中国农业科学 ›› 2014, Vol. 47 ›› Issue (4): 613-621.doi: 10.3864/j.issn.0578-1752.2014.04.001

• 作物遗传育种·种质资源·分子遗传学 •    下一篇

磷转运蛋白基因TaPHT2;1在染色体上定位 及对小麦磷素吸收和利用效率的影响

 郭丽1, 郭程瑾1, 路文静2, 李小娟2, 肖凯1   

  1. 1、河北农业大学农学院,河北保定071001;
    2、河北农业大学生命科学学院,河北保定071001
  • 收稿日期:2013-09-18 出版日期:2014-02-15 发布日期:2013-11-11
  • 通讯作者: 肖凯,E-mail:xiaokai@hebau.edu.cn
  • 作者简介:郭丽,Tel:0312-7528416;E-mail:guoli1212@sina.com
  • 基金资助:

    国家自然科学基金(31371618,31201674)、河北省自然科学基金(C2013204146)和河北省作物生长重点调控实验室基金

Chromosome Localization of Phosphate Transporter Gene TaPHT2;1 and Its Effects on Phosphate Uptake and Utilization in Wheat

 GUO  Li-1, GUO  Cheng-Jin-1, LU  Wen-Jing-2, LI  Xiao-Juan-2, XIAO  Kai-1   

  1. 1、College of Agronomy, Agricultural University of Hebei, Baoding 071001, Hebei;
    2、College of Life Sciences, Agricultural  University of Hebei, Baoding 071001, Hebei
  • Received:2013-09-18 Online:2014-02-15 Published:2013-11-11

摘要: 【目的】以中国春遗传背景的整套B染色体双端体为材料,鉴定磷转运蛋白基因TaPHT2;1的染色体定位特征。解析不同供磷水平下,上述材料和不同磷利用效率小麦品种该基因的表达特征及其与植株干物质生产能力和磷效率特征的联系。【方法】采用溶液培养法水培中国春(CS)及该品种遗传背景的整套B染色体组双端体和不同磷效率小麦品种材料。以B染色体组供试端体为材料进行TaPHT2;1 PCR扩增,鉴定TaPHT2;1在染色体上的定位。采用半定量RT-PCR及qRT-PCR技术分析B染色体组供试端体和小麦品种TaPHT2;1的表达水平。采用常规分析技术,测定供试材料单株干重和磷吸收参数。【结果】①PCR检测发现,在CS及所有供试B染色体组双端体材料中,除缺失1B长臂的1BS外,其它所有材料均能特异扩增出目标基因TaPHT2;1,表明TaPHT2;1定位在1B长臂。②丰、缺磷条件下,TaPHT2;1在CS及除1BS外双端体根、叶中的表达均表现为叶片优势表达特征,且在叶片中表达受到低磷胁迫的诱导。TaPHT2;1在根系中的表达不受低磷逆境调控。表明TaPHT2;1在介导丰磷下磷素吸收、转运及增强低磷下植株体内磷素再度调运中可能发挥重要功能。③丰磷条件下,与CS相比,1BS的单株干重和全磷含量显著降低;缺磷条件下,1BS的单株干重与CS相比也显著下降,但全磷含量增加。表明位于1B染色体长臂后的磷转运蛋白基因TaPHT2;1,对丰、缺磷条件下的植株磷素吸收、转运具有较大影响,进一步对不同供磷水平下的植株干重产生重要调控效应。④丰磷条件下,与CS相比,1BS的单株磷累积量显著增加,磷利用效率没有改变;缺磷条件下,与CS相比,1BS的单株磷累积量没有变化,磷利用效率显著降低。不同磷利用效率品种相比,丰磷条件下,随着品种磷利用效率提高,叶片中TaPHT2;1的表达水平、单株干重、全磷含量和单株磷累积量也随着增加;缺磷条件下,随着小麦品种磷利用效率提高,叶片中TaPHT2;1的表达水平、单株干重和磷利用效率也随之增高,但全磷含量呈下降趋势,单株磷累积量品种间差异较小。因此,TaPHT2;1的表达水平与小麦品种丰、缺磷条件下的磷素吸收、利用和干物质积累能力具有紧密联系。【结论】小麦磷转运蛋白基因TaPHT2;1位于1B染色体长臂。该基因通过其特定对外界供磷水平产生应答,在较大程度上调控植株的磷素吸收和利用能力,对不同供磷水平下的植株干重产生重要影响。TaPHT2;1在调控植株丰磷下磷素吸收和低磷下磷素利用中发挥重要作用,可作为鉴定小麦品种磷效率的评价指标。

