Integrated control of tobacco black shank by combined use of riboflavin and <Emphasis Type="Italic">Bacillus subtilis</Emphasis> strain Tpb55

We investigated the effect of riboflavin on the biocontrol activity of Bacillus subtilis Tpb55 against Phytophthora nicotianae (Pn), which causes tobacco black shank. Riboflavin (0.2 mg ml^?1) significantly improved the biocontrol activity of Tpb55 (2.0 × 10⁸ cfu ml^?1). Riboflavin (0.02–0.5 mg ml^?1) alone could not significantly inhibit Pn growth. However, it enhanced the B. subtilis population, both in vitro and in tobacco roots and significantly increased the activity of defense enzymes, peroxidase, catalase, superoxide dismutase, and β-1,3-glucanase, in the roots of B. subtilis-treated tobacco seedlings. Our results indicate that riboflavin can stimulate the growth of B. subtilis Tpb55 and induce resistance to Pn in tobacco plants. These findings should boost the prospects for practical application of B. subtilis Tpb55 as a biocontrol agent against black shank of tobacco.     

<Emphasis Type="Italic">Rhizophagus irregularis</Emphasis> MUCL 41833 can colonize and improve P uptake of <Emphasis Type="Italic">Plantago lanceolata</Emphasis> after exposure to ionizing gamma radiation in root organ culture

Long-lived radionuclides such as ⁹⁰Sr and ¹³⁷Cs can be naturally or accidentally deposited in the upper soil layers where they emit β/γ radiation. Previous studies have shown that arbuscular mycorrhizal fungi (AMF) can accumulate and transfer radionuclides from soil to plant, but there have been no studies on the direct impact of ionizing radiation on AMF. In this study, root organ cultures of the AMF Rhizophagus irregularis MUCL 41833 were exposed to 15.37, 30.35, and 113.03 Gy gamma radiation from a ¹³⁷Cs source. Exposed spores were subsequently inoculated to Plantago lanceolata seedlings in pots, and root colonization and P uptake evaluated. P. lanceolata seedlings inoculated with non-irradiated AMF spores or with spores irradiated with up to 30.35 Gy gamma radiation had similar levels of root colonization. Spores irradiated with 113.03 Gy gamma radiation failed to colonize P. lanceolata roots. P content of plants inoculated with non-irradiated spores or of plants inoculated with spores irradiated with up to 30.35 Gy gamma radiation was higher than in non-mycorrhizal plants or plants inoculated with spores irradiated with 113.03 Gy gamma radiation. These results demonstrate that spores of R. irregularis MUCL 41833 are tolerant to chronic ionizing radiation at high doses.     

Application of phytohormones during seed hydropriming and heat shock treatment on sunflower (<Emphasis Type="Italic">Helianthus annuus</Emphasis> L.) chilling resistance and changes in soluble carbohydrates

The objective of this study was to analyze the mechanism of some physiological processes accompanying acquisition of sunflower (Helianthus annuus L.) chilling resistance due to seeds hydropriming in the presence of salicylic acid, jasmonic acid, 24-epibrassinolide followed exposition of seeds to short-term heat shock treatment. The seeds were hydroprimed at 25 °C in limited amounts of water or solution of salicylic or jasmonic acid at 10^?2, 10^?3 and 10^?4 M concentration, 24-epibrassinolide at 10^?6, 10^?8 and 10^?10 M concentration. The seeds were incubated for 2 days, subjected to short-term heat shock (45 °C, 2 h) and chilled for 21 days at 0 °C. Sunflower chilling susceptibility and physiological responses were evaluated according to the inhibition of radicle growth, the inhibition of the number of lateral roots formation, the activity of catalase and changes in soluble carbohydrates in seedlings developing for 72 h at 25 °C. Hydropriming and short-term heat shock application explicitly reduced inhibition of roots as well as lateral roots development by allowing the germinating seeds to recover from the growth-inhibiting effects of chilling. Seeds hydropriming in solutions containing salicylic acid, jasmonic acid and 24-epibrassinolide followed heat shock treatment additionally promoted the activity of catalase and sugars metabolism, which stimulated seedlings development and alleviated the decrease of F _v/F _m caused by chilling conditions. These beneficial effects contributed to increased resistance of sunflower seedlings to chilling stress. The present study demonstrated that the most profitable effect on reducing negative effect of chilling may be achieved by short-term heat shock applied during hydropriming in water supplemented with 24-epiBL (10^?8 and 10^?10 M) or salicylic acid (10^?3 and 10^?4 M).     

