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Chromosome Arm Locations of Barley Sucrose Transporter Gene in Transgenic Winter Wheat Lines

机译：大麦蔗糖转运蛋白基因在转基因冬小麦品系中的染色体臂位置

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摘要

Three transgenic HOSUT lines of winter wheat, HOSUT12, HOSUT20, and HOSUT24, each harbor a single copy of the cDNA for the barley sucrose transporter gene HvSUT1 (SUT), which was fused to the barley endosperm-specific Hordein B1 promoter (HO; the HOSUT transgene). Previously, flow cytometry combined with PCR analysis demonstrated that the HOSUT transgene had been integrated into different wheat chromosomes: 7A, 5D, and 4A in HOSUT12, HOSUT20, and HOSUT24, respectively. In order to confirm the chromosomal location of the HOSUT transgene by a cytological approach using wheat aneuploid stocks, we crossed corresponding nullisomic-tetrasomic lines with the three HOSUT lines, namely nullisomic 7A-tetrasomic 7B with HOSUT12, nullisomic 5D-tetrasomic 5B with HOSUT20, and nullisomic 4A-tetrasomic 4B with HOSUT24. We examined the resulting chromosomal constitutions and the presence of the HOSUT transgene in the F2 progeny by means of chromosome banding and PCR. The chromosome banding patterns of the critical chromosomes in the original HOSUT lines showed no difference from those of the corresponding wild type chromosomes. The presence or absence of the critical chromosomes completely corresponded to the presence or absence of the HOSUT transgene in the F2 plants. Investigating telocentric chromosomes occurred in the F2 progeny, which were derived from the respective critical HOSUT chromosomes, we found that the HOSUT transgene was individually integrated on the long arms of chromosomes 4A, 7A, and 5D in the three HOSUT lines. Thus, in this study we verified the chromosomal locations of the transgene, which had previously been determined by flow cytometry, and moreover revealed the chromosome-arm locations of the HOSUT transgene in the HOSUT lines.
机译：冬小麦的三个转基因HOSUT品系HOSUT12，HOSUT20和HOSUT24均具有大麦蔗糖转运蛋白基因HvSUT1（SUT）的cDNA的单个副本，该基因与大麦胚乳特异的大麦醇溶蛋白B1启动子（HO; the HOSUT转基因）。以前，流式细胞仪结合PCR分析表明，HOSUT转基因已被整合到不同的小麦染色体中：分别位于HOSUT12，HOSUT20和HOSUT24中的7A，5D和4A。为了通过细胞学方法使用小麦非整倍体原种来确定HOSUT转基因的染色体位置，我们将对应的无效等位基因-四体系与三个HOSUT系杂交，即无效等位基因7A-四体性7B与HOSUT12，无效等位基因5D-四体性5B与HOSUT20，以及带有HOSUT24的无效4A-四体4B。我们通过染色体谱带和PCR检查了F2子代中的染色体组成和HOSUT转基因的存在。原始HOSUT系中关键染色体的染色体谱带模式与相应的野生型染色体没有差异。关键染色体的存在与否完全对应于F2植物中HOSUT转基因的存在与否。调查端粒位于F2子代，它们分别来自关键的HOSUT染色体，我们发现，HOSUT转基因分别整合在3条HOSUT系的4A，7A和5D染色体的长臂上。因此，在这项研究中，我们验证了转基因的染色体位置，该位置先前已通过流式细胞仪确定，而且还揭示了HOSUT品系中HOSUT转基因的染色体臂位置。

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