Stress treatments and in vitro culture conditions influence microspore embryogenesis and growth of callus from anther walls of sweet pepper (Capsicum annuum L.)

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作者：Verónica Parra-Vega (1) Bego?a Renau-Morata (1) (2) Alicia Sifres (1) José M. Seguí-Simarro (1)
关键词：Androgenesis ; Capsicum annuum ; Heat stress ; Microspore embryogenesis ; In vitro culture
刊名：Plant Cell, Tissue and Organ Culture
出版年：2013
出版时间：March 2013
年：2013
卷：112
期：3
页码：353-360
全文大小：556KB
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作者单位：Verónica Parra-Vega (1) Bego?a Renau-Morata (1) (2) Alicia Sifres (1) José M. Seguí-Simarro (1) 1. COMAV, Universitat Politècnica de València, CPI, Edificio 8E - Escalera I, Camino de Vera, s/n, 46022, Valencia, Spain 2. Departamento de Producción Vegetal, Universitat Politècnica de València, Camino de Vera, s/n, 46022, Valencia, Spain
ISSN：1573-5044

文摘

Production of doubled haploids (DHs) is a convenient tool to obtain pure lines for breeding purposes. Until now, the easiest and most useful approach to obtain pepper DHs is via anther culture. However, this method has an associated possibility of producing calli from anther wall tissues that would be coexisting in the anther locule with embryos derived from microspores. Using two established protocols for anther culture, Dumas de Vaulx et al. (Agronomie 2:983-88, 1981) and Supena et al. (Sci Hort 107:226-32, 2006a; Plant Cell Rep 25:1-0, 2006b) callus and embryo development was assessed in four sweet pepper cultivars. For all genotypes tested, the protocol of Dumas de Vaulx et al. (Agronomie 2:983-88, 1981) promoted both embryo development and callus growth, whereas the protocol of Supena et al. (Sci Hort 107:226-32, 2006a; Plant Cell Rep 25:1-0, 2006b) produced no callus but only embryos. However, differences in embryo production were observed among these genotypes. In parallel, anthers were exposed to a 35?°C inductive heat shock for 4, 8, 12 and 16?days, prior to culture at 25?°C. The duration of the heat shock had significant effects in embryo production, but also in callus generation. Callus generation increased with prolonged exposures to 35?°C. Embryo and callus origin was analyzed by flow cytometry, light microscopy and molecular markers. Tests conducted demonstrated a gametophytic origin for all of the embryos tested, and a sporophytic origin for all of the calli. Together, our results reveal that culture conditions have a significant influence on the presence of calli derived from anther walls, which could be minimized by reducing heat shock exposure and/or using a shed-microspore approach.

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