Diversity of low-molecular weight organic acids synthesized by <Emphasis Type="Italic">Salix</Emphasis> growing in soils characterized by different Cu,Pb and Zn concentrations

The aim of the study was to evaluate the biosynthesis and exudation of 10 low-molecular weight organic acids (LMWOAs) into the rhizosphere with a simultaneous analysis of the acid contents in the roots and leaves of 9 Salix taxa growing on two experimental areas, differing in their concentrations of copper (Cu), lead (Pb) and zinc (Zn) in the soil (Area 1—low, Area 2—high concentration). The obtained results reveal a significant difference in the phytoextraction of the tested Salix taxa for the analysed metals in both areas. The highest contents of Cu, Pb and Zn were observed for all Salix collected from Area 2, especially in S. × smithiana roots (116 ± 8.76, 87.84 ± 7.30 and 203.42 ± 14.62 mg kg^?1 DW, respectively). The results obtained in Area 2 also revealed acidification of the rhizosphere and a higher concentration of acids, mainly oxalic, malic, malonic, acetic and citric acids. Contents of oxalic, malic, acetic and citric acids increased in the roots of Salix taxa from Area 2, while in the leaves formic and succinic acids were also present. S. × smithiana was the taxon with the highest concentration of acids in the rhizosphere and roots (73.48 ± 6.77 and 49.79 ± 2.65 μM 100 g^?1 DW, respectively), while in leaves a higher content was observed for S. alba and S. viminalis ‘PR’ taxa (78.12 ± 3.95 and 71.12 ± 3.75 μM 100 g^?1 DW, respectively).     

The effects of exogenous ABA applied to maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) roots on plant responses to chilling stress

This work aimed to discuss the effects of exogenous abscisic acid (ABA) on the root growth regulation of maize seedlings under chilling stress. The roots of the maize cultivar Zhengdan 958 were irrigated with ABA (10^?7, 10^?6, 10^?5 and 10^?4 M) at the third true leaf stage under chilling duration (0, 2, 4, 6, and 8 days). The biomass, the phenylalanine ammonia lyase (PAL), and polyphenol oxidase (PPO) enzyme activities, total phenolic and flavonoid contents, the ferric reducing ability of plasma (FRAP) antioxidant capacity, and 2,2-azinobis (3-ethlbenzothiazo-line-6-sulfonic acid) diammonium salt radical (ABTS^·+) scavenging capacity of the roots of maize seedlings were measured after the treatment. The results showed that appropriate concentrations of exogenous ABA effectively enhanced root biomass, increased PAL and PPO enzyme activities, and significantly increased total phenolic contents and flavonoid contents. Moreover, the ABA markedly improved the FRAP antioxidant capacity and ABTS^·+ scavenging capacity under low-temperature stress. These results indicate that ABA-treated maize seedlings are resistant to chilling stress and that the optimum concentration of ABA is 10^?5 M. Exogenous applications of ABA have a concentration effect in alleviating chilling stress, in which low concentrations have a promoting effect and high concentrations have an inhibiting effect.     

