Exogenous melatonin enhances the reactive oxygen species metabolism, antioxidant defense-related gene expression, and photosynthetic capacity of Phaseolus vulgaris L. to confer salt stress tolerance.

Melatonin (MT) has been reported to regulate certain plant physiological processes and promote tolerance to different environmental stresses such as salinity. Green bean (Phaseolus vulgaris L. cv. Royal Nel) seedlings were exposed to 200 mM NaCl with or without pre-treatment with 150 µM MT. Salt stress led to a lower chlorophyll content, a reduced photosynthetic activity, increased reactive oxygen species (ROS) contents, and decreased photosystem II (PSII) activity. The application of exogenous MT to green bean seedlings under salt stress improved photosynthetic activity and alleviated the oxidative damages by enhancing the activity of antioxidant enzymes. The expression of catalase (CAT1), glutathione reductase (GR), superoxide dismutase (CuZnSOD1), ascorbate peroxidase (APX), Peroxiredoxin Q (PrxQ), and 2-cysteine peroxiredoxin (2-Cys-Prx) encoding genes was significantly increased under salt stress in green bean seedling compared with the untreated control. However, plants treated with exogenous MT and NaCl had 28.8, 21.1, 26.1, 20, 26.2, and 22.4% higher CuZnSOD, CAT1, APX, GR, PrxQ, and 2-Cys-Prx transcript levels, respectively, compared to NaCl stress alone. Our study revealed the protective mechanisms mediated by exogenous MT application in NaCl stress alleviation and our findings could be used in the management of green bean cultivation in salinity-prone soils.

Exogenous Application of Foliar Salicylic Acid and Propolis Enhances Antioxidant Defenses and Growth Parameters in Tomato Plants.

Salicylic acid (SA) and propolis (PR) are known to regulate the physiological process and to have a relevant role in bioactive compounds content. Our experiment was designed to evaluate the effect of SA and PR application on the growth, yield, and quality parameters of tomato grown for the fresh market in field conditions in Egypt. We studied the effect of twelve treatments where SA (0.50, 1.00, 1.50, 2.00, and 2.50 mM) and PR (1, 2, 10, 20, and 100 mg propolis mL-1) were applied at increasing doses as a sole agent or combined each other (1.50 mM + 10 mg mL-1 for SA and PR, respectively). An untreated control was also considered. Tomato plants treated with SA (0.50, 1.00, and 1.50 mM) showed a significant effect in all traits especially SA1 (0.50 mM) in growth parameters and SA2 (1.00 mM) in pigment and antioxidant content. Propolis foliar application was more effective than SA as it revealed that raising the concentration of aqueous extract enhanced the growth parameters and pigment in tomato. The best result was obtained by the 10 mg mL-1 treatment. The effect of propolis on antioxidant enzymes varied as the 10 mg mL-1 treatment was effective on peroxidases and superoxide dismutase, while 100 mg mL-1 was more effective on catalase. Salicylic acid and propolis have a positive effect on both preserving tomato plants and on nutrient supply, so the mixed intermediate concentration (1.50 mM + 10 mg mL-1) is considered very effective and results in an improvement of all plant traits.

Investigating molecular evolutionary forces and phylogenetic relationships among melatonin precursor-encoding genes of different plant species.

A total of 53 plant species accessions from different geographic regions, including four melatonin precursor-coding genes obtained from Arachis hypogaea (ASMT1, 2, 3 and T5H) underwent extensive molecular evolutionary analyses. Evolutionary relationships were inferred and showed that dichotomous bifurcating trees did not reflect the true phylogeny since reticulate events took place due likely to recombination. Thus, a phylogenetic network was reconstructed for each type of enzyme and highlighted the presence of such incompatibilities. GARD algorithm pointed out that ASMT1, 2, and 3-coding gene sequences contained recombination sites with significant topological incongruence on both sides of the breakpoints (for ASMT1, and 2), while only on one side of the breakpoints for ASMT3. In contrast, no statistically recombination signal was recorded in T5H-coding gene. Furthermore, gene duplication was localized in the ancestor of a monophyletic group of Populus accessions. Selection pressure was assessed using several statistical models incorporated in HyPhy package through the datamonkey web server. It was demonstrated that numerous individual sites and tree branches experienced predominantly purifying selection. In contrast, the BUSTED model evidenced a gene-wide episodic diversifying selection in the phylogeny of only three enzyme-coding genes (ASMT, and 2, and T5H). Likewise, it was shown that Mixed Effects Model of Episodic Selection (MEME) model detected only episodic positively selected sites in all four melatonin enzymes-coding genes; whereas, REL model failed to detect neither positive nor negative selection in tested individual sites of ASMT3-coding gene.

Metabolite profiling at the cellular and subcellular level reveals metabolites associated with salinity tolerance in sugar beet.

