Application of Streptomyces pactum Act12 Enhances Drought Resistance in Wheat

The use of beneficial microbes to improve drought resistance in crops has great application potential in agricultural production, yet the effects of actinomycetes upon crop resistance to drought are rarely reported. Streptomyces pactum Act12 is a known multi-functional biocontrol agent of soil-borne diseases in several horticultural crops and medicinal plants. Here, we systematically analyzed how Act12 treatment affects drought resistance in drought-sensitive wheat (Triticum aestivum L.) cultivar Xinong 979 by considering both its effects and underlying mechanisms. After seed exposure to a cell-free culture filtrate of Act12, we measured several plant growth variables, osmotic adjustment and antioxidant capacity, cell membrane peroxidation, and drought resistance-related gene expression in wheat seedlings under drought stress conditions simulated by polyethylene glycol 6000. Results showed that, under drought stress, wheat seedling exposure to Act12 cell-free filtrate facilitated plant growth, with significant increases in shoot fresh weight (21.3%), shoot length (10.3%), and root length (13.6%). Act12 treatment also significantly increased total soluble sugar content in wheat leaves while decreasing their malondialdehyde content by 20.5%. Under non-drought conditions, Act12 treatment increased the content of both proline and glutathione in wheat leaves; however, both were lowered in Act12-treated plants compared with non-treated plants at 96 h of drought stress. Further analysis revealed that Act12 treatment increased the content of leaf abscisic acid and upregulated the expression levels of several drought resistance-related genes, such as EXPA2, EXPA6, P5CS, and SnRK2. These results suggest that application of S. pactum Act12 can enhance the osmotic adjustment and antioxidant capacity of plants via induction of abscisic acid accumulation and up-regulation of drought resistance-related gene expression, thereby mitigating drought stress impact in wheat.

     

Drought stress amelioration in wheat through inoculation with Burkholderia phytofirmans strain PsJN

Plant growth promoting endophytic bacteria Burkholderia phytofirmans PsJN was used to investigate the potential to ameliorate the effects of drought stress on growth, physiology and yield of wheat (Triticum aestivum L.) under natural field conditions. Inoculated and uninoculated (control) seeds of wheat cultivar Sahar 2006 was sown in the field. The plants were exposed to drought stress at different stages of growth (tillering stage and flowering stage) by skipping the respective irrigation. The results showed that drought stress adversely affected the physiological, biochemical and growth parameters of wheat seedlings. It decreased the CO₂ assimilation, stomatal conductance, relative water content, transpiration rate and chlorophyll contents in wheat. Inoculation of wheat with PsJN significantly diluted the adverse effects of drought on relative water contents and CO₂ assimilation rate thus improving the photosynthetic rate, water use efficiency and chlorophyll content over the uninoculated control. Grain yield was also decreased when plants were exposed to drought stress at the tillering and flowering stage, but inoculation resulted in better grain yield (up to 21 and 18 % higher, respectively) than the respective uninoculated control. Similarly, inoculation improved the ionic balance, antioxidant levels, and also increased the nitrogen, phosphorus, potassium and protein concentration in the grains of wheat. The results suggested that B. phytofirmans strain PsJN could be effectively used to improve the growth, physiology and quality of wheat under drought conditions.     

Plant growth promoting bacteria enhance water stress resistance in green gram plants

Plant growth promoting bacterial (PGPB) strains Pseudomonas fluorescens Pf1 and endophytic Bacillus subtilis EPB5, EPB22, EPB 31 were tested for their capacity to induce water stress related proteins and enzymes in green gram (Vigna radiata) plants. Among the different bacteria used, P. fluorescens Pf1 increased the vigour index, fresh weight and dry weight of green gram seedlings in vitro. Quantitative and qualitative analyses of stress-related enzymes indicated the greater activity of catalase and peroxidase in green gram plants bacterized with P. fluorescens Pf1 against water stress when compared to untreated plants. The greater accumulation of proline was recorded in Pf1 treated plants compared to untreated plants. The pot culture study revealed the greater resistance to water stress by green gram plants treated with P. fluorescens Pf1 compared to untreated plants. The greater activity of stress-related enzymes in green gram plants mediated by PGPB could pave the way for developing drought management strategies.     

