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  • Author or Editor: Hong-di Huang x
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The rapid expansion of Asian populations in the United States presents significant requirements for Asian vegetables. Flowering chinese cabbage (Brassica rapa L. ssp. chinensis var. utilis Tsen et Lee) is one of the most popular vegetables in China. The main factors restricting the progress in its breeding and genetic studies is the time required in generating desired pure line populations. Doubled haploid (DH) populations of flowering chinese cabbage have not been established because of technical difficulties. An appropriate combined protocol for a fast generation cycling system could advance up to seven generations, allowing the production of pure line seeds within 336–420 days among four cultivars and one hybrid of flowering chinese cabbage. The previous six generation cycles were accelerated using the embryo culture plus soil method which bypassed seed maturation through in vitro culture of immature embryos and promoted plant reproduction under stressed conditions, then the seventh generation cycle was accomplished until mature seeds were harvested using the soil method. During the culture of immature embryos, 12-day-old embryos could germinate and develop successfully on a Murashige and Skoog medium (MS) medium () containing 10% young coconut juice. This combined protocol bypasses the current obstacles in constructing DH populations of flowering chinese cabbage and is a possible alternative for producing pure lines. Its wider adoption could facilitate the breeding and biological studies of other Brassicaceae vegetables.

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An efficient biolistic transformation system of banana combined with a liquid medium selection system was developed during this study. An embryogenic cell suspension (ECS) of Musa acuminata cv. Baxi (AAA) was bombarded with a particle delivery system. After 7 days of restoring culture in liquid M2 medium, embryogenic cells were transferred to a liquid selection M2 medium supplemented with 10 μg/mL hygromycin for resistance screening. The untransformed cell clusters were inhibited or killed, and a small number of transformants proliferated in the liquid selection medium. After the 0th, first, second, and third generation of antibiotic screening, there were 0, 65, 212, and 320, respectively, vitality-resistant buds obtained from a 0.5-mL packed cell volume (PCV) of embryogenic cell suspension. The β-glucuronidase (GUS) staining, polymerase chain reaction (PCR) analysis, and Southern blot hybridization results all demonstrated a 100% positive rate of regenerated resistant seedlings. Interestingly, the number of buds obtained through third-generation screening was almost equal to that obtained from the original ECS in M2 medium without antibiotics. These results suggested that the liquid medium selection system facilitated the proliferation of a positive transgenic ECS, which significantly improved the regeneration rate of transformants. This protocol is suitable for the genetic transformation of all banana genotypes and is highly advantageous to varieties with low callusing potential.

Open Access