Role of plant growth-promoting rhizobacteria and their exopolysaccharide in drought tolerance of maize

The present study deals with the isolation and characterization of exopolysaccharides (EPS) produced by the plant growth-promoting rhizobacteria (PGPR) from arid and semiarid regions of Pakistan, and to investigate the drought tolerance potential of these PGPR on maize when used as bioinoculant alone and in combination with their respective EPS. Three bacterial strains Proteus penneri (Pp1), Pseudomonas aeruginosa (Pa2), and Alcaligenes faecalis (AF3) were selected as EPS-producing bacteria on the basis of mucoid colony formation. All these strains were gram negative, motile, and positive for catalase. Strain Pp1 was positive for oxidase test and was phosphate solubilizing, while Pa2 and AF3 were negative. The isolated strains were sequenced using 16SrRNA. Total soluble sugar, protein, uronic acid, emulsification activity, and Fourier-transformed infrared spectroscopy of EPS were determined. Drought stress had significant adverse effects on growth of maize seedlings. Seed bacterization of maize with EPS-producing bacterial strains in combination with their respective EPS improved soil moisture contents, plant biomass, root and shoot length, and leaf area. Under drought stress, the inoculated plants showed increase in relative water content, protein, and sugar though the proline content and the activities of antioxidant enzymes were decreased. The Pa2 strain isolated from semiarid region was most potent PGPR under drought stress. Consortia of inocula and their respective EPS showed greater potential to drought tolerance compared to PGPR inocula used alone.     

Influence of silicon on antioxidant mechanisms and lipid peroxidation in chickpea (Cicer arietinum L.) cultivars under drought stress

Abstract

The effects of exogenous silicon (Si) on leaf relative water content (RWC), and the growth, Si concentrations, lipid peroxidation (MDA), lipoxygenase (LOX) activity, proline and H₂O₂ accumulation, non-enzymatic antioxidant activity (AA) and the activity of some antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; ascorbate peroxidase, APX) in shoots of ten chickpea cultivars grown under drought were investigated. Drought stress decreased the growth of all the cultivars while applied Si improved the growth at least five of the 10 chickpea cultivars. Silicon applied to the soil at 100 mg kg^?1 significantly increased Si concentrations of the cultivars and counteracted the deleterious effects of drought in 5 of the ten chickpea cultivars by increasing their RWC. In most cultivars tested H₂O₂, proline and MDA content and LOX activity were increased by drought whereas application of Si decreased their levels. APX activity was increased by drought but it was depressed by Si. In general, SOD and CAT activities of the cultivars were decreased by drought. Depending on cultivars, the CAT activity was decreased, and increased or unchanged in response to applied Si, while the SOD activity of the cultivars increased or unchanged by Si. The non-enzymatic antioxidant activity of the cultivars was also increased by Si. These observations implied an essential role for Si in minimizing drought stress-induced limitation of the growth and oxidative membrane damage in chickpea plants.     

Effect of elevated carbon-dioxide on plant growth,physiology, yield and seed quality of chickpea (Cicer arietinum L.) in Indo-Gangetic plains

In the present scenario of climate change with constantly increasing CO₂ concentration, there is a risk of altered crop performance in terms of growth, yield, grain nutritional value and seed quality. Therefore, an experiment was conducted in open top chamber (OTCs) during 2017–18 and 2018–19 to assess the effect of elevated atmospheric carbondioxide (e[CO₂]) (600 ppm) on chickpea (cv. JG 14) crop growth, biomass accumulation, physiological function, seed yield and its quality in terms of germination and vigour. The e[CO₂] treatment increased the plant height, leaf and stem biomass over ambient CO₂ (a[CO₂]) treatment. The e[CO₂] increased seed yield by 11–18% which was attributed to an increase in the number of pods (6–10%) and seeds plant⁻¹ (8–9%) over a[CO₂]. However, e[CO₂] reduced the seed protein (7%), total phenol (13%) and thiobarbituric acid reactive substances (12%) and increased the starch (21%) and water uptake rate as compared to seeds harvested from a[CO₂] environment. Exposing chickpea plant to e[CO₂] treatment had no impact on germination and vigour of the harvested seeds. Also, the physical attributes, total soluble sugar and antioxidant enzymes activities of harvested seeds were comparable in a[CO₂] and e[CO₂] treatment. Hence, the experimental findings depict that e[CO₂] upto 600 ppm could add to the growth and productivity of chickpea in a sub-tropical climate with an implication on its nutritional quality of the produce.     

