Confer Drought Resistance Research Articles

The aquaporin gene PvXIP1;2 conferring drought resistance identified by GWAS at seedling stage in common bean.

A whole-genome resequencing-derived SNP dataset used for genome-wide association analysis revealed 12 loci significantly associated with drought stress based on survival rate after drought stress at seedling stage. We further confirmed the drought-related function of an aquaporin gene (PvXIP1;2) located at Locus_10. A variety of adverse conditions, including drought stress, severely affect common bean production. Molecular breeding for drought resistance has been proposed as an effective and practical way to improve the drought resistance of common bean. A genome-wide association analysis was conducted to identify drought-related loci based on survival rates at the seedling stage using a natural population consisting of 400 common bean accessions and 3,832,340 SNPs. The coefficient of variation ranged from 40.90 to 56.22% for survival rates in three independent experiments. A total of 12 associated loci containing 89 significant SNPs were identified for survival rates at the seedling stage. Four loci overlapped in the region of the QTLs reported to be associated with drought resistance. According to the expression profiles, gene annotations and references of the functions of homologous genes in Arabidopsis, 39 genes were considered potential candidate genes selected from 199 genes annotated within all associated loci. A stable locus (Locus_10) was identified on chromosome 11, which contained LEA, aquaporin, and proline-rich protein genes. We further confirmed the drought-related function of an aquaporin (PvXIP1;2) located at Locus_10 by expression pattern analysis, phenotypic analysis of PvXIP1;2-overexpressing Arabidopsis and Agrobacterium rhizogenes-mediated hairy root transformation systems, indicating that the association results can facilitate the efficient identification of genes related to drought resistance. These loci and their candidate genes provide a foundation for crop improvement via breeding for drought resistance in common bean.

Genome-wide identification of microRNAs associated with osmotic stress and elucidation of the role of miR319 in Medicago ruthenica seedlings

Drought is a major environmental stress that affects plant growth, development, and productivity. Medicago ruthenica, a leguminous forage, has garnered attention owing to its resistance to abiotic stress. The purpose of the current study was to explore genes conferring drought resistance to M. ruthenica. MicroRNAs (miRNAs) are post-transcriptional regulators of gene expression in plants and are associated with developmental plasticity and abiotic/biotic stress responses. Here, high-throughput small RNA, mRNA, and degradome sequencing analyses were performed to analyze miRNAs and their potential target genes in the leaves of M. ruthenica seedlings under osmotic stress conditions. In total, 591 miRNAs were identified. A comparison of the expression levels showed that 15 miRNAs (14 upregulated and 1 downregulated) were significantly differentially expressed following PEG6000 treatment compared with those in the control (0 h). Most miRNAs are highly conserved between M. ruthenica and Medicago truncatula. Using TargetFinder, 11 target genes were predicted; the expression of these target genes negatively correlated with that of five miRNAs related to osmotic stress response. miR319 downregulated the expression of teosinte branched/cycloidea/proliferating cell factor 4 (TCP4), which encodes plant-specific transcription factors, more significantly in the leaves than in the roots. These results were confirmed using quantitative real-time polymerase chain reaction, northern blotting, RLM 5ʹRACE, and a Nicotiana benthamiana transient expression system. The miR319-TCP4 module may act as a homeostasis factor in M. ruthenica roots following drought injury, and it is conserved among plant species.