Sugar beet is among the most salt-tolerant crops. This study aimed to investigate the metabolic adaptation of sugar beet to salt stress at the cellular and subcellular levels. Seedlings were grown hydroponically and subjected to stepwise increases in salt stress up to 300 mM NaCl. Highly enriched fractions of chloroplasts were obtained by non-aqueous fractionation using organic solvents. Total leaf metabolites and metabolites in chloroplasts were profiled at 3 h and 14 d after reaching the maximum salinity stress of 300 mM NaCl. Metabolite profiling by gas chromatography-mass spectrometry (GC-MS) resulted in the identification of a total of 83 metabolites in leaves and chloroplasts under control and stress conditions. There was a lower abundance of Calvin cycle metabolites under salinity whereas there was a higher abundance of oxidative pentose phosphate cycle metabolites such as 6-phosphogluconate. Accumulation of ribose-5-phosphate and ribulose-5-phosphate coincided with limitation of carbon fixation by ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). Increases in glycolate and serine levels indicated that photorespiratory metabolism was stimulated in salt-stressed sugar beet. Compatible solutes such as proline, mannitol, and putrescine accumulated mostly outside the chloroplasts. Within the chloroplast, putrescine had the highest relative level and probably assisted in the acclimation of sugar beet to high salinity stress. The results provide new information on the contribution of chloroplasts and the extra-chloroplast space to salinity tolerance via metabolic adjustment in sugar beet.

Assessment of sucrose transporters, metabolites and sucrose phosphate synthase in different sugarcane tissues.

The expression of sucrose-phosphate synthase II (SPSII) and sucrose transporters ShSUT1A and ShSUT4 were determined by RT-PCR and qRT-PCR in the sink and source leaves and in rind and pith of mature internodes of four high-yielding Hawaiian sugarcane cultivars. Expression of SPSII, ShSUT1A, and ShSUT4 was lower in pith than in rind, except in one cultivar, but else quite similar in the cultivars. The strong expression of transporter ShSUT4 in the rind of the internodes may hint to a special role of ShSUT4 in the rind. ShSUT4-expression in the sink and source leaves was similar in all four cultivars, whereas large differences were found for the expression of ShSUT1A and SPSII between the source and sink leaves and between the cultivars. The levels of sucrose precursors were doubled in source leaves compared to sink leaves, whereas they were higher in immature internode compared to mature internode. The role of sucrose transporters and SPSII in leaves and internodes is discussed, but the large differences, which were observed in the transcript levels of SPSII and sucrose transporters between some cultivars, although all the cultivars were similarly high-yielding cultivars, show that SPSII and SUT transcript levels cannot be used as indicators of high-yield cultivars.

Redox and Reactive Oxygen Species Network in Acclimation for Salinity Tolerance in Sugar Beet.

Fine-tuned and coordinated regulation of transport, metabolism and redox homeostasis allows plants to acclimate to osmotic and ionic stress caused by high salinity. Sugar beet is a highly salt tolerant crop plant and is therefore an interesting model to study sodium chloride (NaCl) acclimation in crops. Sugar beet plants were subjected to a final level of 300 mM NaCl for up to 14 d in hydroponics. Plants acclimated to NaCl stress by maintaining its growth rate and adjusting its cellular redox and reactive oxygen species (ROS) network. In order to understand the unusual suppression of ROS accumulation under severe salinity, the regulation of elements of the redox and ROS network was investigated at the transcript level. First, the gene families of superoxide dismutase (SOD), peroxiredoxins (Prx), alternative oxidase (AOX), plastid terminal oxidase (PTOX) and NADPH oxidase (RBOH) were identified in the sugar beet genome. Salinity induced the accumulation of Cu-Zn-SOD, Mn-SOD, Fe-SOD3, all AOX isoforms, 2-Cys-PrxB, PrxQ, and PrxIIF. In contrast, Fe-SOD1, 1-Cys-Prx, PrxIIB and PrxIIE levels decreased in response to salinity. Most importantly, RBOH transcripts of all isoforms decreased. This pattern offers a straightforward explanation for the low ROS levels under salinity. Promoters of stress responsive antioxidant genes were analyzed in silico for the enrichment of cis-elements, in order to gain insights into gene regulation. The results indicate that special cis-elements in the promoters of the antioxidant genes in sugar beet participate in adjusting the redox and ROS network and are fundamental to high salinity tolerance of sugar beet.

Molecular and biochemical characterisation of a novel type II peroxiredoxin (XvPrx2) from the resurrection plant Xerophyta viscosa.

A type II peroxiredoxin gene (XvPrx2) was isolated from a Xerophyta viscosa (Baker) cDNA cold-stress library. The polypeptide displayed significant similarity to other plant type II peroxiredoxins, with the conserved amino acid motif (PGAFTPTCS) proposed to constitute the active site of the enzyme. Northern blot analyses showed that XvPrx2 gene was stress-inducible in response to abiotic stresses while gel analyses revealed that XvPrx2 homologues exist within the X. viscosa proteome. Using a yellow fluorescent reporter protein, the XvPrx2 protein localised to the cytosol. A mutated protein (XvV7) was generated by converting the valine at position 76 to a cysteine and an in vitro DNA protection assay showed that, in the presence of either XvPrx2 or XvV7, DNA protection occurred. In addition, an in vivo assay showed that increased protection was conferred to Escherichia coli cells overexpressing either XvPrx2 or XvV7. The XvPrx2 activity was maximal with DTT as electron donor and H2O2 as substrate. Using E. coli thioredoxin, a 2-15-fold lower enzyme activity was observed. The XvPrx2 activity with glutathione was significantly lower and glutaredoxin had no measurable effect on this reaction. The XvV7 protein displayed significantly lower activity compared with XvPrx2 for all substrates assessed.

