants. The physiological mechanism of sorghum root response to drought was attributed to the elimination of harmful free radicals and to the alleviation of oxidative stress via the synergistic action of antioxidant enzymes, such as superoxide dismutase, peroxidase, and polyphenol oxidase. The high-resolution proteome map demonstrated significant variations in about 65 protein spots detected on Coomassie Brilliant Blue-stained 2-DE gels. Of these, 52 protein spots were identified by matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF-TOF MS) representing 49 unique proteins; the levels of 43 protein spots were increased, and 22 were decreased under drought condition. The proteins identified in this study are involved in a variety of cellular functions, including carbohydrate and energy metabolism, antioxidant and defense response, protein synthesis/processing/degradation, transcriptional regulation, amino acid biosynthesis, and nitrogen metabolism, which contribute jointly to the molecular mechanism of outstanding drought tolerance in sorghum plants. Analysis of protein expression patterns and physiological analysis revealed that proteins associated with changes in energy usage; osmotic adjustment; ROS scavenging; and protein synthesis, processing, and proteolysis play important roles in maintaining root growth under drought stress. This study provides new insight for better understanding of the molecular basis of drought stress responses, aiming to improve plant drought tolerance for enhanced yield.

Arabidopsis mgd mutants with reduced monogalactosyldiacylglycerol contents are hypersensitive to aluminium stress.

Aluminium (Al) is a key element that plays a major role in inhibiting plant growth and productivity under acidic soils. While lipids may be involved in plant tolerance/sensitivity to Al, the role of monogalactosyldiacylglycerol (MGDG) in Al response remains unknown. In this study, Arabidopsis MGDG synthase (AtMGD) mutants (mgd1, mgd2 and mgd3) and wild-type (Col-0) plants were treated with AlCl3; the effect of aluminium on root growth, aluminium distribution, plasma membrane integrity, lipid peroxidation, hydrogen peroxide content and membrane lipid compositions were analysed. Under Al stress, mgd mutants exhibited a more severe root growth inhibition, plasma membrane integrity damage and lipid peroxidation compared to Col-0. Al accumulation in root tips showed no difference between Col-0 and mutants under Al stress. Lipid analysis demonstrated that under Al treatment the MGDG content in all plants and MGDG/DGDG (digalactosyldiacylglycerol) remarkably reduced, especially in mutants impairing the stability and permeability of the plasma membrane. These results indicate that the Arabidopsis mgd mutants are hypersensitive to Al stress due to the reduction in MGDG content, and this is of great significance in the discovery of effective measures for plants to inhibit aluminium toxicity.

IbINH positively regulates drought stress tolerance in sweetpotato.

Invertase inhibitor (INH) post-translationally regulates the activity of invertase, which hydrolyzes sucrose into glucose and fructose, and plays essential roles in plant growth and development. However, little is known about the role of INH in growth and responses to environmental challenges in sweetpotato. Here, we identified and characterized an INH-like gene (IbINH) from sweetpotato. IbINH belongs to the pectin methylesterase inhibitor super family. IbINH transcript was the most abundant in storage roots. IbINH mRNA levels were significantly up-regulated in response to drought, abscisic acid (ABA), salicyclic acid (SA) and jasmonic acid (JA) treatments. Overexpressing IbINH in sweetpotato (SI plants) led to the decrease of plant growth and the increase of drought tolerance, while down-regulation of IbINH (RI plants) by RNAi technology resulted in vigorous growth and drought sensitivity. Furthermore, sucrose was increased and hexoses was decreased in SI plants, but the opposite results were observed in RI plants. Moreover, higher levels of sugars were accumulated in SI plants in comparison to non-transgenic plants (NT plants) and RI plants during water deficit. In addition, ABA biosynthesis-involved and abiotic stress response-involved genes were prominently up-regulated in SI plants under drought stress. Taken together, these results indicate that IbINH mediates plant growth and drought stress tolerance in sweetpotato via induction of source-sink strength and ABA-regulated pathway.

[Compensation effect of re-watering after different drought stresses on source-sink metabolism during tuber expansion period of potato.] / 马铃薯块茎膨大期不同程度干旱后复水的源库补偿效应.