Shoot endophytic plant growth-promoting bacteria reduce cadmium toxicity and enhance switchgrass (<Emphasis Type="Italic">Panicum virgatum</Emphasis> L.) biomass

The aims of the study were to increase the biomass and to alleviate the deleterious effects of cadmium (Cd) in the switchgrass cultivars (Panicum virgatum L.) Alamo and Cave-in-Rock (CIR) under cadmium (Cd) stress using Cd-tolerant shoot endophytic plant growth-promoting bacteria (PGPB). Four shoot endophytic bacterial strains, viz. Bc09, So23, E02, and Oj24, were isolated from the above-ground parts of plants grown in a Cd-polluted soil and were successfully identified by 16S rRNA gene sequencing as Pseudomonas grimontii, Pantoea vagans, Pseudomonas veronii, and Pseudomonas fluorescens, respectively. These four strains were adapted to high CdCl₂ concentrations as they had higher Cd uptake capacities. In addition, they possessed a huge amount of growth regulatory activities e.g., indole acetic acid production, 1-aminocyclopropane-1-carboxylic acid deaminase (ACCD) activity, and phosphate solubilization. Growth particularly the height and biomass of both cultivars increased significantly in response to PGPB inoculation in the 20 µM CdCl₂ stress. The shoot biomass of the PGPB-inoculated Alamo was higher than the CIR under Cd stress. Interestingly, the level of Cd inside PGPB-inoculated plant tissues and the translocation factors were lower compared with the noninoculated Cd control plants. CIR plants exhibited higher Cd content than Alamo plants. Through confocal microscopy, green fluorescence was observed in roots and leaf tissues 2 days after the inoculation of green fluorescent protein (GFP)-labeled bacteria in Alamo, which confirmed the successful colonization of bacteria inside the plant tissues. These shoot endophytic PGPB and switchgrass interactions are useful for the sustainable biomass production of bioenergy crop in a Cd-contaminated environment.     

Selenium (Se) Seed Priming Induced Growth and Biochemical Changes in Wheat Under Water Deficit Conditions

Insufficient stand establishment at early growth stages in wheat (Triticum aestivum L.) due to drought stress is a major problem that limits overall efficiency and yield of crop. Priming of seed is an effective method for raising seed performance and improving tolerance of crops to abiotic stresses especially drought. The seeds of two local wheat cultivars (Kohistan-97 and Pasban-90) were soaked in distilled water or sodium selenate solutions of 25, 50, 75, and 100 μM for 1/2 or 1 h at 25 °C and later re-dried to their original moisture levels before sowing. One-hour priming significantly increased root length stress tolerance index, dry matter stress tolerance index, and total biomass of seedlings; however, no significant effect of changing duration of Se seed priming was observed on plant height stress tolerance index and shoot/root ratio. Among cultivars, Kohistan-97 was found to be more responsive to Se seed treatment as 1 h priming at 100 μM significantly increased its total biomass by 43 % as compared to control treatment. Although biomass of seedlings was not affected with Se seed priming under normal conditions, but it increased significantly with increase in rates of Se under drought stress conditions. One-hour priming at 75 μM increased the total sugar content and total free amino acids in both wheat cultivars. A more significant decrease in soluble proteins of seedlings was observed by 1 h priming than 1/2 h priming under drought stress conditions.     

Metabolic profile and antioxidant responses during drought stress recovery in sugarcane treated with humic acids and endophytic diazotrophic bacteria

Water deficit is the major yield‐limiting factor for sugarcane crop production that can be enhanced by inoculating with plant growth promoting bacteria (PGPB) combined with humic substances. The aim of this work was to examine changes to the metabolic profile and antioxidant enzyme activity of sugarcane treated with PGPB and humic acid (HA) after drought and then rehydration. The drought was imposed by withholding irrigation for 21 days thereby measuring enzyme activity, metabolic profile and photosynthetic rate 1 week after rehydratation. Growth of plants treated with HA, PGPB and with both treatments combined (PGPB + HA) was higher than control plants. The antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) activities remained higher after rehydration only in plants treated with HA. Plants treated with HA and PGPB + HA exhibited increased transpiration, stomatal conductance and net photosynthesis than plants treated with PGPB. The PGPB‐treated plants exhibited drought resistance that resembled ‘delayed stress onset’, which is a new term for preserving water in the plants tissues. Water preservation in plants treated with PGPB was corroborated by higher relative water content (RWC) than control plants at the end of the drought period. Plants treated with HA + PGPB exhibited the highest water potential after rehydration and high RWC. Osmotic adjustment in the other treatments (control, HA and PGPB) was indicated by a new pattern of metabolic response after rehydration, including generally enhanced carbohydrates and proteins and specific changes induced by HA‐enhancing aromatic compounds, whereas PGPB exhibited enhanced fatty acids and other aliphatic H species. Humic acids assist with drought stress recovery by inducing antioxidant enzyme activity whereas PGPB induced preservation of leaf water potential and RWC by closing stomata efficiently, resulting in plant water preservation.     