Impact of <Emphasis Type="Italic">Heterobasidion</Emphasis> root-rot on fine root morphology and associated fungi in <Emphasis Type="Italic">Picea abies</Emphasis> stands on peat soils

We examined differences in fine root morphology, mycorrhizal colonisation and root-inhabiting fungal communities between Picea abies individuals infected by Heterobasidion root-rot compared with healthy individuals in four stands on peat soils in Latvia. We hypothesised that decreased tree vitality and alteration in supply of photosynthates belowground due to root-rot infection might lead to changes in fungal communities of tree roots. Plots were established in places where trees were infected and in places where they were healthy. Within each stand, five replicate soil cores with roots were taken to 20 cm depth in each root-rot infected and uninfected plot. Root morphological parameters, mycorrhizal colonisation and associated fungal communities, and soil chemical properties were analysed. In three stands root morphological parameters and in all stands root mycorrhizal colonisation were similar between root-rot infected and uninfected plots. In one stand, there were significant differences in root morphological parameters between root-rot infected versus uninfected plots, but these were likely due to significant differences in soil chemical properties between the plots. Sequencing of the internal transcribed spacer of fungal nuclear rDNA from ectomycorrhizal (ECM) root morphotypes of P. abies revealed the presence of 42 fungal species, among which ECM basidiomycetes Tylospora asterophora (24.6 % of fine roots examined), Amphinema byssoides (14.5 %) and Russula sapinea (9.7 %) were most common. Within each stand, the richness of fungal species and the composition of fungal communities in root-rot infected versus uninfected plots were similar. In conclusion, Heterobasidion root-rot had little or no effect on fine root morphology, mycorrhizal colonisation and composition of fungal communities in fine roots of P. abies growing on peat soils.     

Efficacy of bacterial seed treatments for the control of <Emphasis Type="Italic">Fusarium verticillioides</Emphasis> in maize

Maize colonization by the phytopathogenic fungi Fusarium verticillioides leads to economical and food quality losses and also implicates potential health risks. In order to control this fungal species different strategies are being considered. In the present work we investigated the in situ effects of the use of two in vitro proven bacterial biological control agents against Fusarium verticillioides, using maize seedlings grown in greenhouse conditions. The treatment of maize seeds with Fusarium verticillioides 10⁷ spores ml^?1 negatively affected the length of the stem and the weight of the root systems of resultant seedlings, and also reduced the numbers of non-rhizospheric organisms with ammonification and nitrification activities. The addition of Bacillus amyloliquefaciens or Microbacterium oleovorans, at a concentration of 10⁹ colony forming units ml^?1, to the seeds significantly reduced Fusarium verticillioides count at the root inner tissues of resultant seedlings. When testing the impact of bacterial treatments on soil populations, no alterations with respect to control numbers of organisms with nitrification, ammonification or cellulolytic potentials were observed. Culturable bacterial richness and diversity calculated at the rhizoplane and root inner tissues of maize seedlings neither changed in bacterized treatments when compared to control treatment. Our study showed that the Fusarium verticillioides in vitro proven antagonists, Bacillus amyloliquefaciens and Microbacterium oleovorans (at 10⁹ colony forming units ml^?1), were also effective at greenhouse conditions without causing major changes in culturable rhizospheric and endophytic microbial richness and diversity.     

Consequences of inoculation with native arbuscular mycorrhizal fungi for root colonization and survival of <Emphasis Type="Italic">Artemisia tridentata</Emphasis> ssp. <Emphasis Type="Italic">wyomingensis</Emphasis> seedlings after transplanting