Effects of exogenously applied salicylic acid and putrescine alone and in combination with rhizobacteria on the phytoremediation of heavy metals and chickpea growth in sandy soil

The present attempt was made to study the role of exogenously applied salicylic acid (SA) and putrescine (Put) on the phytoremediation of heavy metals and on the growth parameters of chickpea grown in sandy soil. The SA and Put were applied alone as well as in combination with plant growth promoting rhizobacteria (PGPR). The PGPRs, isolated from the rhizosphere of chickpea, were characterized on the basis of colony morphology and biochemical traits through gram staining, catalase and oxidase tests, and identified by 16S-rRNA gene sequencing as Bacillus subtilis, Bacillus thuringiensis and Bacillus megaterium. The chickpea seeds were soaked in 24 h old fresh cultures of isolates for 2–3 h prior to sowing. The growth regulators (PGRs), SA and Put (150 mg/L), were applied to the seedlings as foliar spray at three-leaf stage. The result revealed that plants treated with SA and Put alone or in combination with PGPRs, significantly enhanced the accumulation of heavy metals in plant shoot. PGPR induces Ni accumulation in sensitive variety and Pb in both the varieties, the PGR in combination augment the bioremediation effects of PGPR and both sensitive and tolerant variety showed significant accumulation of Ni, Cd, and Pb. SA was more effective in accumulating Ni and Cd whereas, significant accumulation of Pb was recorded in Put. PGPRs further augmented the PGRs induced accumulation of heavy metals and macronutrients in chickpea shoot and in rhizosphere. SA increased the proline content of tolerant variety while decreasing the lipid peroxidation and proline content of the sensitive variety but decreased the stimulating effect of PGPR in proline production. Interactive effects of PGPR and PGRs are recommended for inducing phytoremediation in chickpea plants under drought stress.     

Genetic engineering of chickpea (Cicer arietinum L.) with the P5CSF129A gene for osmoregulation with implications on drought tolerance

Abiotic stresses including water deficit severely limits crop yields in the semi-arid tropics. In chickpea, annual losses of over 3.7 million tones have been estimated to be due to water deficit conditions alone. Therefore, major efforts are needed to improve its tolerance to water deficit, and genetic engineering approaches provide an increasing hope for this possibility. We have used transgenic technology for the introduction of an osmoregulatory gene P5CSF129A encoding the mutagenized Δ¹-pyrroline-5-carboxylate synthetase (P5CS) for the overproduction of proline. A total of 49 transgenic events of chickpea were produced with the 35S:P5CSF129A gene through Agrobacterium tumefaciens-mediated gene transfer through the use of axillary meristem explants. Eleven transgenic events that accumulated high proline (2–6 folds) were further evaluated in greenhouse experiments based on their transpiration efficiency (TE), photosynthetic activity, stomatal conductance, and root length under water stress. Almost all the transgenic events showed a decline in transpiration at lower values of the fraction of transpirable soil water (dryer soil), and extracted more water than their untransformed parents. The accumulation of proline in the selected events was more pronounced that increased significantly in the leaves when exposed to water stress. However, the overexpression of P5CSF129A gene resulted only in a modest increase in TE, thereby indicating that the enhanced proline had little bearing on the components of yield architecture that are significant in overcoming the negative effects of drought stress in chickpea.     

Effects of selection criteria on yield stability in chickpea (Cicer arietinum L.)

Summary Selection in the F₃ generation for seed yield, fruiting branches/plant, effective pods/plant, and seed index (100-seed weight) was carried out in two chickpea crosses. Sixty F₅ lines (15 lines/selection criterion) along with check variety were evaluated for seed yield in three distinct environments. The effects of selection criteria on yield stability was examined using linear regression approach and genotype-grouping technique. There were no differences between selection criteria for linear yield responses of F₅ lines to different environments. Within all four selection criteria the lines showed similar linear responses. The non-linear component was relatively higher for lines selected for effective pods and seed index than lines selected for yield and fruiting branches. On the basis of mean yield and coefficient of variation across environments, the seed index was the least effective selection criterion for developing high yielding and stable lines. When the results of stability parameters and genotype-grouping technique were considered together, selection for yield and fruiting branches was highly effective for isolating stable and high yielding lines.     

Synergistic effect of Pseudomonas putida and Bacillus amyloliquefaciens ameliorates drought stress in chickpea (Cicer arietinum L.)

