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Progress towards onion transformatio...

El-Sharif, Ahmed Hussein.

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Progress towards onion transformation via particle bombardment.

紀錄類型:	書目-語言資料,印刷品 : Monograph/item
正題名/作者:	Progress towards onion transformation via particle bombardment./
作者:	El-Sharif, Ahmed Hussein.
面頁冊數:	151 p.
附註:	Source: Dissertation Abstracts International, Volume: 58-02, Section: B, page: 0461.
Contained By:	Dissertation Abstracts International58-02B.
標題:	Agriculture, Agronomy. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9723756
ISBN:	9780591325843

Progress towards onion transformation via particle bombardment.
El-Sharif, Ahmed Hussein.

Progress towards onion transformation via particle bombardment. - 151 p.

Source: Dissertation Abstracts International, Volume: 58-02, Section: B, page: 0461.

Thesis (Ph.D.)--New Mexico State University, 1997.

The Bio-Rad PDS-1000/HE biolistics system was used to develop a genetic transformation system for onions. The biological materials for bombardment were zygotic embryos and callus and suspension cultures from cell lines with confirmed regeneration ability. Two genotypes from Allium cepa 'Sunlite' and 'Buffalo' and one from Allium fistulosum 'Ishikura' were used in this study. Transient expression assays using the B-glucuronidase (GUS) reporter gene system were used to optimize the transformation conditions.

ISBN: 9780591325843Subjects--Topical Terms:

1018679
Agriculture, Agronomy.

Progress towards onion transformation via particle bombardment.
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245 1 0 $a Progress towards onion transformation via particle bombardment.
300 $a 151 p.
500 $a Source: Dissertation Abstracts International, Volume: 58-02, Section: B, page: 0461.
500 $a Chair: LeRoy A. Daugherty.
502 $a Thesis (Ph.D.)--New Mexico State University, 1997.
520 $a The Bio-Rad PDS-1000/HE biolistics system was used to develop a genetic transformation system for onions. The biological materials for bombardment were zygotic embryos and callus and suspension cultures from cell lines with confirmed regeneration ability. Two genotypes from Allium cepa 'Sunlite' and 'Buffalo' and one from Allium fistulosum 'Ishikura' were used in this study. Transient expression assays using the B-glucuronidase (GUS) reporter gene system were used to optimize the transformation conditions.
520 $a Purified plasmid DNA carrying the GUS gene was used to coat the microcarriers. Three different rupture disks were tested. The most consistent bombardments were obtained when the vacuum in the biolistic chamber reached 23-24 in of Hg, the flow of helium reached 900 psi, microcarriers were loaded at level 2 and the target tissue placed at level 4. After 48 h. incubation, sample tissues were tested for GUS activity. Then the number of blue spots were counted, which indicated the location of GUS enzyme activity. Plasmid-bombarded tissues exhibited vivid blue spots.
520 $a Various promoters combined with the GUS coding sequence were tested. The maize Ubi-1 promoter was the most consistent and strongly expressing promoter across all six genotypes using either callus or suspension cells. The wheat EM promoter was strongly expressed in some genotypes but not others, and generally was stronger in suspension cells.
520 $a Dose response curves for kanamycin and bialaphos were initiated with control callus and embryos cultures to establish a useful positive selection agent. Kanamycin was not a suitable selection agent using the concentrations tested, ranging from 50 to 200 mg/L. Bialaphos gave the best selection control using small cell aggregates to avoid escapes. For callus and embryos, 5 mg/L and for suspension cells 2.5 mg/L bialaphos gave appropriate selection control.
520 $a Some bombarded callus and suspension lines continued to exhibit GUS activity upon assay after several weeks under selection indicating the possibility of stable onion transformation. Other bombarded tissues were no longer GUS positive; either these tissues were bialaphos selection escapes, or they were transformant tissues with silenced GUS expression. A genetic transformation system for onion would be useful to introduce genes into onion aimed at improvement of fungal disease resistance.
590 $a School code: 0143.
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650 4 $a Biology, Genetics. $3 1017730
650 4 $a Agriculture, Plant Culture. $3 1018669
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710 2 $a New Mexico State University. $3 783784
773 0 $t Dissertation Abstracts International $g 58-02B.
790 1 0 $a Daugherty, LeRoy A., $e advisor
790 $a 0143
791 $a Ph.D.
792 $a 1997
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