In arid environments, the propagule density of arbuscular mycorrhizal fungi (AMF) may limit the extent of the plant–AMF symbiosis. Inoculation of seedlings with AMF could alleviate this problem, but the success of this practice largely depends on the ability of the inoculum to multiply and colonize the growing root system after transplanting. These phenomena were investigated in Artemisia tridentata ssp. wyomingensis (Wyoming big sagebrush) seedlings inoculated with native AMF. Seedlings were first grown in a greenhouse in soil without AMF (non-inoculated seedlings) or with AMF (inoculated seedlings). In spring and fall, 3-month-old seedlings were transplanted outdoors to 24-L pots containing soil from a sagebrush habitat (spring and fall mesocosm experiments) or to a recently burned sagebrush habitat (spring and fall field experiments). Five or 8 months after transplanting, colonization was about twofold higher in inoculated than non-inoculated seedlings, except for the spring field experiment. In the mesocosm experiments, inoculation increased survival during the summer by 24 % (p?=?0.011). In the field experiments, increased AMF colonization was associated with increases in survival during cold and dry periods; 1 year after transplanting, survival of inoculated seedlings was 27 % higher than that of non-inoculated ones (p?<?0.001). To investigate possible mechanisms by which AMF increased survival, we analyzed water use efficiency (WUE) based on foliar ¹³C/¹²C isotope ratios (δ ¹³C). A positive correlation between AMF colonization and δ ¹³C values was observed in the spring mesocosm experiment. In contrast, inoculation did not affect the δ ¹³C values of fall transplanted seedlings that were collected the subsequent spring. The effectiveness of AMF inoculation on enhancing colonization and reducing seedling mortality varied among the different experiments, but average effects were estimated by meta-analyses. Several months after transplanting, average AMF colonization was in proportion 84 % higher in inoculated than non-inoculated seedlings (p?=?0.0042), while the average risk of seedling mortality was 42 % lower in inoculated than non-inoculated seedlings (p?=?0.047). These results indicate that inoculation can increase AMF colonization over the background levels occurring in the soil, leading to higher rates of survival.     

Species and Genotype Effects of Bioenergy Crops on Root Production,Carbon and Nitrogen in Temperate Agricultural Soil

Bioenergy crops have a secondary benefit if they increase soil organic C (SOC) stocks through capture and allocation below-ground. The effects of four genotypes of short-rotation coppice willow (Salix spp., ‘Terra Nova’ and ‘Tora’) and Miscanthus (M.?×?giganteus (‘Giganteus’) and M. sinensis (‘Sinensis’)) on roots, SOC and total nitrogen (TN) were quantified to test whether below-ground biomass controls SOC and TN dynamics. Soil cores were collected under (‘plant’) and between plants (‘gap’) in a field experiment on a temperate agricultural silty clay loam after 4 and 6 years’ management. Root density was greater under Miscanthus for plant (up to 15.5 kg m^?3) compared with gap (up to 2.7 kg m^?3), whereas willow had lower densities (up to 3.7 kg m^?3). Over 2 years, SOC increased below 0.2 m depth from 7.1 to 8.5 kg m^?3 and was greatest under Sinensis at 0–0.1 m depth (24.8 kg m^?3). Miscanthus-derived SOC, based on stable isotope analysis, was greater under plant (11.6 kg m^?3) than gap (3.1 kg m^?3) for Sinensis. Estimated SOC stock change rates over the 2-year period to 1-m depth were 6.4 for Terra Nova, 7.4 for Tora, 3.1 for Giganteus and 8.8 Mg ha^?1 year^?1 for Sinensis. Rates of change of TN were much less. That SOC matched root mass down the profile, particularly under Miscanthus, indicated that perennial root systems are an important contributor. Willow and Miscanthus offer both biomass production and C sequestration when planted in arable soil.     

Abscisic Acid-Induced Starch Accumulation in Bioenergy Crop Duckweed <Emphasis Type="Italic">Spirodela polyrrhiza</Emphasis>

Spirodela polyrrhiza, a fast-growing duckweed with high starch and low lignin content, shows promise as a feedstock for bioenergy. Abscisic acid (ABA) is a biological hormone that controls plant growth and stress response. The effects of different ABA concentrations (0, 1.0 × 10^?5, 1.0 × 10^?4, 1.0 × 10^?3, 1.0 × 10^?2, and 1.0 × 10^?1 mg/L) on duckweed biomass growth, carbon dioxide fixation, formation of photosynthetic pigments (Chlorophyll a (Chla), Chlorophyll b (Chlb), and carotenoids), the activities of soluble starch synthase (SSS) and starch branching enzyme (SBE), and the starch content of biomass were investigated in this study. ABA at concentrations lower than 1.0 × 10^?3 mg/L promoted carbon dioxide fixation, whereas it inhibited carbon dioxide fixation at concentrations over 1.0 × 10^?3 mg/L. ABA enhanced SSS and SBE activities at concentrations lower than 1.0 × 10^?2 mg/L. ABA treatment increased the content of Chla, Chlb, and carotenoids and resulted in the enhancement of starch content. Chla content gradually increased with the increasing concentration of ABA (1.0 × 10^?5 to 1.0 × 10^?2 mg/L). After culturing for 10 days, starch content in 1.0 × 10^?2 mg/L ABA medium reached 35.3% of dry weight (DW), which was the highest level in this study. This suggests that there is a great potential to develop a technology to increase starch accumulation in duckweed which can be used as an alternative to corn, sugarcane, or other food crops as a starch source.     

