non-aqueous-isolation-enrichment-glandular-capitate-stalked-sessile

Non-Aqueous Isolation and Enrichment of Glandular Capitate Stalked and Sessile Trichomes from <em>Cannabis sativa</em>

This paper presents a protocol for the convenient and high-throughput isolation and enrichment of glandular capitate stalked and sessile trichomes from ...

ARO-Newe Ya‘ar Center

Taste has been the subject of human selection in the evolution of agricultural crops, and acidity is one of the three major components of fleshy fruit taste, together with sugars and volatile flavour compounds. We identify a family of plant-specific genes with a major effect on fruit acidity by map-based cloning of C. melo PH gene (CmPH) from melon, Cucumis melo taking advantage of the novel natural genetic variation for both high and low fruit acidity in this species. Functional silencing of orthologous PH genes in two distantly related plant families, cucumber and tomato, produced low-acid, bland tasting fruit, showing that PH genes control fruit acidity across plant families. A four amino-acid duplication in CmPH distinguishes between primitive acidic varieties and modern dessert melons. This fortuitous mutation served as a preadaptive antecedent to the development of sweet melon cultigens in Central Asia over 1,000 years ago.

The PH gene determines fruit acidity and contributes to the evolution of sweet melons.

Melon (Cucumis melo) fruits exhibit phenotypic diversity in several key quality determinants such as taste, color and aroma. Sucrose, carotenoids and volatiles are recognized as the key compounds shaping the above corresponding traits yet the full network of biochemical events underlying their synthesis have not been comprehensively described. To delineate the cellular processes shaping fruit quality phenotypes, a population of recombinant inbred lines (RIL) was used as a source of phenotypic and genotypic variations. In parallel, ripe fruits were analyzed for both the quantified level of 77 metabolic traits directly associated with fruit quality and for RNA-seq based expression profiles generated for 27,000 unigenes. First, we explored inter-metabolite association patterns; then, we described metabolites versus gene association patterns; finally, we used the correlation-based associations for predicting uncharacterized synthesis pathways.

Systems approach for exploring the intricate associations between sweetness, color and aroma in melon fruits.

Melon fruit flesh color is primarily controlled by the "golden" single nucleotide polymorhism of the "Orange" gene, CmOr, which dominantly triggers the accumulation of the pro-vitamin A molecule, β-carotene, in the fruit mesocarp. The mechanism by which CmOr operates is not fully understood. To identify cellular and metabolic processes associated with CmOr allelic variation, we compared the transcriptome of bulks of developing fruit of homozygous orange and green fruited F3 families derived from a cross between orange and green fruited parental lines.

A bulk segregant transcriptome analysis reveals metabolic and cellular processes associated with Orange allelic variation and fruit β-carotene accumulation in melon fruit.

Volatile esters contribute to the aroma and flavor of many fruits but are normally absent in grape berries (Vitis vinifera L.). To examine the biosynthetic potential of grape berries to form volatile esters, berry sections were incubated with exogenous L-Phe, L-Leu or L-Met. In general, amino-acid incubation caused the accumulation of the respective aldehydes and alcohols. Moreover, L-Leu incubation resulted in the accumulation of 3-methylbutyl acetate and L-Phe incubation resulted in the accumulation 2-phenylethyl acetate in 'Muscat Hamburg' but not in the other grape accessions. Exogenous L-Met administration did not result in volatile esters accumulation but the accumulation of sulfur volatile compounds such as methional and dimethyl disulfide was prominent. Berry-derived cell-free extracts displayed differential alcohol acetyltransferase activities and supported the formation of 3-methylbutyl acetate and benzyl acetate. 2-Phenylethyl acetate was produced only in 'Muscat Hamburg' cell-free extracts. VvAAT2, a newly characterized gene, was preferentially expressed in 'Muscat Hamburg' berries and functionally expressed in E. coli. VvAAT2 possesses alcohol acetyltransferase activity utilizing benzyl alcohol, 2-phenylethanol, hexanol or 3-methylbutanol as substrates. Our study demonstrates that grape berries have a concealed potential to accumulate volatile esters and this process is limited by substrate availability.

Concealed ester formation and amino acid metabolism to volatile compounds in table grape (Vitis vinifera L.) berries.

Aroma volatiles are essential for plant ecological fitness and reproduction. Plants produce and use volatiles to attract pollinators and seed dispersers, repel herbivores and recruit their natural enemies, and communicate with other plants. Amino acids and their biosynthetic intermediates play key roles as precursors for the biosynthesis of plant volatiles. Different plants utilize different strategies and biosynthetic pathways to meet their specific biological needs. This review focuses on the different biosynthetic pathways that plants utilize to form amino acid-derived aroma volatiles, emphasizing their common and unique aspects and stressing the importance of the limiting enzymes residing in the primary-specialized metabolism interface. We also briefly review how biotechnology has used this interface and point to promising future directions for improving the quality of agricultural produce and the production of key volatiles.

Article	Year
The PH gene determines fruit acidity and contributes to the evolution of sweet melons. Nature communications\| PubMed ID: 24898284	2014
Systems approach for exploring the intricate associations between sweetness, color and aroma in melon fruits. BMC plant biology\| PubMed ID: 25887588	2015
A bulk segregant transcriptome analysis reveals metabolic and cellular processes associated with Orange allelic variation and fruit β-carotene accumulation in melon fruit. BMC plant biology\| PubMed ID: 26553015	2015
Concealed ester formation and amino acid metabolism to volatile compounds in table grape (Vitis vinifera L.) berries. Plant science : an international journal of experimental plant biology\| PubMed ID: 30080607	2018
Amino acids metabolism as a source for aroma volatiles biosynthesis. Current opinion in plant biology\| PubMed ID: 35533493	2022

Efraim Lewinsohn

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