Groundwater subsidizes tree growth and transpiration in sandy humid forests

AbstractAs drought variability increases in forests around the globe, it is critical to evaluate and understand ecosystem attributes that ameliorate drought impacts. Trees in arid and semi‐arid ecosystems can sustain tree growth and transpiration during drought by accessing shallow groundwater, yet the extent to which groundwater influences forest growth and transpiration in humid environments has largely been unexplored. We quantified groundwater's influence on tree growth and transpiration in northern humid forests with sandy soils. We hypothesized that even in wet regions, soil droughts occur relatively frequently in forests with sandy soils and result in water stress and reduced tree growth. Further, we hypothesized these reductions in productivity are ameliorated if the forest can access shallow groundwater during dry conditions. We evaluated tree growth responses using tree cores in Pinus resinosa trees and estimated forest groundwater use from diel water table fluctuations across sites covering a 1‐ to 9‐m depth‐to‐groundwater (DTG) gradient. In areas of shallow groundwater (DTG < 2.5 m), we observed twice as much tree growth and high, frequent groundwater use (up to 81% of non‐rainy summer days). Groundwater's influence on tree growth and transpiration declined as groundwater deepened along the DTG gradient in the range 1–5 m below land surface. These findings suggest that water provided by a shallow water table subsidizes evapotranspiration in humid forests and results in enhanced tree growth. Our research provides a basis for understanding the role of groundwater in conferring drought resistance in humid forests to help guide sustainable water and forest management decisions.

The AP2 transcription factor NtERF172 confers drought resistance by modifying NtCAT.

SummaryDrought stress often limits plant growth and global crop yields. Catalase (CAT)‐mediated hydrogen peroxide (H2O2) scavenging plays an important role in the adaptation of plant stress responses, but the transcriptional regulation of the CAT gene in response to drought stress is not well understood. Here, we isolated an APETALA2/ETHYLENE‐RESPONSIVE FACTOR (AP2/ERF) domain‐containing transcription factor (TF), NtERF172, which was strongly induced by drought, abscisic acid (ABA) and H2O2, from tobacco (Nicotiana tabacum) by yeast one‐hybrid screening. NtERF172 localized to the nucleus and acted as a transcriptional activator. Chromatin immunoprecipitation, yeast one‐hybrid assays, electrophoretic mobility shift assays and transient expression analysis assays showed that NtERF172 directly bound to the promoter region of the NtCAT gene and positively regulated its expression. Transgenic plants overexpressing NtERF172 displayed enhanced tolerance to drought stress, whereas suppression of NtERF172 decreased drought tolerance. Under drought stress conditions, the NtERF172‐overexpressed lines showed higher catalase activity and lower accumulation of H2O2 compared with wild‐type (WT) plants, while the NtERF172‐silenced plants showed the inverse correlation. Exogenous application of amino‐1,2,4‐triazole (3‐AT), an irreversible CAT inhibitor, to the NtERF172‐overexpression lines showed decreased catalase activity and drought tolerance, and increased levels of cellular H2O2. Knockdown of NtCAT in the NtERF172‐overexpression lines displayed a more drought stress‐sensitive phenotype than NtERF172‐overexpression lines. We propose that NtERF172 acts as a positive factor in drought stress tolerance, at least in part through the regulation of CAT‐mediated H2O2 homeostasis.

Arabidopsis thaliana trehalose-6-phosphate phosphatase gene TPPI enhances drought tolerance by regulating stomatal apertures.

Transpiration occurs through stomata. The alteration of stomatal apertures in response to drought stress is an important process associated with water use efficiency (WUE). Trehalose-6-phosphate phosphatase (TPP) family genes have been reported to participate in adjustment of stomatal aperture. However, there have been no reports of the trehalose metabolism pathway genes improving WUE, and the upstream signalling pathway modulating these genes is not clear. Here, we demonstrate that a member of the TPP gene family, AtTPPI, confers drought resistance and improves WUE by decreasing stomatal apertures and improving root architecture. The reduced expression of AtTPPI caused a drought-sensitive phenotype, while its overexpression significantly increased drought tolerance. Abscisic acid (ABA)-induced stomatal closure experiments confirmed that AtTPPI mutation increased the stomatal aperture compared with that of wild-type plants; in contrast, overexpression plants had smaller stomatal apertures than those of wild-type plants. Moreover, AtTPPI mutation also caused stunted primary root length and compromised auxin transport, while overexpression plants had longer primary root lengths. Yeast one-hybrid assays showed that ABA-responsive element-binding factor1 (ABF1), ABF2, and ABF4 directly regulated AtTPPI expression. In summary, the way in which AtTPPI responds to drought stress suggests that AtTPPI-mediated stomatal regulation is an important mechanism to cope with drought stress and improve WUE.