24-Epibrassinolide reduces stress in nematode-infected tomato (<Emphasis Type="Italic">Solanum lycopersicum</Emphasis> L.) plants cultured <Emphasis Type="Italic">in vitro</Emphasis>

24-Epibrassinolide (EBL) is considered the most probable brassinosteroid (BR) candidate that could be used for practical application in agriculture. EBL-induced stress-protective properties were evaluated in in vitro-grown tomato (Solanum lycopersicum L.) varieties Pusa Ruby (susceptible to nematodes) and PNR-7 (resistant to nematodes) during nematode pathogenesis. Sterilized tomato seeds treated with 10^?11, 10^?9, or 10^?7 M EBL and germinated in vitro were inoculated with second-stage juveniles of Meloidogyne incognita [(Kofoid and White) Chitwood]. Whole plant analyses of morphological and biochemical parameters 7 d after inoculation showed significant improvements in plant growth and development for both varieties and a highly significant reduction in the number of galls in the susceptible variety. Increased specific activities of antioxidative enzymes (catalase, ascorbate peroxidase, glutathione reductase, glutathione peroxidase, guaiacol peroxidase, and superoxide dismutase) were observed in EBL-treated seedlings of both varieties, but increases were higher in the resistant variety. A highly significant increase in antioxidants (ascorbic acid content, total flavonoid content, total glutathione content, and total phenolic content) was observed in EBL-treated Pusa Ruby seedlings, whereas in PNR-7, significant increases were found except for total flavonoid content, which increased non-significantly. Confocal microscopic images showed amelioration of stress in roots of EBL-treated seedlings as indicated by the decrease in level of green fluorescence in them as compared to untreated and nematode-inoculated roots.     

Evaluation of a field experiment for the conservation of a <Emphasis Type="Italic">Magnolia stellata</Emphasis> stand using clear-cutting

Magnolia stellata is a rare subcanopy tree species that grows in secondary forests in warm temperate zones. It is now endangered due to habitat degradation by vegetation succession. In an attempt to improve the habitat, a 30 m?×?10 m plot (0.03 ha) was set up with all vegetation including M. stellata being clear-cut in January 2012. The number of sprouts increased for 1–2 years after clear-cutting and then gradually decreased or remained constant. Five years after clear-cutting, the numbers of individuals and stems, and the total basal area (BA), were 87.0, 165.5 and 3.2%, respectively, of the values before clear-cutting. BA was highest for Ilex pedunculosa, followed by M. stellata and Hydrangea paniculata. Some sprouted individuals of M. stellata produced flower buds in the second year after clear-cutting, and flowered and fruited in the spring and summer of the third year, respectively. The densities of potential canopy species were 18,533 ha^?1 (height >?0.5 m) and 7,267 ha^?1 (height >?1.2 m), vastly exceeding the value of the criterion for successful natural regeneration after clear-cutting of warm temperate forests in the region (3,000 ha^?1). Based on this criterion, it is thus considered that the natural regeneration has reached completion. However, 45.1% (height >?0.5 m) and 95.5% (height >?1.2 m) of M. stellata individuals were regenerated by sprouting. Further research is needed into how individuals, regenerated from seedlings, develop and reach sexual maturity, and how successive generations change.     