The compensation effect of re-watering after drought has been widely reported in various crops during different growth stages. It is considered as an important self-regulation mechanism for plants to resist abiotic stresses and also an efficient utilization of limited water resource. In this study, two rounds of re-watering after drought treatments were carried out during tuber expansion period of potato, to investigate the drought threshold of potato and explore the potential mechanisms of compensation effect with source-sink aspect. We used virus-free plantlets of "Atlantic" potato as experimental materials. Four treatments were included: sufficient water supply (W), re-watering after mild drought (D1-W), re-watering after medium drought (D2-W) and re-watering after severe drought (D3-W). The results showed that potato yield exhibited an over-compensation effect after two rounds of D1-W treatment, with water use efficiency and yield being increased by 17.5% and 6.3%, respectively, compared with the sufficient water supply. D2-W treatment had no significant effect on potato yield, but water use efficiency was increased by 8.4%, indicating a near-equivalent compensation effect. On the contrary, D3-W treatment did not show any compensation effect in yield. In addition, leaf chlorophyll content, net photosynthetic rate, and leaf area were all reduced after drought treatment, indicating a reduction in "source" size and activity. After re-watering, D1-W and D2-W treatments showed over-compensation and compensation effects through improving source supply capacity. Meanwhile, re-watering after moderate drought increased the sink activity through significantly enhancing the activities of key enzymes in tubers (sink), thus increased the average weight of tubers. In conclusion, re-watering after moderate drought stress during potato tuber expansion period had compensation and over-compensation effects on both source and sink, and thus could compensate for the drought-induced yield loss and improve water use efficiency.

A single amino acid change at position 96 (Arg to His) of the sweetpotato Orange protein leads to carotenoid overaccumulation.

KEY MESSAGE: IbOr-R96H resulted in carotenoid overaccumulation and enhanced abiotic stress tolerance in transgenic sweetpotato calli. The Orange (Or) protein is involved in the regulation of carotenoid accumulation and tolerance to various environmental stresses. Sweetpotato IbOr, with strong holdase chaperone activity, protects a key enzyme, phytoene synthase (PSY), in the carotenoid biosynthetic pathway and stabilizes a photosynthetic component, oxygen-evolving enhancer protein 2-1 (PsbP), under heat and oxidative stresses in plants. Previous studies of various plant species demonstrated that a single-nucleotide polymorphism (SNP) from Arg to His in Or protein promote a high level of carotenoid accumulation. Here, we showed that the substitution of a single amino acid at position 96 (Arg to His) of wild-type IbOr (referred to as IbOr-R96H) dramatically increases carotenoid accumulation. Sweetpotato calli overexpressing IbOr-WT or IbOr-Ins exhibited 1.8- or 4.3-fold higher carotenoid contents than those of the white-fleshed sweetpotato Yulmi (Ym) calli, and IbOr-R96H overexpression substantially increased carotenoid accumulation by up to 23-fold in sweetpotato calli. In particular, IbOr-R96H transgenic calli contained 88.4-fold higher levels of ß-carotene than those in Ym calli. Expression levels of carotenogenesis-related genes were significantly increased in IbOr-R96H transgenic calli. Interestingly, transgenic calli overexpressing IbOr-R96H showed increased tolerance to salt and heat stresses, with similar levels of malondialdehyde to those in calli expressing IbOr-WT or IbOr-Ins. These results suggested that IbOr-R96H is a useful target for the generation of efficient industrial plants, including sweetpotato, to cope with growing food demand and climate change by enabling sustainable agriculture on marginal lands.

Melatonin promotes plant growth by increasing nitrogen uptake and assimilation under nitrogen deficient condition in winter wheat.

