Research Of Ramie (Bochmeria Nivea L. Gaud) Anther Culture Method

Ramie (Bochmeria nivea L. Gaud.) is an important fiber crops and speciallyproduction in China. The ramie output in China accounts for the world ultimate output95％, the textile processing ability approximately composes 80％. Because of its excellenttextile fiber quality, ramie has the important status in textile industry. Thus ramie has thefine reputation of "China Grass" in the world.But in the application, the questions of its extremely complex heredity backgroundand long traditional breeding method cycle, have seriously limited the scientific researchdevelopment of ramie. Many crops have been obtained dihaploid plant by double thehaploid in present. The anther culture is a simple method to obtain haploid. We haveestablished the complete protocol for ramie anther callus induction and callus ploidyidentify by researching the major effect factors of the anther callus induction frequency in13 ramie varieties or materials. And it was the foundation for developing research. Themain research content and result as follows:The research of anther contaminated factors indicated that it was easy to becontaminated whenthe anther was kept in 4℃for longer than 2 days or cultured inliquid medium. The callus induction frequency of Tanjiahema, Jiangkou, Huangganma,Cuba ramie were highest when sterilized by 1％NaClO solution.We have optimized the ramie anther culture method by selecting the basic media,sucrose concentration, plant growth regulators and genotypes. Anthers cultured in MSNand B5N media indicated higher callus induction frequency than in other media. Whenthe media contained 3％sucrose concentration the anthers had high callus inductionfrequency. 9 media which contained different plant growth regulators and concentrationswere selected. The result shows that major part of the media can induct anther callus. Andthe medium which contains 2,4-D 2.0mg/L and BA 0.2 mg/L had stable effect of callusinduction. And we have selected some materials which have high callus inductionfrequency, such as 94-10 (81.05％). We translated the anther callus of Tanjiahema, Bailizi Qing, 94-10 to differentdifferentiation media. Callus grew slowly and their structure were loose decadent pelletwhen they cultured in the media contains 2,4-D 0.5mg/L and BA 0.5mg/LThe anther callus were identified the ploidy by FACSCalibur and cytology method.No haploid cells were found by the two methods.