Expression of the CCCH-tandem zinc finger protein gene OsTZF5 under a stress-inducible promoter mitigates the effect of drought stress on rice grain yield under field conditions.

SummaryIncreasing drought resistance without sacrificing grain yield remains an ongoing challenge in crop improvement. In this study, we report that Oryza sativa CCCH‐tandem zinc finger protein 5 (OsTZF5) can confer drought resistance and increase grain yield in transgenic rice plants. Expression of OsTZF5 was induced by abscisic acid, dehydration and cold stress. Upon stress, OsTZF5‐GFP localized to the cytoplasm and cytoplasmic foci. Transgenic rice plants overexpressing OsTZF5 under the constitutive maize ubiquitin promoter exhibited improved survival under drought but also growth retardation. By introducing OsTZF5 behind the stress‐responsive OsNAC6 promoter in two commercial upland cultivars, Curinga and NERICA4, we obtained transgenic plants that showed no growth retardation. Moreover, these plants exhibited significantly increased grain yield compared to non‐transgenic cultivars in different confined field drought environments. Physiological analysis indicated that OsTZF5 promoted both drought tolerance and drought avoidance. Collectively, our results provide strong evidence that OsTZF5 is a useful biotechnological tool to minimize yield losses in rice grown under drought conditions.

Drought Resistance by Engineering Plant Tissue-Specific Responses.

Drought is the primary cause of agricultural loss globally, and represents a major threat to food security. Currently, plant biotechnology stands as one of the most promising fields when it comes to developing crops that are able to produce high yields in water-limited conditions. From studies of Arabidopsis thaliana whole plants, the main response mechanisms to drought stress have been uncovered, and multiple drought resistance genes have already been engineered into crops. So far, most plants with enhanced drought resistance have displayed reduced crop yield, meaning that there is still a need to search for novel approaches that can uncouple drought resistance from plant growth. Our laboratory has recently shown that the receptors of brassinosteroid (BR) hormones use tissue-specific pathways to mediate different developmental responses during root growth. In Arabidopsis, we found that increasing BR receptors in the vascular plant tissues confers resistance to drought without penalizing growth, opening up an exceptional opportunity to investigate the mechanisms that confer drought resistance with cellular specificity in plants. In this review, we provide an overview of the most promising phenotypical drought traits that could be improved biotechnologically to obtain drought-tolerant cereals. In addition, we discuss how current genome editing technologies could help to identify and manipulate novel genes that might grant resistance to drought stress. In the upcoming years, we expect that sustainable solutions for enhancing crop production in water-limited environments will be identified through joint efforts.

Plant growth promoting rhizobacteria (PGPR) confer drought resistance and stimulate biosynthesis of secondary metabolites in pennyroyal (Mentha pulegium L.) under water shortage condition

Water deficit is one of the main devastating environmental factors limiting crops productivity. Plant growth promoting rhizobacteria (PGPR) can improve plant growth in stress conditions and enhance plant tolerance to drought stress. The aim of this study was to investigate the role of PGPR to protect pennyroyal (Mentha pulegium L.) as an important industrial and functional plant against drought damages. The greenhouse study was conducted as a factorial experiment in a completely randomized design. The first factor was PGPR inoculation (Azotobacter chroococcum (Ac), Azospirillum brasilense (Ab), Ac + Ab and control (without PGPR)). The second factor was irrigation regime that applied at three levels: field capacity (FC), 0.7 FC and 0.4 FC. Drought stress significantly decreased chlorophyll fluorescence (Fv/Fm) and relative water content (RWC) of pennyroyal. Conversely, antioxidant enzymes activity, osmoprotectants accumulation, membrane lipid peroxidation, abscisic acid (ABA) and secondary metabolites including phenolic, flavonoid and essential oil contents and DPPH radical scavenging activity of the plant extract increased due to water deficit. PGPR inoculation significantly reduced adverse effects of water stress on physio- biochemical characteristics and secondary metabolites production in pennyroyal. The results indicated that the bacteria were different in their ability to enhance plant investigated characteristics, and co-inoculation of the bacteria was more effective on improving of pennyroyal physiological and phytochemical parameters. The highest ABA, proteins and soluble sugars, phenolic, flavonoid and oxygenated monoterpenes contents as well as DPPH radical scavenging activity were observed in the plants treated with Ac + Ab under severe drought stress.

