Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Comparative analysis of stylar canal cells transcriptome in clementine mandarin (Citrus clementina)

ABSTRACT: Compared to what is known in model species, reproductive biology in citrus is still poorly understood. Although in recent years several efforts have been made to study pollen-pistil interaction and self-incompatibility, little information is available about the molecular mechanisms regulating these processes. We performed microarray analysis for the identification of candidate genes involved in pollen-pistil interaction and self-incompatibility in clementine (Citrus clementina Hort. ex Tan.). The analysis was performed comparing the transcriptome of laser-microdissected stylar canal cells isolated from two clementine genotypes differing for self-incompatibility response (‘Comune’, self-incompatible; and ‘Monreal’, a self compatible mutation of ‘Comune’). Styles with stigmas, collected 24 hours after self pollination, were immediately snap-frozen in OCT embedding medium (Sakura Finetek, Zoeterwoude, Netherlands) in Peel-A-Way plastic embedding molds (Polysciences, Polysciences, Warrington, PA, USA). Transversal sections 10 µm thick at the upper part of the style were cut with a Leica CM1900 cryostat (Leica Microsystems, Germany) at -20°C. A Leica AS Laser Microdissection system (Leica Microsystems) was used for the isolation of stylar canals from transversal sections. Canals from the stigma were discarded to avoid contamination with pollen or pollen tubes. Three biological replicates were prepared for each genotype. Each biological replicate consisted of bulks of about 200 microdissected areas (composed of an average of 50 cells) coming from two different molds

ORGANISM(S): Citrus clementina

SUBMITTER: Marco Caruso

PROVIDER: E-GEOD-33014 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Comparative transcriptome analysis of stylar canal cells identifies novel candidate genes implicated in the self-incompatibility response of Citrus clementina.

Caruso Marco M Merelo Paz P Distefano Gaetano G La Malfa Stefano S Lo Piero Angela Roberta AR Tadeo Francisco R FR Talon Manuel M Gentile Alessandra A

BMC plant biology 20120214

<h4>Background</h4>Reproductive biology in citrus is still poorly understood. Although in recent years several efforts have been made to study pollen-pistil interaction and self-incompatibility, little information is available about the molecular mechanisms regulating these processes. Here we report the identification of candidate genes involved in pollen-pistil interaction and self-incompatibility in clementine (Citrus clementina Hort. ex Tan.). These genes have been identified comparing the tr ...[more]

PMID: 22333138

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Project description:Gametophytic self-incompatibility is the primary cause of low fruit set in almond. The mechanism of recognition that determines whether the gametophyte is successfully fertilized between pollen tube SCF (F-box-SSK1-Cul1-Rbx1) protein and pistil S-RNase protein during fertilization is unclear. In this study, the pistils of two almond cultivars 'Wanfeng' and 'Nonpareil' were used as the experimental materials after selfing- and nonselfing/cross-pollination, and pistils from the stamen-removed flowers were used as controls. We used fluorescence microscopy to observe the development of pollen tubes after pollination and 4-dimensional label-free quantitation (4D-LFQ) to detect the protein expression profiles of 'Wanfeng' and 'Nonpareil' pistils and in controls. The results showed that it took 24-36 h for the development of the pollen tube to 1/3 of the pistil, and a total of 7684 differentially accumulated proteins (DAPs) were identified in the pistil after pollinating 36 h, of which 7022 were quantifiable. The up- and down-regulation of the 12 differential protein expressions identified by parallel reaction monitoring (PRM) was the same as that identified by 4D-LFQ, with an average fold-change difference of 14.34%, a maximum of 31.37% and a minimum of 3.68%. Bioinformatics analysis based on the function of the differential proteins, including classification, enrichment, and clustering, identified RNA polymerases (4 DAPs), autophagy (3 DAPs), oxidative phosphorylation (3 DAPs), and homologous recombination (2 DAPs) pathways associated with the self-incompatibility process. The interaction between the serine/threonine kinase (MARK2) protein E3 ubiquitin ligase and the microtubule protein component microtubule-associated protein tau (MAPT/ACT) was found using the STRING database, which demonstrated the involvement of the MARK2 protein in the reaction of pollen tube recognition the nonself- and the self-S-RNase protein. It provides a new way to reveal the mechanism by which almond pollen tubes recognize the self and nonself S-RNase enzyme protein.

Dataset Information

Comparative analysis of stylar canal cells transcriptome in clementine mandarin (Citrus clementina)

Publications

Comparative transcriptome analysis of stylar canal cells identifies novel candidate genes implicated in the self-incompatibility response of Citrus clementina.

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