Eva Casanova

Trichoderma asperellum Strain T34 Controls Fusarium Wilt Disease in Tomato Plants in Soilless Culture Through Competition for Iron

Trichoderma asperellum strain T34 has been reported to control the disease caused by Fusarium oxysporum f.sp. lycopersici (Fol) on tomato plants. To study the importance of iron concentration in the growth media for the activity and competitiveness of T34 and the pathogen, we tested four iron concentrations in the nutrient solution [1, 10, 100, and 1000 µM provided as EDTA/Fe(III)] in a biological control experiment with T34 and Fol in tomato plants. The reduction of the Fusarium-infected shoot by T34 was only significant at 10 µM Fe. We hypothesized that Fe competition is one of the key factors in the biocontrol activity exerted by T34 against Fol, as an increase in Fe concentration over 10 µM would lead to the suppression of T34 siderophore synthesis and thus inhibition of Fe competition with Fol. T34 significantly reduced the populations of Fol at all the doses of Fe assayed. In contrast, Fol enhanced the populations of T34 at 1 and 10 µM Fe. Nevertheless, several plant physiological parameters like net CO2 assimilation (A), stomatal conductance (gs), relative quantum efficiency of PSII (ΦPSII), and efficiency of excitation energy capture by open PSII reactive centers (Fv′/Fm′) demonstrated the protection against Fol damage by treatment with T34 at 100 µM Fe. The first physiological parameter affected by the disease progression was gs. Plant dry weight was decreased by Fe toxicity at 100 and 1,000 µM. T34-treated plants had significantly greater heights and dry weights than control plants at 1,000 µM Fe, even though T34 did not reduce the Fe content in leaves or stems. Furthermore, T34 enhanced plant height even at the optimal Fe concentration (10 µM) compared to control plants. In conclusion, T. asperellum strain T34 protected tomato plants from both biotic (Fusarium wilt disease) and abiotic stress [Fe(III) toxic effects].

Levels and immunolocalization of endogenous cytokinins in thidiazuron-induced shoot organogenesis in carnation.

We evaluated the capacity of the plant growth regulator thidiazuron (TDZ), a substituted phenylurea with high cytokinin-like activity, to promote organogenesis in petals and leaves of several carnation cultivars (Dianthus spp.), combined with 1-naphthaleneacetic acid (NAA). The involvement of the endogenous auxin indole-3-acetic acid (IAA) and purine-type cytokinins was also studied. Shoot differentiation was found to depend on the explant, cultivar and balance of growth regulators. TDZ alone (0.5 and 5.0 micromol/L) as well as synergistically with NAA (0.5 and 5.0 micromol/L) promoted shoot organogenesis in petals, and was more active than N6-benzyladenine. In petals of the White Sim cultivar, TDZ induced cell proliferation in a concentration-dependent manner and, on day 7 of culture, the proportion of meristematic regions in those petals allowed the prediction of shoot regeneration capacity after 30 days of culture. Immunolocalization of CK ribosides, N6-(delta2-isopentenyl)adenosine, zeatin riboside (ZR) and dihydrozeatin riboside (DHZR), in organogenic petals showed them to be highly concentrated in the tips of bud primordia and in the regions with proliferation capacity. All of them may play a role in cell proliferation, and possibly in differentiation, during the organogenic process. After seven days of culture of White Sim petals, NAA may account for the changes found in the levels of IAA and DHZR, whereas TDZ may be responsible for the remarkable increases in N6-(delta2-isopentenyl)adenine (iP) and ZR. ZR is induced by low TDZ concentrations (0.0-0.005 micromol/L), whereas iP, that correlates with massive cell proliferation and the onset of shoot differentiation, is associated with high TDZ levels (0.5 micromol/L). In addition to the changes observed in quantification and in situ localization of endogenous phytohormones during TDZ-induced shoot organogenesis, we propose that TDZ also promotes growth directly, through its own biological activity. To our knowledge, this study is the first to evaluate the effect of TDZ on endogenous phytohormones in an organogenic process.

The suppressive effects of composts used as growth media against Botrytis cinerea in cucumber plants

The incidence/severity of soil-borne plant diseases is often reduced when composts are used as growth media. However, much less information is available about the effects of composts on the development of foliar diseases. Here we studied the suppressive capacity of five composts (from olive marc-cotton gin trash, grape marc, cork, spent mushroom and municipal organic and yard wastes) as growth media against Botrytis cinerea disease in cucumber plants. We also examined the putative correlations of several biotic and abiotic factors involved in disease suppression. The suppressive capacity of the growth media was studied by comparing disease incidence/severity in plants grown in composts with that occurring in plants grown in commercial peats, which are conducive to most soilborne diseases. Correlations were made between the occurrence of disease and leaf nutrient status, as well as electrical conductivity (EC) and microbial activity (measured as β-glucosidase activity) in the growth media. Cucumber plants grown in the peats showed greater severity of B. cinerea during the bioassay than those grown in composts. Mo, Ca and Si content in leaves showed negative correlations with this disease. A negative correlation with disease severity was observed for EC and microbial activity in the growth media. The noticeable reduction in B. cinerea in plants grown in composts was related to the supply of specific chemical elements, a certain degree of salt stress, and the high microbial activity of composts.

Effects of agar concentration and vessel closure on the organogenesis and hyperhydricity of adventitious carnation shoots

Carnation plantlets (Dianthus caryophyllus L.) cultured in vitro often develop morphological and physiological anomalies, a phenomenon called hyperhydricity, which impairs their survival ex vitro. When the agar concentration of the growth medium was increased (from 0 to 12 g dm−3), thereby reducing water availability, the hyperhydricity of those adventitious shoots regenerated from carnation petals decreased. This was accompanied by a progressive fall in the water content of shoots (94.9 to 91.4 %), fresh mass (from 57.2 to 1.8 mg), number of leaf parenchyma cell layers (from 9.3 to 7.7), and the size of these cells (from 968 to 254 µm2). However, the number of regenerated shoots also decreased (17.7 in 2 g dm−3 agar to 4.3 in 12 g dm−3). Similarly, in ventilated tubes, which exhibit a lower relative humidity than tightly closed tubes, shoot organogenesis diminished up to 28 %, in tandem with shoot water content. Thus, relative humidity and water availability in culture vessels do not only influence shoot hyperhydricity in carnations, but also greatly affect adventitious shoot organogenesis.

The rolC gene in carnation exhibits cytokinin- and auxin-like activities

The overexpression of the rolC gene of the Ri plasmid of Agrobacterium rhizogenes in transgenic plants alters their development. Few studies have been performed on adventitious shoot or root formation in rolC-transformed plants. In this study we evaluated a possible cytokinin-like and auxin-like effect on shoot and root regeneration from petals and leaves of four lines of rolC-transgenic carnation plants (Dianthus caryophyllus L. cv. White Sim). rolC was found to enhance shoot regeneration, by increasing either the number of shoots per regenerative explant, or the number of shoot-forming explants. Remarkable root regeneration, in a medium with only auxin, was obtained from rolC explants, due to the increased percentage of root-forming explants. Our results show that rolC exhibits both cytokinin-like and auxin-like activities in rolC-transgenic carnation tissues.