Microspore culture and the performance of microspore derived doubled haploid in <i>Brassica juncea</i> (L.)

Abstract

The experiment was conducted at the Tissue Culture Laboratory as well as in the net house of Oilseed Research Centre, Bangladesh Agricultural Research Institute (BARI), Joydebpur, Gazipur on microspore culture and the performance of microspore derived doubled haploid (DH) in Brassica juncea (L.). The variety BARI Sarisha II of B. junccea was used for microspore culture and 8 DH genotypes viz., BJDH 01, BJDH 05, BJDH ii, BJDH 12, BJDH 14, BJDH 17, BJDH 18, and BJDH 20 along with their parent Rajat collected from the Plant Research International, the Netherlands were used for performance evaluation. Four-day cold treatment at 6 °C along with bud size 3.0-3.1 mm of BARI Sarisha II showed better response in the embryonic development. Significant variation was observed for the different yield contributing characters among the DH genotypes along with their parent Rajat. All the DH genotypes were late both for days to flower and days to maturity compared to the parent. Most of the DH genotypes produced higher seed yield than the parent. The genotypes BJDII 14 (7 g) and BJDH 01(7 g) were bold seeded and almost double or more in 1000-seed weight compared to the parent. Total oil content of the DH genotypes (40.64-42.28) were significantly different from the parent (3 g). Oil content of BJDH 01 was almost 3% higher than the parent could be selected to incorporate in the future breeding program.

Microspore culture protocol for Indonesian Brassica oleracea

“…The optimal size for B. juncea cv. 'BARI Sarisha 11' was 3.0-3.1 mm (Ali et al 2008), or 3.1-3.5 mm in cv. 'Varuna' (Prem et al 2008).…”

“…bud length) and developmental stage of immature pollen grains: most microspores at the late-uninucleate to early binucleate stage can undergo embryogenesis (Pechan and Keller 1988;Hansen and Svinnset 1993;Custers 2003;Prem et al 2005;Abraha et al 2008). The high response of microspore Zaki and Dickinson (1995) B. campestris Canadian Cv-2 5390 embryos/100 buds nr nr Guo and Pulli (1996) B. juncea Couve Algarvia 9 embryos/2 buds nr nr Dias and Correia (2002) B. juncea Rajat 3500 embryos/100 buds 52 nr Chanana et al (2005) Interspecific hybrid of B. napus L 365 embryos/3 buds nr nr Smýkalová et al (2006) HSH nr not reported, a under optimal conditions embryogenesis in the same developmental stage was also recorded in B. campestris (Guo and Pulli 1996), B. carinata (Abraha et al 2008), B. juncea (Chanana et al 2005;Agarwal et al 2006;Ali et al 2008;Prem et al 2008), B. napus cv. 'Topas' (Anandarajah et al 1991;Takahata and Keller 1991;Supena et al 2006), and B. oleracea var.…”

“…In different studies, the density applied in Brassica microspore culture to obtain high embryo formation was 4 9 10 4 cells/ml for B. napus L. cv. 'Topas' (Anandarajah et al 1991;Takahata and Keller 1991;Supena et al 2006), B. juncea (Chanana et al 2005;Agarwal et al 2006;Ali et al 2008;Prem et al 2008), B. oleracea var. costata DC (Dias and Correia 2002), or 5 9 10 4 cells/ml in B. campestris (Guo and Pulli 1996).…”

“…Success in developing such a protocol is affected by the condition and preparation of the donor plant, plant genotype, bud size or microspore developmental stage, microspore density, heat shock treatment, pre-treatment, microspore isolation technique, culture medium, incubation system, among other factors (Takahata and Keller 1991;Guo and Pulli 1996;Tian et al 2004;Chanana et al 2005;Ali et al 2008;Prem et al 2008;Wang et al 2009;Ferrie and Caswell 2010). Various approaches (plant growth regulators, vitamins, sucrose, starvation, cold treatment, or heat shock) have been used to induce pollen embryogenesis with different successful results (Ferrie and Caswell 2010).…”

“…oleifera (Guo and Pulli 1996), B. napus cv. 'Topas' (Tian et al 2004), B. juncea L. (Chanana et al 2005;Ali et al 2008;Prem et al 2005Prem et al , 2008, B. oleracea var. acephala (Zhang et al 2008), and B. campestris ssp.…”