Two plant growth promoting rhizobacteria (PGPR) Pseudomonas putida NBRIRA and Bacillus amyloliquefaciens NBRISN13 with ability to tolerate abiotic stress along with multiple PGP traits like ACC deaminase activity, minerals solubilisation, hormones production, biofilm formation, siderophore activity were evaluated for their synergistic effect to ameliorate drought stress in chickpea. Earlier we have reported both the strains individually for their PGP attributes and stress amelioration in host plants. The present study explains in detail the possibilities and benefits of utilizing these 2 PGPR in consortium for improving the chickpea growth under control and drought stressed condition. In vitro results clearly demonstrate that both the PGPR strains are compatible to each other and their synergistic growth enhances the PGP attributes. Greenhouse experiments were conducted to evaluate the effect of inoculation of both strains individually and consortia in drought tolerant and sensitive cultivars (BG362 and P1003). The growth parameters were observed significantly higher in consortium as compared to individual PGPR. Colonization of both PGPR in chickpea rhizosphere has been visualized by using gfp labeling. Apart from growth parameters, defense enzymes, soil enzymes and microbial diversity were significantly modulated in individually PGPR and in consortia inoculated plants. Negative effects of drought stress has been ameliorated and apparently seen by higher biomass and reversal of stress indicators in chickpea cultivars treated with PGPR individually or in consortia. Findings from the present study demonstrate that synergistic application has better potential to improve plant growth promotion under drought stress conditions.     

Effects of supplemental nitrate and thermal regime on the nitrogen nutrition of chickpea (Cicer arietinum L.)

Summary Nodulated chickpea plants were grown in pots in a glasshouse programmed to simulate either hot (32.5°C day/18°C night) or warm (25°/18°C) thermal regimes characteristic of those experienced by crops grown in different seasons or locations in the semi-arid tropics. The plants were irrigated with nutrient solution either devoid of inorganic nitrogen or containing 0.71, 1.43 or 2.86 mM nitrate. Increasing concentrations of supplemental nitrate stimulated the rate of dry matter production by vegetative plants in both thermal regimes. Differences between vegetative dry weight of plants given nitrate and those relying exclusively on symbiotic dinitrogen fixation were greatest in the hot regime where the durations of vegetative growth were shorter. However, symbiotically-dependent plants and those given 0.71 mM nitrate continued to produce branches throughout the reproductive period, particularly in the warm regime. As they matured, these plants became more comparable in vegetative stature to those which had received greater concentrations of nitrate and had established final branch numbers earlier (i.e prior to main pod-fill). Potential seed yields were determined primarily by the number of potential reproductive sites (nodes) available (i.e. by the extent of branching) which largely determined the number of seeds harvested. Since final branch numbers in all nitrate treatments were greatest in the warm regime, yields were also larger than those at 32.5°C. The implications of these data for the nitrogen economy of chickpea crops is discussed.One of a series of papers resulting from a collaborative project with the International Crops Research Institute for the Semi-Arid Tropics, India; sponsored by the UK Overseas Development Administration.     

Cobalt stress affects nitrogen metabolism,photosynthesis and antioxidant system in chickpea (Cicer arietinum L.)

Abstract

Plants of chickpea were exposed to varied levels of cobalt (Co) and sampled at the 60-day stage. Cobalt at concentration <100 µM significantly increased the number of nodules, their dry mass, leghemoglobin concentration and the activity of nitrogenase. Similarly, the activities of glutamate dehydrogenase, glutamine synthetase and glutamate synthase also exhibited an increase in the presence of Co <100 µM, in nodules and leaves, respectively. The various photosynthetic attributes in leaves and the activity of antioxidative enzymes both in nodules and leaves were inhibited by Co in a concentration-dependent manner. However, the lipid peroxidation and the content of proline exhibited a significant increase in response to Co and were at a maximum in the plants exposed to 250 µM concentration of cobalt. Since most of the parameters showed a significant increase in response to 50 µM cobalt, this concentration may be regarded as a threshold concentration.     

