Phoma medicaginis Isolate Differences Determine Disease Severity and Phytoestrogen Production in Annual Medicago spp.

Phoma black stem and leaf spot disease of annual Medicago spp., caused by Phoma medicaginis, not only can devastate forage and seed yield but can reduce herbage quality by inducing production of phytoestrogens (particularly coumestrol and 4'-O-methylcoumestrol), which can also reduce the ovulation rates of animals grazing infected forage. We determined the consequent phytoestrogen levels on three different annual Medicago species/cultivars (Medicago truncatula cultivar Cyprus, Medicago polymorpha var. brevispina cultivar Serena, and Medicago murex cultivar Zodiac) after inoculation with 35 isolates of P. medicaginis. Across the isolate × cultivar combinations, leaf disease incidence, petiole/stem disease incidence, leaf disease severity, petiole disease severity, and leaf yellowing severity ranged up to 100, 89.4, 100, 58.1, and 61.2%, respectively. Cultivars Cyprus and Serena were the most susceptible and cultivar Zodiac was the most resistant to P. medicaginis. Isolates WAC3653, WAC3658, and WAC4252 produced the most severe disease. Levels of phytoestrogens in stems ranged from 25 to 1,995 mg/kg for coumestrol and from 0 to 418 mg/kg for 4'-O-methylcoumestrol. There was a significant positive relationship of disease incidence and severity parameters with both coumestrol and 4'-O-methylcoumestrol contents, as noted across individual cultivars and across the three cultivars overall, where r = 0.39 and 0.37 for coumestrol and 4'-O-methylcoumestrol, respectively (P < 0.05). Although cultivar Serena was most susceptible to P. medicaginis and produced the highest levels of phytoestrogens in the presence of P. medicaginis, cultivar Zodiac contained the highest levels of phytoestrogens in comparison with other cultivars in the absence of P. medicaginis. There was a 15-fold increase in coumestrol in cultivar Serena but only a 7-fold increase in cultivar Zodiac from infection of P. medicaginis. The study highlights that the intrinsic ability of a particular cultivar to produce phytoestrogens in the absence of the pathogen, and its comparative ability to produce phytoestrogens in the presence of the P. medicaginis, are both important and highly relevant to developing new annual Medicago spp. cultivars that offer improved disease resistance and better animal reproductive outcomes.

Effects of plant growth-promoting bacteria on EDTA-assisted phytostabilization of heavy metals in a contaminated calcareous soil.

The objective of this research was to determine the combined effects of ethylenediaminetetraacetic acid (EDTA) and plant growth-promoting rhizobacteria (PGPR) on the phytostabilization of Cd, Pb, and Zn by corn and chemical fractionation of these elements in soil. Three heavy metal-resistant bacteria (P18, P15, and P19) were selected. All strains, belonging to the fluorescent pseudomonads, exhibited plant growth-promoting properties, including phosphorus solubilization and production of siderophore, indole acetic acid, and 1-aminocyclopropane-1-carboxylic acid deaminase. Applying EDTA individually or in combination with bacterial strains (P18 and P15) significantly increased shoot biomass. The highest dry shoot biomass was recorded in the combined treatment of EDTA and P15-inoculated pots. Application of EDTA in PGPR-inoculated pots increased concentrations of heavy metals in corn shoots and roots compared to the control. The highest concentration of Zn in corn root and shoot was observed in P15 + EDTA treatment, which were 2.0-fold and 1.3-fold higher than those in the untreated soil. Results of chemical speciation showed that the co-application of EDTA and fluorescent pseudomonads strains increased the bioavailability of Zn, Pb, and Cd by their redistribution from less soluble fractions to water-soluble forms. It was concluded that bacterial inoculation could improve the efficiency of EDTA in phytostabilization of heavy metals from multi-metal contaminated soils.