摘要
本研究以毛叶铁线莲(Clematis lanuginose)的花粉为试验材料,采用正交设计法研究毛叶铁线莲花粉离体条件下,蔗糖、硼酸、温度三个因素对其萌芽率及花粉管生长的影响,筛选出适宜毛叶铁线莲花粉离体萌发的最佳培养组合,并用该组合测定出的其花粉萌发率与0.5%氯化三苯基四氮唑法(TTC法)、碘-碘化钾法(I2-KI法)2种快速染色法对毛叶铁线莲花粉活力测定值进行比较研究。结果显示,蔗糖、硼酸、温度三个因素对花粉萌发率影响的主次关系依次为硼酸>温度>蔗糖,毛叶铁线莲花粉离体萌发的最佳培养组合为蔗糖10%、硼酸0.01%、温度20℃。I2-KI法测定的花粉活力为38.67%,花粉活力偏低;离体萌发法与0.5%TTC法测定的花粉活力分别为82.67%和87.00%,0.5%TTC溶液测定的活力值最高,比离体萌发法高4.33%,0.5%TTC法和离体培养法方差分析结果表明两者无显著差异,均能准确反映毛叶铁线莲的花粉活力。因此0.5%TTC染色法可选作快速测定毛叶铁线莲花粉活力的最优方法,它具有快速、简便等优点,为铁线莲育种、花粉储存、花粉活力检测等相关研究提供理论基础。
In order to screen o ut the optimum culture combination of pollen germination in vitro, the pollen of Clematis lanuginose was used as the experimental material, and using orthogonal design to study the effects of sucrose, boric acid concentration and temperature on the pollen of Clematis lanuginose in vitro. What＇s more, we used this pollen germination rate, 0.5% TTC（2, 3, 5-Triphenyte-trazoliumchloride）, and I2-KI and vitro germination to determine the viability of C. lanuginose. The results showed that boric acid concentration has the highest effect on pollen germination, while sucrose concentration the lowest. The optimal combination of vitro conditions for C.lanuginose pollen germination was 10% sucrose, 0.01% boric acid and 20℃. The pollen viability of C. lanuginose determined by I2-KI method was 38.67%, which was low; meanwhile, the pollen germination in vitro and TTC method were 82.67% and 87% respectively, TTC method was more than 4.33% in vitro methods, showing no significant difference. Therefore, both TTC and in vitro methods could accurately test the pollen vitality of Clematis lanuginose, while the TTC had its own unique advantages, such as simplicity, efficiency and so on, so it could be the most suitable method for determining the pollen vitality of C. Lanuginose. This research laid theoretical basis for the research on Clematis breeding, pollen storage and pollen viability test.
出处
《分子植物育种》
CAS
CSCD
北大核心
2017年第11期4667-4672,共6页
Molecular Plant Breeding
基金
江苏省科技计划--产学研前瞻性联合研究项目(BY2015074-01)
江苏省中国科学院植物研究所青年基金(SQ201301)共同资助
关键词
毛叶铁线莲
花粉活力
正交设计
0.5%TTC
Clematis lanuginose, Pollen viability, Orthogonal design, 0.5% TTC method
