Herbaspirillum seropedicae promotes maize growth but fails to control the maize leaf anthracnose.

Herbaspirillum seropedicae is an endophytic diazotrophic bacterium and a plant growth promoting bacteria. Colletotrichum graminicola causes the anthracnose, one of the most destructive maize diseases worldwide. The main objective of this work was to evaluate the effects of H. seropedicae SmR1 strain on the plant growth and leaf anthracnose of maize plants grown in substrate amended or not amended with humic substances. In the first assay, plants were pre-treated with H. seropedicae and inoculated with C. graminicola at 7, 14 and 21 days after treatment (DAT). In the second assay, plants were treated with H. seropedicae, grown in substrate amended with humic substances and inoculated at 3 and 7 DAT. The anthracnose severity was assessed by measurement of necrotic and chlorotic leaf area, and bacteria were quantified in leaves by quantitative PCR. H. seropedicae did not affect the disease severity in maize leaves, although it efficiently colonized the leaf tissues and it promoted maize leaf growth. Humic substances improved H. seropedicae colonization in maize.

Defense responses induced by ulvan in wheat against powdery mildew caused by Blumeria graminis f. sp. tritici.

Ulvan is a water-soluble sulfated heteropolysaccharide extracted from the cell walls of the green seaweeds Ulva spp. This polysaccharide is known to induce resistance and protect plants against a broad range of plant pathogenic fungi, such as Blumeria graminis f. sp. tritici (Bgt), the causal agent of powdery mildew in wheat. We aimed to study the defense mechanisms induced by ulvan against Bgt in susceptible wheat by investigating the defense-related gene expression, enzymes activity, accumulation of phenolic compounds and hydrogen peroxide (H2O2), as well as the development of Bgt infection structures in vitro and in planta. Symptoms were reduced by 42% in ulvan-treated plants. In vitro, ulvan did not inhibit conidial germination of Bgt but in planta, increased the appressorial germ tubes without haustorium. Ulvan increased the presence of fluorescent papillae and accumulation of H2O2 at the penetration sites of Bgt, as well as the content of phenolic compounds. POX, PAL and LOX activities were stimulated in ulvan-treated plants during the first 48 h after inoculation. However, few of defense-related genes studied were differentially expressed in infected plants after ulvan treatment. By contrast, in non-infected conditions, ulvan up-regulated the expression of genes involved in phenylpropanoid metabolism, i.e. PAL, CHS, COMT, ANS and FLS, genes encoding pathogenesis-related proteins, i.e. PR1, PR9, PR15, and LOX during the first 96 h after treatment. This study provides new insights about the multiple ulvan effects on wheat defense responses, and especially the elicitation of the phenylpropanoid pathway leading to phenolic compounds accumulation, which could be involved in cell wall reinforcement.

Real-time PCR quantification of the plant growth promoting bacteria Herbaspirillum seropedicae strain SmR1 in maize roots.

The plant growth promoting bacteria Herbaspirillum seropedicae SmR1 is an endophytic diazotroph found in several economically important crops. Considering that methods to monitor the plant-bacteria interaction are required, our objective was to develop a real-time PCR method for quantification of PGPB H. seropedicae in the rhizosphere of maize seedlings. Primer pairs were designed, and their specificity was verified using DNA from 12 different bacterial species. Ten standard curves of qPCR assay using HERBAS1 primers and tenfold serial dilutions of H. seropedicae SmR1 DNA were performed, and PCR efficiency of 91 % and correlation coefficient of 0.99 were obtained. H. seropedicae SmR1 limit of detection was 10(1) copies (corresponding to 60.3 fg of bacterial DNA). qPCR assay using HERBAS1 was used to detect and quantify H. seropedicae strain SmR1 in inoculated maize roots, cultivated in vitro and in pots, harvested 1, 4, 7, and 10 days after inoculation. The estimated bacterial DNA copy number per gram of root was in the range 10(7)-10(9) for plants grown in vitro and it was around 10(6) for plants grown in pots. Primer pair HERBAS1 was able to quantify H. seropedicae SmR1, and this assay can be useful for monitoring plant-bacteria interaction.