Use of fluorescence in situ hybridization for gross mapping of transgenes and screening for homozygous plants in transgenic barley (Hordeum vulgare L.)

被引:19
作者
Choi, HW [1 ]
Lemaux, PG [1 ]
Cho, MJ [1 ]
机构
[1] Univ Calif Berkeley, Dept Plant & Microbial Biol, Berkeley, CA 94720 USA
关键词
barley; fluorescence in situ hybridization; transgene localization; transgene expression; homozygosity;
D O I
10.1007/s00122-002-0997-y
中图分类号
S3 [农学(农艺学)];
学科分类号
0901 ;
摘要
Introduced transgenes, uidA, sgfp (S65T) and/ or bar, were localized using fluorescence in situ hybridization (FISH) on metaphase chromosomes of transgenic barley produced by microparticle bombardment of immature embryos. Of the 19 independent transgenic lines (eight diploid and 11 tetraploid), nine had uidA and ten had sgfp (S65T). All lines tested had three or more copies of the transgenes and 18 out of 19 lines had visibly different integration sites. At a gross level, it appeared that no preferential integration sites of foreign DNA among chromosomes were present in the lines tested; however, a distal preference for transgene integration was observed within the chromosome. In diploid To plants that gave a 3:1 segregation ratio of transgene expression in the T-1, only single integration sites were detected on one of the homologous chromosomes. Homozygous diploid plants had doublet signals on a pair of homologous chromosomes. All tetraploid T-0 plants that gave a 3:1 segregation ratio in the T-1 generation had only a single integration site on one of the homologous chromosomes. In contrast, the single tetraploid T-0 plant with a 35:1 segregation ratio in the T-1 generation had doublet signals on a pair of homologous chromosomes. In the one tetraploid To line, which had a homozygote-like segregation ratio (45:0), there were doublet signals at two loci on separate chromosomes. We conclude that the application of FISH for analysis of transgenic plants is useful for the gross localization of transgene(s) and for early screening of homozygous plants.
引用
收藏
页码:92 / 100
页数:9
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共 43 条
  • [1] Widely separated multiple transgene integration sites in wheat chromosomes are brought together at interphase
    Abranches, R
    Santos, AP
    Wegel, E
    Williams, S
    Castilho, A
    Christou, P
    Shaw, P
    Stoger, E
    [J]. PLANT JOURNAL, 2000, 24 (06) : 713 - 723
  • [2] DETECTION OF A 17-KB UNIQUE SEQUENCE (T-DNA) IN PLANT CHROMOSOMES BY INSITU HYBRIDIZATION
    AMBROS, PF
    MATZKE, MA
    MATZKE, AJM
    [J]. CHROMOSOMA, 1986, 94 (01) : 11 - 18
  • [3] LOCALIZATION OF AGROBACTERIUM-RHIZOGENES T-DNA IN PLANT CHROMOSOMES BY INSITU HYBRIDIZATION
    AMBROS, PF
    MATZKE, AJM
    MATZKE, MA
    [J]. EMBO JOURNAL, 1986, 5 (09) : 2073 - 2077
  • [4] Visual screening of microspore-derived transgenic barley (Hordeum vulgare L.) with green-fluorescent protein
    Carlson, AR
    Letarte, J
    Chen, J
    Kasha, KJ
    [J]. PLANT CELL REPORTS, 2001, 20 (04) : 331 - 337
  • [5] Transformation of recalcitrant barley cultivars through improvement of regenerability and decreased albinism
    Cho, MJ
    Jiang, W
    Lemaux, PG
    [J]. PLANT SCIENCE, 1998, 138 (02) : 229 - 244
  • [6] Inheritance of tissue-specific expression of barley hordein promoter-uidA fusions in transgenic barley plants
    Cho, MJ
    Choi, HW
    Buchanan, BB
    Lemaux, PG
    [J]. THEORETICAL AND APPLIED GENETICS, 1999, 98 (08) : 1253 - 1262
  • [7] Endosperm-specific expression of green fluorescent protein driven by the hordein promoter is stably inherited in transgenic barley (Hordeum vulgare) plants
    Cho, MJ
    Choi, HW
    Jiang, W
    Ha, CD
    Lemaux, PG
    [J]. PHYSIOLOGIA PLANTARUM, 2002, 115 (01) : 144 - 154
  • [8] Production of transgenic tall fescue and red fescue plants by particle bombardment of mature seed-derived highly regenerative tissues
    Cho, MJ
    Ha, CD
    Lemaux, PG
    [J]. PLANT CELL REPORTS, 2000, 19 (11) : 1084 - 1089
  • [9] Increased chromosomal variation in transgenic versus nontransgenic barley (Hordeum vulgare L.) plants
    Choi, HW
    Lemaux, PG
    Cho, MJ
    [J]. CROP SCIENCE, 2000, 40 (02) : 524 - 533
  • [10] Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants
    Christensen, AH
    Quail, PH
    [J]. TRANSGENIC RESEARCH, 1996, 5 (03) : 213 - 218