Terminal drought and seed priming improves drought tolerance in wheat

Plants retain the preceding abiotic stress memory that may aid in attainment of tolerance to subsequent stresses. This study was conducted to evaluate the influence of terminal drought memory (drought priming) and seed priming in improving drought tolerance in wheat (Triticum aestivum L.). During first growing season, wheat was planted in field under optimal (well-watered) and drought stress imposed at reproductive stage (BBCH growth stage 49) until maturity (BBCH growth stage 83). Seeds collected from both sources were subjected to hydropriming or osmopriming (with 1.5% CaCl₂ solution); while, dry seed was taken as control. Treated and control seeds, from both sources, were sown in soil filled pots. After the completion of seedling emergence, pots were maintained at 50% water holding capacity (drought) or 100% water holding capacity (well-watered). Drought stress suppressed the plant growth (2–44%), perturbed water relations (1–18%) and reduced yield (192%); however, osmolytes accumulation (3–14%) and malondialdehyde contents (26–29%) were increased under drought. The crop raised from the seeds collected from terminal drought stressed plants had better growth (5–63%), improved osmolyte accumulation (13–45%), and lower lipid peroxidation (3%) than the progeny of well-watered crop. Seed priming significantly improved the crop performance under drought stress as compared to control. However, osmopriming was more effective than hydropriming in this regard as it improved leaf area (9–43%), tissue water status (2–47%), osmolytes accumulation (6–48%) and grain yield (14–79%). In conclusion, terminal drought induced modifications in seed composition and seed priming improved transgenerational drought tolerance through improvement in tissue water status and osmolytes accumulation, and decrease in lipid peroxidation.     

Drought tolerance through over-expression of the expansin gene TaEXPB23 in transgenic tobacco

Expansins are proteins that are the key regulators of wall extension during plant growth. To investigate the role of TaEXPB23, a wheat expansin gene, we analyzed TaEXPB23 mRNA expression levels in response to water stress in wheat and examined the drought resistance of transgenic tobaccos over-expressing TaEXPB23. We found that the expression of TaEXPB23 corresponded to wheat coleoptile growth and the response to water stress. The results also indicated that the transgenic tobacco lines lost water more slowly than the wild-type (WT) plants under drought stress; their cells could sustain a more integrated structure under water stress than that of WT. Other physiological and biochemical parameters under water stress, such as electrolyte leakage, malondialdehyde (MDA) level, photosynthetic rate, F_v/F_m and ΦPSII, also suggested that the transgenic tobaccos were more drought resistant than WT plants.     

Isolation and characterization of plant growth promoting endophytic diazotrophic bacteria from Korean rice cultivars

We have isolated 576 endophytic bacteria from the leaves, stems, and roots of 10 rice cultivars and identified 12 of them as diazotrophic bacteria using a specific primer set of nif gene. Through 16S rDNA sequence analysis, nifH genes were confirmed in the two species of Penibacillus, three species of Microbacterium, three Bacillus species, and four species of Klebsiella. Rice seeds treated with these plant growth-promoting bacteria (PGPB) showed improved plant growth, increased height and dry weight and antagonistic effects against fungal pathogens. In addition, auxin and siderophore producing ability, and phosphate solubilizing activity were studied for the possible mechanisms of plant growth promotion. Among 12 isolates tested, 10 strains have shown higher auxin producing activity, 6 isolates were confirmed as strains with high siderophore producing activity while 4 isolates turned out to have high phosphate-solubilizing activity. These results strongly suggest that the endophytic diazotrophic bacteria characterized in this study could be successfully used to promote plant growth and inducing fungal resistance in plants.     

