High frequency production of embryos from liquid flask cultures of oilseed rape

✍ Scribed by Chiang-Shiong Loh; Wendy Shu; Eugene Khor

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 266 KB
• Volume: 54
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/(sici)1097-0290(19970505)54:3<231::aid-bit4>3.0.co;2-g

✦ Synopsis

As part of the program to scale-up the produc- . In a haploid plant, all mutation and tion of artificial seeds of winter oilseed rape, Brassica recessive genes are expressed and can be conveniently napus ssp. oleifera, we established a liquid flask culture and rapidly detected ). The significance system that enables the high frequency production of of such haploids and their double haploid forms have freely suspended embryos. As many as 4000 embryos been obvious for geneticists and plant breeders for the could be obtained from 1 mL packed-cell-volume of cells. For initiation of liquid flask cultures, four different types production of inbred lines and for obtaining homozyof callus tissues were used. Among them, the most emgous lines without lengthy inbreeding . bryogenic cell suspension cultures were obtained from Appearance of haploid forms in natural populations spontaneous callus developed on the surface of secondis at best infrequent . In oilseed rape, ary embryos precultured in medium supplemented with the frequencies of naturally occurring haploids ranged 2,4-dichlorophenoxyacetic acid (4.52 M) and kinetin (0.46 M) (type B callus). Growth curves of the cell susfrom 0.008 to 0.685% (Renard and Dosba, 1980; Thomppension were determined and the cell suspension was son, 1975).

able to grow in medium without plant growth regulators.

The possible use of secondary embryos for the pro-Embryos were observed to develop directly from the cells duction of large numbers of haploid plants throughout without going through an obvious callus phase. When subcultured to agar medium containing 44.38 M benzythe year for breeding purposes has been described (Inlaminopurine, about 43% of the embryos developed into