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Characterisation of the phytase gene in trifoliate orange (Poncirus trifoliata (L.) Raf.) seedlings

文献类型： 外文期刊

作者： Shu, Bo 1 ; Wang, Peng 1 ; Xia, Ren-Xue 1 ;

作者机构： 1.Huazhong Agr Univ, Key Lab Hort Plant Biol, Minist Educ, Wuhan 430070, Peoples R China

2.Chinese Acad Trop Agr Sci, South Subtrop Crops Res Inst, Zhanjiang 524091, Peoples R China

3.Minist Agr, Key Lab Trop Fruit Biol, Zhanjiang 524091, Peoples R China

关键词： Organic phosphate;Acid phosphatase;Phytase;Gene expression;Enzyme activity

期刊名称：SCIENTIA HORTICULTURAE （ 影响因子：3.463； 五年影响因子：3.672 ）

ISSN：

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收录情况： SCI

摘要： Phosphate (P) deficiency decreases the yield and quality of citrus fruits. The P concentration, P uptake, and the expression of three predicted acid phosphatase genes in trifoliate orange seedlings were measured among inorganic phosphate (Pi), organic phosphate (Po), and no phosphate ( P) treatments to assess the ability of Po to improve trifoliate orange P deficiency. The three genes were expressed in Pichia pastoris X-33 to characterise the trifoliate orange phytase gene. Our analysis showed that trifoliate orange could utilise phytin without microorganisms in P-deficient conditions by promoting phytase and acid phosphatase activity; however, Po had a smaller effect on the improvement of trifoliate orange P deficiency than Pi fertiliser. The expression of the three genes in roots and leaves with Pi treatment did not respond to P solutions, but nearly all three genes were up-regulated in roots or leaves with Po and P treatment after P solutions were supplied. Further, the PtPAP3 recombinant protein (r-PtPAP3), which was expressed in P. pastoris, showed phytase activity that was as high as that of cereal phytases. The r-PtPAP3 K-m values were 46.2 mu M for phytate and 1631 mu M for rho-nitrophenylphosphate. Incubation with Mn2+ increased the specific activity of r-PtPAP3 for phytate by approximately 1.9-fold, and Fe2+ increased it by approximately 1.2-fold. In summary, the function and expression pattern of PtPAP3 allowed a detailed understanding of the significance of phytate metabolism of a woody perennial. (C) 2015 Elsevier B.V. All rights reserved.

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