Early plant growth and biochemical responses induced by <Emphasis Type="Italic">Azospirillum brasilense</Emphasis> Sp245 lipopolysaccharides in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) seedlings are attenuated by procyanidin B2

This study analyzes the effects of procyanidin B2 on early wheat plant growth and plant biochemical responses promoted by lipopolysaccharides (LPS) derived from the rhizobacteria Azospirillum brasilense Sp245. Measurements of leaf, root length, fresh weight, and dry weight showed in vitro plant growth stimulation 4 days after treatment with A. brasilense as well as LPS. Superoxide anion (O₂ ^·?) and hydrogen peroxide (H₂O₂) levels increased in seedling roots treated with LPS (100 μg mL^?1). The chlorophyll content in leaf decreased while the starch content increased 24 h after treatment in seedling roots. The LPS treatment induced a high increase in total peroxidase (POX) (EC 1.11.1.7) activity and ionically bound cell wall POX content in roots, when compared to respective controls. Early plant growth and biochemical responses observed in wheat seedlings treated with LPS were inhibited by the addition of procyanidin B2 (5 μg mL^?1), a B type proanthocyanidin (PAC), plant-derived polyphenolic compound with binding properties of LPS. All results suggest first that the ionically bound cell wall POX enzymes could be a molecular target of A. brasilense LPS, and second that the recognition or association of LPS by plant cells is required to activate plant responses. This last event could play a critical role during plant growth regulation by A. brasilense LPS.     

Ultraviolet radiation reduces lichen growth rates

The symbiosis of Leguminosae with arbuscular mycorrhizal fungi (AMF) and N₂-fixing nodulating bacteria (NFNB) can occur simultaneously, forming a tripartite symbiosis. In particular, AMF can colonize root nodules, although this interaction is not yet well elucidated, especially with regard to nodule activity and to the influence of external factors, such as biostimulants. In this study, we hypothesized that the application of the flavonoid formononetin, used to stimulate root colonization by native AMF, increases the AMF colonization of soybean (Glycine max) root nodules, especially under low availability of phosphorus (P). To test this hypothesis, we performed a field experiment in randomized blocks in a 4 × 3 factorial design, with 4 treatments of formononetin (0, 0.46, 0.92 and 1.84 g per kg seed) and 3 of P (0, 60 and 120 kg ha^?1) with 5 replicates. Nodules and roots were collected during the R2 stage (full flowering) and evaluated with respect to AMF colonization. Formononetin stimulated mycorrhizal fungi colonization of active nodules, especially when no P was applied, as also observed for AMF root colonization; however, it had no effect with 60 and 120 kg P ha^?1. Thus, the application of formononetin increases surface AMF colonization of active nodules and roots, but its effect disappears with an increase in P and the inactivity of the nodule.     

Effects of 28-homobrassinoloid on key physiological attributes of <Emphasis Type="Italic">Solanum lycopersicum</Emphasis> seedlings under cadmium stress: Photosynthesis and nitrogen metabolism

Heavy metal accumulation due to environmental pollution, especially in agricultural ecosystem can cause serious deterioration of crop yield and quality. In present study we assessed the effect of exogenous 28-homobrassinoloid (HBL; 10^?8 M) on growth, photosynthesis, indices of chlorophyll a fluorescence and nitrogen metabolism in Solanum lycopersicum seedlings grown under two doses (Cd₁: 3 mg kg^?1 sand and Cd₂: 9 mg kg^?1 sand) of cadmium. Accumulation of Cd in root tissues was considerably higher than shoot hence, Cd declined the growth, pigment contents, and photosynthetic O₂ yield in its concentration dependent manner. Chlorophyll a fluorescence due to Cd stress was negatively affected as shown by decreased Q_A ^? reoxidation kinetics: φP₀, ψ₀, φE₀ and PI__ABS and increased energy flux parameters: ABS/RC, TR₀/RC, ET₀/RC and DI₀/RC. HBL application under Cd stress improved the photochemistry of photosystem II (PS II) by affecting these parameters positively. Treatment of Cd in test seedlings resulted into significant decrease in nitrate reductase, nitrite reductase, glutamine synthetase and glutamate synthase activities, and induced enhancing effect on ammonium content and glutamate dehydrogenase activity. Exogenous HBL treatment alleviated the negative effect of Cd on growth, photosynthesis, contents of protein, carbohydrate and inorganic nitrogen and nitrogen assimilating enzymes. The data indicate that exogenous HBL protects the test seedlings during the early growth phase against Cd phytotoxicity by regulating Cd accumulation in tissues and two key metabolic processes; photosynthesis and nitrogen metabolism.     