Melatonin (MEL) has been widely reported to be beneficial to plant growth and development, but few studies have combined investigations of the performance and function of MEL with detailed physiologically based analyses of nitrogen (N) uptake and metabolism in staple crops. In this study, the effect of MEL application on winter wheat seedling growth and grain yield were investigated in hydroponic and pot experiments at different N levels. The result showed that application of 1 µM MEL in hydroponic solution significantly improved the wheat seedling growth under both N sufficient and deficient conditions, but the effect of MEL on promoting seedling growth was prominent under N deficient condition. Meanwhile, MEL-treated plants maintained higher N contents and nitrate nitrogen levels in shoot under N deficient condition, and also maintained higher nitrate nitrogen levels in root. Further investigation showed that nitrate reductase (NR) and glutamine synthetase (GS) activities were higher in MEL-treated plants than that of MEL-untreated plants under N deficiency. The N absorption calculated based on N contents and biomass showed that MEL could promote the N absorption under N deficient condition. In pot experiment, pre-soaking of seeds with 100 µM MEL enhanced per-plant yield by 16% under N sufficient condition and 23% under N deficient condition. Taken together, the results of this study indicate that MEL is involved in promoting N uptake and assimilation through up-regulating the activities of N uptake and metabolism related enzymes and, ultimately, promotes the plant growth and yield, especially under N deficient condition.

Down-regulation of lycopene ε-cyclase expression in transgenic sweetpotato plants increases the carotenoid content and tolerance to abiotic stress.

Carotenoids are required for many biological processes in plants and humans. Lycopene ε-cyclase (LCY-ε) catalyzes the conversion of lycopene into lutein via the α-branch carotenoid biosynthesis pathway. Down-regulation of IbLCY-ε by RNAi increases carotenoid accumulation and salt stress tolerance in transgenic sweetpotato calli. As the role of IbLCY-ε in carotenoid biosynthesis and environmental stress responses in whole plants is poorly understood, transgenic sweetpotato (RLE plants) with reduced expression of IbLCY-ε were developed. RLE plants contained higher levels of total carotenoid and ß-carotene, due to an elevated ß-carotene/lutein ratio rather than increased de novo biosynthesis. RLE plants showed high reactive oxygen species/radical-scavenging activity. They also exhibited an enhanced tolerance of both salt and drought stress, which was associated with lower membrane permeability and a higher photosynthetic rate, respectively. Elevated carotenoid accumulation in RLE plants mitigated the reductions in leaf photosystem II efficiency and chlorophyll induced by abiotic stress. Expression of the carotenoid cleavage genes 9-cis-epoxycarotenoid dioxygenase, carotenoid cleavage dioxygenase 1 (CCD1) and CCD4 was higher in RLE plants, as was abscisic acid accumulation. IbLCY-ε silencing thus offers an effective approach for developing sweetpotato plants with increased tolerance to abiotic stress that will grow on global marginal lands with no reduction in nutritional value.

Overexpression of alfalfa Orange gene in tobacco enhances carotenoid accumulation and tolerance to multiple abiotic stresses.

The multifunctional Orange (Or) protein plays crucial roles in carotenoid homeostasis, photosynthesis stabilization, and antioxidant activity in plants under various abiotic stress conditions. The Or gene has been cloned in several crops but not in alfalfa (Medicago sativa L.). Alfalfa is widely cultivated across the world; however, its cultivation is largely limited by various abiotic stresses, including drought. In this study, we isolated the Or gene from alfalfa (MsOr) cv. Xinjiang Daye. The amino acid sequence of the deduced MsOr protein revealed that the protein contained two trans-membrane domains and a DnaJ cysteine-rich zinc finger domain, and showed a high level of similarity with the Or protein of other plants species. The MsOr protein was localized in leaf chloroplasts of tobacco. The expression of MsOr was the highest in mature leaves and was significantly induced by abiotic stresses, especially drought. To perform functional analysis of the MsOr gene, we overexpressed MsOr gene in tobacco (Nicotiana benthamiana). Compared with wild-type (WT) plants, transgenic tobacco lines showed higher carotenoid accumulation and increased tolerance to various abiotic stresses, including drought, heat, salt, and methyl viologen-mediated oxidative stress. Additionally, contents of hydrogen peroxide and malondialdehyde were lower in the transgenic lines than in WT plants, suggesting superior membrane stability and antioxidant capacity of TOR lines under multiple abiotic stresses. These results indicate the MsOr gene as a potential target for the development of alfalfa cultivars with enhanced carotenoid content and tolerance to multiple environmental stresses.

Melatonin Mitigates Salt Stress in Wheat Seedlings by Modulating Polyamine Metabolism.