干旱胁迫对不同耐旱性玉米杂交种产量和根系生理特性的影响

通过盆栽人工模拟干旱试验,研究了全生育期中度干旱胁迫对不同耐旱性玉米杂交种（耐旱：京科628;不耐旱：农大95）产量及根系生理特性的影响.结果表明：干旱胁迫下,耐旱玉米产量比对照减少33.7%,不耐旱玉米则比对照减少62.3%.干旱胁迫下,玉米根系生物量降低且最大值出现时间提前,与对照相比,不耐旱玉米根冠比升高,耐旱玉米根冠比前期升高后期降低;根系活力降低,不耐旱玉米根系活力降低幅度大于耐旱玉米;根系超氧化物歧化酶（SOD）活性前期高于对照后期低于对照,耐旱玉米根系SOD活性开始低于对照的时间比不耐旱玉米晚;根系丙二醛（MDA）含量升高,随干旱胁迫处理时间的延长,不耐旱玉米MDA含量比对照升高的幅度大于耐旱玉米;根系可溶性蛋白含量降低,不耐旱玉米的降低幅度大于耐旱玉米.干旱胁迫下耐旱玉米杂交种根系活力﹑根系SOD活性及可溶性蛋白含量较高,减缓了根系的衰老进程,延长了根系功能期,这可能是耐旱玉米杂交种在干旱胁迫下仍能获得较高产量的重要原因之一.     

园艺植物水分胁迫生理及耐旱机制研究进展

概述了园艺植物在水分胁迫下的生理生化,分子反应及耐旱机制研究进展,并指出尚需进一步研究的问题。     

Identification of operative dose of NPK on yield enhancement of desi and kabuli chickpea (Cicer arietinum L.) in diverse milieu

The effect of fertilizer on yield and yield related traits studied in two consecutive years at two different locations. Three different doses of fertilizer (NPK) applied at the rate of 9:23:0, 18:46:12 and 27:69:25 respectively on sixteen chickpea, genotypes (Desi and Kabuli advance lines and commercial varieties). Data recorded for days to 50% flowering, plant height, primary and secondary branches, pods per plant, 100-grain weight and grain yield (kg/ha). In Desi chickpea highest grain yield (kg/ha) in both years was produced by advance line D-12026 and in Kabuli advance line K-70005 at Faisalabad location. Grain yield kg/ha had significant positive correlation with all the considered parameters except days to 50% flowering and days to 50% maturity. The treatment comparison manifested that fertilizer doses 9:23:0 enhanced grain yield. The high dose of fertilizer is not recommendable. The grain yield of Desi and Kabuli chickpea at two locations Pulses Research Institute (PRI) Faisalabad and GBRSS (Gram Breeding Research Sub Station) Kallurkot had significant variation. The grain yield (kg ha⁻¹) was significant high in research area of PRI, Faisalabad in both years. The NPK 9:23:0 found operative dose of fertilizer for chickpea.     

Pseudomonas putida modulates the expression of miRNAs and their target genes in response to drought and salt stresses in chickpea (Cicer arietinum L.)

MicroRNAs are small non-coding regulatory RNA molecules that play an important role in the modulation of gene expression during various environmental stresses. Pseudomonas putida RA, a plant growth promoting rhizobacteria (PGPR) colonizes the root surface of plants improving their growth and development during abiotic stresses modulating the expression of stress-responsive genes; however, the impact of RA on stress responsive-miRNA remains elusive. The present study was an attempt to delineate the role of PGPR in modulating stress responsive-miRNAs in a tolerant desi chickpea genotype exposed to drought and salt stresses. The existence of variable expression patterns of individual miRNAs and their target genes under these stresses at different time points indicate a distinct miRNA-mediated perception and response mechanisms operating under these stresses in the presence or absence of RA in chickpea.     

丛枝菌根真菌对植物耐旱性的影响研究进展

丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)能与植物根系形成互惠共生体,对植物的生长发育和抗逆性有积极的影响,在改善植物水分代谢和提高植物耐旱性中发挥了重要作用.本文综述了近年来AMF与植物水分代谢关系的研究进展,从植物的光合作用、蒸腾与气孔导度、水分利用效率、水力导度、渗透调节、内源激素和抗氧化系统等方面说明AMF对植物水分代谢的影响.从4个方面介绍了AMF提高植物耐旱性的机理:1)菌丝网络增加植物根系吸收范围;2)增强植物保水能力和抗氧化能力;3)稳定和改善土壤团聚体;4)促进植物养分吸收.并提出今后研究需注意的问题和建议.     

Abundance and polymorphism of di-, tri-and tetra-nucleotide tandem repeats in chickpea (Cicer arietinum L.)

