Purification and Properties of Acid Phosphatase-1 from a Nematode Resistant Tomato Cultivar

Abstract

In tomato the acid phosphatase-1 isozyme (Apase-1) is inherited as a single locus linked to the nematode resistance gene (Mi)

Purification and properties of acid phosphatase secreted from lupin roots under phosphorus-deficiency conditions

“…indicate that the native enzyme is a homo-dimer with a molecular weight of 72,000 for the subunit. This molecular weight of the homo-dimer was larger than that of other acid phosphatases purified from seeds of triticale (45,700 for monomer, Ching et al 1987), cell cultures of tomato (51,000 for dimer, Paul and Williamson 1987), barley roots (79,000 and 77,600 for dimer, Panara et al 1990), soybean axes and cotyledons (i00,000 for dimer, Kaneko et al 1990), seeds of sunflower (103,000 for dimer, Park and Etten 1986), tuberous root of sweet potato (110,000 and 105,000 for dimer, Uehara et al 1974), and cell culture of soybean (130,000 for dimer, LeBansky et al 1992), but smaller than that from cotton seedlings (200,000 for tetramer, Bhargava and Sachar 1987), and peanut seeds (240,000 for hexomer, Basha et al 1984). The isoelectric points (pI) of crude acid phosphatases from secretion, roots and leaves were determined by I E F (Fig.…”

Isolation and Characterization of a Tomato Acid Phosphatase Complementary DNA Associated with Nematode Resistance

“…The procedure developed here for purification of Apase-11 is considerably faster than previously reported methods (14,22). Because the procedure eliminates the ammonium sulfate precipitation and lengthy dialysis steps the purification can be accomplished in 3 d. Each chromatography step can be carried out in <2.5 h with minimal sample manipulation between steps.…”

“…Approximately 25 to 50 ,ug of pure Apase-1' was recovered from 1.5 kg of packed cells. The presence of other interfering acid phosphatases, which account for most of the activity present in cell extracts, prevents the determination of overall recoveries and fold purifications (14). The concentration of Apase-l1 in crude extracts was estimated by comparing the Apase-1 enzyme activity of crude and purified samples using the cellulose acetate gel electrophoresis procedure.…”

“…Purification of Apase-11 from Tomato Suspension Cells Tomato, Lycopersicon esculentum cv VFNT, cells were grown in 6-L flasks for 7 to 10 d with 1 L ofcomplete medium containing sucrose (14). Six flasks yield 1.5 (2).…”

“…Apase-1' activity was determined by a cellulose acetate gel electrophoresis procedure using f3-3-napthylphosphate as the substrate (14) and by a microtiter plate assay using p-nitrophenylphosphate as the substrate. Other conditions for enzyme assays were as previously described (14).…”