马蹄莲花粉活力鉴定与贮藏影响因子研究

以马蹄莲不同品种的花粉为材料,研究马蹄莲花粉活力检测的适宜方法和贮藏影响因子。比较了TTC、Lugol’s 染色液、亚甲基蓝、醋酸洋红4种染色法和离体培养法鉴定马蹄莲花粉活力的效果,以及花粉干燥时间（含水量）、贮藏温度、预冻方法、解冻方法对花粉贮藏的影响。结果表明：所选用的4种染色法中,TTC法能将马蹄莲花粉染成深红、浅红、粉红,从而区分出花粉有无活力及活力的强弱;Lugol’s 染色液、醋酸洋红不能使花粉染色;亚甲基蓝将所有花粉染成同一颜色;所以除TTC法外均不适合马蹄莲花粉活力的鉴定。离体培养法培养24 h可以较好鉴定马蹄莲花粉活力,其最佳培养基配方为80 g/L蔗糖+0.2 g/L硼酸+0.3 g/L硝酸钙。马蹄莲花粉的适宜干燥条件是室温下干燥27 h,之后不需任何预冻处理直接放入冰箱,最适解冻方式是37 ℃水浴2 min。不同贮藏温度下马蹄莲花粉活力变化存在明显差异,试验温度中以-80 ℃的贮藏效果最好,贮藏61 d后仍能保持40%以上的活力,其次是-20 ℃贮藏,然后是室温,4 ℃贮藏效果最差。综上所述,TTC染色法和离体培养法是马蹄莲花粉活力测定的适宜方法,-80 ℃的低温可明显延长马蹄莲花粉的贮藏时间,研究结果为马蹄莲花粉离体保存提供了科学依据。

Identification of Pollen Vitality and Factors Influencing Storage Quality of Zantedeschia spp.

The pollen of different varieties of Zantedeschia spp.was selected as the experimental material to determine the optimal method to detect pollen viability and factors which influence pollen storage quality.The effects of TTC,Lugol’s solution,methylene blue and acetic acid magenta to detect pollen vitality were compared.The influence of pollen drying time(mean water content),storage temperature,pre-cooling method and thawing method on pollen storage was explored.The results showed that the TTC method could dye the pollen of Zantedeschia into deep red,light red,and pink.However,the Lugol’s solution method and ammonium acetate method could not dye the pollen,and methylene blue could dye all pollen blue.So only TTC method was suitable to identify Zantedeschia pollen vitality.The most suitable culture medium for pollen germination was consisted of 80 g/L sucrose,0.2 g/L boric acid,0.3 g/L calcium nitrate,and the best detection time for pollen vitality was 24 h cultured in the medium.The treatment of drying pollen for 27 h,then putting into the refrigerator directly,combined with water bath at 37℃for 2 min could work with low temperature to maintain the vitality of the pollen well.Temperature had great impact on the vitality of Zantedeschia pollen stored.Among the experimental temperature,-80℃was the best one and still maintained more than 40%viability after storage for 61 days,then-20℃was second,room temperature was the third,and 4℃was the worst.Taken together,TTC staining and cultured in vitro media were the suitable methods for identifying the vitality of Zant edeschia pollen.Low temperature-80℃could prolong the storage time of Zantedeschia pollen significantly.The results would provide a scientific basis for preserving Zantedeschia pollen.