Changes in Root Anatomy of Peanut (Arachis hypogaea L.) under Different Durations of Early Season Drought

Changes in the anatomical structure of peanut roots due to early season drought will likely affect the water acquiring capacity of the root system. Yet, as important as these changes are likely to be in conferring drought resistance, they have not been thoroughly investigated. The objective of this study was to investigate the effects of different durations of drought on the root anatomy of peanut in response to early season drought. Plants of peanut genotype ICGV 98305 were grown in rhizoboxes with an internal dimension of 50 cm in width, 10 cm in thickness and 120 cm in height. Fourteen days after emergence, water was withheld for periods of 0, 7, 14 or 21 days. After these drought periods, the first and second order roots from 0–20 cm below soil surface were sampled for anatomical observation. The mean xylem vessel diameter of first- order lateral roots was higher than that of second- order lateral roots. Under early season drought stress root anatomy changes were more pronounced in the longer drought period treatments. Twenty-one days after imposing water stress, the drought treatment and irrigated treatment were clearly different in diameter, number and area of xylem vessels of first- and second-order lateral roots. Plants under drought conditions had a smaller diameter and area of xylem vessels than did the plants under irrigated control. The ability of plants to change root anatomy likely improves water uptake and transport and this may be an important mechanism for drought tolerance. The information will be useful for the selection of drought durations for evaluation of root anatomy related to drought resistance and the selection of key traits for drought resistance.

Overexpression of the vascular brassinosteroid receptor BRL3 confers drought resistance without penalizing plant growth

Drought represents a major threat to food security. Mechanistic data describing plant responses to drought have been studied extensively and genes conferring drought resistance have been introduced into crop plants. However, plants with enhanced drought resistance usually display lower growth, highlighting the need for strategies to uncouple drought resistance from growth. Here, we show that overexpression of BRL3, a vascular-enriched member of the brassinosteroid receptor family, can confer drought stress tolerance in Arabidopsis. Whereas loss-of-function mutations in the ubiquitously expressed BRI1 receptor leads to drought resistance at the expense of growth, overexpression of BRL3 receptor confers drought tolerance without penalizing overall growth. Systematic analyses reveal that upon drought stress, increased BRL3 triggers the accumulation of osmoprotectant metabolites including proline and sugars. Transcriptomic analysis suggests that this results from differential expression of genes in the vascular tissues. Altogether, this data suggests that manipulating BRL3 expression could be used to engineer drought tolerant crops.

The foliar change from seedlings to adults in Allocasuarina (Casuarinaceae): the evolutionary and ecological aspects of leaf reduction, xeromorphy and scleromorphy

The morpho‐anatomical structure of the adult photosynthetic organs of the Casuarinaceae is unique in plants. We examine the shoot structure of five species from Allocasuarina ‐ the most derived genus in the family. While the cotyledons of seedlings are similar to those of many other plants the subsequent leaves form a whorl of chlorenchymatic tissue fused to the cortex of the articles of segmented branchlets with only the leaf tips free at the apex of each article. Each leaf in the whorl is called a phyllichnium and consists of a vascular trace below the photosynthetic chlorenchyma with sclerenchyma developed within the mesophyll. The branchlets of Allocasuarina are considered xeromorphic as the stomata of most species are restricted to the sides of the furrows formed by the closely appressed edges of the phyllichnia. The scleromorphic foliage of the ancestral Casuarinaceae may have developed from adaptation to nutrient poor soils but was also preadapted to confer drought resistance. The narrow foliage with abundant sclerenchyma, restricted stomatal area and minimal distance from the atmosphere to xylem tissues ensured that the genus could become competitively successful on impoverished soils and be a distinctive component of arid and even mesic woodlands and forests in Australia.

Root bacterial endophytes confer drought resistance and enhance expression and activity of a vacuolar H+ -pumping pyrophosphatase in pepper plants.

