Distribution of type I phytochrome (phyA) RNA1 and RNA2 in etiolated pea seedlings

Four-day-old etiolated pea seedlings were divided into 11 parts along the axis, from which poly(A)+ RNA and DNA were extracted. Using a slot-blot hybridization assay, the abundance of pea type I phytochrome (phyA) poly(A)+ RNA was measured in each portion of the etiolated pea seedling. For the quantification of the phyA RNA, pea phyA RNA synthesized in vitro was used as an RNA standard. The hook region contained the highest abundance of phyA RNA (approximately 0.3 ng phyA RNA per microgram DNA) in the etiolated seedling. Two mRNAs of different length (the shorter designated as RNA1 and the longer RNA2) are produced in detectable amounts from single pea phyA in the etiolated seedling; the ratio of the abundance of phyA RNA1 to phyA RNA2 was determined in each of the 11 parts by a primer extension assay. The abundance of phyA RNA1 in the plumule and hook regions was 3-5-fold higher than that of RNA2, whereas the ratio of their abundance was approximately unity in other regions. A time course study of the abundance of both RNAs was carried out during the imbibition of seeds and indicated that the accumulation of phyA RNA1 occurred more rapidly in the cotyledons than that of RNA2 during the first day of imbibition, whereas the accumulation of phyA RNA2 increased rapidly during the second day and became as high as that of phyA RNA1 by the third day.