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Titolo:
Agrobacterium tumefaciens-mediated transformation of Allium cepa L.: the production of transgenic onions and shallots
Autore:
Zheng, SJ; Khrustaleva, L; Henken, B; Sofiari, E; Jacobsen, E; Kik, C; Krens, FA;
Indirizzi:
Univ Wageningen & Res Ctr, Plant Res Int, NL-6700 AA Wageningen, Netherlands Univ Wageningen & Res Ctr Wageningen Netherlands NL-6700 AA Netherlands Res Inst Vegetables, AARD, Lembang, Indonesia Res Inst Vegetables Lembang Indonesia etables, AARD, Lembang, Indonesia Univ Wageningen & Res Ctr, Dept Plant Sci, NL-6700 AJ Wageningen, Netherlands Univ Wageningen & Res Ctr Wageningen Netherlands NL-6700 AJ Netherlands
Titolo Testata:
MOLECULAR BREEDING
fascicolo: 2, volume: 7, anno: 2001,
pagine: 101 - 115
SICI:
1380-3743(2001)7:2<101:ATTOAC>2.0.ZU;2-Y
Fonte:
ISI
Lingua:
ENG
Soggetto:
MANIHOT-ESCULENTA CRANTZ; ORYZA-SATIVA-L; GENE-TRANSFER; INDICA RICE; T-DNA; TRANSIENT EXPRESSION; SUSPENSION-CULTURES; PLANT-REGENERATION; IMMATURE EMBRYOS; REPORTER GENE;
Keywords:
Agrobacterium tumefaciens; Allium cepa; FISH (fluorescence in situ hybridization); transformation; transgenic plant characterization;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
49
Recensione:
Indirizzi per estratti:
Indirizzo: Kik, C Univ Wageningen & Res Ctr, Plant Res Int, POB 16, NL-6700 AA Wageningen, Netherlands Univ Wageningen & Res Ctr POB 16 Wageningen Netherlands NL-6700 AA
Citazione:
S.J. Zheng et al., "Agrobacterium tumefaciens-mediated transformation of Allium cepa L.: the production of transgenic onions and shallots", MOL BREED, 7(2), 2001, pp. 101-115

Abstract

This paper describes the development of a reliable transformation protocolfor onion and shallot (Allium cepa L.) which can be used year-round. It isbased on Agrobacterium tumefaciens as a vector, with three-week old callus, induced from mature zygotic embryos, as target tissue. For the development of the protocol a large number of parameters were studied. The expressionof the uidA gene coding for beta -glucuronidase was used as an indicator in the optimization of the protocol. Subspecies (onion and shallot) and cultivar were important factors for a successful transformation: shallot was better than onion and for shallot cv. Kuning the best results were obtained. Also, it was found that constantly reducing the size of the calli during subculturing and selection by chopping, thus enhancing exposure to the selective agent hygromycin, improved the selection efficiency significantly. Furthermore, callus induction medium and co-cultivation period showed a significant effect on successful stable transformation. The usage of different Agrobacterium strains, callus ages, callus sources and osmotic treatments during co-cultivation did not influence transformation efficiency. The highest transformation frequency (1.95%), was obtained with shallot cv. Kuning. A total of 11 independent transformed callus lines derived from zygotic embryos were produced: seven lines from shallot and four lines from onion. Large differences in plantlet production were observed among these lines. The best line produced over 90 plantlets. Via PCR the presence of the uidA and hpt(hygromycin phosphotransferase) genes could be demonstrated in these putative transformed plants. Southern hybridization showed that most lines originated from one transformation event. However, in one line plants were obtained indicating the occurrence and rescue of at least three independent transformation events. This suggested that T-DNA integration occurred in different cells within the callus. Most transgenic plants only had one copy of T-DNA integrated into their genomes. FISH performed on 12 plants from two different lines representing two integration events showed that original T-DNAintegration had taken place on the distal end of chromosomes 1 or 5. A total of 83 transgenic plants were transferred to the greenhouse and these plants appeared to be diploid and normal in morphology.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 04/12/20 alle ore 08:49:41