The abundance and polymorphism of 38 different simple-sequence repeat motifs was studied in four accessions of cultivated chickpea (Cicer arietinum L.) by in-gel hybridization of synthetic oligonucleotides to genomic DNA digested with 14 different restriction enzymes. Among 38 probes tested, 35 yielded detectable hybridization signals. The abundance and level of polymorphism of the target sequences varied considerably. The probes fell into three broad categories: (1) probes yielding distinct, polymorphic banding patterns; (2) probes yielding distinct, monomorphic banding patterns, and (3) probes yielding blurred patterns, or diffused bands superimposed on a high in lane background. No obvious correlation existed between abundance, fingerprint quality, and the sequence characteristics of a particular motif. Digestion with methyl-sensitive enzymes revealed that simple-sequence motifs are enriched in highly methylated genomic regions. The high level of intraspecific polymorphism detected by oligonucleotide fingerprinting suggests the suitability of simple-sequence repeat probes as molecular markers for genome mapping.     

Isolation and characterisation of endophytic actinobacteria and their effect on the growth and nodulation of chickpea (Cicer arietinum)

Plant and Soil - Phyllostachys praecox is one of the bamboo species that cultivated extensively in southern China. An organic material mulching technique is commonly adopted in winter to increase...     

The effect of soil pH on chickpea (Cicer arietinum) genotype sensitivity to isoxaflutole

A glasshouse experiment was conducted to investigate the effect of soil pH on chickpea (Cicer arietinum) tolerance to isoxaflutole applied pre-emergence at 0, 75 (recommended rate) and 300 g a.i. ha⁻¹. For this study, the variables examined were two desi chickpea genotypes (97039-1275 as a tolerant line and 91025-3021 as a sensitive line) and four pH levels (5.1, 6.9, 8.1, and 8.9). The results demonstrated differential tolerances among chickpea genotypes to isoxaflutole at different rates and soil pH levels. Isoxaflutole applied pre-emergence resulted in increased phytotoxicity with increases in soil pH and herbicide rate. Even the most tolerant chickpea genotype was damaged when exposed to higher pH and herbicide rates, as indicated by increased leaf chlorosis and significant reductions in plant height, and shoot and root dry weight. The effects were more severe with the sensitive genotype. The susceptibility of chickpea to this herbicide depends on genotype and soil pH which should be taken into account in breeding new lines, and in the agronomy of chickpea production.     

Diversity of pollinators and their role in the pollination biology of chickpea,Cicer arietinum L. (Fabaceae)

Chickpea (Cicer arietinum L.) is an important source of food for people worldwide. In the current study, we studied its pollination biology with special reference to floral visitors along with their visitation rate, frequency and pollen load during 2012 and 2013. We also explored the effect of floral visitors on the capsule weight, seed weight, seed numbers and seed germination. Results revealed three bees, two wasps, five flies, one moth and three butterfly species on the flowers of chickpea. Apis dorsata, A. florea, Amegilla sp. and Eristalinus aeneus were the major species with 434–474, 223–311, 69–74 and 81–136 individuals, respectively in both years. Floral visitors differed significantly in term of visitation frequency with A. florea as the most frequent visitor (9.13–9.86 visits/flower/5 min) followed by E. aeneus (5.43–5.58 visits/flower/5 min) and A. dorsata (1.72–2.31 visits/flower/5 min) in both years. Similarly, A. florea had statistically highest visitation rate (16.85–19.99 flowers visited/min) followed by E. aeneus (9.73–10.68 flowers visited/min). A. dorsata had significantly higher pollen load on its body (84629–85,104 pollen grains) followed by A. florea (64940–65,135 pollen grains) and Amegilla sp. (64020–65,120 pollen grains). The open-pollinated flowers had significantly higher capsule weight (0.27 ± 0.01 g), seed weight (0.18 ± 0.01 g), seed numbers (1.67 ± 0.07 seeds) and seed germination (95 ± 1.38%) as compared to flowers deprived of pollinators in cages. The results suggested A. florea, A. dorsata and E. aeneus could be effective pollinators of chickpea. Hence these three species can be properly utilized on commercial scale to increase crop yield.     

Effect of organic amendments on root rot of gram (Cicer arietinum L.) and their influence on plant growth

Summary Rhizoctonia root rot of gram caused byR. bataticola was significantly controlled by the amendment of soil with wheat straw, maize straw and sorghum straws. Amendment of soil with sarson straw was almost ineffective. Organic amendments with urd, mung and Trifolium straws increased the disease. It appears that perhaps a comparatively high C/N ratio of the organic amendment controls the root rot disease of gram. Evidence is also presented which suggests that certain organic amendments increase growth of gram.