Impacts of simulated drought stress and artificial damage on concentrations of flavonoids in <Emphasis Type="Italic">Jatropha curcas</Emphasis> (L.), a biofuel shrub

We studied the possible roles of flavonoids in the antioxidant and antiherbivore chemistry in Jatropha curcas (L.), a Latin American shrub that holds great potential as a source of biofuel. Changes in flavonoid concentrations in the leaves of J. curcas seedlings exposed to artificial damage and to different rainfall patterns were assessed by applying a 3²-factorial experiment in a greenhouse. The concentrations of different flavonoids in the leaves of seedlings were significantly affected by interaction effects of artificial damage, drought stress and age of the seedling. The highest flavonoid concentrations were obtained in seedlings imposed to the highest percentage of artificial damage (50 %) and grown under extreme drought stress (200 mm year^?1). In this treatment combination, flavonoid concentrations were three-fold as compared to seedlings exposed to the same level of artificial damage but grown in 1900 mm year^?1 rainfall application. Without artificial damage, the concentration of flavonoids in the seedlings grown in 200 mm year^?1 rainfall application was still two-fold compared to seedlings grown in higher (>800 mm year^?1) rainfall applications. Thus, the observed flavonoid concentration patterns in the leaves of J. curcas seedlings were primarily triggered by drought stress and light rather than by artificial damage, suggesting that drought causes oxidative stress in J. curcas.     

Efficient direct plant regeneration from immature leaf roll explants of sugarcane (<Emphasis Type="Italic">Saccharum officinarum</Emphasis> L.) using polyamines and assessment of genetic fidelity by SCoT markers

A rapid and efficient method for in vitro direct plant regeneration from immature leaf roll explants of Saccharum officinarum L. (sugarcane) cv. Co 86032 was developed by the application of exogenous polyamines (PA). The effect of explant source from apical meristems and pre-culture of explants in the dark on shoot regeneration was studied. Adventitious shoot regeneration occurred on the proximal regions of immature leaf roll explants when pre-incubated in the dark for 2 wk and the regeneration response was decreased from the middle to distal end. A higher number of direct shoots (130 primary shoots explant^?1) and multiple shoots (657 secondary shoots explant^?1), were obtained with a combination of spermidine (103.27 μM), spermine (49.42 μM), and putrescine (31.04 μM) along with plant growth regulators. Shoot induction was increased up to twofold and multiplication was increased up to threefold in the medium supplemented with PA. Profuse rooting was observed in putrescine (93.12 μM), spermidine (68.84 μM), and spermine (24.71 μM), with mean number of 57 roots. A twofold increase in the number of roots was observed in medium supplemented with PA with respect to control cultures, which facilitated the successful transplantation and acclimatization process of in vitro propagated sugarcane plants. Histology and scanning electron microscopy analyses supported adventitious direct shoot regeneration from immature leaf roll explants. The genetic stability of in vitro regenerated plants was confirmed using start codon targeted polymorphism marker system.     

Improved production of <Emphasis Type="Italic">Oroxylum indicum</Emphasis> (L.) Kurz by altering the salts of the medium

The present study was conducted to test the effects of KNO₃, KH₂PO₄, and CaCl₂ on shoot multiplication, root proliferation, and accumulation of phytochemicals in in vitro cultures of Oroxylum indicum. The results indicate that modifying the MS salt formulation in relation to particular inorganic nutrients highly affected shoot multiplication, root proliferation, and accumulation of flavonoids in in vitro cultures. A concentration of 0.60 g L^?1 CaCl₂ resulted in the highest frequency of shoot regeneration (5.6 shoots per explant). A concentration of 0.40 g L^?1 CaCl₂ resulted in the highest frequency of root regeneration (7.8 roots per shoot). Modifications of the concentrations of inorganic salts were also found to be advantageous for production media for both multiple shoots and shoot-derived root in vitro cultures. Multiple shoots generated on shoot induction medium with a concentration of 0.60 g L^?1 CaCl₂ and roots generated on root induction medium with a concentration of 1.5 g L^?1 KNO₃ yielded about a five times higher flavonoid level than cultures generated on control medium respectively.     

