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

Abstract

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.

Production of doubled haploid plants from anther cultures of borage (Borago officinalis L.) by the application of chemical and physical stress

“…Flower buds were removed from donor plants, surface sterilized and cultured according to Chardoli Eshaghi et al (2015). Briefly, 5-7 mm-long buds, containing mid-to-late microspores, were excised and surface sterilized in a laminar flow hood by immersion in 70% (v/v) ethanol for 30 s, followed by immersion in 2.5% (v/v) sodium hypochlorite solution for 10 min, and finally rinsed in sterile distilled water three times.…”

“…Despite their advantages, DH technology has not been fully implemented in borage yet. In a previous study, we demonstrated that production of haploids in borage is possible, since we showed for the first time the production of borage embryo-like structures (ELSs) from anther cultures (Chardoli Eshaghi et al 2015). However, optimization of the procedure and regeneration of haploid and DH plants remained pending.…”

Improvement in androgenic response of borage (Borago officinalis L.) cultured anthers using antibrowning agents and picloram

“…This phenomenon leads to a decrease in culture competence (Benson, 2000). In our previous report (Chardoli Eshaghi et al, 2015), we first reported an anther culture protocol for borage under the influence of temperature pretreatments and some hormone treatments. We successfully obtained a limited number of gametic embryos in that study.…”

“…Borage buds (5-7 mm) containing anthers with microspores at the mid-to-late uninucleate stage were collected from donor plants of 50-60 days old and disinfected as follows: surface sterilization with 70% (v/v) ethanol for 30 s, followed by 2.5% (v/v) cold (4 °C) sodium hypochlorite for 10 min, and three rinses of 5 min each in cold (4 °C) sterile distilled water. Anthers with microspores at the mid-to-late uninucleate stage (Chardoli Eshaghi et al, 2015) were dissected and plated with 20 anthers per petri dish (90-100 mm in diameter) on 20 mL of induction medium comprising the macroelements and microelements of B5 (Gamborg et al, 1968), vitamins of NLN medium (Lichter, 1982), 0.8% agar, 3% maltose, 2 mg L -1 2,4-D, and 1 mg L -1 BAP (Chardoli Eshaghi et al, 2015). Dishes were placed at 4 °C in the dark for 5 days and then transferred to 25 °C in a growth room under a 16/8 photoperiod with a light intensity of 40 µmol m -2 s -1 for 2 weeks.…”

“…2.6. Determination of chromosome number and ploidy level Cytological analyses of the diploid root of borage seedling and anther-derived calli and embryos were carried out by the method described by Chardoli Eshaghi et al (2015). In order to determine the ploidy level of anther-derived calli and embryos and also the borage seedlings, flow cytometry was carried out with a ploidy analyzer (Partec GmbH, Germany).…”