磷酸盐转运蛋白OsPT4影响水稻氮磷积累与利用的机理研究

doi:10.16819/j.1001-7216.2021.200803

中国水稻科学 ›› 2021, Vol. 35 ›› Issue (6): 565-572.DOI: 10.16819/j.1001-7216.2021.200803

磷酸盐转运蛋白OsPT4影响水稻氮磷积累与利用的机理研究

孙雅菲^#, 宋科^#, 秦秦, 孙丽娟, 薛永^*

上海市农业科学院生态环境保护研究所/上海市农业环境保护监测站/上海市设施园艺技术重点实验室,上海 201403

收稿日期:2020-08-03 修回日期:2020-12-04 出版日期:2021-11-10 发布日期:2021-11-10
通讯作者: 薛永
作者简介:
^#共同第一作者
基金资助:
上海市青年英才扬帆计划资助项目(19YF1443200);上海市科技兴农重点攻关项目[沪农科推字(2020)2-1];国家青年自然科学基金资助项目(31902102);上海市农业科学院国家自然科学基金匹配项目[生科创-GT2020（匹配-3）];上海市农业科学院卓越团队计划资助项目[2017（A-03）]

Research on the Mechanisim of OsPT4 Regulating the Accumulation and Utilization of Nitrogen and Phosphorus in Rice

Yafei SUN^#, Ke SONG^#, Qin QIN, Lijuan SUN, Yong XUE^*

Institute of Eco-Environment and Plant Protection, Shanghai Academy of Agricultural Sciences/Shanghai Monitor of Agricultural Environmental Protection/Shanghai Key Laboratory of Facility Horticulture Technology, Shanghai 201403, China

Received:2020-08-03 Revised:2020-12-04 Online:2021-11-10 Published:2021-11-10
Contact: Yong XUE
About author:
^#These authors contributed equally to this work

摘要/Abstract

摘要：

【目的】磷酸盐(Pi)转运蛋白OsPT4是水稻Pht1家族成员之一,负责Pi吸收以及向地上部的转运。探究OsPT4超表达对不同Pi条件下水稻氮(N)和磷(P)积累与利用的影响及其机理具有重要意义。【方法】以日本晴背景的OsPT4超表达株系为研究材料,通过设置正常供Pi与缺Pi的水培与桶培实验,检测生殖生长阶段不同组织中OsPT4的相对表达量,探究不同Pi处理条件下不同组织(叶片、叶鞘、茎秆、稻壳、穗柄和糙米)中的N和P浓度,并计算Pi吸收率及N和P利用效率,同时分析株高、单株产量、千粒重和结实率等产量构成因素。【结果】OsPT4在水稻生殖生长阶段的根系中相对表达丰度较高,OsPT4超表达使水稻剑叶、叶鞘、茎秆、稻壳、穗柄和糙米中的总P浓度不同程度提高,并显著提高了不同Pi处理条件下的Pi吸收与利用效率。同时,OsPT4的超表达可显著提高正常供Pi与缺Pi土壤条件下的单株产量与千粒重,以及缺Pi土壤中生长的结实率。除此之外,OsPT4的超表达使缺Pi条件下瘪壳与糙米中总N浓度平均升高16.8%和19.8%,N利用效率平均升高6.6%。【结论】OsPT4超表达不仅显著提高Pi吸收与利用效率,同时对不同Pi条件下的生理氮素利用率起促进作用。

关键词: 磷, 氮, OsPT4, 磷素利用率, 氮素利用率

Abstract:

【Objective】 The phosphate (Pi) transporter, OsPT4, is a member of the Pht1 family of rice, responsible for Pi uptake and transportation. It is important to explore the effect of OsPT4 overexpression on nitrogen (N) and phosphorous (P) accumulation and utilization in rice under different Pi conditions and its mechanism. 【Method】The OsPT4-overexpressing lines with Nipponbare background were used as material. Through the hydroponic and pot-cultivated experiments with (+Pi) or without Pi (-Pi) supply, the relative expression of OsPT4 in different tissues during reproductive growth stage was detected. The N and P concentrations in different tissues (leaf blade, leaf sheath, stem, rice hull, panicle axis and brown rice) under different Pi treatments were also measured. In addition, the Pi absorption rate and N/P use efficiency were calculated, and the plant height, yield per plant, 1000-grain weight and seed setting rate were analyzed. 【Result】The relative expression of OsPT4 was higher in rice roots during reproductive growth stage. Overexpression of OsPT4 increased the total P concentration in rice flag leaf blade, leaf sheaths, stems, rice hull, panicle axis and brown rice to varying degrees, and significantly improved the Pi absorption and use efficiency under different Pi regimes. Meanwhile, OsPT4 overexpression could significantly increase the yield per plant, 1000-grain weight and seed setting rate in +Pi and -Pi soils. In addition, the overexpression of OsPT4 also increased the total N concentration in unfilled rice hull and brown rice by 16.8% and 19.8%, and the N use efficiency by 6.6% under Pi-deficiency. 【Conclusion】The overexpression of OsPT4 not only promotes the Pi uptake and use efficiency, but also facilitates the physiology N use efficiency in different Pi conditions.

