An in vitro method for obtaining bamboo mosaic virus (BaMV)-free plantlets of Bambusa oldhamii Munro was developed. BaMV-free meristems were incubated on MS basal medium supplemented with 0.45 μm thidiazuron (TDZ) to induce the development of multiple shoots. Multiple shoot proliferation was higher in stationary liquid culture than on semisolid medium. Cytokinin was the key component for inducing proliferation, and TDZ was the stable and effective cytokinin for proliferation in long-term subcultures. Multiple shoots rooted after 1 month in MS basal medium containing 10.74 to 26.85 μm α-naphthaleneacetic acid with a rooting efficiency of 83%. Healthy, well-developed plantlets were transferred to soil in pots and raised in a greenhouse. Those plants derived from tissue culture were more vigorous than the ones derived from the traditional in vivo vegetative propagation method, air layering. The tissue culture-derived plants could produce the culms after 15 months. Fifteen of 38 plants flowered 2 years after being transplanted to the field.
Choun-Sea Lin, Krishnan Kalpana, Wei-Chin Chang and Na-Sheng Lin
Choun-Sea Lin, Nien-Tzu Liu, De-Chih Liao, Jau-Song Yu, Chuang-Hwei Tsao, Chao-Hsiung Lin, Chih-Wen Sun, Wann-Neng Jane, Hsing Sheng Tsay, Jeremy Jian-Wei Chen, Erh-Min Lai, Na-Sheng Lin, Wei-Chin Chang and Chung-Chih Lin
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.