Mass spectrometry screening reveals widespread diversity in trichome specialized metabolites of tomato chromosomal substitution lines

Glandular secreting trichomes of cultivated tomato (Solanum lycopersicum) and close relatives produce a variety of structurally diverse volatile and non-volatile specialized (‘secondary’) metabolites, including terpenes, flavonoids and acyl sugars. A genetic screen is described here to profile leaf trichome and surface metabolite extracts of nearly isogenic chromosomal substitution lines covering the tomato genome. These lines contain specific regions of the Solanum pennellii LA0716 genome in an otherwise ‘wild-type’ M82 tomato genetic background. Regions that have an impact on the total amount of extractable mono- and sesquiterpenes (IL2-2) or only sesquiterpenes (IL10-3) or specifically influence accumulation of the monoterpene α-thujene (IL1-3 and IL1-4) were identified using GC-MS. A rapid LC-TOF-MS method was developed and used to identify changes in non-volatile metabolites through non-targeted analysis. Metabolite profiles generated using this approach led to the discovery of introgression lines producing different acyl chain substitutions on acyl sugar metabolites (IL1-3/1-4 and IL8-1/8-1-1), as well as two regions that influence the quantity of acyl sugars (IL5-3 and IL11-3). Chromosomal region 1-1/1-1-3 was found to influence the types of glycoalkaloids that are detected in leaf surface extracts. These results show that direct chemical screening is a powerful way to characterize genetic diversity in trichome specialized metabolism

Pyrosequencing of the Camptotheca acuminata transcriptome reveals putative genes involved in camptothecin biosynthesis and transport

Background: Camptotheca acuminata is a Nyssaceae plant, often called the "happy tree", which is indigenous in Southern China. C. acuminata produces the terpenoid indole alkaloid, camptothecin (CPT), which exhibits clinical effects in various cancer treatments. Despite its importance, little is known about the transcriptome of C. acuminata and the mechanism of CPT biosynthesis, as only few nucleotide sequences are included in the GenBank database.Results: From a constructed cDNA library of young C. acuminata leaves, a total of 30,358 unigenes, with an average length of 403 bp, were obtained after assembly of 74,858 high quality reads using GS De Novo assembler software. Through functional annotation, a total of 21,213 unigenes were annotated at least once against the NCBI nucleotide (Nt), non-redundant protein (Nr), Uniprot/SwissProt, Kyoto Encyclopedia of Genes and Genomes (KEGG), and Arabidopsis thaliana proteome (TAIR) databases. Further analysis identified 521 ESTs representing 20 enzyme genes that are involved in the backbone of the CPT biosynthetic pathway in the library. Three putative genes in the upstream pathway, including genes for geraniol-10-hydroxylase (CaPG10H), secologanin synthase (CaPSCS), and strictosidine synthase (CaPSTR) were cloned and analyzed. The expression level of the three genes was also detected using qRT-PCR in C. acuminata. With respect to the branch pathway of CPT synthesis, six cytochrome P450s transcripts were selected as candidate transcripts by detection of transcript expression in different tissues using qRT-PCR. In addition, one glucosidase gene was identified that might participate in CPT biosynthesis. For CPT transport, three of 21 transcripts for multidrug resistance protein (MDR) transporters were also screened from the dataset by their annotation result and gene expression analysis.Conclusion: This study produced a large amount of transcriptome data from C. acuminata by 454 pyrosequencing. According to EST annotation, catalytic features prediction, and expression analysis, novel putative transcripts involved in CPT biosynthesis and transport were discovered in C. acuminata. This study will facilitate further identification of key enzymes and transporter genes in C. acuminata

Durian (<i>Durio zibethinus</i> L.): Nutritional Composition, Pharmacological Implications, Value-Added Products, and Omics-Based Investigations

Durian (Durio zibethinus L.), a tropical fruit celebrated in Southeast Asia for its distinct flavor, is the focus of this comprehensive review. The fruit’s pulp is rich in high-value bioactive compounds, including gamma-glutamylcysteine, a precursor to the essential antioxidant glutathione. With durian cultivation gaining prominence in Southeast Asia due to its economic potential through cultivar enhancement, an in-depth examination of durian-related research becomes crucial. This review explores the health benefits of durian, analyzing the nutritional compositions and bioactive compounds present in the pulp, peel, and seed. It also underscores durian-based food products and the potential for valorizing durian waste. This review encapsulates the significant advancements made in omics-based research, aimed at deciphering the molecular complexities of durian fruit post-harvest ripening and the metabolic shifts impacting its sensory attributes. It is the first to summarize studies across genome, transcriptome, and metabolome levels. Future research should prioritize the development of molecular markers to accelerate the breeding of elite cultivars with preferred traits. It also proposes the exploration of durian waste valorization, including underexplored parts like flowers and leaves for their bioactive compounds, to promote a sustainable bioeconomy. Finally, it suggests the development of innovative durian products catering to the tastes of health-conscious consumers

Gene expression analysis, subcellular localization, and in planta antimicrobial activity of rice (Oryza sativa L.) defensin 7 and 8

Defensins are a group of plant antimicrobial peptides. In a previous study, it was reported that two recombinant rice (Oryza sativa L.) defensin (OsDEF) genes (OsDEF7 and OsDEF8) produced heterologously by bacteria inhibited the growth of several phytopathogen. Here, we analyzed gene expression patterns in Thai jasmine rice (O. sativa L. ssp. indica ‘KDML 105’) using quantitative reverse transcription-polymerase chain reaction and compared them with those in Japanese rice (O. sativa L. ssp. japonica ‘Nipponbare’). Although the cultivars exhibited similar gene expression patterns at the developmental stages examined, the expression levels differed between organs. Upon Xanthomonas oryzae pv. oryzae infection in the leaves, both OsDEFs were highly upregulated at 8 days post-infection, suggesting that they play a role in pathogen defense. Moreover, in silico analyses revealed that OsDEF expression levels were affected by drought, cold, imbibition, anoxia, and dehydration stress. Using green fluorescent protein (GFP) fusions, we found that both OsDEFs were in the extracellular compartment, confirming their functions against pathogen infection. However, when recombinant OsDEFs (without GFP) were produced in tobacco BY-2 cells or Nicotiana benthamiana leaves, they could not be detected in either the culture medium or the cells. Yet, N. benthamiana leaves infiltrated with OsDEF7 or OsDEF8 constructs exhibited in planta inhibitory activity against the phytopathogen Xanthomonas campestris pv. glycines, suggesting that recombinant OsDEFs were present. Additionally, when targeting them to the ER compartment, recombinant OsDEFs could be detected. Lower inhibitory activity was observed when recombinant OsDEFs were targeted to the ER. These results suggest that OsDEFs play a role in controlling plant diseases