关键词: 小麦(Triticum aestivum L.) , 磷转运蛋白基因 , 磷素供应 , 磷素吸收利用 , 干物质生产

Abstract: 【Objective】 In this study, the chromosome localization of TaPHT2;1, a phosphate transporter gene in wheat, was determined by using Chinese spring (CS) and its chromosome-based ditelosimic lines of B genome. Moreover, the expression pattern of TaPHT2;1 as well as its relationship with plant dry matter production and P use efficiency was studied under high- and low-Pi conditions.【Method】The cultivar CS together with its chromosome-based ditelosimic lines of B genome as well as wheat cultivars with varied P use efficiencies was hydroponically cultured. The chromosomal localization of TaPHT2;1 was detected by PCR amplification using specific primers with genome DNA of the tested materials as the template. The expression patterns of TaPHT2;1 in CS, its chromosome-based ditelosimic lines of B genome, and wheat cultivars with different P use efficiencies were determined by semi-quantitative RT-PCR and real time PCR. The plant dry weight and P acquisition parameters of the tested materials were assayed by following the conventional approach. 【Result】 For the CS and its chromosome-based ditelosimic lines of B genome, only 1BS that a line lacking the long arm of 1B was failed to detect the transcripts of TaPHT2;1, indicating that TaPHT2;1 is located in the long arm of 1B. Under P sufficience and P deprivation, the expression of TaPHT2;1 in roots and leaves of CS and other ditelosimic lines of B genome other than 1BS exhibited to be predominant in leaves and the expression levels were induced by Pi deprivation stress. The expression of TaPHT2;1 in roots was not regulated by Pi deprivation stress. These results suggest that TaPHT2;1 is involved in mediating plant Pi acquisition and cellular Pi translocation under Pi sufficience and in regulating re-transportation of cellular Pi under Pi deprivation. Under P sufficience, the plant dry weight and total Pi content in 1BS were significantly decreased in comparison with those in CS; under Pi deprivation, the plant dry weight of 1BS was also significantly lower than that of CS, but 1BS had higher total Pi content than CS. Therefore, the phosphate transporter gene TaPHT2;1 that located on the long arm of 1 B exerts dramatic effects on plant dry mass production under various Pi-supply conditions through its regulation of plant P acquisition and cellular Pi translocation. Under Pi sufficience, the accumulative Pi amount per plant of 1BS was significantly increased compared with that of CS, but there were no variations in Pi use efficiency between them. Under Pi deprivation, the Pi use efficiency of 1BS was significantly decreased compared with that of CS, but there were no variations in accumulative Pi amount per plant between them. In addition, under Pi sufficience, the expression level of TaPHT2;1, plant dry weight, total Pi content, and accumulative Pi amount per plant were all increased along with the increase of Pi use efficiencies across the tested wheat cultivars that displayed varied Pi use utilization properties. Under Pi deprivation, the expression level, plant dry weight, and Pi use efficiency were also increased along with the increase of the Pi use efficiencies in the tested wheat cultivars. However, the total Pi content exhibited a decrease tendency and the accumulative Pi amount per plant showed little variation among the wheat cultivars. Taken together, these results clearly indicate that there is a close association between the expression level of TaPHT2;1 and plant Pi acquisition, Pi utilization, and dry matter production in wheat cultivars with varied Pi use efficiencies under the conditions of Pi sufficience and Pi deprivation.【Conclusion】The wheat phosphate transporter gene TaPHT2;1 is located on the long arm of 1B. TaPHT2;1 plays an important role in regulation of plant dry mass production through its distinct response to external Pi conditions, which further modifies to most extent the plant Pi acquisition and utilization. This study confirms that TaPHT2;1 is a critical component in regulating plant Pi acquisition under Pi sufficience and Pi utilization under Pi deprivation. It can act as a molecular reference in evaluating Pi use efficiencies in wheat.

Key words: wheat (Triticum aestivum L.) , phosphate transporter gene , Pi supply , phosphate acquisition and utilization , dry mass production