TRANSGENIC TOBACCO PLANTS AND THEIR PROGENY DERIVED BY MICROPROJECTILE BOMBARDMENT OF TOBACCO-LEAVES

被引：65

作者：

TOMES, DT ^{[1
]}

WEISSINGER, AK ^{[1
]}

ROSS, M ^{[1
]}

HIGGINS, R ^{[1
]}

DRUMMOND, BJ ^{[1
]}

SCHAAF, S ^{[1
]}

MALONESCHONEBERG, J ^{[1
]}

STAEBELL, M ^{[1
]}

FLYNN, P ^{[1
]}

ANDERSON, J ^{[1
]}

HOWARD, J ^{[1
]}

机构：

[1] N CAROLINA STATE UNIV,DEPT CROP SCI,RALEIGH,NC 27695

来源：

PLANT MOLECULAR BIOLOGY | 1990年 / 14卷 / 02期

关键词：

direct gene transfer; gene rearrangements; Nicotiana tabacum; particle gun; transformation;

D O I：

10.1007/BF00018566

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Transgenic tobacco plants and progeny carrying coding sequences for neomycin phosphotransferase II (NPTII) and beta-glucuronidase (GUS) were recovered following microprojectile bombardment of tobacco leaves. Transgenic plants were regenerated from bombarded leaf pieces of tobacco cvs. 'Xanthi' and 'Ky 17' which were cultured in the presence of 100 or 200 μg/ml kanamycin for six to eight weeks. Among 160 putative transgenic plants from at least 16 independent transformation events 76% expressed NPTII, and 50% expressed GUS. Southern analysis of plants expressing either one or both of the enzymes indicated DNA in high molecular weight DNA in 8 of 9 independent transformants analyzed. Two independent transformants and their progeny were analyzed in detail. Analysis of progeny for quantitative enzyme levels of NPTII and GUS, and Southern analysis of parents and progeny clearly demonstrated that the genes were transmitted to progeny. One transformant demonstrated Mendelian ratios for seed germination on kanamycin-containing medium while the other transformant had non-Mendelian ratios. DNA analysis of progeny indicate complex integration of the plasmid DNA, and suggest that rearrangements of this DNA has occurred. These results are consistent with other methods of direct DNA uptake into cells, and verify that the microprojectile bombardment method is capable of DNA delivery into intact plant cells which can give rise to transgenic plants and progeny. © 1990 Kluwer Academic Publishers.

引用

页码：261 / 268

页数：8

共 25 条

[11] STABLE GENETIC-TRANSFORMATION OF INTACT NICOTIANA CELLS BY THE PARTICLE BOMBARDMENT PROCESS [J].