It has been previously shown that the transgenic overexpression of the plant root vacuolar proton pumps H+ -ATPase (V-ATPase) and H+ -PPase (V-PPase) confer tolerance to drought. Since plant-root endophytic bacteria can also promote drought tolerance, we hypothesize that such promotion can be associated to the enhancement of the host vacuolar proton pumps expression and activity. To test this hypothesis, we selected two endophytic bacteria endowed with an array of in vitro plant growth promoting traits. Their genome sequences confirmed the presence of traits previously shown to confer drought resistance to plants, such as the synthesis of nitric oxide and of organic volatile organic compounds. We used the two strains on pepper (Capsicuum annuum L.) because of its high sensitivity to drought. Under drought conditions, both strains stimulated a larger root system and enhanced the leaves' photosynthetic activity. By testing the expression and activity of the vacuolar proton pumps, H+ -ATPase (V-ATPase) and H+ -PPase (V-PPase), we found that bacterial colonization enhanced V-PPase only. We conclude that the enhanced expression and activity of V-PPase can be favoured by the colonization of drought-tolerance-inducing bacterial endophytes.

The Maize WRKY Transcription Factor ZmWRKY40 Confers Drought Resistance in Transgenic Arabidopsis

Abiotic stresses restrict the growth and yield of crops. Plants have developed a number of regulatory mechanisms to respond to these stresses. WRKY transcription factors (TFs) are plant-specific transcription factors that play essential roles in multiple plant processes, including abiotic stress response. At present, little information regarding drought-related WRKY genes in maize is available. In this study, we identified a WRKY transcription factor gene from maize, named ZmWRKY40. ZmWRKY40 is a member of WRKY group II, localized in the nucleus of mesophyll protoplasts. Several stress-related transcriptional regulatory elements existed in the promoter region of ZmWRKY40. ZmWRKY40 was induced by drought, high salinity, high temperature, and abscisic acid (ABA). ZmWRKY40 could rapidly respond to drought with peak levels (more than 10-fold) at 1 h after treatment. Overexpression of ZmWRKY40 improved drought tolerance in transgenic Arabidopsis by regulating stress-related genes, and the reactive oxygen species (ROS) content in transgenic lines was reduced by enhancing the activities of peroxide dismutase (POD) and catalase (CAT) under drought stress. According to the results, the present study may provide a candidate gene involved in the drought stress response and a theoretical basis to understand the mechanisms of ZmWRKY40 in response to abiotic stresses in maize.

The Flowering Repressor SVP Confers Drought Resistance in Arabidopsis by Regulating Abscisic Acid Catabolism

Terrestrial plants must cope with drought stress to survive. Under drought stress, plants accumulate the phytohormone abscisic acid (ABA) by increasing its biosynthesis and decreasing its catabolism. However, the regulatory pathways controlling ABA catabolism in response to drought remain largely unclear. Here, we report that the flowering repressor SHORT VEGETATIVE PHASE (SVP) is induced by drought stress and associates with the promoter regions of the ABA catabolism pathway genes CYP707A1, CYP707A3 and AtBG1, causing decreased expression of CYP707A1 and CYP707A3 but enhanced expression of AtBG1 in Arabidopsis leaves. Loss-of-function mutations in CYP707A1 and CYP707A3 or overexpression of AtBG1 could rescue the drought-hypersensitive phenotype of svp mutant plants by increasing cellular ABA levels. Collectively, our results suggest that SVP is a central regulator of ABA catabolism and that a regulatory pathway involving SVP, CYP707A1/3, and AtBG1 plays a critical role in plant response to water deficit and plant drought resistance.

