Saponins, a group of glycosidic compounds present in several plant species, have aglycone moieties that are formed using triterpenoid or steroidal skeletons. In spite of their importance as antimicrobial compounds and their possible benefits for human health, knowledge of the genetic control of saponin biosynthesis is still poorly understood. In the Medicago genus, the hemolytic activity of saponins is related to the nature of their aglycone moieties. We have identified a cytochrome P450 gene (CYP716A12) involved in saponin synthesis in Medicago truncatula using a combined genetic and biochemical approach. Genetic loss-of-function analysis and complementation studies showed that CYP716A12 is responsible for an early step in the saponin biosynthetic pathway. Mutants in CYP716A12 were unable to produce hemolytic saponins and only synthetized soyasaponins, and were thus named lacking hemolytic activity (lha). In vitro enzymatic activity assays indicate that CYP716A12 catalyzes the oxidation of b-amyrin and erythrodiol at the C-28 position, yielding oleanolic acid. Transcriptome changes in the lha mutant showed a modulation in the main steps of triterpenic saponin biosynthetic pathway: squalene cyclization, b-amyrin oxidation, and glycosylation. The analysis of CYP716A12 expression in planta is reported together with the sapogenin content in different tissues and stages. This article provides evidence for CYP716A12 being a key gene in hemolytic saponin biosynthesis.

BackgroundOlive (Olea europaea L.) fruits contain numerous secondary metabolites, primarily phenolics, terpenes and sterols, some of which are particularly interesting for their nutraceutical properties. This study will attempt to provide further insight into the profile of olive phenolic compounds during fruit development and to identify the major genetic determinants of phenolic metabolism.ResultsThe concentration of the major phenolic compounds, such as oleuropein, demethyloleuropein, 3–4 DHPEA-EDA, ligstroside, tyrosol, hydroxytyrosol, verbascoside and lignans, were measured in the developing fruits of 12 olive cultivars. The content of these compounds varied significantly among the cultivars and decreased during fruit development and maturation, with some compounds showing specificity for certain cultivars. Thirty-five olive transcripts homologous to genes involved in the pathways of the main secondary metabolites were identified from the massive sequencing data of the olive fruit transcriptome or from cDNA-AFLP analysis. Their mRNA levels were determined using RT-qPCR analysis on fruits of high- and low-phenolic varieties (Coratina and Dolce d’Andria, respectively) during three different fruit developmental stages. A strong correlation was observed between phenolic compound concentrations and transcripts putatively involved in their biosynthesis, suggesting a transcriptional regulation of the corresponding pathways. OeDXS, OeGES, OeGE10H and OeADH, encoding putative 1-deoxy-D-xylulose-5-P synthase, geraniol synthase, geraniol 10-hydroxylase and arogenate dehydrogenase, respectively, were almost exclusively present at 45 days after flowering (DAF), suggesting that these compounds might play a key role in regulating secoiridoid accumulation during fruit development.ConclusionsMetabolic and transcriptional profiling led to the identification of some major players putatively involved in biosynthesis of secondary compounds in the olive tree. Our data represent the first step towards the functional characterisation of important genes for the determination of olive fruit quality.

Background and Aims Postveraison limitation of canopy photosynthesis delays grape berry ripening and reduces sugar accumulation, thus lowering the alcohol content of the subsequent wines. This study was designed to evaluate whether similar results could be obtained by defoliation apical to the bunch zone using a leaf‐plucking machine when berry sugar content was approximately 16–17°Brix. Methods and Results In 2011 and 2012, defoliation treatments were applied postveraison to cv. Sangiovese vines (D) on either side of each row using a mechanical leaf remover, and these D vines were compared to a nondefoliated control (C). The machine removed 35% of the leaves on the vine and created a 50‐cm vertical window without leaves above the bunch area, but retained a few leaves at the canopy apex (about 0.50 m2/vine). In both years, leaf removal reduced the rate of berry sugar accumulation and led to a 1.2 lower harvest °Brix and consequently, a lower wine alcohol (−0.6%) content in D relative to that of C vines. In 2012, sugar content of D vines, monitored in a group of vines that was not harvested, had recovered to that of C vines 2 weeks after harvest. The concentration of total phenolic compounds in the grapes, the chemical and chromatic characteristics of the wines and the replenishment of soluble sugars, starch and total nitrogen in the canes and roots were similar in the D and C vines. Conclusion To achieve an effective delay in sugar accumulation in the berries, leaves should be removed at 16–17°Brix, and at least 30–35% of vine leaf area should be removed. Significance of the Study Mechanical removal of leaves postveraison above the bunch zone of Sangiovese can be an easy and economically viable technique for delaying sugar accumulation in the berries and for limiting the alcohol content of wines with no negative impact on desirable composition of either berries or wines.

