The Effect of Centrifugal Treatment on the Emergence of Plantlet from Cultured Anther of Tobacco

Androgenesis-inducing stress treatments change phytohormone levels in anthers of three legume species (Fabaceae)

“…Auxin is involved in many plant growth functions including flowering and fertilization (Aloni et al 2006;Hirano et al 2008;Tromas and Perrot-Rechenmann 2010;Zhao 2010). Tanaka (1973) was the first to report that centrifugation of tobacco anthers in the presence of IAA improved androgenesis induction. In contrast, Sangwan-Norreel (1977) found a positive effect of centrifugation without the addition of hormones to Datura innoxia anthers.…”

“…Centrifugation increases androgenesis induction of cultured anthers in Datura innoxia and in tobacco (Nicotiana tabacum L.) (Sangwan-Norreel 1977;Tanaka 1973). Similarly, centrifugation positively affects androgenesis induction in chickpea and pea Ochatt et al 2009).…”

Microspore Embryogenesis

“…In tobacco anthers, a four-fold increase in the regeneration of haploid plants was reported following centrifugation of the anthers prior to culture, although the maturity stage of the microspores determined the degree of success (Tanaka 1973). Experiments with chick pea (Cicer arietinum) also appear to suggest that centrifugation may have a positive impact on embryo initiation from cultured anthers, even though the response is variety specific (Grewal et al 2009).…”

“…For example, in rapeseed, severe heat stress (41 0 C) is required to induce efficient embryogenic development in late bi-nucleate microspores (Binarova et al 1997), mild heat stress (32 0 C) to induce late uni-nucleate to early bi-nucleate staged microspores (Ferrie and Keller 1995), whereas uni-nucleate microspores at an earlier stage of development corresponding to vacuolated stage are induced more effectively with colchicine (Zhao et al 1996). Similarly, in tobacco bi-cellular pollen is easily induced under sucrose and nitrogen starvation pre-treatment conditions (Garrido et al 1995, Kyo andHarada 1986) whereas microspores at the uni-nucleate stage are induced under heat shock (Touraev et al 1996a), and centrifugal pre-treatment (Tanaka 1973), although the latter two pre-treatments have little influence in inducing embryogenesis from bi-cellular pollen. Accordingly, through careful experimentation and judicious choice of pre-treatments, it would be possible to induce microspores of a wider developmental range than anticipated before, towards successful embryogenic re-programming.…”

Flow cytometry enables identification of sporophytic eliciting stress treatments in gametic cells

“…Improved androgenic response after modification of osmotic pressure has been observed in a number of species (Jain et al, 1996(Jain et al, -1997Lionneton et al, 2001), including legumes (Croser et al, 2006;Grewal et al, 2009;Ochatt et al, 2009). Centrifugation is applied as part of many microspore isolation protocols, but has also been used as a stress treatment for induction of androgenesis from intact anther culture in chickpea , tobacco (Tanaka, 1973), Datura innoxia Mill. (Sangwan-Norreel, 1977) and rapeseed (Aslam et al, 1990).…”