Kim Moonza, Jang In-Chang, Kim Jin-Ae, Park Eun-Joon, Yoon Michung, Lee Youngwon
Department of Life Sciences, Mokwon University, Seo-Gu, Taejon, South Korea.
Plant Cell Rep. 2008 Mar;27(3):425-34. doi: 10.1007/s00299-007-0442-4. Epub 2007 Sep 13.
We report high frequencies of embryo production and plant regeneration through isolated microspore culture of hot pepper (Capsicum annuum L.). Microspores cultured in modified NLN medium (NLNS) divided and developed to embryos. Globular and heart-shaped embryos were observed from 3 weeks after the beginning of culture, and many embryos reached the cotyledonary stage after 4 weeks of culture. These cotyledonary embryos developed to plantlets after transfer to solid B5 basal medium. We also optimized conditions for embryo production by varying the pretreatment media, the carbon sources, and culture densities. Heat shock treatment in sucrose-starvation medium was more effective than in B5 medium. Direct comparisons of sucrose and maltose as carbon sources clearly demonstrated the superiority of sucrose compared to maltose, with the highest frequency of embryo production being obtained in 9% (w/v) sucrose. Microspore plating density was critical for efficient embryonic induction and development, with an optimal plating density of 8 x 10(4)-10 x 10(4)/ml. Under our optimized culture conditions, we obtained over 54 embryos, and an average of 5.5 cotyledonary embryos when 10 x 10(4) microspores were grown on an individual plate.
我们报道了通过辣椒（Capsicum annuum L.）游离小孢子培养获得高频胚胎生产和植株再生的情况。在改良的NLN培养基（NLNS）中培养的小孢子分裂并发育成胚胎。培养开始3周后观察到球形和心形胚胎，培养4周后许多胚胎达到子叶期。将这些子叶期胚胎转移到固体B5基本培养基后发育成小植株。我们还通过改变预处理培养基、碳源和培养密度优化了胚胎生产条件。在蔗糖饥饿培养基中进行热激处理比在B5培养基中更有效。直接比较蔗糖和麦芽糖作为碳源清楚地表明，与麦芽糖相比，蔗糖具有优越性，在9%（w/v）蔗糖中获得的胚胎生产频率最高。小孢子接种密度对于高效胚胎诱导和发育至关重要，最佳接种密度为8×10⁴ - 10×10⁴/ml。在我们优化的培养条件下，当在单个平板上培养10×10⁴个小孢子时，我们获得了超过54个胚胎，平均有5.5个处于子叶期的胚胎。
