DDG1 and G Protein α Subunit RGA1 Interaction Regulates Plant Height and Senescence in Rice (<i>Oryza sativa</i>)

Many studies have already shown that dwarfism and moderate delayed leaf senescence positively impact rice yield, but the underlying molecular mechanism of dwarfism and leaf senescence remains largely unknown. Here, using map-based cloning, we identified an allele of DEP2, DDG1, which controls plant height and leaf senescence in rice. The ddg1 mutant displayed dwarfism, short panicles, and delayed leaf senescence. Compared with the wild-type, ddg1 was insensitive to exogenous gibberellins (GA) and brassinolide (BR). DDG1 is expressed in various organs, especially in stems and panicles. Yeast two-hybrid assay, bimolecular fluorescent complementation and luciferase complementation image assay showed that DDG1 interacts with the α-subunit of the heterotrimeric G protein. Disruption of RGA1 resulted in dwarfism, short panicles, and darker-green leaves. Furthermore, we found that ddg1 and the RGA1 mutant was more sensitive to salt treatment, suggesting that DDG1 and RGA1 are involved in regulating salt stress response in rice. Our results show that DDG1/DEP2 regulates plant height and leaf senescence through interacting with RGA1.     

Conventional Breeding and Molecular Markers for Blast Disease Resistance in Rice (<i>Oryza sativa</i> L.)

Monogenic lines, which carried 23 genes for blast resistance were tested and used donors to transfer resistance genes by crossing method. The results under blast nursery revealed that 9 genes from 23 genes were susceptible to highly susceptible under the three locations (Sakha, Gemmeza, and Zarzoura in Egypt); Pia, Pik, Pik-p, Piz-t, Pita, Pi b, Pi, Pi 19 and Pi 20. While, the genes Pii, Pik-s, Pik-h, Pi z, Piz-5, Pi sh, Pi 3, Pi 1, Pi 5, Pi 7, Pi 9, Pi 12, Pikm and Pita-2 were highly resistant at the same locations. Clustering analysis confirmed the results, which divided into two groups; the first one included all the susceptible genes, while the second one included the resistance genes. In the greenhouse test, the reaction pattern of five races produced 100% resistance under artificial inoculation with eight genes showing complete resistance to all isolates. The completely resistant genes: Pii, Pik-s, Piz, Piz-5 (=bi2) (t), Pita (=Pi4) (t), Pita, Pi b and Pi1 as well as clustering analysis confirmed the results. In the F₁ crosses, the results showed all the 25 crosses were resistant for leaf blast disease under field conditions. While, the results in F₂ population showed seven crosses with segregation ratio of 15 (R):1 (S), two cross gave segregated ratio of 3 R:1 S and one gave 13:3. For the identi- fication of blast resistance genes in the parental lines, the marker K3959, linked to Pik-s gene and the variety IRBLKS-F5 carry this gene, which was from the monogenic line. The results showed that four genotypes; Sakha 105, Sakha 103, Sakha 106 and IRBLKS-F5 were carrying Pik-s gene, while was absent in the Sakha 101, Sakha 104, IRBL5-M, IRBL9-W, IRBLTACP1 and IRBL9-W(R) genotypes. As for Pi 5 gene, the results showed that it was present in Sakha 103 and Sakha 104 varieties and absent in the rest of the genotypes. In addition, Pita-Pita- 2 gene was found in the three Egyptian genotypes (Sakha 105, Sakha 101 and Sakha 104) plus IRBLTACP1 monogenetic. In F2 generation, six populations were used to study the inheritance of blast resistance and specific primers to confirm the ratio and identify the resistance genes. However, the ratios in molecular markers were the same of the ratio under field evaluation in the most population studies. These findings would facilitate in breeding programs for gene pyramiding and gene accumulation to produce durable resistance for blast using those genotypes.     