The secoiridoids are the main class of specialized metabolites present in olive (Olea europaea L.) fruit. In particular, the secoiridoid oleuropein strongly influences olive oil quality because of its bitterness, which is a desirable trait. In addition, oleuropein possesses a wide range of pharmacological properties, including antioxidant, anti-inflammatory, and anti-cancer activities. In accordance, obtaining high oleuropein varieties is a main goal of molecular breeding programs. Here we use a transcriptomic approach to identify candidate genes belonging to the secoiridoid pathway in olive. From these candidates, we have functionally characterized the olive homologue of iridoid synthase (OeISY), an unusual terpene cyclase that couples an NAD (P)H-dependent 1,4-reduction step with a subsequent cyclization, and we provide evidence that OeISY likely generates the monoterpene scaffold of oleuropein in olive fruits. OeISY, the first pathway gene characterized for this type of secoiridoid, is a potential target for breeding programs in a high value secoiridoid-accumulating species.Olive (Olea europaea L.) produces a range of secondary metabolites that strongly affect the taste and nutritional properties of olive oil and fruits. The most abundant of these secondary metabolites are the secoiridoids, monoterpenoids with a 3,4-dihydropyran skeleton. These compounds are present as oleosidic secoiridoids or oleosides that have an exocyclic olefinic functionality (1) and possess a tyrosine-derived component (see Fig.

Abstract:The effectiveness of a postveraison application of the film-forming antitranspirant Vapor Gard (VG, a.i. di-1-p-menthene) was investigated as a technique to delay grape ripening and reduce sugar accumulation in the berry. The study was carried out over the 2010-2011 seasons in a nonirrigated vineyard of cv. Sangiovese in central Italy.Vapor Gard was applied at 2% concentration to the upper two-thirds of the canopy (most functional leaves) and it significantly lowered leaf assimilation and transpiration rates and increased intrinsic water use efficiency. The F v / F m ratio was not modified, emphasizing that photoinhibition did not occur at the photosystem II complex, whereas the reduction of pool size of plastoquinone matched well with reduced CO 2 fixation found in VG-treated vines. In both years VG treatment reduced the pace of sugar accumulation in the berry as compared to control vines, scoring a -1.2 Brix at harvest and wine alcohol content at -1% without compromising the recovery of concentrations of carbohydrates and total nitrogen in canes and roots. Concurrently, organic acids, pH, and phenolic richness of grapes and wines were unaffected, whereas there was a decrease in anthocyanin content in the berry (-19% compared to control vines) and in the wine (-15% compared to control vines). The application of VG at postveraison above the cluster zone is an effective, simple, and viable technique to hinder berry sugaring and obtain less alcoholic wines.To be effective the spraying should be performed at ~14 to 15 Brix, making sure that the lower leaf epidermis is fully wetted by the chemical.

Calderini, O., Donnison, I.S., Polegri, L., Panara, F., Thomas, A., Arcioni, S., Pupilli, F. (2011). Partial isolation of the genomic region linked with apomixis in Paspalum simplex. Molecular Breeding, 28 (2), 265-276. IMPF: 02.85 Sponsorship: BBSRC RONO: BBS/E/W/00003134AApomixis is a form of asexual reproduction through seed and has the potential to be applied, to great benefit, to agriculture. Understanding the genetic control of apomixis has proven to be a challenging task because the trait is mainly present in wild species and genetic mapping is often impaired by a block of recombination. A physical mapping approach has therefore been undertaken to unlock the genetic control of apomixis in Paspalum simplex Morong, a species with a relatively small genome and which exhibits a degree of genetic synteny with rice. In this paper, we report on the construction of a bacterial artificial chromosome library for Paspalum simplex with a coverage of approximately three genome equivalents and an average insert size of 94 kb. The BAC library was screened with 19 sequence characterized amplified region markers which were 100% linked to apomixis and a recombinant SCAR marker, all developed through a bulked segregant analysis strategy. A mini-sequencing procedure reported in the literature greatly aided the direct development of SCAR markers from amplified fragment length polymorphism bands. Several BAC clones linked to apomixis were identified and assembled into seven contigs and 18 singletons. Two of the BAC clones identified contained independently isolated markers. This is the first such report in an apomictic model that lacks recombination at the locus. We believe that extension of the contigs coupled to high-throughput sequencing will help the understanding of the genomic structure of the apomixis locus in P. simplex.Peer reviewe

Background: Medicago truncatula is a model species for legumes. Its functional genomics have been considerably boosted in recent years due to initiatives based both in Europe and US. Collections of mutants are becoming increasingly available and this will help unravel the genetic control of important traits for many species of legumes.

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