Molecular Characterization and Functional Analysis of OsPHY1, a Purple Acid Phosphatase (PAP)-Type Phytase Gene in Rice (Oryza sativa L.)

doi:10.1016/S1671-2927(00)8650

Journal of Integrative Agriculture ›› 2012, Vol. 12 ›› Issue (8): 1217-1226.DOI: 10.1016/S1671-2927(00)8650

• 论文 • 下一篇

Molecular Characterization and Functional Analysis of OsPHY1, a Purple Acid Phosphatase (PAP)-Type Phytase Gene in Rice (Oryza sativa L.)

LI Rui-juan, LU Wen-jing, GUO Cheng-jin, LI Xiao-juan, GU Jun-tao, XIAO Kai

1.College of Life Sciences, Agricultural University of Hebei, Baoding 071001, P.R.China
2.College of Agronomy, Agricultural University of Hebei, Baoding 071001, P.R.China

收稿日期:2011-04-01 出版日期:2012-08-01 发布日期:2012-09-09
通讯作者: Correspondence XIAO Kai, Tel: +86-312-7528115, Fax: +86-312-7528400, E-mail: xiaokai3@yahoo.com
基金资助:
This work was supported by the National Natural Science Foundation of China (30871466) and the Key Laboratory of Crop Growth Regulation of Hebei Province, China.

Molecular Characterization and Functional Analysis of OsPHY1, a Purple Acid Phosphatase (PAP)-Type Phytase Gene in Rice (Oryza sativa L.)

LI Rui-juan, LU Wen-jing, GUO Cheng-jin, LI Xiao-juan, GU Jun-tao, XIAO Kai

1.College of Life Sciences, Agricultural University of Hebei, Baoding 071001, P.R.China
2.College of Agronomy, Agricultural University of Hebei, Baoding 071001, P.R.China

Received:2011-04-01 Online:2012-08-01 Published:2012-09-09
Contact: Correspondence XIAO Kai, Tel: +86-312-7528115, Fax: +86-312-7528400, E-mail: xiaokai3@yahoo.com
Supported by:
This work was supported by the National Natural Science Foundation of China (30871466) and the Key Laboratory of Crop Growth Regulation of Hebei Province, China.

摘要/Abstract

摘要： As a specific type of acid phosphatses, phytases play diverse roles in plants by catalazing the degradation of phytic acid and its derivatives. In this study, a rice phytase gene referred to OsPHY1 has been functionally characterized. OsPHY1 contains a 1 620 bp of open reading frame, encoding a 539-aa polypeptide. A conserve domain metallophosphatase (MPP) (MPP_PAPs), generally harbored in phytase and purple acid phosphatases (PAP), was identified in OsPHY1 (residue 194- 398). Phylogenetic analysis revealed that OsPHY1 shares high similarities with phytase genes and PAP-type genes that derived from diverse plant species. The OsPHY1 transcripts were detected to be abundant in germinating seeds, suggesting that this gene plays potential roles on degradation of seed phytic acid and its derivatives during the germination process. Biochemical analysis confirmed that OsPHY1 possesses strong catalytic activities on phytic acid-Na2, with optimal temperature of 57°C and suitable pH of 3.5. Based on transgene analysis, the putative role of OsPHY1 in plants on utilization of phytate was assessed. Under the condition that phytic acid-Na2 was used as sole P source, the OsPHY1- overexpressing tobacco plants behaved higher phytase activities, higher concentrations of Pi, more accumulative amount of total phosphorus, and much more improved growth traits than those of the control plants. Therefore, OsPHY1 is acted as an important component on degradation of the phytins during the seed germination process in rice. Also, OsPHY1 has a potential use on generation of elite crop germplasms with improved use efficiencies on phytate and its derivatives.

关键词: rice (Oryza sitava L.), phytase, expression, transgenic tobacco, phosphorus utilization efficiency

Abstract: As a specific type of acid phosphatses, phytases play diverse roles in plants by catalazing the degradation of phytic acid and its derivatives. In this study, a rice phytase gene referred to OsPHY1 has been functionally characterized. OsPHY1 contains a 1 620 bp of open reading frame, encoding a 539-aa polypeptide. A conserve domain metallophosphatase (MPP) (MPP_PAPs), generally harbored in phytase and purple acid phosphatases (PAP), was identified in OsPHY1 (residue 194- 398). Phylogenetic analysis revealed that OsPHY1 shares high similarities with phytase genes and PAP-type genes that derived from diverse plant species. The OsPHY1 transcripts were detected to be abundant in germinating seeds, suggesting that this gene plays potential roles on degradation of seed phytic acid and its derivatives during the germination process. Biochemical analysis confirmed that OsPHY1 possesses strong catalytic activities on phytic acid-Na2, with optimal temperature of 57°C and suitable pH of 3.5. Based on transgene analysis, the putative role of OsPHY1 in plants on utilization of phytate was assessed. Under the condition that phytic acid-Na2 was used as sole P source, the OsPHY1- overexpressing tobacco plants behaved higher phytase activities, higher concentrations of Pi, more accumulative amount of total phosphorus, and much more improved growth traits than those of the control plants. Therefore, OsPHY1 is acted as an important component on degradation of the phytins during the seed germination process in rice. Also, OsPHY1 has a potential use on generation of elite crop germplasms with improved use efficiencies on phytate and its derivatives.