The benefits of foliar inoculation with <Emphasis Type="Italic">Azospirillum brasilense</Emphasis> in soybean are explained by an auxin signaling model

Azospirillum sp. is one of the most studied genera of plant growth-promoting rhizobacteria (PGPR). The ability of Azospirillum sp. to promote plant growth has been associated with its ability to produce several phytohormones, such as auxins, gibberellins and cytokinins, but mainly indole-3-acetic acid (IAA). It has been propoosed that the production of IAA explains the positive effects of co-inoculation with Azospirillum sp. on the rhizobia-legume symbiosis. In this study, we constructed an IAA-deficient mutant of A. brasilense Az39 (ipdC ^? ) by using a restriction-free cloning method. We inoculated soybean seeds with 1·10⁶ cfu·seed^?1 of Bradyrhizobium japonicum E109 and co-inoculating leaves at the V3 stage with 1·10⁸ cfu.plant^?1 of A. brasilense Az39 wt or ipdC ^? or inoculated leaves with 20 μg.plant^?1 synthetic IAA. The results confirmed soybean growth promotion as there was increased total plant and root length, aerial and root dry weight, number of nodules on the primary root, and an increase in the symbiosis established with B. japonicum E109. Nodule weight also increased after foliar co-inoculation with the IAA- producer A. brasilense Az39. The exogenous application of IAA decreased aerial and root length, as well as the number of nodules on primary roots in comparison with the Az39 wt strain. These results allow us to propose a biological model of response to foliar co-inoculation of soybean with IAA-producing rhizobacteria. This model clearly shows that both the presence of microorganism as part of the colonization process and the production of IAA in situ are co-responsible, via plant signaling molecules, for the positive effects on plant growth and symbiosis establishment.     

Endophytic <Emphasis Type="Italic">Alcaligenes</Emphasis> Isolated from Horticultural and Medicinal Crops Promotes Growth in Okra (<Emphasis Type="Italic">Abelmoschus</Emphasis><Emphasis Type="Italic">esculentus</Emphasis>)

The potential of endophytic bacteria to act as biofertilizers and bioprotectants has been demonstrated, and considerable progress has been made in explaining their role in plant protection. In the present study, three endophytic bacterial strains (BHU 12, BHU 16 isolated from the leaves of Abelmoschus esculentus, and BHU M7 isolated from the leaves of Andrographis paniculata) were used which displayed high sequence similarity to Alcaligenes faecalis. The biofilm formation ability of these endophytic strains in the presence of okra root exudates confirms their chemotactic ability, an initial step for successful endophytic colonization. Further, reinoculation of spontaneous rifampicin-tagged mutants into okra seedlings revealed a CFU count above 10⁵ cells g^?1 of all three endophytic strains in root samples during the first 15 days of plant growth. The CFU count increased up to 10¹³ by 30 days of plant growth, followed by a gradual decline to approximately 10¹⁰ cells g^?1 at 45 days of plant growth. Systemic endophytic colonization was further supported by 2, 3, 5-triphenyl tetrazolium chloride staining and fluorescence imaging of ds-RED expressing conjugants of the endophytic strains. The strains were further assessed for their plausible in vivo and in vitro plant growth-promoting and antagonistic abilities. Our results demonstrated that the endophytic strains BHU 12, BHU 16, and BHU M7 augmented plant biomass by greater than 40 %. Root and shoot lengths of okra plants when primed by BHU 12, BHU 16, and BHU M7 increased up to 34 and 14.5 %, respectively. The endophytic isolates also exhibited significant in vitro antagonistic potential against the collar rot pathogen Sclerotium rolfsii. In summary, our results demonstrate excellent potential of the three endophytic bacterial strains as biofertilizers and biocontrol agents, indicating the possibility for use in sustainable agriculture.