Tianjin Key Laboratory of Protein Sciences, Department of Plant Biology and Ecology, College of Life Sciences, Nankai University, Tianjin, China.
Plant Biotechnol J. 2020 Aug;18(8):1749-1762. doi: 10.1111/pbi.13338. Epub 2020 Jan 30.
Amino acid transport via phloem is one of the major source-to-sink nitrogen translocation pathways in most plant species. Amino acid permeases (AAPs) play essential roles in amino acid transport between plant cells and subsequent phloem or seed loading. In this study, a soybean AAP gene, annotated as GmAAP6a, was cloned and demonstrated to be significantly induced by nitrogen starvation. Histochemical staining of GmAAP6a:GmAAP6a-GUS transgenic soybean revealed that GmAAP6a is predominantly expressed in phloem and xylem parenchyma cells. Growth and transport studies using toxic amino acid analogs or single amino acids as a sole nitrogen source suggest that GmAAP6a can selectively absorb and transport neutral and acidic amino acids. Overexpression of GmAAP6a in Arabidopsis and soybean resulted in elevated tolerance to nitrogen limitation. Furthermore, the source-to-sink transfer of amino acids in the transgenic soybean was markedly improved under low nitrogen conditions. At the vegetative stage, GmAAP6a-overexpressing soybean showed significantly increased nitrogen export from source cotyledons and simultaneously enhanced nitrogen import into sink primary leaves. At the reproductive stage, nitrogen import into seeds was greatly enhanced under both sufficient and limited nitrogen conditions. Collectively, our results imply that overexpression of GmAAP6a enhances nitrogen stress tolerance and source-to-sink transport and improves seed quality in soybean. Co-expression of GmAAP6a with genes specialized in source nitrogen recycling and seed loading may represent an interesting application potential in breeding.
氨基酸通过韧皮部运输是大多数植物物种中源到库氮转移的主要途径之一。氨基酸转运体(AAPs)在植物细胞之间的氨基酸运输和随后的韧皮部或种子装载中发挥重要作用。在这项研究中,克隆了大豆 AAP 基因,注释为 GmAAP6a,并证实其对氮饥饿有显著诱导作用。GmAAP6a:GmAAP6a-GUS 转基因大豆的组织化学染色表明,GmAAP6a主要在韧皮部和木质部薄壁细胞中表达。使用有毒氨基酸类似物或单一氨基酸作为唯一氮源的生长和运输研究表明,GmAAP6a 可以选择性地吸收和运输中性和酸性氨基酸。在拟南芥和大豆中过表达 GmAAP6a 导致对氮限制的耐受性提高。此外,在低氮条件下,转基因大豆中氨基酸的源到库转移明显改善。在营养生长阶段,过表达 GmAAP6a 的大豆表现出明显增加的氮从源子叶的输出,同时增强了氮向库初生叶的输入。在生殖阶段,在充足和有限的氮条件下,氮向种子的输入大大增强。总之,我们的结果表明,过表达 GmAAP6a 增强了氮胁迫耐受性和源到库的运输,并提高了大豆的种子质量。与专门用于源氮回收和种子装载的基因共同表达可能代表了在育种中的一个有趣的应用潜力。