Microspore culture protocol for Indonesian Brassica oleracea

Abstract

A microspore culture protocol for Brassica oleracea of Indonesian origin (cv. 'Kemeh') has been successfully established. A high number of embryos formed with high microspore density i.e. 15 9 10 4 cells/ ml. Embryo formation was improved by using flower buds (4.5-4.6 mm in length) as explants, a temperature treatment at 30.5°C for 48 h and then transfer to 25°C continuously until embryos formed. A total of 295 embryos were obtained from 189 buds, 30% of which were abnormal (i.e. with an abnormal cotyledon or lacking hypocotyls). All normal embryos that grew and survived, 165 in total, were successfully transferred to soil and grew well in plastic bags (15 cm in diameter) containing a mixture of burnedrice husk and organic manure (1:1, v/v).

Microspore embryogenesis and production of haploid and doubled haploid plants in carrot (Daucus carota L.)

“…With DH production systems established, homozygous lines can be achieved in one generation, which would allow to reduce the time and cost of cultivar development (Dunwell 2010;Ferrie and Caswell 2010;Germanà 2011). The system has been effectively used in barley, wheat (Jauhar et al 2009;Ferrie and Caswell 2010), maize (Smith et al 2008;Chang and Coe 2009), and various Brassica species (Belmonte et al 2006;Fang et al 2006;Zhang et al 2008;Takahira et al 2011;Winarto and Teixeira da Silva 2011). The genotype is one of the crucial factors evaluated in many plants (Ferrie et al 1995;Dunwell 2010).…”

“…The benefits of various starvation and stress techniques have been reported in several plant species (Gu et al 2004;Zoriniants et al 2005;Dunwell 2010;Zhuang et al 2010;Bhowmik et al 2011;Chen et al 2011;Seguí-Simarro et al 2011;Takahira et al 2011;Winarto and Teixeira da Silva 2011). A 28-day treatment of spikes at 4°C was shown to lead to a much increased yield of microspore calli in cereals (Dunwell 2010).…”

Effects of histone deacetylase inhibitors on microspore embryogenesis and plant regeneration in Pakchoi ( Brassica rapa ssp. chinensis L.)

“…Microspore embryogenesis is influenced by many factors such as donor plant genotype and growth status, microspore vigor, culture conditions, pretreatments, and culture medium (Babbar et al, 2004;Bhowmik et al, 2011;Ferrie and Caswell, 2011;Winarto and Teixeira da Siva, 2011). Stress has been frequently used to induce embryogenesis, and treatments used in efficient microspore embryogenesis at present are the application of a brief heat-shock (Zhao et al, 1996), cold pretreatment (Pechan and Smykal, 2001;Touraev et al, 1997), and carbon starvation (Ahmadi et al, 2014a) that modified the microspore's external environmental condition to induce them to produce embryos.…”

Microspore embryogenesis and plant regeneration in Brussels sprouts (Brassica oleracea L. var. gemmifera)

“…gongylodes, Yan et al, 1999), ornamental kale (B. oleracea L. var. acephala, Zhang et al, 2008), and Indonesian B. oleracea morphotypes (Winarto and Teixeira da Silva, 2011) subsequently. Moreover, many studies have been carried out to improve and to utilize this technique, such as the characterization of microspore embryogenesis process in vitro, the identification of different factors influencing embryogenesis and the utilization of this protocol in breeding process (Touraev et al, 1997;Takahata, 1997;Soriano et al, 2013).…”