MAIN CONCLUSION: The work provides the first-time evidence of tissue-specific expression of a phytase gene in the germinating seeds of Phaseolus vulgaris. Phytase enzyme plays a major role in germinating seeds. It is also active during N2 fixation within nodules of legumes. The effect of phosphorus (P) deficiency on phytase gene expression and localization in N2-fixing root nodules has been recently studied in hydroaeroponic culture of Phaseolus vulgaris. In this study, phytase gene transcripts within the germinating seed tissues of the P-inefficient P. vulgaris recombinant inbred line RIL147 were in situ localized with a similar RT-PCR recipe as that used for nodules. Our results show that the phytase gene expression was mainly localized in the outer layers, vascular cells and parenchyma of germinating seeds whereas it was localized in the inner and middle cortex of nodules. Image analysis quantified higher fluorescence intensity of the phytase transcript signal in the seed embryo than in radicles, cotyledons or the nodule cortex. Furthermore, the phytase activity was 22-fold higher in cotyledons (43 nmol min(-1) g(-1) dry weight) than in nodules (2 nmol min(-1) g(-1) dry weight). The K m and V m values of phytase activity in cotyledons were also significantly higher than in nodules. Interestingly, the amplified sequence of cDNA phytase exhibited highest homology with the Glycine max purple acid phosphatase (NM_001289274) 90 % for germinating seed as compared to nodule phytase cDNA displaying 94 % homology with the Glycine max phytase (GQ422774.1). It is concluded that phytase enzymes are likely to vary from seeds to nodules and that phytase enzymes play key roles in the use of organic P or N2 fixation, as it is well known for germination.
MAIN CONCLUSION: The work provides the first-time evidence of tissue-specific expression of a phytase gene in the germinating seeds of Phaseolus vulgaris. Phytase enzyme plays a major role in germinating seeds. It is also active during N2 fixation within nodules of legumes. The effect of phosphorus (P) deficiency on phytase gene expression and localization in N2-fixing root nodules has been recently studied in hydroaeroponic culture of Phaseolus vulgaris. In this study, phytase gene transcripts within the germinating seed tissues of the P-inefficient P. vulgaris recombinant inbred line RIL147 were in situ localized with a similar RT-PCR recipe as that used for nodules. Our results show that the phytase gene expression was mainly localized in the outer layers, vascular cells and parenchyma of germinating seeds whereas it was localized in the inner and middle cortex of nodules. Image analysis quantified higher fluorescence intensity of the phytase transcript signal in the seed embryo than in radicles, cotyledons or the nodule cortex. Furthermore, the phytase activity was 22-fold higher in cotyledons (43 nmol min(-1) g(-1) dry weight) than in nodules (2 nmol min(-1) g(-1) dry weight). The K m and V m values of phytase activity in cotyledons were also significantly higher than in nodules. Interestingly, the amplified sequence of cDNA phytase exhibited highest homology with the Glycine max purple acid phosphatase (NM_001289274) 90 % for germinating seed as compared to nodule phytase cDNA displaying 94 % homology with the Glycine maxphytase (GQ422774.1). It is concluded that phytase enzymes are likely to vary from seeds to nodules and that phytase enzymes play key roles in the use of organic P or N2 fixation, as it is well known for germination.