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- Author or Editor: Chung-Chih Lin x
A xylem mutant (vse) was isolated from a Bambusa edulis (Odashima) Keng plantlet following vegetative micropropagation and subculture for 7 consecutive years and induced to proliferate in medium supplemented with 0.1 mg·L-1 (0.5 μm) thidiazuron (TDZ) and to develop roots in medium supplemented with 5 mg·L-1 (26.9 μm) α-naphthaleneacetic acid (NAA). Subsequent investigations comparing the growth habits of mutant plantlets with those of the wild type indicated that the growth of the former was retarded in a greenhouse. Several morphological abnormalities were observed in the vse mutant: it had thinner stems with fewer trichromes on the surface; the xylem vessels were smaller in diameter and contained crystal-like structures in the pith; the leaves were shorter and narrower with a sharp leaf blade angle; the roots were thinner and contained fewer xylem cells. The cation concentrations of both the mutant and wild type were similar in the in vitro analysis, except for those of iron and potassium, which were lower in mutant leaves in vivo. In 2-month-old mutant plants, iron chlorosis was observed on young leaves and a potassium deficiency was observed on older leaves. After 1 year of growth in the greenhouse, all of the wild-type plants had survived, but only 27% (16/60) of the mutant vse plants were alive.
The chloroplast genome of an albino mutant isolated from tissue culture of the bamboo Bambusa edulis Munro was examined to identify aberrations. A number of the chloroplast genes encoding ATP synthases, photosystem II subunits, NADH dehydrogenase, and ribosomal proteins had been deleted, at least partially, in the albino mutant. Comparison of the two-dimensional electrophoresis profiles of albino and green bamboos revealed three spots of reduced intensity, indicating repression of these proteins in the albino mutants. Mass spectroscopic analysis subsequently revealed that two of these proteins are 33-kDa subunits of the photosystem II oxygen-evolving protein complex (PsbO) and one is a 23-kDa subunit of photosystem II oxygen-evolving protein complex (PsbP). The genes encoding these two proteins were cloned from B. edulis, and were denoted BePsbO (accession no. EF669513) and BePsbP (accession no. EF669512). Reverse transcription polymerase chain reaction and two-dimensional gel analyses of BePsbO and BePsbP in green and albino bamboos grown in the light or dark revealed that the albino mutant, similar to its green counterpart, sensed the light signal, resulting in the induction of BePsbO and BePsbP transcription, but it did not accumulate the protein products. We conclude that the repression of protein-expressing BePsbO and BePsbP is because of a defect in post-transcriptional regulation in the albino mutant.