Identification of a Novel <i>OsCYP2</i> Allele that Was Involved in Rice Response to Low Temperature Stress

Cyclophilin (CYP) plays an important role in plant response to stress, and OsCYP2, one gene of cyclophlilin family, is involved in auxin signal transduction and stress signaling in rice. However, the mechanism that OsCYP2 is involved in rice response to low temperature is still unclear. We identified a new OsCYP2 allelic mutant, lrl3, with fewer lateral roots, and the differences in shoot height, primary root length and adventitious root length increased with the growth process compared to the wild-type plant. Auxin signaling pathway was also affected and became insensitive to gravity. The transgenic rice plants with over-expression of OsCYP2 were more tolerant to low temperature than the wild-type plants, suggesting that OsCYP2 was involved in the low temperature response in rice. In addition, OsCYP2 negatively regulated the expression of OsTPS38, a terpene synthase gene, and was dependent on the OsCDPK7-mediated pathway in response to low temperature stress. OsTPS38- overexpressed transgenic line ox-2 was more sensitive to low temperature. Therefore, OsCYP2 may negatively regulate OsTPS38 through an OsCDPK7-dependent pathway to mediate the response to low temperature in rice. These results provide a new basis for auxin signaling genes to regulate rice response to low temperature stress.     

Phenotype Analysis and Fine Mapping of the Male Sterile Mutant <i>ms10</i> in Rice (<i>Oryza sativa</i> L.)

There is a positive correlation between fertility and yield, and the decrease of fertility is bound to a greatly reduced crop yield. Male sterile mutants can be used in hybrid rice. Therefore, rice male sterility has an important value in research and application, and the study of related mutants is also very vital. The mutant ms10 (male sterile 10) reported in this study was induced by ethyl methane sulfonate (EMS) in the indica maintainer line Xinong 1B. There was no significant difference between the ms10 and wild type in the vegetative growth stage. However, in the reproductive growth stage, ms10 showed that the plant became shorter, the anther became smaller and the color became lighter, and finally showed the phenotype of male sterility in comparison to the wild type. I₂-KI staining showed that the pollen was malformed and only a little was active. Scanning electron microscopy observation showed that the exine waxy layer of the ms10 anther decreased, suggesting that the protective effect on pollen was decreased. This may be one of the reasons leading to the phenotype of male sterility. Finally, the pollen showed shrinkage and collapsed, and the structure of germinating pore cover disappeared. This may be the result of sterility. Genetic analysis showed that the male sterility phenotype of the mutant was controlled by a single recessive nuclear gene. MS10 was mapped between the molecular markers IND37 and IND51 on chromosome 4, with a physical distance of 178.6 kb. These results lay the foundation for further studies on MS10.     

<i>In vitro</i> Evaluation of Seed Germination in Twelve Alfalfa Cultivars under Salt Stress

Alfalfa (Medicago sativa L.), when exposed to abiotic stress such as salinity, suffers significant losses in yield and productivity. The present study evaluated the salinity tolerance of 12 alfalfa cultivars in vitro using five concentrations of sodium chloride (NaCl), ranging from 0 to 250 mmol L⁻¹ . The results obtained in the current study revealed that the Saudi cultivars, Kasimi and Hassawi, and the German cultivar (Berlin) had the highest salinity tolerance in terms of germination percentage (GP), corrected germination rate index (CGRI), days to reach 50% germination (GT50), and ability to form cotyledonary and true leaves. Under mmol L⁻¹ NaCl, the Saudi cultivar Kasimi cultivar showed GP, CGRI, and GT50 of 55.20%, 123.15, and 3.77 days, respectively. Similarly, the German cultivar (Berlin) showed GP, CGRI, and GT50 of 50.06%, 86.61, and 5.17 days, respectively. These findings might reveal a pivotal aspect in salt tolerance in alfalfa. Our results will help to select salt-tolerant alfalfa cultivars that could thrive in arid and semi-arid areas with salinity problems.     

