Abstract

Phosphorus deficiency reduces forage yield and stand persistence of alfalfa (Medicago sativa L.). Our objectives were to isolate and characterize a high-affinity phosphate-transporter (P-transporter) from alfalfa roots (Medicago sativa L.); determine how phosphorus (P) nutrition impacts P-uptake, growth, and carbohydrate and protein metabolism of alfalfa cells; and learn how expression of the P-transporter is influenced by P nutrition. An 1087-base pair (bp) sequence was isolated using RT-PCR that possessed high nucleotide and amino acid sequence similarity to high-affinity P-transporters. Cultured cells were sampled at 3-day intervals for 9 days while growing in media containing P concentrations ranging from 0 to 10 mM. Media P concentrations declined rapidly in all P treatments by day 6. Low media P concentrations (0, 0.1 and 0.5 mM) reduced cell growth rates compared to higher media P levels (2.5, 5 and 10 mM). Suspension cell cultures supplied 0.5, 2.5, 5, and 10 mM P had lower starch concentrations by day 3 compared to cells cultured in media containing 0 and 0.1 mM P. Steady-state transcript levels for the high-affinity P- transporter were high in P-deprived cells, but declined within 1 day when cells were provided 10 mM P.

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