Key words: rice (Oryza sitava L.), phytase, expression, transgenic tobacco, phosphorus utilization efficiency

LI Rui-juan, LU Wen-jing, GUO Cheng-jin, LI Xiao-juan, GU Jun-tao, XIAO Kai. Molecular Characterization and Functional Analysis of OsPHY1, a Purple Acid Phosphatase (PAP)-Type Phytase Gene in Rice (Oryza sativa L.)[J]. Journal of Integrative Agriculture, 2012, 12(8): 1217-1226.

参考文献

[1]Ames B N. 1966. Assay of inorganic phosphate, total phosphate and phosphatases. In: Neufeld E, Ginsburg V, eds., Methods in Enzymology. Academic Press, New York, USA. pp. 115-118.

[2]Asmar F. 1997. Variation in activity of root extracellular phytase between genotypes of barley. Plant Soil, 195, 61-64.

[3]Dionisio G, Holm P B, Brinch-Pedersen H. 2007. Wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.) multiple inositol polyphosphate phosphatases (MINPPs) are phytases expressed during grain filling and germination. Plant Biotechnology Journal, 5, 325-338.

[4]Flanagan J U, Cassady A I, Schenk G, Guddat L W, Hume D A. 2006. Identification and molecular modeling of a novel, plant-like, human purple acid phosphatase. Gene, 377, 12-20.

[5]Goldstein A H, Baertlein D A, McDaniel R G. 1988. Phosphate starvation inducible metabolism in Lycopersicon esculentum. 1. Excretion of acidphosphatase by tomato plants and suspension-cultured cells. Plant Physiology, 87, 711-715.

[6]Guo L, Zhao Y X, Zhang S H, Zhang H N, Xiao K. 2009. Improvement of organic phosphate acquisition in transgenic tobacco plants by overexpression of a soybean phytase gene Sphy1. Frontiers of Agriculture in China, 3, 259-265.

[7]Hayes J E, Richardson A E, Simpson R J. 1999. Phytase and acid phosphatase activities in extracts from roots of temperate pasture grass and legume seedlings. Australian Journal of Plant Physiology, 26, 801-809.

[8]Hegeman C E, Grabau E A. 2001. A novel phytase with sequence similarity to purple acid phosphatases is expressed in cotyledons of germinating soybean seedlings. Plant Physiology, 126, 1598-1608.

[9]Lambers H, Shane M W, Cramer M D, Pearse S J, Veneklaas E J. 2006. Root structure and functioning for efficient acquisition of phosphorus: matching morphological and physiological traits. Annals of Botany, 98, 693-713.

[10]Li D, Zhu H, Liu K, Liu X, Leggewie G, Udvardi M, Wang D. 2002. Purple acid phosphatases of Arabidopsis thaliana: comparative analysis and differential regulation by phosphate deprivation. Journal of Biological Chemistry, 277, 27772-27781.

[11]Li M, Osaki M, Rao I M, Tadano T. 1997. Secretion of phytase from the roots of several plant species under phosphorus-deficient conditions. Plant Soil, 195, 161-169.

[12]Loewus F A, Murthy P P N. 2000. Myo-inositol metabolism in plants. Plant Science, 150, 1-19.

[13]Lott J N A, Greenwood J S, Batten B D. 1995. Mechanisms and regulation of mineral nutrient storage during seed development. In: Kigel J, Galili G, eds., Seed Development and Germination. Marcel Dekker, New York, USA. pp. 215-235.

[14]Lung S C, Leung A, Kuang R, Wang Y, Leung P, Lim B L. 2008. Phytase activity in tobacco (Nicotiana tabacum) root exudates is exhibited by a purple acid phosphatase. Phytochemistry, 69, 365-373.

[15]Maugenest S, Martinez I, Lescure A M. 1997. Cloning and characterization of a cDNA encoding a maize seedling phytase. Biochemical Journal, 322, 511-517.

