Fazli Mabood

The class IId bacteriocin thuricin-17 increases plant growth.

The mechanisms by which many plant growth promoting rhizobacteria (PGPR) affect plants are unknown. We recently isolated a rhizosphere bacterium (Bacillus thuringiensis NEB17), that promotes soybean growth and screened the liquid growth medium in which it grew for plant growth stimulating materials. We have also shown that it produces a bacteriocin (named by us as thuricin-17 and a member of the recently described class IId bacteriocins). Here we show that application of this bacteriocin to leaves (spray) or roots (drench) directly stimulates the growth of both a C3 dicot (soybean) and a C4 monocot (corn). This growth stimulation is similar in nature to that previously seen when plants are treated with Nod factors. Strain NEB17 contains three copies of the gene for thuricin 17 that code for identical amino acid sequences. These two lines of evidence suggest that the dual functions of these proteins may have constrained their evolution. This is the first report of direct plant growth enhancement by a bacteriocin.

Induction of defense-related enzymes in soybean leaves by class IId bacteriocins (thuricin 17 and bacthuricin F4) purified from Bacillus strains.

We have recently discovered a new class of bacteriocin (class IId) which stimulates plant growth in a way similar to Nod factors. Nod factors have been shown to provoke aspects of plant disease resistance. We investigated the effects of bacteriocins [thuricin 17 (T17) and bacthuricin F4 (BF4)] on the activities of phenylalanine ammonia lyase (PAL), guaiacol peroxidase (POD), ascorbate peroxidase (APX), superoxide dismutase (SOD), and polyphenol oxidase (PPO). Bacteriocin solutions were fed into the cut stems of soybean (Glycine max L. Merr. cv. OAC Bayfield) seedlings at the first trifoliate stage. PAL activity in T17 treated leaves was the highest at 72h after treatment and was 75.5% greater than the control at that time. At 72h after treatment POD activities in T17 and BF4 treated leaves increased by 72.7 and 91.3%, respectively, as compared with the control treatment. APX activity was 52.3 and 49.6% respectively, greater than the control in T17 and BF4 treated leaves at 72h after treatment. SOD activity in T17 treated leaves was the highest at 72h after treatment and was 26.0% greater than the control at that time. SOD activity was 70.5 and 60.2% greater, respectively, than the control in T17 and BF4 treated leaves, at 72h. Using PAGE we found that one APX isozyme (28kDa isoform) showed the strongest induction in all bacteriocin treated leaves at 72h. Activity of the seven SOD isozymes was increased by both bacteriocins, relative to the control treatment. The 33kDa PPO isozyme was induced strongly by both bacteriocins, relative to the control treatment. These results indicate that class IId bacteriocins can act as an inducer of plant disease defense-related enzymes and may be acting through mechanisms similar to Nod factors.

The Role of Salicylates in RHIZOBIUM -Legume Symbiosis and Abiotic Stresses in Higher Plants

Salicylic acid (SA) is an endogenous plant growth regulator. SA is involved in various physiological processes of plant growth and development and plays an active role in plant defense responses. SA also plays a major role during the early stages of Rhizobium-legume symbiosis. Nod factors produced by rhizobia, in response to legume produced flavonoids, affect SA content of the host plant during the early stages of nodulation. On the other hand, SA inhibits bacterial growth and the production of Nod factors by rhizobia. Exogenous application of SA delays nodule formation and decreases the number of nodules at the roots of the host plant. SA protects plants under abiotic stresses such as drought, salinity, low and high temperatures, and the damaging action of heavy metals. The ability of SA to protect plants exposed to abiotic stresses is due to the induction of a series of signal transduction cascades leading to the expression of genes responsible for the protection of plants from the stress.

Signals in the Underground: Microbial Signaling and Plant Productivity

Green plants are the vehicle by which virtually all energy enters the terrestrial biosphere. The rhizosphere is the first place where other organisms have a chronic access to this energy source and therefore an area of intense biological activity. Plants exude about 40% of photosynthates into the rhizosphere which makes it energy rich (Lynch and Whipps 1991). Due to the availability of substrates for metabolism, the rhizosphere is able to support large populations of microbes such as bacteria, actinomyctes, fungi, protozoa, algae, and viruses, etc. These rhizosphere inhabiting microorganisms compete for water, nutrients and space and sometimes improve their competitiveness by developing an intimate association with the plant. The rhizosphere can also be a battlefield among these microorganisms, with continuous competition and hostility among its inhabitants. The victors sometimes affect plant growth and development. However, it is becoming increasingly clear that establishment of the association between rhizobacteria and plants depends upon an exchange of signal molecules and it is during this initial signal exchange that plants and bacteria sometimes accept or reject each other. PGPR, first defined by Joseph W. Kloepper, are soil bacteria that are able to colonize plant roots and promote plant growth and development (Kloepper and Schroth 1978). These include intracellular PGPR (iPGPR) and extracellular PGPR (ePGPR) that promote plant growth and development via diverse mechanisms (Gray and Smith 2005). Inoculants of iPGPR (rhizobia – Rhizobium, Bradyrhizoum, Sinorhizobium) are currently used as commercial products to enhance nodulation and nitrogen fixation by legumes (Vessey 2003). The use of ePGPR in commercial inoculants, to promote plant growth and development, has also reached a reasonable degree of sophistication (Haas and Defago 2005; Vessey 2003).