Melatonin, a small molecular weight indoleamine molecule, is involved in various biological processes and responses to environmental cues in plants. However, its function in abiotic stress response and the underlying mechanisms is less clear. In this study, we investigated the effect of melatonin on wheat seedlings growth under salt stress condition. Exogenous melatonin pretreatment partially mitigated the salt-induced inhibition of whole-plant growth as judged from shoot dry weight, IAA content, leaf photosynthesis rate, maximum photochemistry efficiency of photosystem II, and chlorophyll. The mitigation was also observed in reduced accumulation of H2O2 in melatonin-pretreated wheat seedlings exposed to salt stress. Exogenous melatonin increased endogenous melatonin content by evaluating the levels of TaSNAT transcript, which encodes a key regulatory enzyme in the melatonin biosynthetic pathway. Furthermore, melatonin increased polyamine contents by accelerating the metabolic flow from the precursor amino acids arginine and methionine to polyamines; melatonin also decreased the degradation of salt-induced polyamines. Taken together, these results provide the evidence that melatonin mitigates salt stress mainly through its regulation on polyamine metabolism of wheat seedlings.

Suppression of the ß-carotene hydroxylase gene increases ß-carotene content and tolerance to abiotic stress in transgenic sweetpotato plants.

ß-carotene, a carotenoid that plays a key photo-protective role in plants is converted into zeaxanthin by ß-carotene hydroxylase (CHY-ß). Previous work showed that down-regulation of IbCHY-ß by RNA interference (RNAi) results in higher levels of ß-carotene and total carotenoids, as well as salt stress tolerance, in cultured transgenic sweetpotato cells. In this study, we introduced the RNAi-IbCHY-ß construct into a white-fleshed sweetpotato cultivar (cv. Yulmi) by Agrobacterium-mediated transformation. Among the 13 resultant transgenic sweetpotato plants (referred to as RC plants), three lines were selected for further characterization on the basis of IbCHY-ß transcript levels. The RC plants had orange flesh, total carotenoid and ß-carotene contents in storage roots were 2-fold and 16-fold higher, respectively, than those of non-transgenic (NT) plants. Unlike storage roots, total carotenoid and ß-carotene levels in the leaves of RC plants were slightly increased compared to NT plants. The leaves of RC plants also exhibited tolerance to methyl viologen (MV)-mediated oxidative stress, which was associated with higher 2,2-diphenyl-1- picrylhydrazyl (DPPH) radical-scavenging activity. In addition, RC plants maintained higher levels of chlorophyll and higher photosystem II efficiency than NT plants after 250 mM NaCl stress. Yield of storage roots did not differ significantly between RC and NT plants. These observations suggest that RC plants might be useful as a nutritious and environmental stress-tolerant crop on marginal lands around the world.

IbOr Regulates Photosynthesis under Heat Stress by Stabilizing IbPsbP in Sweetpotato.

The Orange (Or) protein regulates carotenoid biosynthesis and environmental stress in plants. Previously, we reported that overexpression of the sweetpotato [Ipomoea batatas (L.) Lam] Or gene (IbOr) in transgenic Arabidopsis (referred to as IbOr-OX/At) increased the efficiency of photosystem II (PSII) and chlorophyll content after heat shock. However, little is known about the role of IbOr in PSII-mediated protection against abiotic stress. In this study, comparative proteomics revealed that expression of PsbP (an extrinsic subunit of PSII) is up-regulated in heat-treated IbOr-OX/At plants. We then identified and functionally characterized the PsbP-like gene (IbPsbP) from sweetpotato. IbPsbP is predominantly localized in chloroplast, and its transcripts are tissue-specifically expressed and up-regulated in response to abiotic stress. In addition, IbOr interacts with IbPsbP and protects it from heat-induced denaturation, consistent with the observation that transgenic sweetpotato overexpressing IbOr maintained higher PSII efficiency and chlorophyll content upon exposure to heat stress. These results indicate that IbOr can protect plants from environmental stress not only by controlling carotenoid biosynthesis but also by directly stabilizing PSII.

Down-regulation of GIGANTEA-like genes increases plant growth and salt stress tolerance in poplar.

