《植物生理学报》 2017, 53(4): 545-554

草莓钼转运蛋白基因MOT1的克隆与表达以及对氮代谢的影响

刘利, 孙明岳, 张蕊, 杨超, 李玲, 高东升, 付喜玲^*

作物生物学国家重点实验室, 山东果蔬优质高效生产协同创新中心, 山东农业大学园艺科学与工程学院, 山东泰安271018

通信作者：付喜玲；E-mail: xilingfu@sdau.edu.cn

摘　要：

从‘章姬’草莓中克隆了一个MOT1家族成员的全长, 测序发现其CDS序列与公布的森林草莓基因组序列一致, 将其命名为FaMOT1。进化树分析表明FaMOT1所编码的氨基酸与苹果和桃亲缘关系最近。荧光定量PCR结果表明, FaMOT1基因在草莓的根、茎、叶、花和10 d幼果中有不同程度的表达。对草莓幼苗喷施不同浓度的钼酸钠, 结果发现, 各处理叶片的钼浓度随着施钼量的增加而显著提高。2.4 mmol·L^-1 Na₂MoO₄处理显著诱导根部和叶片的FaMOT1的表达, 处理2和4 h时4.8 mmol·L^-1 Na₂MoO₄处理的茎部FaMOT1表达水平最高, 处理6和12 h时2.4 mmol·L^-1 Na₂MoO₄处理的茎部FaMOT1表达水平最高。2.4 mmol·L^-1 Na₂MoO₄处理叶片的硝态氮浓度显著低于其它处理, 氨态氮浓度显著高于其它处理; 4.8 mmol·L^-1 Na₂MoO₄处理根部和叶片的氨态氮浓度最低。以上结果表明FaMOT1基因的表达受到钼的诱导和调节, 合适浓度的钼处理有利于硝态氮的吸收和向氨态氮的转化以及氨态氮向有机氮的转化。

关键词：草莓; 钼; MOT1; 基因克隆; 表达分析

收稿：2016-10-17   修定：2017-03-15

资助：国家自然科学基金(31672137)和山东省现代农业产业技术体系果品创新团队-栽培与土肥岗(SDAIT-06-01)。

Molecular cloning, expression analysis and its effect on nitrogen metabolism of molybdenum transporter gene MOT1 in strawberry (Fragaria ×ananassa)

LIU Li, SUN Ming-Yue, ZHANG Rui, YANG Chao, LI Ling, GAO Dong-Sheng, Fu Xi-Ling^*

State Key Laboratory of Crop Biology, Shandong Collaborative Innovation Center for Fruit and Vegetable Production with High Quality and Efficiency, College of Horticulture Science and Engineering, Shandong Agricultural University, Taian, Shandong 271018, China

Corresponding author: Fu Xi-Ling; E-mail: xilingfu@sdau.edu.cn

Abstract:

The MOT1 homologue named FaMOT1 was isolated from strawberry (Fragaria ×ananassa). The obtained CDS was consistent with the ones released from F. vesca genome. Phylogenetic tree analysis showed that its encoded amino acids sequence had a relatively close evolutionary relationship with Malus domestica and Prunus persica. Quantitative real-time PCR analysis showed that FaMOT1 had different levels of expression in strawberry roots, stems, leaves, flowers and young fruits of 10 d. The effects of different concentrations of Na₂MoO₄ foliar spraying on seedlings and FaMOT1 expression were examined in this experiment. The molybdenum content of leaves increased with the increase of molybdenum sprays. FaMOT1 gene was significantly induced in roots and leaves by 2.4 mmol·L^-1 Na₂MoO₄. FaMOT1 was higher expressed in stems after treatment of 4.8 mmol·L^-1 Na₂MoO₄ for 2 and 4 h, and higher expressed in stems after treatment of 2.4 mmol·L^-1 Na₂MoO₄ for 6 and 12 h. The treatment of 2.4 mmol·L^-1 Na₂MoO₄ showed lowest leaf nitrate content and highest leaf ammonia nitrogen content. The lowest ammonia nitrogen contents of leaf and root were got in the seedlings sprayed with 4.8 mmol·L^-1 Na₂MoO₄. All these results indicated that the expression of FaMOT1 was induced and regulated by molybdenum, and appropriate molybdenum spraying could not only enhance nitrate uptake and transformation into ammonia nitrogen, but also be in favor of the transformation of ammonia nitrogen into organic nitrogen.

Key words: strawberry (Fragaria ×ananassa); molybdenum; MOT1; molecular cloning; expression analysis