Response of Alfalfa Genotypes to Saline Water Irrigation

The influence of saline water (4, 8, 12 dS m^–1) irrigation on gas exchange and growth response of alfalfa genotypes Anand-2, T-9 and IL-112 was studied. T-9 and IL-112 showed a significant increase in net photosynthetic rate (P_N) at low salinity (4 dS m^–1) compared to the control whereas Anand-2 maintained an unaltered P_N. Reduction in P_N at higher salinities was primarily due to reduction of stomatal conductance. There was a greater reduction in transpiration rate as compared to P_N rate, which resulted in an increase in water use efficiency (WUE). High WUE may serve as one of the strategies of the plant to withstand saline environment. However, the slight increase in WUE in Anand-2 could not help in maintaining its growth. Increase in Na⁺ concentration in comparison to K⁺ concentration may also contribute to the inhibition in growth.     

Characterization and Expression of Ammonium Transporter in Peach (<i>Prunus persica</i>) and Regulation Analysis in Response to External Ammonium Supply

As the preferred nitrogen (N) source, ammonium (NH₄⁺ ) contributes to plant growth and development and fruit quality. In plants, NH₄⁺ uptake is facilitated by a family of NH₄⁺ transporters (AMT). However, the molecular mechanisms and functional characteristics of the AMT genes in peach have not been mentioned yet. In this present study, excess NH₄⁺ stress severely hindered shoot growth and root elongation, accompanied with reduced mineral accumulation, decreased leaf chlorophyll concentration, and stunned photosynthetic performance. In addition, we identified 14 putative AMT genes in peach (PpeAMT). Expression analysis showed that PpeAMT genes were differently expressed in peach leaves, stems and roots, and were distinctly regulated by external NH₄⁺ supplies. Putative cis-elements involved in abiotic stress adaption, Ca²⁺ response, light and circadian rhythms regulation, and seed development were observed in the promoters of the PpeAMT family genes. Phosphorylation analysis of residues within the C-terminal of PpeAMT proteins revealed many conserved phosphorylation residues in both the AMT1 and AMT2 subfamily members, which could potentially play roles in controlling the NH₄⁺ transport activities. This study provides gene resources to study the biological function of AMT proteins in peach, and reveals molecular basis for NH₄⁺ uptake and N nutrition mechanisms of fruit trees.     

Effects of Barium Stress in <i>Brassica juncea</i> and <i>Cakile maritima</i>: The Indicator Role of Some Antioxidant Enzymes and Secondary Metabolites

Soil contamination by toxic trace metal elements, like barium (Ba), may stimulate various undesirable changes in the metabolic activity of plants. The plant responses are fast and with, direct or indirect, generation of reactive oxygen species (ROS). To cope with the stress imposed by the ROS production, plants developed a dual cellular system composed of enzymatic and non-enzymatic players that convert ROS, and their by-products, into stable nontoxic molecules. To assess the Ba stress response of two Brassicaceae species (Brassica juncea, a glycophyte, and Cakile maritime, a halophyte), plants were exposure to different Ba concentrations (0, 100, 200, 300 and 500 μM). The plants response was evaluated through their morphology and development, the determination of plant leaves antioxidant enzymatic activities and by the production of plants secondary metabolites. Results indicated that the two Brassicaceae species have the ability to survive in an environment containing Ba (even at 500 μM). The biomass production of C. maritima was slightly affected whereas an increase in biomass B. juncea was noticed. The stress imposed by Ba activated the antioxidant defense system in the two species, noticed by the changes in the leaves activity of catalase (CAT), ascorbate peroxidase (APX) and guaicol peroxidase (GPX), and of the secondary metabolites, through the production of total phenols and flavonoids. The enzymatic response was not similar within the two plant species: CAT and APX seem to have a more important role against the oxidative stress in C. maritima while in B. juncea is GPX. Overall, total phenols and flavonoids production was more significant in the plants aerial part than in the roots, of the both species. Although the two Brassicaceae species response was different, in both plants catalytic and non-catalytic transformation of ROS occurs, and both were able to overcome the Ba toxicity and prevent the cell damage.     