[16]Mehta B D, Jog S P, Johnson S C, Murthy P P. 2006. Lily pollen alkaline phytase is a histidine phosphatase similar to mammalian multiple inositol polyphosphate phosphatase (MINPP). Phytochemistry, 67, 1874-1886.

[17]Mimura T. 1999. Regulation of phosphate transport and homeostasis in plant cells. International Review of Cytology, 191, 149-200.

[18]Morgan M A. 1997. The Behaviour of Soil and Fertilizer Phosphorus. CAB International, Oxon, UK. Mudge S R, Rae A L, Diatloff E, Smith F W. 2002. Expression analysis suggests novel roles for members of the Pht1 family of phosphate transporters in Arabidopsis. Plant Journal, 31, 341-353.

[19]Mullaney E J, Ullah A H. 2003. The term phytase comprises several different classes of enzymes. Biochemical and Biophysical Research Communications, 312, 179-184.

[20]Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15, 473-497.

[21]Oelkers E H, Valsami-Jones E. 2008. Phosphate mineral reactivity and global sustainability. Elements, 4, 83-87.

[22]Raboy V. 1997. Accumulation and storage of phosphate and minerals. In: Larkins B A, Vasil I K, eds., Cellular and Molecular Biology of Plant Seed Development. Kluwer Academic Publishers, Dordrecht, The Netherlands. pp. 441-477.

[23]Raghothama K G. 1999. Phosphate acquisition. Annual Review of Plant Physiology and Plant Molecular Biology, 50, 665-693.

[24]Richardson A E, Hadobas P A, Hayes J E. 2001. Extracellular secretion of Aspergillus phytase from Arabidopsis roots enables plants to obtain phosphorus from phytate. Plant Journal, 25, 641-649.

[25]Schachtman D P, Reid R J, Ayling S M. 1998. Phosphorus uptake by plants: from soil to cell. Plant Physiology, 116, 447-453.

[26]Sharpley A N, Chapra S C, Wedepohl R, Sims J T, Daniel T C, Reddy K R. 1994. Managing agricultural phosphorus for protection of surface waters-issues and options. Journal of Environmental Quality, 23, 437-451.

[27]Shenoy V V, Kalagudi G M. 2005. Enhancing plant phosphorus use efficiency for sustainable cropping. Biotechnology Advances, 23, 501-513.

[28]Vance C P, Uhde-stone C, Allan D L. 2003. Phosphorus acquisition and use: critical adaptations by plants for securing a nonrenewable resource. New Phytologist, 157, 423-447.

[29]Vincent J B, Crowder M W, Averill B A. 1992. Hydrolysis of phosphate monoesters: a biological problem with multiple chemical solutions. Trends in Biochemical Sciences, 17, 105-110.

[30]Wodzinski R J, Ullah A H J. 1996. Phytases. Advances in Applied Microbiology, 42, 263-302.

[31]Xiao K, Harrison M J, Wang Z Y. 2005. Transgenic expression of a novel M. truncatula phytase gene results in improved acquisition of organic phosphorus by Arabidopsis. Planta, 222, 27-36.

[32]Xiao K, Katagi H, Harrison M, Wang Z Y. 2006. Improved phosphorus acquisition and biomass production in Arabidopsis by transgenic expression of a purple acid phosphatase gene from M. truncatula. Plant Science, 170, 191-202.

[33]Zhang W Y, Gruszewski H A, Chevone B I, Nessler C L. 2007. An Arabidopsis purple acid phosphatase with phytase activity increases foliar ascorbate. Plant Physiology, 146, 431-440.

Molecular Characterization and Functional Analysis of OsPHY1, a Purple Acid Phosphatase (PAP)-Type Phytase Gene in Rice (Oryza sativa L.)

Molecular Characterization and Functional Analysis of OsPHY1, a Purple Acid Phosphatase (PAP)-Type Phytase Gene in Rice (Oryza sativa L.)