Exploiting inter-organismal chemical communication for improved inoculants

The combination of rising fossil fuel prices and a need to reduce greenhouse gas emissions will lead to expanded use of crop inoculants (bio-fertilizers) both for increased production of biomass (for bio-fuels and soil C storage) and to reduce production of nitrous oxide, through increased reliance on biological nitrogen fixation. Over the last century inoculants have been improved through strain selection, improved carriers (including sterile carriers), and increased cell densities. During the last few decades our understanding of signalling between symbiotic bacteria and plants has expanded enormously, with the signalling between rhizobia and their legume hosts being the model system. Recent work has shown that adverse environmental conditions can inhibit this signalling and that addition of plant-to-microbe signals into inoculants can help overcome this. This is also true of addition of the microbe-to-plant signals that act as the return signals of this system; however, they have also been shown to cau...

Effect of chitin hexamer and thuricin 17 on lignification-related and antioxidative enzymes in Soybean Plants

Inducers of disease resistance in crop plants have a role in sustainable agriculture. We describe a set of bacteriocins that can potentially improve plant growth by controlling specific pathogens and inducing generalized resistance. Solutions of the bacteriocin thuricin 17 and/or a chitin hexamer (a known inducer and positive control) were applied to leaves of two-week-old soybean plants, and levels of lignification-related and antioxidative enzymes were monitored. Phenyl ammonia lyase (PAL) activity in thuricin 17-treated leaves was highest at 60 h after treatment, being 61.8% greater than the control. PAL activity also was increased 18.1% at 72 h after treatment with the chitin hexamer. Tyrosine ammonia lyase (TAL) activity in leaves was 57.0% higher than the control at 48 h after treatment with thuricin 17, while such activity in chitin hexamer-treated leaves was increased by 23.8% at 72 h. At 36 h after treatment with the chitin hexamer or chitin hexamer + thuricin 17, the total concentration of phenolic compounds was 15.3 or 19.3%, respectively, greater than the control. At 72 h, total phenolic concentrations increased by 23.2 and 19%, respectively, in response to thuricin 17 and chitin hexamer + thuricin 17. POD activity in thuricin 17-treated leaves increased by 74.6 and 81.2% at 48 and 72 h, respectively, whereas SOD activity increased by 24.9 and 79.9%, respectively, in chitin hexamer- and thuricin 17-treated leaves at 48 h. A peroxidase isozyme (31 kDa isomer) was induced in thuricin 17-treated leaves at 60 h, while catalase (59 kDa isomer) was induced in chitin hexamer-treated leaves. PAGE showed that two major SOD bands (Fe-SODs) were produced by both types of treatment. Collectively, these results indicate that the bacteriocin thuricin 17 can act as an inducer of plant disease defenses (i.e., activated lignification-related enzymes, antioxidative enzymes, and related isozymes) and that this induction is similar, but not identical, to that of the chitin hexamer elicitor. Although treatment with thuricin 17 + chitin hexamer also induced those responses, it did not present a clear pattern of additivity or synergy.

Low root zone temperature effects on bean (Phaseolus vulgaris L.) plants inoculated with Rhizobium leguminosarum bv. phaseoli pre-incubated with methyl jasmonate and/or genistein

Abstract Methyl jasmonate (MeJA) has recently been shown to act as a plant-to-bacteria signal. We tested the hypothesis that pre-induction of Rhizobium leguminosarum bv. phaseoli cells with genistein and/or MeJA would at least partially overcome the negative effects of low root zone temperature (RZT) on bean nodulation, nitrogen fixation and plant growth. Otebo bean plants were grown at constant air temperature (25oC) and two RZT regimes (25 and 17oC) and inoculated with R. leguminosarum bv. phaseoli pre-induced with MeJA and/or genistein. Our results indicate that low RZT inhibited nodulation, nitrogen fixation and plant growth. The plants growing at low RZT began fixing nitrogen seven days later compared to those at higher RZT. The low RZT plants had fewer nodules, lower nodule weight, less N fixation, slower plant growth, fewer leaves, smaller leaf area, and less dry matter accumulation comared to plants at a higher RZT. Rhzobium leguminosarum bv. phaseoli cells induced with genistein and/or MeJA enhanced bean nodulation, nitrogen fixation and growth at both optimum and suboptimum RZTs. The results of this study indicate that MeJA improves bean nitrogen fixation and growth at both optimum and suboptimum RZTs, and can be used alone or in combination with genistein to partially overcome the low RZT induced inhibitory effects on nodulation and nitrogen fixation.

Effect of Nod factor sprays on soybean growth and productivity under field conditions

Abstract A field experiment was conducted at Ste. Anne de Bellevue, Quebec during the growing seasons of 2002 and 2003, to study the effect of Nod factor treatments, applied at specific growth stages, on photosynthesis and biomass accumulation by soybean grown under two tillage systems (conventional tillage, no-tillage). Spray application of Nod factors increased photosynthesis at the four fully expanded trifoliate leaves and full bloom growth stages under no-tillage and conventional tillage; they increased plant biomass at the beginning seed filling stage. Nod factor treatment did not affect yield in 2002, due to severe drought during the reproductive period. In 2003, Nod factor application increased yield under conventional tillage but not under no-tillage, and this may have been due to the negative effect of no-tillage on plant growth on the clay-loam soil at the experimental site. This study indicated that responses to Nod factors are affected by environmental conditions, and that Nod factors may be useful when applied to soybean grown under conventional tillage.