The flowering time regulator GIGANTEA (GI) connects networks involved in developmental stage transitions and environmental stress responses in Arabidopsis. However, little is known about the role of GI in growth, development and responses to environmental challenges in the perennial plant poplar. Here, we identified and functionally characterized three GI-like genes (PagGIa, PagGIb and PagGIc) from poplar (Populus alba × Populus glandulosa). PagGIs are predominantly nuclear localized and their transcripts are rhythmically expressed, with a peak around zeitgeber time 12 under long-day conditions. Overexpressing PagGIs in wild-type (WT) Arabidopsis induced early flowering and salt sensitivity, while overexpressing PagGIs in the gi-2 mutant completely or partially rescued its delayed flowering and enhanced salt tolerance phenotypes. Furthermore, the PagGIs-PagSOS2 complexes inhibited PagSOS2-regulated phosphorylation of PagSOS1 in the absence of stress, whereas these inhibitions were eliminated due to the degradation of PagGIs under salt stress. Down-regulation of PagGIs by RNA interference led to vigorous growth, higher biomass and enhanced salt stress tolerance in transgenic poplar plants. Taken together, these results indicate that several functions of Arabidopsis GI are conserved in its poplar orthologues, and they lay the foundation for developing new approaches to producing salt-tolerant trees for sustainable development on marginal lands worldwide.

Overexpressing Arabidopsis ABF3 increases tolerance to multiple abiotic stresses and reduces leaf size in alfalfa.

Arabidopsis ABSCISIC ACID-RESPONSIVE ELEMENT-BINDING FACTOR 3 (ABF3), a bZIP transcription factor, plays an important role in regulating multiple stress responses in plants. Overexpressing AtABF3 increases tolerance to various stresses in several plant species. Alfalfa (Medicago sativa L.), one of the most important perennial forage crops worldwide, has high yields, high nutritional value, and good palatability and is widely distributed in irrigated and semi-arid regions throughout the world. However, drought and salt stress pose major constraints to alfalfa production. In this study, we developed transgenic alfalfa plants (cv. Xinjiang Daye) expressing AtABF3 under the control of the sweetpotato oxidative stress-inducible SWPA2 promoter (referred to as SAF plants) via Agrobacterium tumefaciens-mediated transformation. After drought stress treatment, we selected two transgenic lines with high expression of AtABF3, SAF5 and SAF6, for further characterization. Under normal conditions, SAF plants showed smaller leaf size compared to non-transgenic (NT) plants, while no other morphological changes were observed. Moreover, SAF plants exhibited enhanced drought stress tolerance and better growth under drought stress treatment, which was accompanied by a reduced transpiration rate and lower reactive oxygen species contents. In addition, SAF plants showed an increased tolerance to salt and oxidative stress. Therefore, these transgenic AtABF3 alfalfa plants might be useful for breeding forage crops with enhanced tolerance to environmental stress for use in sustainable agriculture on marginal lands.

Molecular characterization of tocopherol biosynthetic genes in sweetpotato that respond to stress and activate the tocopherol production in tobacco.

Tocopherol (vitamin E) is a chloroplast lipid that is presumed to be involved in the plant response to oxidative stress. In this study, we isolated and characterized five tocopherol biosynthetic genes from sweetpotato (Ipomoea batatas [L.] Lam) plants, including genes encoding 4-hydroxyphenylpyruvate dioxygenase (IbHPPD), homogentisate phytyltransferase (IbHPT), 2-methyl-6-phytylbenzoquinol methyltransferase (IbMPBQ MT), tocopherol cyclase (IbTC) and γ-tocopherol methyltransferase (IbTMT). Fluorescence microscope analysis indicated that four proteins localized into the chloroplast, whereas IbHPPD observed in the nuclear. Quantitative RT-PCR analysis revealed that the expression patterns of the five tocopherol biosynthetic genes varied in different plant tissues and under different stress conditions. All five genes were highly expressed in leaf tissues, whereas IbHPPD and IbHPT were highly expressed in the thick roots. The expression patterns of these five genes significantly differed in response to PEG, NaCl and H2O2-mediated oxidative stress. IbHPPD was strongly induced following PEG and H2O2 treatment and IbHPT was strongly induced following PEG treatment, whereas IbMPBQ MT and IbTC were highly expressed following NaCl treatment. Upon infection of the bacterial pathogen Pectobacterium chrysanthemi, the expression of IbHPPD increased sharply in sweetpotato leaves, whereas the expression of the other genes was reduced or unchanged. Additionally, transient expression of the five tocopherol biosynthetic genes in tobacco (Nicotiana bentamiana) leaves resulted in increased transcript levels of the transgenes expressions and tocopherol production. Therefore, our results suggested that the five tocopherol biosynthetic genes of sweetpotato play roles in the stress defense response as transcriptional regulators of the tocopherol production.