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	RONG Rui-juan, WU Peng-cheng, LAN Jin-ping, WEI Han-fu, WEI Jian, CHEN Hao, SHI Jia-nan, HAO Yu-jie, LIU Li-juan, DOU Shi-juan, LI Li-yun, WU Lin, LIU Si-qi, YIN Chang-cheng, LIU Guo-zhen. Western blot detection of PMI protein in transgenic rice[J]. Journal of Integrative Agriculture, 2016, 15(4): 726-734.
[2]	LI Bi-feng, ZHU Ya-xin, GU Zhao-bing, CHEN Yuan, LENG Jing, GOU Xiao, FENG Li, LI Qing, XI Dong-mei, MAO Hua-ming, YANG Shu-Li. Screening and characterization of a novel ruminal cellulase gene (Umcel-1) from a metagenomic library of gayal (Bos frontalis)[J]. Journal of Integrative Agriculture, 2016, 15(4): 855-861.
[3]	ZHAO Yong-ying, WANG Xiang, WEI Li, WANG Jing-xuan, YIN Jun. Characterization of Ppd-D1 alleles on the developmental traits and rhythmic expression of photoperiod genes in common wheat[J]. Journal of Integrative Agriculture, 2016, 15(3): 502-511.
[4]	XU Yu-chao, HOU Xi-lin, XU Wei-wei, SHEN Lu-lu, Lü Shan-wu, ZHANG Shi-lin, HU Chun-mei. Isolation and characterization of an ERF-B3 gene associated with flower abnormalities in non-heading Chinese cabbage[J]. Journal of Integrative Agriculture, 2016, 15(3): 528-536.
[5]	ZHANG Jiao, XING Yan-ru, HOU Bo-feng, YUAN Zhu-ting, LI Yao, JIE Wen-cai, SUN Yang, LI Fei. Amplification and function analysis of N6-adenine-specific DNA methyltransferase gene in Nilaparvata lugens[J]. Journal of Integrative Agriculture, 2016, 15(3): 591-599.
[6]	TIAN Jing-jing, CHEN Xiang-ning, XIE Yuan-hong, LU Yong, XU Wen-tao, XU Li, DU Bin. Expression and characterization of a codon-optimized butyrylcholinesterase for analysis of organophosphate insecticide residues[J]. Journal of Integrative Agriculture, 2016, 15(3): 684-693.
[7]	WANG Fei-bing, ZHAI Hong, AN Yan-yan, SI Zeng-zhi, HE Shao-zhen, LIU Qing-chang. Overexpression of IbMIPS1 gene enhances salt tolerance in transgenic sweetpotato[J]. Journal of Integrative Agriculture, 2016, 15(2): 271-281.
[8]	TIAN Jia, ZENG Bin, LUO Shu-ping, LI Xiu-gen, WU Bin, LI Jiang. Cloning, localization and expression analysis of two fw2.2-like genes in small- and large-fruited pear species[J]. Journal of Integrative Agriculture, 2016, 15(2): 282-294.
[9]	LIU Jie-ying, ZHANG Chong, SHAO Qi, TANG Yu-fan, CAO Song-xiao, GUO Xiao-ou, JIN Ya-zhong, QI Hong-yan. Effects of abiotic stress and hormones on the expressions of five 13-CmLOXs and enzyme activity in oriental melon (Cucumis melo var. makuwa Makino)[J]. Journal of Integrative Agriculture, 2016, 15(2): 326-338.
[10]	LI Hui-fang, SHU Jing-ting, SHAN Yan-ju, CHEN Wen-feng, SONG Chi, XU Wen-juan. Myofiber development during embryonic to neonatal development in duck breeds differing in muscle growth rates[J]. Journal of Integrative Agriculture, 2016, 15(2): 403-413.
[11]	ZHANG Wei, LI Bei, YU Bin. Genome-wide identification, phylogeny and expression analysis of the SBP-box gene family in maize (Zea mays)[J]. Journal of Integrative Agriculture, 2016, 15(1): 29-41.
[12]	ZHANG Bai-zhong, KONG Fan-chao, WANG Hua-tang, GAO Xi-wu, ZENG Xin-nian, SHI Xue-yan. Insecticide induction of O-demethylase activity and expression of cytochrome P450 genes in the red imported fire ant (Solenopsis invicta Buren)[J]. Journal of Integrative Agriculture, 2016, 15(1): 135-144.
[13]	ZOU Xi-ling, ZENG Liu, LU Guang-yuan, CHENG Yong, XU Jin-song, ZHANG Xue-kun. Comparison of transcriptomes undergoing waterlogging at the seedling stage between tolerant and sensitive varieties of Brassica napus L.[J]. Journal of Integrative Agriculture, 2015, 14(9): 1723-1734.
[14]	TAN Wan-qin, Phang Chiun Yee, Sieo Chin Chin, Yiap Beow Chin, Clemente Michael Wong Vui Ling, Norhani Abdullah, Son Radu, Ho Yin Wan. Cloning of a novel phytase from an anaerobic rumen bacterium, Mitsuokella jalaludinii, and its expression in Escherichia coli[J]. Journal of Integrative Agriculture, 2015, 14(9): 1816-1826.
[15]	HAN Ji-long, YANG Min, GUO Ting-ting, YUE Yao-jing, LIU Jian-bin, NIU Chun-e, WANG Chao-feng, YANG Bo-hui. Molecular characterization of two candidate genes associated with coat color in Tibetan sheep (Ovis arise)[J]. Journal of Integrative Agriculture, 2015, 14(7): 1390-1397.