Journal of Integrative Agriculture ›› 2025, Vol. 24 ›› Issue (5): 1646-1655.DOI: 10.1016/j.jia.2024.09.009

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TaPHT1;6磷高效优异等位变异位点的挖掘及功能标记的开发

  

  • 收稿日期:2024-05-27 修回日期:2024-09-14 接受日期:2024-08-04 出版日期:2025-05-20 发布日期:2025-04-14

Identification of Pi-efficient elite allele of the TaPHT1;6 gene and development of its functional marker in common wheat (Triticum aestivum L.)

Huanting Shi1, 2, Chuang Lou1, Jinfeng Wang1, Dianqi Dong1, Longfei Yang1, Gezi Li1, Zhiqiang Tian1, Qiaoxia Han1#, Pengfei Wang1#, Guozhang Kang1, 2, 3#   

  1. 1 National Engineering Research Centre for Wheat, Henan Agricultural University, Zhengzhou 450046, China

    2 Shennong Laboratory of Henan Province, Zhengzhou 450002, China

    3 State Key Laboratory of High-Efficiency Production of Wheat–Maize Double Cropping, Henan Agricultural University, Zhengzhou 450046, China

  • Received:2024-05-27 Revised:2024-09-14 Accepted:2024-08-04 Online:2025-05-20 Published:2025-04-14
  • About author:#Correspondence Qiaoxia Han, E-mail: hqxia@henau.edu.cn; Pengfei Wang, E-mail: wangpf@henau.edu.cn; Guozhang Kang, E-mail: guozhangkang@henau.edu.cn
  • Supported by:

    This work was financially supported by the Shennong Laboratory Project of Henan Province, China (SN01-2022-01), the China Postdoctoral Science Foundation (2023M731006), and the Project of Science and Technology of Henan Province, China (232102111104).

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摘要：

磷（Pi）利用效率低是农业生产面临的一个重要挑战，不仅导致生产成本的增加和环境问题，同时也造成了磷矿资源的短缺。高亲和磷转运蛋白在作物磷吸收转运过程中发挥了重要功能，然而编码这些蛋白基因的分子标记甚少被开发。本研究扩增了167个小麦品种的高亲和磷转运蛋白基因（TaPHT1;6-5A、5B、5D）的启动子和编码区序列，发现TaPHT1;6-5A和TaPHT1;6-5D无等位变异位点，TaPHT1;6-5B启动子上存在16个等位变异位点，形成三种单倍型Hap1、Hap2和Hap3。在连续两年田间试验中，测定了三种单倍型小麦品种的生物量、籽粒产量、磷含量、磷肥吸收效率及磷肥利用效率，发现Hap3属于磷高效优异单倍型；在其机理上，通过LUC assay发现Hap3启动子具有更强的基因表达驱动能力，在Hap3小麦品种内TaPHT1;6-5B表达水平显著高于其它两个单倍型；在应用上，基于TaPHT1;6-5B启动子上的等位变异位点，开发了一个用于区分Hap3和其它两种单倍型的功能标记dCAPS-571，可用于磷高效小麦品种的选育。

Abstract:

One of agriculture’s major challenges is the low efficiency of phosphate (Pi) use, which leads to increased costs, harmful environmental impacts, and the depletion of phosphorus (P) resources.  The TaPHT1;6 gene, which encodes a high-affinity Pi transporter (PHT), plays a crucial role in Pi absorption and transport.  In this study, the promoter and coding regions of three TaPHT1;6 gene copies on chromosomes 5A, 5B, and 5D were individually amplified and sequenced from 167 common wheat (Triticum aestivum L.) cultivars.  Sequence analysis revealed 16 allelic variation sites within the promoters of TaPHT1;6-5B among these cultivars, forming three distinct haplotypes: Hap1, Hap2, and Hap3.  Field trials were conducted over two years to compare wheat genotypes with these haplotypes, focusing on assessing plant dry weight, grain yield, P content, Pi fertilizer absorption efficiency, and Pi fertilizer utilization efficiency.  Results indicated that Hap3 represented the favored Pi-efficient haplotype.  Dual-luciferase reporter assay demonstrated that the Hap3 promoter, carrying the identified allelic variation sites, exhibited higher gene-driven capability, leading to increased expression levels of the TaPHT1;6-5B gene.  We developed a distributed cleaved amplified polymorphic site marker (dCAPS-571) to distinguish Hap3 from the other two haplotypes based on these allelic variation sites, presenting an opportunity for breeding Pi-efficient wheat cultivars.  This study successfully identified polymorphic sites on TaPHT1;6-5B associated with Pi efficiency and developed a functional molecular marker to facilitate future breeding endeavors.



Key words: Triticum aestivum L. , high-affinity Pi transporter ,  Pi use efficiency ,  Pi-efficient molecular marker