Participation of Auxin Transport in the Early Response of the <i>Arabidopsis</i> Root System to Inoculation with <i>Azospirillum brasilense</i>

The potential of Plant Growth Promoting Rhizobacteria (PGPR) has been demonstrated in the case of plant inoculation with bacteria of the genus Azospirillum which improves yield. A. brasilense produces a wide variety of molecules, including the natural auxin indole-3-acetic acid (IAA), as well as other phytoregulators. However, several studies have suggested that auxin induces changes in plant development during their interaction with the bacteria. The effects of A. brasilense Sp245 on the development of Arabidopsis thaliana root were investigated to help explain the molecular basis of the interaction. The results obtained showed a decrease in primary root length from the first day and remained so throughout the exposure, accompanied by a stimulation of initiation and maturation of lateral root primordia and an increase of lateral roots. An enhanced auxin response was evident in the vascular tissue and lateral root meristems of inoculated plants. However, after five days of bacterization, the response disappeared in the primary root meristems. The role of polar auxin transport (PAT) in auxins relocation involved the PGP1, AXR4-1, and BEN2 proteins, which apparently mediated A. brasilense-induced root branching of Arabidopsis seedlings.     

Alterations in Growth and Yield of Camelina Induced by Different Planting Densities under Water Deficit Stress

Camelina (Camelina sativa L.) is famous for its oil quality and unique fatty acid pattern. Growth and yield of crops reduced under water deficit conditions. Environmental threat such as drought or water deficit condition is the emerging problem which creates the negative impact on the growth of plants. Based upon the current situation a pot study was performed in rain out-shelter to explore the effect of different plant densities (15, 10 and 5 plants per pot) on growth and seed yield of two camelina genotypes under normal (100% WHC) and water deficit (60% WHC) conditions by using completely randomized design with factorial arrangement having three replicates. Results indicated that individual effects of plant densities and water deficit stress levels considerably influenced the growth and seed yield of camelina but interaction effects did not indicate any significant variation. Maximum values of leaf area index (LAI) and crop growth rate (CGR) were recorded in P3 treatment (15 plants per pot). However, maximum values of leaf area duration (LAD), net assimilation rate (NAR), yield and yield components were observed in the treatment P₁ (5 plants per pot). Water deficit condition (60% WHC) significantly minimized the growth, seed yield (0.82 g/m² ) and yield components of camelina genotypes. Both camelina genotypes (611 and 618) did not differ significantly under water deficit conditions.     

Identification of Resistance to Pathogenesis Related Protein <i>GmPR1L</i> in Tobacco <i>Botrytis cinerea</i> Infection

Soybean (Glycine max (Linn.) Merr.) annual leguminous crop is cultivated all over the world. The occurrence of diseases has a great impact on the yield and quality of soybean. In this study, based on the RNA-seq of soybean variety M18, a complete CDS (Coding sequence) GmPR1L of the pathogenesis-related protein 1 family was obtained, which has the ability to resist fungal diseases. The overexpression vector and interference expression vector were transferred into tobacco NC89, and the resistance of transgenic tobacco (Nicotiana tabacum L.) to Botrytis cinerea infection was identified. The results show that: Compared with the control, the activities of related defense enzymes SOD (Superoxide dismutase), POD (Peroxidase), PAL (L-phenylalanine ammonia-lyase) and PPO (Polyphenol oxidase) in the over-expressed transgenic tobacco OEA1 and OEA2 increased to different degrees, and increased significantly at different infection time points. The activities of defense enzymes in the interfering strains IEA1 and IEA2 were significantly lower than those in the control strains. The results of resistance level identification showed that the disease spot rate of OEA1 was significantly lower than that of the control line, and the disease spot rate of OEA2 was significantly lower than that of the control line. The plaque rate of the interfering expression line IEA1-IEA2 was significantly higher than that of the control line. It is preliminarily believed that the process related protein GmPR1L can improve the resistance of tobacco to B. cinerea.     

