<Emphasis Type="Italic">Gr</Emphasis> and <Emphasis Type="Italic">hp</Emphasis>-<Emphasis Type="Italic">1</Emphasis> tomato mutants unveil unprecedented interactions between arbuscular mycorrhizal symbiosis and fruit ripening

Main conclusion

Systemic responses to an arbuscular mycorrhizal fungus reveal opposite phenological patterns in two tomato ripening mutants depending whether ethylene or light reception is involved. The availability of tomato ripening mutants has revealed many aspects of the genetics behind fleshy fruit ripening, plant hormones and light signal reception. Since previous analyses revealed that arbuscular mycorrhizal symbiosis influences tomato berry ripening, we wanted to test the hypothesis that an interplay might occur between root symbiosis and fruit ripening. With this aim, we screened seven tomato mutants affected in the ripening process for their responsiveness to the arbuscular mycorrhizal fungus Funneliformis mosseae. Following their phenological responses we selected two mutants for a deeper analysis: Green ripe (Gr), deficient in fruit ethylene perception and high-pigment-1 (hp-1), displaying enhanced light signal perception throughout the plant. We investigated the putative interactions between ripening processes, mycorrhizal establishment and systemic effects using biochemical and gene expression tools. Our experiments showed that both mutants, notwithstanding a normal mycorrhizal phenotype at root level, exhibit altered arbuscule functionality. Furthermore, in contrast to wild type, mycorrhization did not lead to a higher phosphate concentration in berries of both mutants. These results suggest that the mutations considered interfere with arbuscular mycorrhiza inducing systemic changes in plant phenology and fruits metabolism. We hypothesize a cross talk mechanism between AM and ripening processes that involves genes related to ethylene and light signaling.

     

Expression of ethylene biosynthetic and signaling genes in relation to ripening of banana fruit after cold storage

Banana fruit are highly sensitive to chilling injury (CI), while the effect of different degrees of CI on the subsequent fruit ripening is largely unknown. In the present work, ripening characteristic of banana fruit after storage at 7 °C for 3 days or for 8 days, and expression levels of eight genes associated with ethylene biosynthetic and signaling, including MaACS1, MaACO1, MaERS1, MaERS3, and MaEIL1–4, were investigated. The results showed that banana fruit stored at 7 °C for 8 days exhibited more severe chilling symptoms than those at 7 °C for 3 days. Compared with banana fruit stored at 7 °C for 8 days, which showed abnormal ripening, more decrease in fruit firmness, while higher increase in ethylene production and hue angle were observed in banana fruit stored at 7 °C for 3 days, which could ripening normally. Moreover, gene expression profiles during ripening revealed that ethylene biosynthetic and signaling genes were differentially expressed in peel and pulp of banana fruit after storage at 7 °C for 3 days and 7 °C for 8 days. In the peel of fruit storage at 7 °C for 3 days, expression levels of MaACS1, MaACO1, MaEIL1, and MaEIL2 increased remarkably while MaERS3, MaEIL1, and MaEIL4 were enhanced in the fruit after storage at 7 °C for 8 days. In the pulp, with the exception of MaACO1 and MaERS3, expression levels of other genes did not exhibit a significant difference, between the banana fruit storage at 7 °C for 3 days and 7 °C for 8 days. Taken together, our results suggest that differential expression of ethylene biosynthetic and signaling genes such as MaERS3, MaACO1, and MaEIL2, may be related to ripening behavior of banana fruit with different degrees of CI after cold storage.     

The effects of condensed tannins derived from senescing <Emphasis Type="Italic">Rhizophora mangle</Emphasis> leaves on carbon,nitrogen and phosphorus mineralization in a <Emphasis Type="Italic">Distichlis spicata</Emphasis> salt marsh soil

Background and aims

Low nitrogen negatively affects soil fertility and plant productivity. Glucose-6-phosphate dehydrogenase (G6PDH) and Epichloë gansuensis endophytes are two factors that are associated with tolerance of Achnatherum inebrians to abiotic stress. However, the possibility that E. gansuensis interacts with G6PDH in enhancing low nitrogen tolerance of host grasses has not been examined.

Methods

A. inebrians plants with (E+) and without E. gansuensis (E?) were subjected to different nitrogen concentration treatments (0.1, 1, and 7.5 mM). After 90 days, physiological studies were carried out to investigate the participation of G6PDH in the adaption of host plants to low nitrogen availability.

