2018年12月16日 星期日
甜菊自交胚挽救及其自交S1代的主要性状分析
Rescue of self-crossing embryo of Stevia rebaudiana and analysis on major characters of its selfcrossing S1 progeny
2018年 第27卷 第4期 页码[81-89]
摘要

对甜菊(Stevia rebaudiana Bertoni)品种‘中山5号’(‘Zhongshan No. 5’)、‘中山6号’(‘Zhongshan No. 6’)、‘中山8号’(‘Zhongshan No. 8’)、‘江甜2号’(‘Jiangtian No. 2’)、‘谱星1号’(‘PC Star No. 1’)和‘守田3号’(‘Morita No. 3’)自交授粉后的有胚率进行了统计,进一步筛选出‘中山8号’的胚挽救方法,并对‘中山8号’自交S1代植株的主要性状进行了分析。结果表明:甜菊不同品种间的有胚率差异明显,其中,自交授粉后5 d‘中山8号’的有胚率最高(16.0%),自交授粉后10 d以上6个品种的有胚率基本为0.0%。随自交授粉后天数的增加,‘中山8号’胚珠和幼胚在MS培养基、愈伤诱导培养基(添加2.0 mg·mL-1 6-BA和2.0 mg·mL-1 NAA的MS培养基)和分化诱导培养基(添加2.0 mg·mL-1 6-BA和0.2 mg·mL-1 NAA的MS培养基)上的萌发时间逐渐缩短,萌发率逐渐升高。自交授粉后7~9 d‘中山8号’幼胚在MS培养基上的萌发率高达80%以上。短日照处理下,‘中山8号’自交S1代植株间现蕾期的差异较大,短日照处理10~46 d现蕾,其中,短日照处理15 d现蕾的植株最多,占植株总数的16.5%。现蕾期‘中山8号’103个自交S1代植株叶片中莱鲍迪苷A(RA)、甜菊苷(ST)、莱鲍迪苷C(RC)、杜尔可苷A(DA)和总甜菊糖苷(TSG)的含量及RA、ST、RC和DA含量占TSG含量的比例的变化范围均较大,RA、ST、RC、DA和TSG的含量分别为0.26%~11.43%、0.36%~6.45%、1.52%~9.18%、0.05%~1.40%和5.80%~16.90%,RA、ST、RC和DA含量占TSG含量的比例分别为3.19%~72.06%、3.20%~46.98%、12.59%~74.27%和0.53%~12.33%。在‘中山8号’及其103个自交S1代植株中,‘中山8号’和9个自交S1代植株为高RA型,7个自交S1代植株为高RA-RC型,11个自交S1代植株为高RC型,1个自交S1代植株为高RA-RC-DA型。上述研究结果表明:胚挽救技术可克服甜菊自交不亲和受精后障碍,获得的自交S1代植株的现蕾期和甜菊糖苷含量变化范围较大,为甜菊的种质创制和新品种培育提供了有效途径。

Abstract

Embryo rate of cultivar ‘Zhongshan No. 5’, ‘Zhongshan No. 6’, ‘Zhongshan No. 8’, ‘Jiangtian No. 2’, ‘PC Star No. 1’, and ‘Morita No. 3’ of Stevia rebaudiana Bertoni after selfcrossing pollination was compared, and furthermore, embryo rescue method of ‘Zhongshan No. 8’ was screened. The main characters of selfcrossing S1 progeny plants of ‘Zhongshan No. 8’ were also analyzed. The results show that there is an obvious difference in embryo rate among different cultivars of S. rebaudiana, in which, embryo rate of ‘Zhongshan No. 8’ is the highest (16.0%) after self-crossing pollination for 5 d, and those of six cultivars are basically 0.0% after self-crossing pollination for longer than 10 d. With the increase of days after selfcrossing pollination, germination times of ovule and embryo of ‘Zhongshan No. 8’ are gradually shortened on MS medium, callus induction medium (MS medium added with 2.0 mg·mL-1 6-BA and 2.0 mg·mL-1 NAA), and differentiation induction medium (MS medium added with 2.0 mg·mL-1 6-BA and 0.2 mg·mL-1 NAA), and their germination rates gradually increase. Germination rate of embryo of ‘Zhongshan No. 8’ on MS medium is above 80% after self-crossing pollination for 7-9 d. Under short-day treatment, there is an obvious difference in budding stage of self-crossing S1 progeny plants of ‘Zhongshan No. 8’, and bud appears after short-day treatment for 10-46 d, in which, number of budding plants is the biggest after short-day treatment for 15 d, accounting for 16.5% of number of total plants. Variation ranges of contents of rebaudioside A (RA), stevioside (ST), rebaudioside C (RC), dulcoside A (DA) and total steviol glycosides (TSG), and percentages of contents of RA, ST, RC and DA to TSG content are relatively large in leaves of 103 self-crossing S1 progeny plants of ‘Zhongshan No. 8’ at budding stage. Contents of RA, ST, RC, DA, and TSG are 0.26%-11.43%, 0.36%-6.45%, 1.52%-9.18%, 0.05%-1.40%, and 5.80%-16.90%, respectively, and percentages of contents of RA, ST, RC, and DA to TSG content are 3.19%-72.06%, 3.20%-46.98%, 12.59%-74.27%, and 0.53%-12.33%, respectively. Among ‘Zhongshan No. 8’ and its 103 self-crossing S1 progeny plants, ‘Zhongshan No. 8’ and 9 self-crossing S1 progeny plants belong to high RA type, 7 self-crossing S1 progeny plants belong to high RA-RC type, 11 selfcrossing S1 progeny plants belong to high RC type, and 1 self-crossing S1 progeny plant belongs to high RA-RC-DA type. It is suggested that the technique of embryo rescue can overcome post-crossing barrier of S. rebaudiana self-incompatibility, and variation ranges of budding stage and steviol glycoside content of self-crossing S1 progeny plants obtained are relatively large, which provides an effective approach for germplasm innovation and breeding of new cultivars.
 

关键词甜菊; 自交; 胚挽救; S1代; 现蕾期; 甜菊糖苷含量
Key wordsStevia rebaudiana Bertoni; self-crossing; embryo rescue; S1 progeny; budding stage; steviol glycoside content
作者杨永恒, 侯孟兰, 张永侠, 徐晓洋, 孙玉明, 张婷, 原海燕
所在单位江苏省中国科学院植物研究所(南京中山植物园), 江苏 南京 210014
点击量58
基金项目国家自然科学基金青年基金项目(31601371); 江苏省自然科学基金青年基金项目(BK20160600)