Transcriptome Profiling of Barley Cultivar Hua 30 MDEC in Response to <i>Agrobacterium</i> Infection

Agrobacterium-mediated transformation has been widely used in plants. However, the mechanism in plant cells’ response to Agrobacterium infection was very complex. The mechanism of the determinants in host cell remains obscure, especially in barley, which is recalcitrant for Agrobacterium-mediated transformation. In the present study, microspore-derived embryogenic calli (MDEC) from barley elite cultivar were employed as unique subjects to characterize the mechanisms during the Agrobacterium infection process. Hua 30 MDEC can be successfully infected by Agrobacterium. RNA-sequencing at different infection points (0, 2, 6, 12, 24 hpi) was performed. The average expressional intensity of the whole genomics increased from 0 to 2 hpi, and then decreased subsequently. More upregulated than downregulated differentially expressed genes (DEGs) were counted at the same time. GO enrichment analysis showed that protein modification was significantly overrepresented in upregulated DEGs. Chromosome-related biological processes, gene expression and cellular metabolic processes were significantly overrepresented in downregulated DEGs. KEGG analysis showed that plant defense responses, phenylpropanoid biosynthesis and biosynthesis of amino acids were significantly enriched across the infection time course. Nine DEGs related to defense responses were identified. All DEGs were upregulated from 2 to 24 hpi. We speculate that these genes are possibly related to Agrobacterium infection. These findings will provide deep insights into the molecular events occurring during the process of Agrobacterium-mediated transformation.     

Microalgae Improve the Photosynthetic Performance of Rice Seedlings (<i>Oryza sativa</i> L.) under Physiological Conditions and Cadmium Stress

The aim of this study was to assess the impact of the microalgae Chlorella vulgaris on the rice seedlings at physiological conditions and under cadmium (Cd) stress. We examined the effects of C. vulgaris in the nutrient solution on rice seedlings grown hydroponically in the presence and the absence of 150 μM CdCl₂, using the low (77 K) temperature and pulse amplitude modulated (PAM) chlorophyll fluorescence, P700 photooxidation measurements, photochemical activities of both photosystems, kinetic parameters of oxygen evolution, oxidative stress markers (MDA, H₂O₂ and proline), pigment content, growth parameters and Cd accumulation. Data revealed that the application C. vulgaris not only stimulates growth and improves the functions of photosynthetic apparatus under physiological conditions, but also reduces the toxic effect of Cd on rice seedlings. Furthermore, the presence of the green microalgae in the nutrient solution of the rice seedlings during Cd exposure, significantly improved the growth, photochemical activities of both photosystems, the kinetic parameters of the oxygen-evolving reactions, pigment content and decreased lipid peroxidation, H₂O₂ and proline content. Data showed that the alleviation of Cd-induced effects in rice seedlings is a result of the Cd sorption by microalgae, as well as the reduced Cd accumulation in the roots and its translocation from the roots to the shoots.     

Interaction of <i>Acaena elongata</i> L. with Arbuscular Mycorrhizal Fungi under Phosphorus Limitation Conditions in a Temperate Forest

The aim of this study was to analyze the performance of Acaena elongata colonized by arbuscular mycorrhizal fungi (AMF) to different phosphorus (P) concentrations, as a measure of AMF dependency. A. elongata, is a species from soils where P availability is limited, such as temperate forests. Our research questions were: 1) How do different P concentrations affect the AMF association in Acaena elongata, and 2) How does the AMF association influence A. elongata’s growth under different P concentrations? A. elongata’s growth, P content in plant tissue, AMF colonization and dependency were measured under four P concentrations: control (0 g P kg⁻¹ ), low (0.05 g P kg⁻¹ ), intermediate (0.2 g P kg⁻¹ ) and high (2 g P kg⁻¹ ) in different harvests. A complete randomized block design was applied. A. elongata’s growth was higher under -AMF in intermediate and high P concentrations, and the lowest growth corresponded to +AMF in the low and intermediate P concentration. We observed a negative effect on the root biomass under +AMF in intermediate P concentration, while the P concentration had a positive effect on the leaf area ratio. The AMF colonization in A. elongata decreased in the highest P concentration and it was favored under intermediate P concentration; while the low and the high concentrations generated a cost-benefit imbalance. Our results suggest that the performance of some plant species in soils with low P availability may not be favored by their association with AMF, but a synergy between AMF and intermediate P concentrations might drive A. elongata’s growth.     

