Immunological assay of phytochrome in small sections of roots and other organs of maize (Zea mays L.) seedlings

Phytochrome was determined in small sections of maize (Zea mays L.) seedlings by means of a highly specific double sandwich enzyme immunoassay which uses a monoclonal anti-phytochrome antibody for binding phytochrome and anti-phytochrome serum to detect the bound phytochrome. The distribution of phytochrome in maize seedlings was followed from germination to the 7th d after soaking the caryopses. Regions of high phytochrome accumulation were found in the coleoptile tip, the root cap and the shoot apex: the values for 5-d-old seedlings were 120, 80 and 70 g phytochrome per g fresh weight (or 0.91, 0.61 and 0.53 nmol·g^-1), respectively. The mesocotyl and the leaves contained relatively low amounts of phytochrome (less than 10 g·g^-1FW), which were almost uniformly distributed throughout these organs. As might be expected, regions of these organs adjacent to the shoot apex showed higher levels. The root, other than root tip, was almost devoid of phytochrome (0.2 to 0.5 g·g^-1). The general distribution of phytochrome in organs did not change during the development of seedlings. The amount of phytochrome, however, did fluctuate: up to the 5th or 6th d after soaking the caryopses, the levels increased in the regions of high phytochrome accumulation but thereafter decreased. After the 6th d the roots were 15 cm or longer and the coleoptiles became prone to penetration by primary leaves. The tips of adventitious roots, emerging after the 6th d, were also found to contain phytochrome. When the root cap was illuminated (4.3 W·m^-1), phytochrome was degraded as in illuminated shoots. Degradation of phytochrome in coleoptile, mesocotyl and shoot apex started with a lag phase but phytochrome degradation in the root cap and the leaves started without a lag. In contrast to shoot phytochrome, which was almost completely degraded under continuous illumination, about 3% of initial phytochrome was measured in root caps after 24 h continuous illumination. Some of the data, obtained by immunological measurements, may indicate differences between phytochrome, or its synthesis or degradation, in the root cap and shoots. The results are discussed with a view to different red-light-mediated responses of grass seedlings.Abbreviations ABTS 2,2-azino-bis(3-ethylbenz-thiazoline)-sulfonic acid - EIA enzyme immunoassay - PBS phosphatebuffered saline - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis