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Yu Takahashi,Yoshihito Takahata,Shuji Yokoi 한국식물생명공학회 2012 Plant biotechnology reports Vol.6 No.4
A number of factors influencing microspore embryogenesis and plant regeneration were examined in five subspecies (rapa, oleifera, niposinica, perviridis,broccoletto) of B. rapa. Addition of 6-benzylaminopurine (BA) in 1/2 NLN-10 medium improved the embryo yield by 2–12 fold. Addition of activated charcoal (AC) in the medium was not effective for microspore embryogenesis. Moreover, AC canceled the positive effect of BA, when the medium containing both BA and AC was used. Of 24genotypes examined for microspore embryogenesis, 22genotypes of all five subspecies produced embryos ranging from 0.02 to 15.0 per 2 9 105 microspores, but two genotypes were not responsive. Low temperature pretreatment of flower buds significantly improved the microspore embryogenesis. When cotyledonary embryos were subcultured on a filter paper placed on top of 0.8 %agar-solidified B5-2 medium and 1.6 % agar B5-2 medium,plant regenerations were increased 4–8 fold compared to 0.8 % agar medium. The ploidy levels of regenerated plants in three genotypes were determined by flow cytometry, revealing that 66–100 % of them were diploid. The results enable the advancement of breeding programs and genetic studies in B. rapa.
Choi, Pilson,Mano, Yoshiro,Ishikawa, Atsuko,Odashima, Masashi,Umezawa, Taishi,Fujimura, Tatsuhito,Takahata, Yoshihito,Komatsuda, Takao The Korean Society of Plant Biotechnology 2010 Plant biotechnology reports Vol.4 No.1
Quantitative trait loci (QTLs) controlling ability of somatic embryogenesis were identified in soybean. A frame map with 204-point markers was developed using an RI population consisting of 117 $F_{11}$ lines derived from a cross between cultivar 'Keburi' and a weedy soybean 'Masshokutou Kou 502'. The parents differed greatly in their abilities of somatic embryogenesis using immature cotyledons as explants. The ability of somatic embryogenesis was evaluated in five different experiments: the $F_{11}$ (evaluated in 1998) and $F_{15}$ (2002) generations cultured on basal media supplemented with $40\;mg\;l^{-1}$ 2,4-D (2,4-D1998 and 2,4-D2002), $F_{14}$ (2001) generation on medium with $40\;mg\;l^{-1}$ 2,4-D and high sucrose concentration [2,4-D2001 ($30\;g\;l^{-1}$ sucrose)], and the $F_{11}$ (1998) and $F_{12}$ (1999) generations on medium with $10\;mg\;l^{-1}$ NAA (NAA1998 and NAA1999). The RILs showed wide and continuous variations in each of the five experiments. In the composite interval mapping analysis, 2 QTLs were found in group 8 (D1b + W, LOD = 5.42, $r^2$ = 37.5) in the experiment of 2,4-D1998 and in group 6 (C2, LOD = 6.03, $r^2$ = 26.0) in the experiment of 2,4-D2001 (high concentration sucrose). In both QTLs, alleles of 'Masshokutou Kou 502' with high ability of somatic embryogenesis contributed to the QTLs. For the other three experiments, no QTL was detected in the criteria of LOD >3.0, suggesting the presence of minor genes.