Improved Tolerance of Three Saudi Pearl Millet Cultivars (<i>Pennisetum spicatum</i>) to Salt Stress by Mycorrhiza

Seeds of three Saudi pearl millet cultivars (Pennisetum spicatum) from three regions (Madinah, Khulais and Jaizan) were inoculated with arbuscular mycorrhizal fungus Glomus mosseae obtained from the Agriculture Research Center of Giza, Egypt to enhance their salt tolerance. Five different NaCl concentrations (0, 30, 60, 90, and 120 mM) were used for treating cultivars with and without mycorrhiza. Growth rates, chlorophyll content, chlorophyll fluorescence (Fv/Fm), proline content and gas exchange were measured to determine the effect of salinity on these cultivars. The results indicated that compared to cultivars without mycorrhiza, all cultivars with mycorrhiza had enhanced growth and physiological parameters including shoot and root length, area and number of leaves, fresh and dry weights of shoots and roots, chlorophyll contents and gas exchanges at 0 and 30 mM of salinity. In addition, the measurements of the different growth rates showed higher growth performance of the cultivars from Madinah and Khulais than the cultivar from Jaizan. However, all cultivars with and without mycorrhiza showed significant reductions in growth rates, chlorophyll contents and gas exchanges at a salinity of 60 mM than those grown at 0 and 30 mM. Moreover, the values of Fv/Fm were significantly reduced in all cultivars with and without mycorrhiza grown at 60 mM than in those grown at 0 mM and 30 mM. Proline contents indicated a progressive increase with the elevation of NaCl concentration stress. The proline contents in the mycorrhiza-inoculated cultivars were significantly higher than those in the non-inoculated cultivars. On the other hand, all cultivars with and without mycorrhiza underwent senescence within four weeks of growth at salinity concentrations of 90 mM and 120 mM. Therefore, relatively low salinity must be maintained to achieve high growth rates and gas exchanges of these inoculated cultivars.     

Physiological Responses of <i>Dendrobium officinale</i> under Exposure to Cold Stress with Two Cultivars

This study aimed to explore the cold tolerance of two cultivars of Dendrobium officinale (MG1, MG2) grown in different regions of China. Under -2°C incubation, cultivar MG1 remained active after 3 d, and continued to grow after returning to room temperature. However, MG2 could only maintain its activity after 2 d treatment at −2°C, and the seedlings died with the low temperature treatment time. Investigation of the characteristics of the plants grown in the south (Hangzhou) or north (Zhengzhou) of China indicated that the leaves of MG1 also had reduced stomatal density, the highest thickness, and a compact microstructure. The contents of proline and soluble sugars were higher in MG1 than those in MG2. The cultivar MG1 had higher SOD enzyme activity than MG2, while CAT and POD activities in samples from Zhengzhou were higher than those from Hangzhou. The contents of polysaccharides and alkaloids in stems of in MG1 were higher than those in MG2, while the content of flavonoids in the Zhengzhou samples was higher than that in the Hangzhou samples. In addition, plant heights, stem diameters, and chlorophyll content were higher in MG1. Overall, MG1 had better cold resistance than MG2. MG1 is a cold tolerant cultivar with thick leaves and reduced stomatal density, higher contents of soluble sugars, proline, CAT, POD, polysaccharides, flavonoids and alkaloids, which together make it more adaptable to low temperatures. Thus, the cultivar MG1, with its demonstrated cold tolerance, can accordingly be grown on a large scale in cold regions, thereby expanding the available planting area for this important traditional medicinal plant to meet the increasing commercial demand for it.