罗汉果脱毒苗的快速繁殖研究

Rapid propagation of Momordica grosvenori virus-free plantlets

目的筛选适宜的罗汉果脱毒苗茎段增殖培养基,研究培养温度与光照对茎段增殖培养的影响.方法采用正交试验设计和完全随机实验设计筛选罗汉果脱毒苗茎段增殖培养基.正交L9(34)实验以在MS+BA 0.1 mg/L+NAA 0.1 mg/L上培养30 d的罗汉果脱毒苗单芽姊妹系的单节茎段为外植体,以培养基中BA,NAA,IBA及蔗糖为试验因子,各因子设3个水平,以在不同处理得到的增殖系数为测定指标;在此基础上进一步研究了培养基中BA和蔗糖质量浓度对增殖效果的影响;采用完全随机实验设计研究温度,光照对茎段增殖培养生成试管苗的影响.结果通过对正交实验结果的极差分析及方差分析表明,在4个因子中,对罗汉果茎段增殖系数影响最大的是BA,其次是蔗糖的NAA,IBA的影响最小;BA,NAA,IBA及蔗糖对增殖系数的影响都达到极显著.根据新复极差检验的结果,当前研究得到的罗汉果离体茎段的最佳增殖培养基为MS+BA 0.5 mg/L+NAA 0.01 mg/L+IBA 0.1 mg/L+蔗糖40 g/L,罗汉果茎段在此培养基上培养30 d后增殖系数为13.13;不同浓度BA与蔗糖对茎段培养的研究结果表明,BA在1.0 mg/L增殖系数最大.考虑为保持试管苗的遗传稳定性,BA为0.5 mg/L为宜.增殖培养基中的蔗糖适宜质量浓度为4.0%;罗汉果茎段培养适宜条件为培养温度25℃,培养光强1 000～3 000 lx.结论增殖培养基的成功筛选及培养温度与光强的选择,有助于实现罗汉果脱毒苗的工厂化生产.

Objective To sift the fitting shoot segment multiplying culture media of Momorclica grosvenori virus-free plantlets and to study the effect of temperature and light on the shoot segment multiplying culture. Methods The selection of the fitting shoot segment multiplying culture media was studied by orthogonal test design and completely random experimental design. Single nodes of virus-free plantlets from single-bud sibling-line cultured on MS＋BA 0. 1 mg/L＋NAA 0.1 mg/L for one month were used as explants, multiplying coefficient of different treatment as index of the experiment, and the effects of four factors, namely BA, NAA, IBA, and sucrose were evaluated by L9（3^4） orthogonal design. On the basis of the result of orthogonal test the different concentration BA and sucrose affecting on the growth of regenerated plantlets were studied, then the effect of temperature and light on the growth of regenerated plantlets were studied. Results The extreme deviation analysis and the variance analysis of the orthogonal test results showed that BA, NAA, IBA, and sucrose were very considerable. BA had the greatest effect, followed by sucrose, NAA, and IBA. The further analysis of SSR test appeared the best medium was the combination of A3B1C3D3 （MS supplemented with 0.5 mg/L BA, 0.01 mg/L NAA, 0. 1 mg/L IBA, 40 g/L sucrose, on which M. grosvenori could multiply 13.13 times ofter 30 d. The results of BA affecting on the growth of regenerated plantlet showed that 1.0 mg/L is the most, followed by 0.5 mg/L. To keep the hereditary stability in regenerated plantlets, the optimized concentration of BA should be 0.5 mg/L. The optimized concentration of sucrose was 4%. The optimized temperature and light are 25 C, 1 000-3 000lx Conclusion The selection of shoot segment multiplying culture media, temperature, and light helps to produce large-scale virus-free seedlings of M. grosvenori.