Characterization of a plant-growth-promoting non-nodulating endophytic bacterium (Stenotrophomonas maltophilia) from the root nodules of Mucuna utilis var. capitata L. (Safed Kaunch).

Nonrhizobial root nodule endophytic bacteria are known to have beneficial effects on host plants and are also considered contaminants or opportunists. They grow either individually or as a co-occupant of the root nodules of legumes. In this study, a nonrhizobial endophytic bacterial strain was isolated from the root nodules of the medicinal legume Mucuna utilis var. capitata L.; phenotypic, genotypic, and agricultural characterization was performed using a HiMedia kit and 16S rRNA sequencing. This strain showed tremendous seedling growth potential (30%), compared with the control, as well as a strong antagonistic nature against the plant pathogenic fungus Fusarium udum when plant growth parameters were analyzed. The strain, identified by 16S rRNA as Stenotrophomonas maltophilia, showed a multitude of plant-growth-promoting attributes both direct (IAA, phosphate solubilization) and indirect (ACC deaminase, siderophore) and enhanced the growth of host plant in field trials. This is the first report of the plant-growth-promoting potential of this endophytic bacterium from the nodules of M. utilis var. capitata L.; hence, it has potential for use in various biotechnological applications in various industries.

Effects of Rhizophagus clarus and P availability in the tolerance and physiological response of Mucuna cinereum to copper.

Arbuscular mycorrhizal fungi (AMF) improve plant ability to uptake P and tolerate heavy metals. This study aimed to evaluate the effect of available P and the inoculation of Rhizophagus clarus in a Cu-contaminated soil (i) on the activity of acid phosphatases (soil and plant), the presence of glomalin, and (ii) in the biochemical and physiological status of Mucuna cinereum. A Typic Hapludalf soil artificially contaminated by adding 60 mg kg-1 Cu was used in a 3 × 2 factorial design with three replicates. Treatments consisted of three P levels: 0, 40, and 100 mg kg-1 P. Each P treatment level was inoculated (+AMF)/non-inoculated (-AMF) with 200 spores of R. clarus per pot, and plants grown for 45 days. The addition of at least 40 mg kg-1 P and the inoculation of plants with R. clarus proved to be efficient to reduce Cu phytotoxicity and increase dry matter yield. Mycorrhization and phosphate fertilization reduced the activity of enzymes regulating oxidative stress (SOD and POD), and altered the chlorophyll a fluorescence parameters, due to the lower stress caused by available Cu. These results suggest a synergism between the application of P and the inoculation with R. clarus, favoring the growth of M. cinereum in a Cu-contaminated soil. This study shows that AMF inoculation represents an interesting alternative to P fertilization to improve plant development when exposed to excess Cu.

Determination of aflatoxins in medicinal plants by high-performance liquid chromatography-tandem mass spectrometry.

PURPOSE: The intention of the proposed work is to study the presence of the aflatoxins B1, B2, G1 and G2 in medicinal plants, namely Mucuna pruriens, Delphinium denudatum and Portulaca oleraceae. METHODOLOGY: The aflatoxins were extracted, purified by immunoaffinity column chromatography and analysed by high-performance liquid chromatography-tandem quadrupole mass spectrometry with electrospray ionisation (HPLC-MS/MS). Fungal count was carried out in PDA media. RESULTS: A good linear relationship was found for AFB1, AFB2, AFG1 and AFG2 at 1-10 ppb (r>0.9995). The analyte accuracy under three different spiking levels was 86.7-108.1 %, with low per cent relative standard deviations in each case. The aflatoxins can be separated within 5 to7 min using an Agilent XDB C18-column. We found that AFB1 and AFB2 were in trace amounts below the detection limit in M. pruriens whilst they were not detected in D. denudatum. P. oleraceae was found to be contaminated with AFB1 and AFB2. AFG1 and AFG2 were not detected in M. pruriens, P. oleraceae and were below the detection limit in D. denudatum. This was consistent with very low numbers of fungal colonies observed after 6 hr of incubation. CONCLUSION: The analytical method developed is simple, precise, accurate, economical and can be effectively used to determine the aflatoxins in medicinal plants and therefore to control the quality of products. The aflatoxin levels in the plant extracts examined were related to the minimal fungal load in the medicinal plants examined.

Practical use of CMC-amended rhizobial inoculant for Mucuna pruriens cultivation to enhance the growth and protection against Macrophomina phaseolina.

In many parts of the world Mucuna pruriens is used as an important medicinal, forage and green manure crop. In the present investigation the effect of the addition of CMC in carrier during development of bioformulation on shelflife, plant growth promotive and biocontrol activity against Macrophomina phaseolina was screened taking M. pruriens as a test crop. Ensifer meliloti RMP6(Ery+Kan+) and Bradyrhizobium sp. BMP7(Tet+Kan+) (kanamycin resistance engineered by Tn5 transposon mutagenesis) used in the study showed production of siderophore, IAA, solubilizing phosphate and biocontrol of M. phaseolina. RMP6(Ery+Kan+) also showed ACC deaminase activity. The survival of both the strains in sawdust-based bioformulation was enhanced with an increase in the concentration of CMC from 0 to 1%. At 0% CMC Bradyrhizobium sp. BMP7(Tet+Kan+) showed more increase in nodule number/plant (500.00%) than E. meliloti RMP6(Ery+Kan+) (52.38%), over the control in M. phaseolina-infested soil. There was 185.94% and 59.52% enhancement in nodule number/plant by RMP6(Ery+Kan+) and BMP7(Tet+Kan+) with an increase in the concentration of CMC from 0% to 1% in the bioformulations. However further increase in concentration of CMC did not result in enhancement in survival of either the strains or nodule number/plant.