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Small RNA Solexa sequencing of multiple tissues of papaya, Carica papaya

ABSTRACT: The tissues of male flowers, female flowers and hermaphrodite are collected and stored in liquid nitrigen immediately. Total RNA was isolated from male, hermaphrodite, and female flowers using the Trizol reagent (Invitrogen USA). Low molecular weight RNA was enriched from the total RNA by precipitating with 0.4M NaCl and polyethylene glycol (PEG). The enriched low molecular weight RNA was then separated on a denaturing 15% polyacrylamide gel. The band corresponding to the RNA standard of 18-30 nucleotides was then excised from the gel and eluted overnight in 0.4M NaCl at 4 degrees Celsius, and subsequently ligated with 5’ and 3’ Illumina small RNA adapters. The adapter ligated library was converted into cDNA and amplified by using PCR and sequenced by Illumina Genome Analyzer II.

ORGANISM(S): Carica papaya

PROVIDER: GSE54097 | GEO | 2014/12/31

SECONDARY ACCESSION(S): PRJNA235179

REPOSITORIES: GEO

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Project description:Papaya (Carica papaya) is a trioecious species, with female, male, and hermaphrodite plants. Because of sex segregation, selecting hermaphroditic plants is vital for orchard establishment due to their higher commercial value. In addition to the costly sexing step, environmental stresses can result in abnormal flower development. However, molecular mechanisms that regulate sex differentiation in papaya are still poorly known. Thus, this study aimed to identify proteins associated with sex development in female and hermaphrodite flowers of papaya through comparative proteomic analysis. Proteins from flower buds at the early and late developmental stages of three papaya genotypes (UENF-CALIMAN 01, JS12, and Sunrise Solo 72/12) were studied via proteomic analysis via the combination of the shotgun method and nanoESI-HDMSE technology. In buds at an early stage of development, 496 proteins exhibited significantly different abundances between sexes for the SS72/12 genotype, 139 for the JS12 genotype, and 165 for the UC-01 genotype. At the final stage of development, there were 181 for SS72/12, 113 for JS12, and 125 for UC-01. The large group of differentially accumulated proteins (DAPs) between the sexes was related to metabolism, as shown by the observation of only the proteins that exhibited the same pattern of accumulation in the three genotypes. Specifically, carbohydrate metabolism proteins were up-regulated in hermaphrodite flower buds early in development, while those linked to monosaccharide and amino acid metabolism increased during late development. Enrichment of sporopollenin and phenylpropanoid biosynthesis pathways characterizes hermaphrodite samples across developmental stages, with predicted protein interactions highlighting the crucial role of phenylpropanoids in sporopollenin biosynthesis for pollen wall formation. Most of the DAPs played key roles in pectin, cellulose, and lignin synthesis and were essential for cell wall formation and male flower structure development, notably in the pollen coat. These findings suggest that hermaphrodite flowers require more energy for development, likely due to complex pollen wall formation. Overall, these insights illuminate the molecular mechanisms of papaya floral development, revealing complex regulatory networks and energetic demands in the formation of male reproductive structures.

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Small RNA Solexa sequencing of multiple tissues of papaya, Carica papaya

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