狼尾草属象草PpPAPs基因的克隆及响应低磷胁迫的表达模式分析

doi:10.11733/j.issn.1007-0435.2021.03.002

草地学报 ›› 2021, Vol. 29 ›› Issue (3): 425-433.DOI: 10.11733/j.issn.1007-0435.2021.03.002

狼尾草属象草PpPAPs基因的克隆及响应低磷胁迫的表达模式分析

黄睿¹, 罗佳佳¹, 吴远航², 刘攀道¹, 刘国道¹

1. 中国热带农业科学院热带作物品种资源研究所, 农业部华南作物基因资源与种质创制重点实验室, 海南海口 571101;
2. 海南大学热带作物学院, 海南海口 570228

收稿日期:2020-09-27 修回日期:2020-11-09 出版日期:2021-03-15 发布日期:2021-04-02
通讯作者: 刘攀道, 刘国道
作者简介:黄睿(1989-),男,研究实习员,硕士,主要从事草业科学方面研究,E-mail:bluesing@126.com
基金资助:
海南省自然科学基金项目（No.319QN309）；国家牧草产业技术体系岗位科学家项目（No.CARS-34）；中国科协青年人才托举工程第五届项目（No.2019QNRC001）；滇桂黔石漠化地区特色作物产业发展关键技术集成示范项目（No.SMH2019-2021）资助

Molecular Cloning of PpPAP Genes and Their Expression Responses to Phosphorus Deficiency in Elephant Grass (Pennisetum purpureum)

HUANG Rui¹, LUO Jia-jia¹, WU Yuan-hang², LIU Pan-dao¹, LIU Guo-dao¹

1. Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agriculture Sciences, Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture and Rural Affairs, Haikou, Hainan Province 571101, China;
2. College of Tropical Crops Hainan University, Haikou, Hainan Province 571101, China

Received:2020-09-27 Revised:2020-11-09 Online:2021-03-15 Published:2021-04-02

摘要/Abstract

摘要： 为解析象草(Pennisetum purpureum)适应低磷胁迫机理，本研究通过水培试验，分析了不同磷源处理对象草生长的影响，并利用同源克隆和cDNA末端快速扩增(rapid amplification of cDNA ends，RACE)技术，从象草中克隆了3个可能参与ATP利用的PAP基因。结果表明，在腺嘌呤核苷三磷酸处理(+ATP)下，象草的干重和全磷含量显著高于低磷处理(-P)，表明象草对ATP具有较强的利用能力。荧光定量PCR分析表明，与正常供磷处理(+P)相比，+ATP处理10 d使3个可能参与ATP利用的PAP基因在象草根系中的表达量提高170%~403%，并伴随着根系胞内和分泌的酸性磷酸酶(acid phosphatase,ACP)活性显著增加。综上所述，在低磷胁迫条件下，象草根系中PpPAPs基因上调表达，可能促进了根系ACP的合成与分泌，这有助于象草对ATP的活化利用。

关键词: 紫色酸性磷酸酶, 低磷胁迫, 有机磷, 象草

Abstract: To insight into mechanisms of elephant grass adapting to low phosphorus (P) stress,the effects of different P sources on elephant grass growth were investigated by hydroponic experiments. Three PpPAP genes related to ATP utilization were cloned from elephant grass by homologous cloning and rapid amplification of cDNA ends (RACE) technology. Results showed that dry weight and total P content at the application of adenosine triphosphate (+ATP) treatment were significantly higher than those at low P (-P) treatment,suggesting that elephant grass has a strong ability to utilize ATP. QRT-PCR analysis showed that after 10 days of growth,the expression of these three PpPAPs associated with ATP utilization in the roots was 170%~403% greater than that of in +ATP treated plants compared to those grown under normal P (+P) treatment. Furthermore,+ATP treatment led to significant increases in the activities of root intracellular and secreted acid phosphatase (ACP),compared to +P treatment. Taken together,the results herein suggest that the expression of PpPAP genes in roots of elephant grass were up-regulated by P deficiency,which may promote the synthesis and secretion of ACP. These adaptive responses of elephant grass may contribute to the utilization of ATP under low P stress.

Key words: Purple acid phosphatase, Phosphorus deficiency, Organic phosphorus, Elephant grass

中图分类号:

S543

黄睿, 罗佳佳, 吴远航, 刘攀道, 刘国道. 狼尾草属象草PpPAPs基因的克隆及响应低磷胁迫的表达模式分析[J]. 草地学报, 2021, 29(3): 425-433.

HUANG Rui, LUO Jia-jia, WU Yuan-hang, LIU Pan-dao, LIU Guo-dao. Molecular Cloning of PpPAP Genes and Their Expression Responses to Phosphorus Deficiency in Elephant Grass (Pennisetum purpureum)[J]. Acta Agrestia Sinica, 2021, 29(3): 425-433.

