Induction of androgenesis and production of haploid embryos in anther cultures of borage (Borago officinalis L.)

[EN] Borage (Borago officinalis L.) is an important medicinal plant with different culinary, pharmaceutical and industrial properties. Unfortunately, there are no published reports on the establishment of protocols to produce DHs in this species up to now. In this work, we show for the first time the induction of borage microspores to become embryogenic calli, from which haploid embryos are produced. In addition, we evaluated the effect of using different flower bud sizes, carbon sources, concentrations of 2,4-D and BAP, cold (4 A degrees C) pretreatments and heat shock treatments. Production of total calli, embryogenic calli and callus-derived embryos was differently affected by the different parameters studied. Our results showed that the use of 5-7 mm-long flower buds, a cold (4 A degrees C) pretreatment during 4 days, a 32 A degrees C heat shock for 3 days, and the addition of 3 % maltose and 2 mgl(-1) 2,4-D and 1 mgl(-1) BAP to the culture medium, was beneficial for embryo production. Overall, this work demonstrates that DH technology is possible in borage, and opens the door for future improvements needed to finally obtain borage DH plants.Eshaghi, ZC.; Abdollahi, MR.; Moosavi, SS.; Deljou, A.; Seguí-Simarro, JM. (2015). Induction of androgenesis and production of haploid embryos in anther cultures of borage (Borago officinalis L.). Plant Cell, Tissue and Organ Culture. 122:321-329. doi:10.1007/s11240-015-0768-5S321329122Abdollahi MR, Moieni A, Javaran MJ (2004) Interactive effects of shock and culture density on embryo induction in isolated microspore culture of Brassica napus L. cv. 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Associação de fases meióticas e estádios dos micrósporos com características morfológicas de botões florais de pimentão Association of meiotic phases and microspore stages with morphological characters of floral buds of pepper

Fases meióticas e estádios de micrósporos de pimentão (Capsicum annuum L. cv. Azeth) foram determinados e associados com características morfológicas adotadas para a seleção de botões florais a serem utilizados na indução de androgênese. Plantas foram mantidas em casa-de-vegetação para coleta dos botões florais, que foram separados em seis classes de acordo com a relação de tamanho entre cálice e corola e presença de pigmentos nas anteras. As anteras foram fixadas em metanol: ácido acético na proporção de 3:1 e armazenadas a -20º C. Preparações citogenéticas desse material foram montadas pela técnica de dissociação e secagem ao ar e coradas com solução de Giemsa. As observações dos botões foram realizadas sob lupa e as preparações citogenéticas em microscópio ótico. Imagens dos botões florais, das anteras e das fases meióticas foram digitalizadas em computador para documentação. Variações de fases meióticas dentro de cada classe de botão floral foram observadas. Embora o critério de presença de antocianina na extremidade das anteras tenha sido aplicado para outras variedades, o mesmo não se mostrou adequado para a determinação do estádio de micrósporo neste estudo. As fases meióticas foram citogeneticamente identificadas; contudo, não foi possível estabelecer sua associação com as classes dos botões florais. Entretanto, botões com o tamanho de cálice coincidindo com o da corola apresentaram maior número de micrósporos em estádio adequado para a cultura de anteras.<br>In the present study, morphological characters adopted for floral bud selection used for androgenesis induction were associated with pepper (Capsicum annuum L. cv. Azeth) meiotic phases and microspore stages. Floral buds were harvested from greenhouse-grown plants and separated into six classes according to size relationships between calyx and corolla, and anthocyanin pigmentation in anthers. After sorting by size, buds were fixed using a methanol: acetic acid (3:1) solution at -20ºC. Cytogenetic preparations were mounted using an adaptation of an air-drying technique and staining through a phosphate buffer Giemsa solution. Bud observations were accomplished under a stereo-microscope and the cytogenetic preparations in an optic microscope. Recordings of floral buds and meiotic phases were performed with an image digitizing computer system. Variations on the meiotic phases were observed within each class. Although the approach based on anthocyanin pigmentation on anthers has been applied to another pepper varieties, it was not a reliable trait for determination of the microspore stage. Meiotic phases were citogeneticaly identified, although bud stage could not be associated with it. On the other hand, buds having the same calyx and corolla sizes presented a larger number of microspores in a suitable stage for anther culture