Transgenic lines of creeping bent grass were generated by Agrobacterium-mediated transformation with the VuNCED1 which was cloned from cow pea has a homology to 9-cis-epoxycarotenoid dioxygenase, which is supposed to be involved in abscisic acid (ABA) biosynthesis. ABA, a cleavage product of carotenoids, is involved in stress responses in plants. The limiting step of ABA biosynthesis in plants is presumably the cleavage of 9-cis-epoxycarotenoids, the first committed step of ABA biosynthesis. Molecular analyses of transgenic lines as performed by Southern hybridization genomic DNA-PCR revealed integration of the VuNCED1. Challenge studies performed with transgenic plants by exposure to salt stress (up to 10 dS m À1 ) and water stress (up to 75%) for 10 weeks, revealed that more than 50% of the transgenic plants could survive NaCl and drought stress whereas wild-type was not. ABA levels were measured under drought and normal conditions, endogenous ABA was dramatically increased by drought and NaCl stress in transgenic plants. These results indicate that it is possible to manipulate ABA levels in plants by over expressing the key regulatory gene in ABA biosynthesis and that stress tolerance can be improved by increasing ABA levels.
A new selection system for onion transformation that does not require the use of antibiotics or herbicides was developed. The selection system used the Escherichia coli gene that encodes phosphomannose isomerase (pmi). Transgenic plants carrying the manA gene that codes for pmi can detoxify mannose-6-phosphate by conversion to fructose-6-phosphate, an intermediate of glycolysis, via the pmi activity. Six-week-old embryogenic callus initiated from seedling radicle was used for transformation. Transgenic plants were produced efficiently with transformation rates of 27 and 23% using Agrobacterium and biolistic system, respectively. Untransformed shoots were eliminated by a stepwise increase from 10 g l(-1) sucrose with 10 g l(-1) mannose in the first selection to only 10 g l(-1) mannose in the second selection. Integrative transformation was confirmed by PCR, RT-PCR and Southern hybridization.
Lilies are one of the most valuable bulbous ornamental cut flower crops and are commercially propagated by conventional scaling, which results in varied bulb quality. This article demonstrates the effectiveness of using specific organ sources in conjunction with the thin cell layer (TCL) system to establish successful organogenic and somatic embryogenic pathways in Lilium longiflorum. TCLs derived from different explant sources, such as receptacle, leaf, young stem, stem node, bulblets, pseudo-bulblets, shoot and somatic embryos, respectively, can be manipulated to form different organs. Furthermore, the choice of plant growth regulator and medium additives such as activated charcoal and sucrose strongly affect the success of the process. This article further highlights that transient transgene expression derived from either particle bombardment or Agro-infection is both strong and repeatable, as a result of the direct exposure of competent cells to the gene introduction method, resulting in a reduction of escapes. The ability to manipulate and control organogenesis through a TCL system, coupled with repeatable, efficient transformation opens up the possibility of micropropagating members of this genus with new and available characteristics, producing superior plants of high quality.
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