Abstract

SummaryThe phytase purple acid phosphatase (HvPAPhy_a) expressed during barley seed development was evaluated as transgene for overexpression in barley. The phytase was expressed constitutively driven by the cauliflower mosaic virus 35S‐promoter, and the phytase activity was measured in the mature grains, the green leaves and in the dry mature vegetative plant parts left after harvest of the grains. The T2‐generation of HvPAPhy_a transformed barley showed phytase activity increases up to 19‐fold (29 000 phytase units (FTU) per kg in mature grains). Moreover, also in green leaves and mature dry straw, phytase activities were increased significantly by 110‐fold (52 000 FTU/kg) and 57‐fold (51 000 FTU/kg), respectively. The HvPAPhy_a‐transformed barley plants with high phytase activities possess triple potential utilities for the improvement of phosphate bioavailability. First of all, the utilization of the mature grains as feed to increase the release of bio‐available phosphate and minerals bound to the phytate of the grains; secondly, the utilization of the powdered straw either directly or phytase extracted hereof as a supplement to high phytate feed or food; and finally, the use of the stubble to be ploughed into the soil for mobilizing phytate‐bound phosphate for plant growth.

Highlights

  • Phytases are phosphatases that can hydrolyze phytic acid (InsP6, myo-inositol-(1,2,3,4,5,6)-hexakisphosphate), the most important phosphorous (P) storage compound in plant seeds, representing 40%–80% of total seed P (Eeckhout and De Paepe, 1994; Lott, 1984)

  • The purpose of this study is to evaluate the transgenic potential of the barley grain phytase gene HvPAPhy_a constitutively expressed by the 35S-promoter for (I) increased mature grain a 2016 The Authors

  • As T0 grains are segregating for the transgene, mature grain phytase activity (MGPA) measurements were performed on pools of 20–25 grains from each T0-plant

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Summary

Introduction

Phytases (myo-inositol hexakisphosphate 3- and 6-phosphohydrolase; EC 3.1.3.8 and EC 3.1.3.26) are phosphatases that can hydrolyze phytic acid (InsP6, myo-inositol-(1,2,3,4,5,6)-hexakisphosphate), the most important phosphorous (P) storage compound in plant seeds, representing 40%–80% of total seed P (Eeckhout and De Paepe, 1994; Lott, 1984). Focus has not been on using the plants own phytases but on using microbial enzymes belonging to the group of histidine acid phosphatases. In addition to increasing seed phytase activity, microbial phytase has been expressed in green vegetative tissues of tobacco, alfalfa and potato. The purpose of this was to use green leaves for extraction of phytase (Ullah et al, 1999, 2002, 2003). Accumulation of functional phytase in mature dead vegetative plant parts left after seeds harvest has so far not been reported but constitutes a potential valuable alternative for phytase production. With a yearly estimated production of mature barley straw, on around 3 tons per hectare or 55 kg straw per 100 kg grain (Statistic Denmark; http://www.dst.dk/en) valorization of this tissue is highly attractive

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