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参考文献

[1] Zhang Z,Liao H,Lucas W J. Molecular mechanisms underlying phosphate sensing,signaling,and adaptation in plants[J]. Journal of Integrative Plant Biology,2014,56(3):192-220
[2] Zhu J,Li M,Whelan M. Phosphorus activators contribute to legacy phosphorus availability in agricultural soils:a review[J]. Science of the Total Environment,2018,612:522-537
[3] López-Arredondo D L,Leyva-González M A,González-Morales S I,et al. Phosphate nutrition:improving low-phosphate tolerance in crops[J]. Annual Review of Plant Biology,2014,65(1):95-123
[4] Weeks J J,Hettiarachchi G M. A review of the latest in phosphorus fertilizer technology:possibilities and pragmatism[J]. Journal of Environmental Quality,2019,48:1300-1313
[5] 丁广大,陈水森,石磊,等. 植物耐低磷胁迫的遗传调控机理研究进展[J]. 植物营养与肥料学报,2013,19(3):733-744
[6] 梁翠月,廖红. 植物根系响应低磷胁迫的机理研究[J]. 生命科学,2015,27(3):389-397
[7] Shen J,Yuan L,Zhang J,et al. Phosphorus dynamics:from soil to plant [J]. Plant Physiology,2011,156:997-1005
[8] Sun L,Wang L,Zheng Z,et al. Identification and characterization of an Arabidopsis phosphate starvation-induced secreted acid phosphatase as a vegetative storage protein[J]. Plant Science,2018,277:278-284
[9] Wang L,Liu D. Functions and regulation of phosphate starvation-induced secreted acid phosphatases in higher plants[J]. Plant Science,2018,271:108-116
[10] Zhu S,Chen M,Liang C,et al. Characterization of purple acid phosphatase family and functional analysis of GmPAP7a/7b involved in extracellular ATP utilization in soybean[J]. Frontiers in Plant Science,2020,11:661
[11] Deng S,Lu L,Li J,et al. Purple acid phosphatase 10c encodes a major acid phosphatase that regulates plant growth under phosphate-deficient conditions in rice[J]. Journal of Experimental Botany,2020,71:4321-4332
[12] 陈志彤,何水林,黄毅斌. 狼尾草属牧草研究进展[J]. 草地学报,2010,18(5):740-748
[13] 张霞,顾洪如,丁成龙,等. 象草若干生物学性状与产量的关系分析[J]. 草地学报,2009,17(5):670-674
[14] Murphy J,Riley J P. A modified single solution method for the determination of phosphate in natural waters[J]. Analytica Chimica Acta,1962,27:31-36
[15] Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding [J]. Analytical Biochemistry,1976,72:248-254
[16] Liang C,Tian J,Lam H M,et al. Biochemical and molecular characterization of PvPAP3,a novel purple acid phosphatase isolated from common bean enhancing extracellular ATP utilization[J]. Plant Physiology,2010,152:854-865
[17] Irving G C J,McLaughlin M J. A rapid and simple field-test for phosphorus in Olsen and Bray No.1 extracts of soil[J]. Communications in Soil Science and Plant Analysis,1990,21:2245-2255
[18] 黄睿,赵兴坤,虞道耿,等热研4号王草对不同有机磷活化利用能力的评价[J]. 热带作物学报,2019,40(5):850-856
[19] Larionov A,Krause A,Miller W. A standard curve based method for relative real time PCR data processing[J]. BMC Bioinformatics,2005,6:62
[20] George T S,Giles C D,Menezes-Blackburn D,et al. Organic phosphorus in the terrestrial environment:a perspective on the state of the art and future priorities[J]. Plant and Soil,2018,427:191-208
[21] Duff S M G,Sarath G,Plaxton W C. The role of acid phosphatases in plant phosphorus metabolism[J]. Physiologia Plantarum,1994,90:791-800
[22] 刘攀道,黄睿,许文茸,等. 植物紫色酸性磷酸酶的研究进展[J]. 热带作物学报,2019,40(2):410-416
[23] 黄宇,张海伟,徐芳森. 植物酸性磷酸酶的研究进展[J]. 华中农业大学学报,2008,27(1):148-154
[24] Tran H T,Hurley B A,Plaxton W C. Feeding hungry plants:the role of purple acid phosphatases in phosphate nutrition[J]. Plant Science,2010,179:14-27
[25] 魏铭,王鑫伟,陈博,等. 植物紫色酸性磷酸酶基因家族功能研究进展[J]. 植物学报,2019,54(1):93-101
[26] Tian J,Liao H. The role of intracellular and secreted purple acid phosphatases in plant phosphorus scavenging and recycling[M]. In:Plaxton WC,Lambers H,eds. Annual Plant Reviews. Hoboken,NJ:John Wiley & Sons,2015,265-287
[27] Wang L,Lu S,Zhang Y,et al. Comparative genetic analysis of Arabidopsis purple acid phosphatases AtPAP10,AtPAP12,and AtPAP26 provides new insights into their roles in plant adaptation to phosphate deprivation[J]. Journal of Integrative Plant Biology,2014,56:299-314
[28] Kong Y B,Li X H,Wang B,et al. The soybean purple acid phosphatase GmPAP14 predominantly enhances external phytate utilization in plants[J]. Frontiers in Plant Science,2018,9:292
[29] Liu P D,Xue Y B,Chen Z J,et al. Characterization of purple acid phosphatases involved in extracellular dNTP utilization in Stylosanthes[J]. Journal of Experimental Botany,2016,67:4141-4154
[30] Mehra P,Pandey B K,Giri J. Improvement in phosphate acquisition and utilization by a secretory purple acid phosphatase (OsPAP21b) in rice[J]. Plant Biotechnology Journal,2017,15:1054-1067

狼尾草属象草PpPAPs基因的克隆及响应低磷胁迫的表达模式分析

Molecular Cloning of PpPAP Genes and Their Expression Responses to Phosphorus Deficiency in Elephant Grass (Pennisetum purpureum)

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