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《Molecular Plant Breeding》 2004-05
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Developing Practicable System of Chromosomal Doubling for the Construction of Doubled Haploid Population in Tobacco

Zhu Huiqin1,2 Zhang Xianyin1 Xue Qingzhong1*1 Department of Agronomy, College of Agriculture and Biotechnologe, Zhejiang University, Hangzhou, 3100292 College of Agriculture and Stockbreeding, Qinghai University, Xining, 810003 * Corresponding author, qzhxue@hotmail.com  
A comparative analysis of effects of four methods on chromosome doubling in tobacco indicated that the efficiency of colchicines (4g/L) applied directly to anther excised from the flower buds of tobacco just before anther culture was the highest (47.5%~75%), followed by that of leaves pre-treated by colchicines before regeneration was 30%~36.67%; that of the haploid plantlets was soaked in colchicine was 16.67%~32.35%; and the efficiency of colchicine soaked the auxiliary bud was the lowest (17.78%). The former both the methods brought about higher score in doubling, but had the higher abnormal plantlets ratio and the more tedious working procedure. Therefore, the method of soaking young plantlets in colchicine was highly effective, quickly and saves, when considering of the construction DH population. It was obvious that the doubling haploids plantlets of tobacco increase with soaking duration, when application of 4g/L of colchicine solution mixed with 20g/L of DMSO. The highest doubling haploid plantlets were obtained in colchicines soaking for 48h. With soaking plantlets in colchicine more than 12 h, the frequency of doubling in two tobacco crosses was raised to 2.30~2.93. Also, the application of DMSO to colchicine at low concentrations could lead to the higher doubling efficiency. Through completing the technical procedure of chromosome doubling, we have constructed two DH populations for the gene localization research.
【CateGory Index】: S572
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