A novel gene of Kalanchoe daigremontiana confers plant drought resistance

Kalanchoe (K.) daigremontiana is important for studying asexual reproduction under different environmental conditions. Here, we describe a novel KdNOVEL41 (KdN41) gene that may confer drought resistance and could thereby affect K. daigremontiana development. The detected subcellular localization of a KdN41/Yellow Fluorescent Protein (YFP) fusion protein was in the nucleus and cell membrane. Drought, salt, and heat stress treatment in tobacco plants containing the KdN41 gene promoter driving β-glucuronidase (GUS) gene transcription revealed that only drought stress triggered strong GUS staining in the vascular tissues. Overexpression (OE) of the KdN41 gene conferred improved drought resistance in tobacco plants compared to wild-type and transformed with empty vector plants by inducing higher antioxidant enzyme activities, decreasing cell membrane damage, increasing abscisic acid (ABA) content, causing reinforced drought resistance related gene expression profiles. The 3,3′-diaminobenzidine (DAB) and nitroblue tetrazolium (NBT) staining results also showed less relative oxygen species (ROS) content in KdN41-overexpressing tobacco leaf during drought stress. Surprisingly, by re-watering after drought stress, KdN41-overexpressing tobacco showed earlier flowering. Overall, the KdN41 gene plays roles in ROS scavenging and osmotic damage reduction to improve tobacco drought resistance, which may increase our understanding of the molecular network involved in developmental manipulation under drought stress in K. daigremontiana.

Knockdown of Rice MicroRNA166 Confers Drought Resistance by Causing Leaf Rolling and Altering Stem Xylem Development.

MicroRNAs are 19- to 22-nucleotide small noncoding RNAs that have been implicated in abiotic stress responses. In this study, we found that knockdown of microRNA166, using the Short Tandem Target Mimic (STTM) system, resulted in morphological changes that confer drought resistance in rice (Oryza sativa). From a large-scale screen for miRNA knockdown lines in rice, we identified miR166 knockdown lines (STTM166); these plants exhibit a rolled-leaf phenotype, which is normally displayed by rice plants under drought stress. The leaves of STTM166 rice plants had smaller bulliform cells and abnormal sclerenchymatous cells, likely causing the rolled-leaf phenotype. The STTM166 plants had reduced stomatal conductance and showed decreased transpiration rates. The STTM166 lines also exhibited altered stem xylem and decreased hydraulic conductivity, likely due to the reduced diameter of the xylem vessels. Molecular analyses identified rice HOMEODOMAIN CONTAINING PROTEIN4 (OsHB4), a member of HD-Zip III gene family, as a major target of miR166; moreover, rice plants overexpressing a miR166-resistant form of OsHB4 resembled the STTM166 plants, including leaf rolling and higher drought resistance. The genes downstream of miR166-OsHB4 consisted of polysaccharide synthesis-related genes that may contribute to cell wall formation and vascular development. Our results suggest that drought resistance in rice can be increased by manipulating miRNAs, which leads to developmental changes, such as leaf rolling and reduced diameter of the xylem, that mimic plants' natural responses to water-deficit stress.

Coordination of rooting depth and leaf hydraulic traits defines drought-related strategies in the campos rupestres, a tropical montane biodiversity hotspot

The survival and coexistence of plants in water-limited environments are related to their ability to coordinate water acquisition and regulation of water loss. To assess the coordination among below and aboveground hydraulic traits and the diversity of water-use strategies, we evaluated rooting depth and several leaf hydraulic traits of 15 species in campos rupestres, a seasonally-dry biodiversity hotspot in central Brazil. We assessed the depth of plant water acquisition by excavating roots and analyzing the stable isotope composition of hydrogen (δD) and oxygen (δ18O) in the xylem and soil water. We also measured mid-morning stomatal conductance, leaf-water potential at turgor loss point (ѰTLP) and pre-dawn leaf water potentials (ѰPD) during wet and dry seasons. We demonstrated that rooting depth is a good predictor of seasonal variations in stomatal conductance and ѰPD. Shallow-rooted plants had greater variation in stomatal conductance and ѰPD than deep-rooted plants. Woody plants with shallower roots also had lower ѰTLP than deep-rooted plants, revealing higher drought resistance. We demonstrate that shallow-rooted species, more exposed to variation in water availability, have mechanisms to confer drought resistance through turgor maintenance. Our results support the theory of hydrological niche segregation and its underlying trade-offs related to drought resistance.

Overexpression of the OsERF71 Transcription Factor Alters Rice Root Structure and Drought Resistance.

