Soybean [Glycine max (L.) Merr.] embryogenic cultures were transformed by particle bombardment with the feedback-insensitive tobacco anthranilate synthase (AS) gene ASA2 driven by the CaMV 35S promoter and selected using hph as the selectable marker gene. Only one of eight regenerated lines that set seed and contained ASA2 expressed the gene highly and contained increased free tryptophan (Trp) levels in leaves, seeds and embryogenic cultures. Leaf extracts of the ASA2 expressing line contained about twice as much AS enzyme activity as the untransformed control and this activity was only slightly more feedback-insensitive. Amino acid analysis showed that both leaves and embryogenic tissue cultures of the ASA2 expressing line had four to five-times the normal levels of free Trp and slightly higher free tyrosine and phenylalanine. The seed total Trp content was only slightly increased. Metabolic profiling-analysis by GC-MS detected no other consistent differences. These studies show that the ASA2 gene can be expressed in soybean and that modest changes in Trp synthesis occurs.

译文

大豆 [Glycine max (L.) Merr。] 用反馈不敏感的烟草邻氨基苯甲酸合酶 (AS) 基因ASA2进行粒子轰击转化,该基因由CaMV 35s启动子驱动,并使用hph作为选择标记基因进行选择。结实种子并含有ASA2的八个再生品系中只有一个高度表达该基因,并在叶片,种子和胚发生培养物中含有增加的游离色氨酸 (Trp) 水平。ASA2表达系的叶片提取物的酶活性是未转化对照的酶活性的两倍,并且该活性仅对反馈不敏感。氨基酸分析表明,ASA2表达系的叶片和胚发生组织培养物的游离Trp水平是正常水平的4至5倍,游离酪氨酸和苯丙氨酸略高。种子总Trp含量仅略有增加。通过gc-ms进行的代谢分析没有发现其他一致的差异。这些研究表明,ASA2基因可以在大豆中表达,并且Trp合成发生适度变化。

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