Key words: phosphorus, nitrogen, OsPT4, phosphorus use efficiency, nitrogen use efficiency

孙雅菲, 宋科, 秦秦, 孙丽娟, 薛永. 磷酸盐转运蛋白OsPT4影响水稻氮磷积累与利用的机理研究[J]. 中国水稻科学, 2021, 35(6): 565-572.

Yafei SUN, Ke SONG, Qin QIN, Lijuan SUN, Yong XUE. Research on the Mechanisim of OsPT4 Regulating the Accumulation and Utilization of Nitrogen and Phosphorus in Rice[J]. Chinese Journal OF Rice Science, 2021, 35(6): 565-572.

图/表 8

表1 OsActin和OsPT4的定量PCR序列

Table 1 qRT-PCR primers for OsActin and OsPT4.

基因名称 Gene name	引物序列(5＇-3＇) Prime sequence(5＇-3＇)	退火温度 Annealing temperature/℃
OsPT4	F: TTCTGCTAGTGTACCAAACAAAATTACA R: GTAAGTGGCATTTATAATATCAACAGTAACC	55
OsActin	F: GGGTTCACAAGTCTGCCTATTGT	55
OsActin	R: ACGGGACACGACCAAGGA	55

图1 不同生长期OsPT4在不同组织中的相对表达量数据均为平均数（n = 3）。

Fig. 1. Relative expression of OsPT4 in different organs at different growth stages of rice. Data are shown as mean (n = 3).

图2 OsPT4超表达材料分子鉴定 A–OsPT4超表达材料拷贝数鉴定;B–超表达材料中OsPT4的相对表达量。M代表标记物,P代表阳性对照。不同字母表示在5%水平上差异显著。WT–野生型;Ox1~ Ox3–三个超表达株系。数据均为平均数±标准差(n = 3);不同字母表示同一时期内各株系在0.05水平上差异显著。

Fig. 2. Molecular identification of OsPT4 overexpressing lines. A, Identification of copy number in OsPT4 overexpressing lines; B, Relative expression of OsPT4 in the overexpressing lines. Different letters indicate significant difference at 0.05 level. Data are shown as mean (n = 3); Different letters indicate significant difference in the same stage at the level of 0.05. WT, Wild type; Ox1-Ox3, Three OsPT4-overexpression lines.

图3 OsPT4的超表达提高营养生长期正常供磷和缺磷条件下生物量及总磷含量数据均为平均数（n = 4）;不同字母表示同一时期内各株系在0.05水平上差异显著。下同。

Fig. 3. Overexpression of OsPT4 increases the biomass and total P concentration under both +P and -P conditions during vegetative growth stage. Data are shown as mean (n = 4); Different letters indicate significant difference in the same stage at the level of 0.05. The same below.

图4 OsPT4的超表达促进生殖生长期正常供磷和缺磷土壤条件下水稻不同组织中的磷积累

Fig. 4. Overexpression of OsPT4 facilitates the P accumulation of rice under both +P and -P soil conditions during reproductive growth stage.

图5 OsPT4的超表达提高正常供磷和缺磷土壤条件下水稻磷吸收及利用效率

Fig. 5. Overexpression of OsPT4 facilitates the Pi uptake and use efficiency of rice under both +P and -P soil conditions.

图6 OsPT4超表达材料在正常供磷与缺磷土壤中的农艺性状分析

Fig. 6. Phenotype of OsPT4 overexpressing lines grown in +P and -P soil conditions.

图7 正常供磷和缺磷土壤中OsPT4超表达材料繁育器官总氮含量及水稻生理氮素利用率

Fig. 7. Total N concentration in reproductive tissues and physiology N use efficiency of OsPT4 overexpressing lines under both +P and -P soil conditions.

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磷酸盐转运蛋白OsPT4影响水稻氮磷积累与利用的机理研究

Research on the Mechanisim of OsPT4 Regulating the Accumulation and Utilization of Nitrogen and Phosphorus in Rice

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