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placenta seeds

Placenta seeds

Metabolomics is defined as the comprehensive analysis of all low molecular weight organic compounds (<1500 Da) in a biological system [32]. Mass spectrometry has become the most widely applied platform for metabolomics, due to the wide range of molecules that can be analyzed on a single run [33]. Global profiling or non-targeted mass spectrometry-based metabolomics have gained importance in the study of crop species and have been applied to investigate potato, tomato, rice, wheat, strawberry, cucumber, and tobacco [34,35,36,37]. In the field of plant metabolomics, liquid chromatography coupled with electrospray ionization high resolution mass spectrometry (UPLC-ESI-HRMS) has emerged as the technique of choice for the putative identification of metabolites in complex matrices. This technique has been widely used, due to its sensitivity, selectivity, and analysis capability [38,39]. Nevertheless, metabolite identification for unknown compounds still remains a big challenge to overcome. In that respect, the recommendations by the Metabolomics Standards Initiative (MSI) recognize five different levels for metabolite confidence annotations. Level 0 requires the full compound 3D structure and stereochemistry information. Levels which are more common include: Level 1 identifications need a confirmation by two orthogonal parameters such as retention time and MS/MS spectrum, normally with match reference standards; and Level 2 requires at least two orthogonal pieces of information, including evidence that excludes all other candidates. Data for Level 2 should describe probable structure and be matched to literature data or databases by diagnostic evidence [40].

2. Results

The global metabolic comparison between the tissues and seeds of C. frutescens showed several feature differences between the pericarp, placenta, and seed. The Level 1 and 2 confidence metabolite annotations allowed us to assign a putative identification to these ions. Around 30% of metabolites were shared between all three parts. Compounds related to the primary metabolism showed few significant differences, they included amino acid related compounds, fatty acids, and phospholipids. As shown in the Venn diagram ( Figure 5 ) and Table S1, placenta and pericarp have the biggest compound class diversity. Significantly, the seeds presented a higher number of putative identifications, and these were primarily saponins, terpenes, and fatty acids.

3. Discussion

In summary, the non-targeted LC-MS metabolomics method that was developed in this study is shown to be a powerful tool for the putative identification of tissue-specific secondary metabolites at the red mature stage of chili pepper fruit. The use of databases available online gave rise to a faster comprehensive elucidation of global characteristics of a complex matrix than more traditional phytochemical studies. Nutraceutical, aroma, flavor, and new compounds that have not been reported before were putatively identified and related to pericarp, placenta or seeds of C. frutescens. As presented here, some of these compounds have been reported with bioactivity properties, supporting empirical properties of pepper fruit that have been known for centuries. The procedure developed here will be utilized for further studies in our laboratory, including to enable the exploration of comparisons between wild cultivars of chili pepper fruit with their cultivated counterparts and for the further understanding of secondary metabolism in this crop. We recommend that complementary analysis should be carried out to confirm structural elucidation. In addition, compound isolation and bioactivity properties should be considered in future studies.

Cherry tomato ( Lycopersicon esculentum Mill.) seeds harvested from fruits at four stages of development [2, 4, 6 and 8 weeks after flowering (WAF)] were washed (1% HCl) and germinated after 72 h of desiccation at 20°C, and 76%, 49% or 12% RH. Seed α-amylase activity was determined at each stage of development and correlated with seed germination. Desiccation at 76% and 49% RH had no significant effect on the germination of seeds at 4, 6 and 8 WAF, whereas it improved germination of seeds at 2 WAF. Low RH (12%) significantly reduced seed germination at all stages of development. There was a positive correlation ( r = 0.68, P ≤ 0.05) between germination and α-amylase activity during seed development. Scanning electron microscopy also revealed a correlation between starch grain occurrence in the endosperm and α-amylase activity. Placenta water potential decreased with fruit and seed maturation, and correlated negatively with improved seed germination and α-amylase activity. Promotion of seed germination by desiccation in developing cherry tomato seeds was lost 4 weeks prior to mass maturity (80 ± 3% seed water content and –2.3 ± 0.1 MPa placenta water potential). It is proposed that the water potential differential between the placenta and the seeds influences α-amylase activity and germination behaviour during development.