Differential expression profiles of poplar MAP kinase kinases in response to abiotic stresses and plant hormones,and overexpression of PtMKK4 improves the drought tolerance of poplar

Mitogen-activated protein kinase (MAPK) cascades are universal signal transduction modules that play essential roles in plant growth, development and stress response. MAPK kinases (MAPKKs), which link MAPKs and MAPKK kinases (MAPKKKs), are integral in mediating various stress responses in plants. However, to date few data about the roles of poplar MAPKKs in stress signal transduction are available. In this study, we performed a systemic analysis of poplar MAPKK gene family expression profiles in response to several abiotic stresses and stress-associated hormones. Furthermore, Populus trichocarpa MAPKK4 (PtMKK4) was chosen for functional characterization. Transgenic analysis showed that overexpression of the PtMKK4 gene remarkably enhanced drought stress tolerance in the transgenic poplar plants. The PtMKK4-overexpressing plants also exhibited much lower levels of H₂O₂ and higher antioxidant enzyme activity after exposure to drought stress compared to the wide type lines. Besides, some drought marker genes including PtP5CS, PtSUS3, PtLTP3 and PtDREB8 exhibited higher expression levels in the transgenic lines than in the wide type under drought conditions. This study provided valuable information for understanding the putative functions of poplar MAPKKs involved in important signaling pathways under different stress conditions.     

ACC Deaminase-Containing Bacillus subtilis Reduces Stress Ethylene-Induced Damage and Improves Mycorrhizal Colonization and Rhizobial Nodulation in Trigonella foenum-graecum Under Drought Stress

This study was aimed at protecting Trigonella plants by reducing stress ethylene levels through ACC (1-aminocyclopropane-1-carboxylic acid) deaminase-containing Bacillus subtilis (LDR2) and promoting plant growth through improved colonization of beneficial microbes like Ensifer meliloti (Em) and Rhizophagus irregularis (Ri) under drought stress. A plant growth-promoting rhizobacterium strain possessing high levels of ACC deaminase characterized as B. subtilis was selected. Application of this strain considerably protected Trigonella plants under severe drought stress conditions; this protection was correlated with reduced levels of ACC (responsible for generation of stress ethylene). The experiment consisted of eight inoculation treatments with different combinations of ACC deaminase-containing rhizobacteria LDR2, Ri, and Em under three water regimes. The tripartite combination of LDR2 + Ri + Em acted synergistically to induce protective mechanisms against decreased soil water availability in Trigonella plants and improved plant weight by 56 % with lower ACC concentration (39 % less than stressed noninoculated plants) under severe drought conditions. Drought-induced changes in biochemical markers like reduced chlorophyll concentration, increased proline content, and higher lipid peroxidation were monitored and clearly indicated the protective effects of LDR2 under drought stress. Under drought conditions, apart from alleviating ethylene-induced damage, LDR2 enhanced nodulation and arbuscular mycorrhizal fungi colonization in the plants resulting in improved nutrient uptake and plant growth.     

Study on Mechanisms of Colonization of Nitrogen-Fixing PGPB, <Emphasis Type="Italic">Klebsiella pneumoniae</Emphasis> NG14 on the Root Surface of Rice and the Formation of Biofilm

Plant growth-promoting bacteria (PGPB) refer to the bacteria beneficial to plants, and they may affect the growth and development of plants directly or indirectly. This article studied the activities of nitrogen fixation and colonization of a strain of PGPB, Klebsiella pneumoniae NG14, which was isolated from the rice root surface. The results showed that NG14 harbouring the nifH gene had nitrogenase activity, ¹⁵N₂-fixing activity, and was able to colonize on the root surface and within the cavity of root vascular tissues of rice. Using proteomics technology to study the differences and changes of membrane proteins (MP) of NG14 bacterial biofilm in non-biological surface, 28 proteins showing significant differences before and after the formation of bacterial biofilm have been identified, in which the precursors of membrane pore protein OmpC relevant to osmotic stress resistance was up-regulated. This study would have positive significance on further understanding of the direct and indirect promotion effects of PGPB and related mechanisms.