Sexual Plant Reproduction

, Volume 14, Issue 4, pp 233–238 | Cite as

Analysis of gene expression during flowering in apomeiotic mutants of Medicago spp.: cloning of ESTs and candidate genes for 2n eggs

  • Gianni Barcaccia
  • Serena Varotto
  • Stefano Meneghetti
  • Emidio Albertini
  • Andrea Porceddu
  • Paolo Parrini
  • Margherita Lucchin
Original Article

Abstract

Mutants showing features of apomixis have been documented in alfalfa (Medicago sativa L.), a natural outcrossing sexual species. A differential display of mRNAs that combines cDNA-AFLP markers and bulked segregant analysis was carried out with the aim of selecting expressed sequence tags (ESTs) and cloning candidate genes for apomeiosis in mutants of alfalfa characterized by 2n egg formation at high frequencies. The approach enabled us to select either mutant- or wild type-specific transcript derived-fragments and to detect transcriptional changes potentially related to 2n eggs. Sequence alignments of a subset of 40 polymorphic clones showed significant homologies to genes of known function. An EST with identity to a β-tubulin gene, highly expressed in the wild type and poorly expressed in the apomeiotic mutants, and an EST with identity to a Mob1-like gene, qualitatively polymorphic between pre- and post-meiotic stages, were selected as candidate genes for apomeiosis because of their putative roles in the cell cycle. A number of clone-specific primers were designed for performing both 5′ and 3′ rapid amplification of cDNA ends to obtain the full-length clones. Southern blot hybridization revealed that both clones belong to a multi-gene family with a minimum of three genomic DNA members each. Northern blot hybridization of total RNA samples and in situ hybridization of whole buds enabled the definition of their temporal and spatial expression patterns in reproductive organs. Experimental achievements towards the elucidation of apomeiotic megasporogenesis in alfalfa are presented and discussed

Differential display cDNA-AFLP Bulked segregant analysis Diplospory Tubulins Mob (Mps-One-Binder) 

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Copyright information

© Springer-Verlag 2001

Authors and Affiliations

  • Gianni Barcaccia
    • 1
  • Serena Varotto
    • 1
  • Stefano Meneghetti
    • 1
  • Emidio Albertini
    • 2
  • Andrea Porceddu
    • 3
  • Paolo Parrini
    • 1
  • Margherita Lucchin
    • 1
  1. 1.Dipartimento di Agronomia Ambientale e Produzioni Vegetali, University of Padua, Agripolis, Via Romea 16, 35020 Legnaro, PaduaItaly
  2. 2.Dipartimento di Biologia Vegetale e Biotecnologie Agroambientali, University of Perugia, Borgo XX Giugno 74, 06121 PerugiaItaly
  3. 3.Istituto per il Miglioramento Genetico delle Piante da Orto e da Fiore, CNR, Viale dell'Università 133, 80055 Portici, NaplesItaly

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