Results

Low nitrogen retarded the growth of A. inebrians. E+ plants had higher total dry weight, chlorophyll a and b contents, net photosynthesis rate, G6PDH activity, and GSH content, while having lower plasma membrane (PM) NADPH oxidase activity, NADPH/NADP⁺ ratios, and MDA and H₂O₂ than in E? A. inebrians plants under low nitrogen concentration.

Conclusions

The presence of E. gansuensis played a key role in maintaining the growth of the A. inebrians plants under low nitrogen concentration by regulating G6PDH activity and the NADPH/NADP⁺ ratio and improving net photosynthesis rate.

     

Morphology engineering of basidiomycetes for improved laccase biosynthesis

Objective

This work is the first application of a morphological engineering technique called microparticle-enhanced cultivation (MPEC) aimed at the facilitation of laccase production in the submerged cultures by two basidiomycetes species Cerrena unicolor and Pleurotus sapidus.

Results

The positive effect of the applied 10 μm Al₂O₃ microparticles at concentrations from 5 to 30 g Al₂O₃ l^?1 was shown. Laccase activity increased 3.5-fold for C. unicolor and 2-fold for P. sapidus at 15 g Al₂O₃ l^?1 on 9 and 14 day of the cultivation, respectively, compared to the control culture without microparticles. The increase of laccase activity in the cultivation broths was caused by the action of Al₂O₃ microparticles on the agglomeration of hyphae. It led to the decrease of the size of the pellets, (on average by 2 mm for C. unicolor), the change of their shape (star-shaped pellets for C. unicolor) and the change of their structure (more compact pellets for P. sapidus).

Conclusions

Application of MPEC for the submerged cultures of two laccase-producing basidiomycetes proved successful in increasing of enzyme production.

     

Peroxide-dependent oxidation reactions catalyzed by CYP191A1 from <Emphasis Type="Italic">Mycobacterium smegmatis</Emphasis>

Objectives

To find the catalytic activities of CYP191A1 from Mycobacterium smegmatis, in which functions of most P450s are unknown, by using a set of reductase systems, peroxides, and various substrates including fatty acids and human drugs.

Results

CYP191A1 was functionally expressed in Escherichia coli and purified. Its catalytic activities were examined with fatty acids, chromogenic and fluorogenic substrates, and several human P450 substrates, in the presence of six different types of electron transfer systems, such as rat NADPH-P450 reductase, Candida NADPH-P450 reductase, ferredoxin/ferredoxin reductase, putidaredoxin/putidaredoxin reductase, and peroxides (H₂O₂ and t-butyl hydroperoxide). The reactions catalyzed by CYP191A1 included the hydroxylation and O-dealkylation of several substrates.

Conclusions

CYP191A1 preferentially catalyzes the peroxide-dependent oxidation of various substrates over the reductase-dependent reaction. Its peroxygenase activity may be used an effective biocatalytic tool to synthesize the metabolites of drugs.

     

Enhanced biomass and oxidative stress tolerance of <Emphasis Type="Italic">Synechococcus elongatus</Emphasis> PCC 7942 overexpressing the <Emphasis Type="Italic">DHAR</Emphasis> gene from <Emphasis Type="Italic">Brassica juncea</Emphasis>

Objectives

To improve the oxidative stress tolerance, biomass yield, and ascorbate/dehydroascorbate (AsA/DHA) ratio of Synechococcus elongatus PCC 7942 in the presence of H₂O₂, by heterologous expression of the dehydroascorbate reductase (DHAR) gene from Brassica juncea (BrDHAR).

Results

Under H₂O₂ stress, overexpression of BrDHAR in the transgenic strain (BrD) of S. elongatus greatly increased the AsA/DHA ratio. As part of the AsA recycling system, the oxidative stress response induced by reactive oxygen species was enhanced, and intracellular H₂O₂ level decreased. In addition, under H₂O₂ stress conditions, the BrD strain displayed increased growth rate and biomass, as well as higher chlorophyll content and deeper pigmentation than did wild-type and control strains.

Conclusion

By maintaining the AsA pool and redox homeostasis, the heterologous expression of BrDHAR increased S. elongatus tolerance to H₂O₂ stress, improving the biomass yield under these conditions. The results suggest that the BrD strain of S. elongatus, with its ability to attenuate the deleterious effects of ROS caused by environmental stressors, could be a promising platform for the generation of biofuels and other valuable bioproducts.