Biology and management of sugarcane yellow leaf virus: an historical overview.

Sugarcane yellow leaf virus (SCYLV) is one of the most widespread viruses causing disease in sugarcane worldwide. The virus has been responsible for drastic economic losses in most sugarcane-growing regions and remains a major concern for sugarcane breeders. Infection with SCYLV results in intense yellowing of the midrib, which extends to the leaf blade, followed by tissue necrosis from the leaf tip towards the leaf base. Such symptomatic leaves are usually characterized by increased respiration, reduced photosynthesis, a change in the ratio of hexose to sucrose, and an increase in starch content. SCYLV infection affects carbon assimilation and metabolism in sugarcane, resulting in stunted plants in severe cases. SCYLV is mainly propagated by planting cuttings from infected stalks. Phylogenetic analysis has confirmed the worldwide distribution of at least eight SCYLV genotypes (BRA, CHN1, CHN3, CUB, HAW, IND, PER, and REU). Evidence of recombination has been found in the SCYLV genome, which contains potential recombination signals in ORF1/2 and ORF5. This shows that recombination plays an important role in the evolution of SCYLV.

Molecular evolutionary history of Sugarcane yellow leaf virus based on sequence analysis of RNA-dependent RNA polymerase and putative aphid transmission factor-coding genes.

RNA-dependent RNA polymerase (RdRp) encoded by ORF2 and putative aphid transmission factor (PATF) encoded by ORF5 of Sugarcane yellow leaf virus (SCYLV) were detected in six sugarcane cultivars affected by yellow leaf using RT-PCR and real-time RT-PCR assays. Expression of both genes varied among infected plants, but overall expression of RdRp was higher than expression of PATF. Cultivar H87-4094 from Hawaii yielded the highest transcript levels of RdRp, whereas cultivar C1051-73 from Cuba exhibited the lowest levels. Sequence comparisons among 25 SCYLV isolates from various geographical locations revealed an amino acid similarity of 72.1-99.4 and 84.7-99.8 % for the RdRp and PATF genes, respectively. The 25 SCYLV isolates were separated into three (RdRp) and two (PATF) phylogenetic groups using the MEGA6 program that does not account for genetic recombination. However, the SCYLV genome contained potential recombination signals in the RdRp and PATF coding genes based on the GARD genetic algorithm. Use of this later program resulted in the reconstruction of phylogenies on the left as well as on the right sides of the putative recombination breaking points, and the 25 SCYLV isolates were distributed into three distinct phylogenetic groups based on either RdRp or PATF sequences. As a result, recombination reshuffled the affiliation of the accessions to the different clusters. Analysis of selection pressures exerted on RdRp and PATF encoded proteins revealed that ORF 2 and ORF 5 underwent predominantly purifying selection. However, a few sites were also under positive selection as assessed by various models such as FEL, IFEL, REL, FUBAR, MEME, GA-Branch, and PRIME.

Putative recombination signature and significance of insertion/deletion events in the RNA-dependent RNA polymerase coding region of sugarcane yellow leaf virus.

The 5898 nucleotide single-strand RNA genome of Sugarcane yellow leaf virus (SCYLV) contains one long open reading frame, which is translated into a 120.6 kDa polyprotein. The sequences of SCYLV isolates from the two SCYLV-susceptible cultivars from Hawaii had a deletion of 48-51 nt in ORF1. SCYLV from 12 sugarcane hybrid cultivars from different origins were tested by RT-PCR using a specific set of primers, to investigate the genome segment for this deletion. Only three cultivars were found not to have the deletion (H87-4319, JA-605 and CP52-43), while SCYLV from nine cultivars (H73-6110, H87-4094, H78-7750, GT54-9, G84-47, H78-4153, H65-7052, C1051-73, Ph-8013) along with aphid (Melanaphis sacchari), which fed on SCYLV-infected H73-6110, contained a deletion of about 50 nt. The deleted sequence was located in the overlap frameshift of ORF1 and ORF2. Thus, ORFs 1 and 2 of SCYLV are translated via ribosomal frameshift and yield the 120.6 kDa viral replicase. ORF1 plays most likely a role in the replication and is a source of large variability among the virus population. To identify possible recombination events located in the RdRp domain of the Hawaiian isolates, two programs were used: RDP v.4.3 and RECCO. It is noteworthy that according both methods Haw73-6110 was found as a potential recombinant. On the other hand, opposed to the RDP package, RECCO revealed that Haw87-4094 isolate was also a recombinant whereas Haw87-4319 was not.