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Identification, characterization and expression analysis of passion fruit (Passiflora edulis) microRNAs.

ABSTRACT: microRNAs (miRNAs) are highly conserved, short (~?21-nucleotide), endogenous, non-coding RNA molecules that play major roles in post-transcriptional silencing by guiding target mRNA cleavage or translational inhibition. In this study, applying high-stringent genome-wide computational-based approaches, a total of 28 putative miRNAs belonging to 17 miRNA families were identified from an antioxidant-rich medicinal plant passion fruit (Passiflora edulis). Inter-tissue (leaves and fruits) and inter-varietal (yellow and purple fruit varieties) quantitative study of six putative passion fruit miRNAs (ped-miR160, ped-miR164, ped-miR166, ped-miR393, ped-miR394, and ped-miR398) showed differential expression. Using psRNATarget tool, a total of 25 potential target proteins of the characterized passion fruit miRNAs were also identified. Most of the target proteins identified in this study, including SQUAMOSA promoter binding, Class III HD-Zip, NAC, Scarecrow, APETALA2, Auxin response factor, MYB, and superoxide dismutase, were found to be involved in development, metabolism, and defense/stress response signaling.

SUBMITTER: Paul S

PROVIDER: S-EPMC6938847 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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Identification, characterization and expression analysis of passion fruit (<i>Passiflora edulis</i>) microRNAs.

Paul Sujay S de la Fuente-Jiménez Juan Luis JL Manriquez Camila Garibay CG Sharma Ashutosh A

3 Biotech 20200102 1

microRNAs (miRNAs) are highly conserved, short (~ 21-nucleotide), endogenous, non-coding RNA molecules that play major roles in post-transcriptional silencing by guiding target mRNA cleavage or translational inhibition. In this study, applying high-stringent genome-wide computational-based approaches, a total of 28 putative miRNAs belonging to 17 miRNA families were identified from an antioxidant-rich medicinal plant passion fruit (<i>Passiflora edulis</i>). Inter-tissue (leaves and fruits) and ...[more]

PMID: 31938684

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Project description:High temperature in summer is an unfavorable factor for passion fruit (Passiflora edulis), which can lead to restricted growth, short flowering period, few flower buds, low fruit setting rate, severe fruit drop, and more deformed fruit. To explore the molecular physiology mechanism of passion fruit responding to high-temperature stress, we use 'Zhuangxiang Mibao', a hybrid passion fruit cultivar, as the test material. Several physiological indicators were measured and compared between high-temperature (average temperature 38 °C) and normal temperature (average temperature 25 °C) conditions, including photosynthesis, chlorophyll fluorescence parameters, peroxidase activity (POD), superoxide dismutase activity (SOD) and malondialdehyde content. We performed RNA-seq analysis combined with biochemistry experiment to investigate the gene and molecular pathways that respond to high-temperature stress. The results showed that some physiological indicators in the high-temperature group, including the net photosynthetic rate, stomatal conductance, intercellular CO2 concentration, transpiration rate, and the maximum chemical quantum yield of photosystemII (PSII), were significantly lower than those of the control group. Malondialdehyde content was substantially higher than the control group, while superoxide dismutase and superoxide dismutase activities decreased to different degrees. Transcriptome sequencing analysis showed that 140 genes were up-regulated and 75 genes were down-regulated under high-temperature stress. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation analysis of differentially expressed genes revealed many metabolic pathways related to high-temperature stress. Further investigation revealed that 30 genes might be related to high-temperature stress, such as chlorophyllide a oxygenase (CAO), glutathione (GSH), WRKY transcription factors (WRKY), and heat shock protein (HSP), which have also been reported in other species. The results of real-time fluorescence quantitative PCR and RNA-seq of randomly selected ten genes are consistent, which suggests that the transcriptome sequencing results were reliable. Our study provides a theoretical basis for the mechanism of passion fruit response to high-temperature stress. Also, it gives a theoretical basis for the subsequent breeding of new heat-resistant passion fruit varieties.

Project description:BackgroundPassion fruit (Passiflora edulis Sims) is an important horticultural crop in the tropics and subtropics, where it has great commercial potential due to high demand for fresh edible fruits and processed juice as well as source of raw materials in cosmetic industries. Genetic engineering shows great potential in passion fruit improvement and can compensate for the limitations of conventional breeding. Despite the success achieved in genetic modification of few passion fruit varieties, transgenic passion fruit production is still difficult for farmer-preferred cultivars. Therefore, it is important to establish a simple and fast Agrobacterium-mediated cell transformation of commercial hybrid passion fruit KPF4 (Passiflora edulis f. edulis × Passiflora edulis f. flavicarpa).ResultsIn the present study, we have developed a simple and fast Agrobacterium-mediated transformation system for hybrid passion fruit KPF4 using leaf disc explants. Factors affecting the rate of transient beta (β)-glucuronidase (gusA) expression and consequently transformation efficiency were optimized as follows: Agrobacterium cell density with an OD600 of 0.5, 30 min infection time, 3 days of co-cultivation duration and the incorporation of 200 µM acetosyringone into Agrobacterium infection suspension medium. Using the optimized conditions, transgenic plants of KPF4 were produced within 2 months with an average transformation efficiency of 0.67%. The β-glucuronidase (GUS) histochemical staining confirmed the expression and integration of an intron-containing gusA gene into transformed leaf discs and transgenic plant lines of KPF4. The presence of gusA gene in the transgenic plants was confirmed by polymerase chain reaction (PCR). The results confirmed that the gusA gene was efficiently integrated into the passion fruit genome.ConclusionsThe developed transformation protocol is simple and rapid and could be useful for functional genomic studies and transferring agronomically important traits into passion fruit hybrid KPF4. This study developed a method that can be used to transfer traits such as resistance to viral diseases, low fruit quality and short storage life. To the best of our knowledge, this is the first report on genetic transformation system for commercial passion fruit hybrid KPF4.

Dataset Information

Identification, characterization and expression analysis of passion fruit (Passiflora edulis) microRNAs.

Publications

Identification, characterization and expression analysis of passion fruit (<i>Passiflora edulis</i>) microRNAs.

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