Transgenic poplar expressing codA exhibits enhanced growth and abiotic stress tolerance.

Glycine betaine (GB), a compatible solute, effectively stabilizes the structure and function of macromolecules and enhances abiotic stress tolerance in plants. We generated transgenic poplar plants (Populus alba × Populus glandulosa) expressing a bacterial choline oxidase (codA) gene under the control of the oxidative stress-inducible SWPA2 promoter (referred to as SC plants). Among the 13 SC plants generated, three lines (SC4, SC14 and SC21) were established based on codA transcript levels, tolerance to methyl viologen-mediated oxidative stress and Southern blot analysis. Growth was better in SC plants than in non-transgenic (NT) plants, which was related to elevated transcript levels of auxin-response genes. SC plants accumulated higher levels of GB under oxidative stress compared to the NT plants. In addition, SC plants exhibited increased tolerance to drought and salt stress, which was associated with increased efficiency of photosystem II activity. Finally, SC plants maintained lower levels of ion leakage and reactive oxygen species under cold stress compared to the NT plants. These observations suggest that SC plants might be useful for reforestation on global marginal lands, including desertification and reclaimed areas.

Transgenic alfalfa plants expressing the sweetpotato Orange gene exhibit enhanced abiotic stress tolerance.

Alfalfa (Medicago sativa L.), a perennial forage crop with high nutritional content, is widely distributed in various environments worldwide. We recently demonstrated that the sweetpotato Orange gene (IbOr) is involved in increasing carotenoid accumulation and enhancing resistance to multiple abiotic stresses. In this study, in an effort to improve the nutritional quality and environmental stress tolerance of alfalfa, we transferred the IbOr gene into alfalfa (cv. Xinjiang Daye) under the control of an oxidative stress-inducible peroxidase (SWPA2) promoter through Agrobacterium tumefaciens-mediated transformation. Among the 11 transgenic alfalfa lines (referred to as SOR plants), three lines (SOR2, SOR3, and SOR8) selected based on their IbOr transcript levels were examined for their tolerance to methyl viologen (MV)-induced oxidative stress in a leaf disc assay. The SOR plants exhibited less damage in response to MV-mediated oxidative stress and salt stress than non-transgenic plants. The SOR plants also exhibited enhanced tolerance to drought stress, along with higher total carotenoid levels. The results suggest that SOR alfalfa plants would be useful as forage crops with improved nutritional value and increased tolerance to multiple abiotic stresses, which would enhance the development of sustainable agriculture on marginal lands.

Transgenic poplar expressing Arabidopsis YUCCA6 exhibits auxin-overproduction phenotypes and increased tolerance to abiotic stress.

YUCCA6, a member of the YUCCA family of flavin monooxygenase-like proteins, is involved in the tryptophan-dependent IAA biosynthesis pathway and responses to environmental cues in Arabidopsis. However, little is known about the role of the YUCCA pathway in auxin biosynthesis in poplar. Here, we generated transgenic poplar (Populus alba × P. glandulosa) expressing the Arabidopsis YUCCA6 gene under the control of the oxidative stress-inducible SWPA2 promoter (referred to as SY plants). Three SY lines (SY7, SY12 and SY20) were selected based on the levels of AtYUCCA6 transcript. SY plants displayed auxin-overproduction morphological phenotypes, such as rapid shoot growth and retarded main root development with increased root hair formation. In addition, SY plants had higher levels of free IAA and early auxin-response gene transcripts. SY plants exhibited tolerance to drought stress, which was associated with reduced levels of reactive oxygen species. Furthermore, SY plants showed delayed hormone- and dark-induced senescence in detached leaves due to higher photosystem II efficiency and less membrane permeability. These results suggest that the conserved IAA biosynthesis pathway mediated by YUCCA family members exists in poplar.