Plant responses to drought stress require the regulation of transcriptional networks via drought-responsive transcription factors, which mediate a range of morphological and physiological changes. AP2/ERF transcription factors are known to act as key regulators of drought resistance transcriptional networks; however, little is known about the associated molecular mechanisms that give rise to specific morphological and physiological adaptations. In this study, we functionally characterized the rice (Oryza sativa) drought-responsive AP2/ERF transcription factor OsERF71, which is expressed predominantly in the root meristem, pericycle, and endodermis. Overexpression of OsERF71, either throughout the entire plant or specifically in roots, resulted in a drought resistance phenotype at the vegetative growth stage, indicating that overexpression in roots was sufficient to confer drought resistance. The root-specific overexpression was more effective in conferring drought resistance at the reproductive stage, such that grain yield was increased by 23% to 42% over wild-type plants or whole-body overexpressing transgenic lines under drought conditions. OsERF71 overexpression in roots elevated the expression levels of genes related to cell wall loosening and lignin biosynthetic genes, which correlated with changes in root structure, the formation of enlarged aerenchyma, and high lignification levels. Furthermore, OsERF71 was found to directly bind to the promoter of OsCINNAMOYL-COENZYME A REDUCTASE1, a key gene in lignin biosynthesis. These results indicate that the OsERF71-mediated drought resistance pathway recruits factors involved in cell wall modification to enable root morphological adaptations, thereby providing a mechanism for enhancing drought resistance.

Assessment the Response of Chickpea Genotypes to Agrobacterium -Mediated Transformation System

Transformation protocol based on the inoculation of chickpea mature embryos with Agrobacterium suspension was carried out. Four chickpea lines and one Iraqi local variety were used as recipient to the foreign gene of Agrobacterium tumefaciens strain (AgL1). Three plasmids were already inserted in the bacteria cells. The first plasmid carries the bar gene coding for phosphinothricin acetyle transferase (PAT), which confers resistance to the herbicide phosphinothricin or glofosinate ammonium(PPT) and uidA (gusA) gene coding for β-glucuronidase (GUS). The other two plasmids carried the LeEREBP gene which confers drought resistance and bar gene coding for phosphinothricin acetyle transferase (PAT). Successfully regenerated explants were subjected to selection pressure on 10 mg /l of phosphinothricin PPT and the putative transgenic explants were rooted on root induction medium consisting of MS basal medium with B5 medium vitamins supplemented with 2.5 ml of 1mg /ml IBA in addition to grafting on 7 days old non- transformed rootstock. PCR approved transgenic chickpea. 600 mg/l of PPT was used by painting the leaves of surviving plants to detect the expression of bar gene which encodes for phosphinothricin acetyle transferase and confirmed herbicide resistance in transgenic plants.

ZmNAC55, a maize stress-responsive NAC transcription factor, confers drought resistance in transgenic Arabidopsis

Abiotic stress has been shown to significantly limit the growth and productivity of crops. NAC transcription factors play essential roles in response to various abiotic stresses. However, only little information regarding stress-related NAC genes is available in maize. Here, we cloned a maize NAC transcription factor ZmNAC55 and identified its function in drought stress. Transient expression and transactivation assay demonstrated that ZmNAC55 was localized in the nucleus and had transactivation activity. Expression analysis of ZmNAC55 in maize showed that this gene was induced by drought, high salinity and cold stresses and by abscisic acid (ABA). Promoter analysis demonstrated that multiple stress-related cis-acting elements exist in promoter region of ZmNAC55. Overexpression of ZmNAC55 in Arabidopsis led to hypersensitivity to ABA at the germination stage, but enhanced drought resistence compared to wild-type seedlings. Transcriptome analysis identified a number of differentially expressed genes between 35S::ZmNAC55 transgenic and wild-type plants, and many of which are involved in stress response, including twelve qRT–PCR confirmed well-known drought-responsive genes. These results highlight the important role of ZmNAC55 in positive regulates of drought resistence, and may have potential applications in transgenic breeding of drought-tolerant crops.