酿酒葡萄桂葡一号花粉原生质体分离条件初探  

Isolation of protoplasts from pollens of wine grape variety Guipu 1

作　　者：廖芬[1,2] 唐文忠[3] 黄茂康[1] 周主贵[4] 

机构地区：[1]广西农业科学院农产品加工研究所 [2]广西作物遗传改良生物技术重点实验室 [3]广西农业科学院园艺研究所 [4]广西农业科学院,南宁530007

出　　处：《南方农业学报》2011年第10期1181-1184,共4页Journal of Southern Agriculture

基　　金：广西自然科学基金项目(桂科自0640023)

摘　　要：【目的】探索葡萄成熟花粉粒原生质体分离条件,为下一步原生质体杂交及遗传改良提供技术参考。【方法】使用低温—酶解法对酿酒葡萄桂葡一号花粉原生质体进行分离,探讨低温处理不同时间、不同花序位置和不同甘露醇浓度对原生质体分离的影响。【结果】随着低温处理时间的延长,原生质体分离得率越高,在0.6mol/L甘露醇条件下,以处理7d的原质体分离得率最高,达18.94%,其次是处理5d。对于不同部位花粉,以中上部花药的原生质体分离得率较高,而以中部花序的花粉原生质体分离得率最高,达19.11%;中下部花药及已开始散粉的花药原生质体分离得率较低。相对于1.2mol/L甘露醇,以添加0.6mol/L甘露醇作为渗透压处理的花粉原生质体分离得率较高。【结论】低温处理时间、花粉成熟度及甘露醇浓度均对酿酒葡萄花粉原生质体的分离有明显影响。选用即将开花的中上部花药为材料,经0～4℃低温预处理5～7d,并以0.6mol/L甘露醇作为渗透压调节剂可获得大量有活性的花粉原生质体。[Objective]The aim of this experiment was to find out suitable conditions for isolation of mature protoplasts from wine grape pollens in order to facilitate protoplast hybridization for improving wine grape cultivars.[Method]The mature protoplasts of pollens taken from grape wine variety Guipu 1 were isolated using cold pretreatment and enzyme decomposition methods.The effects of different duration of low temperatures,different inflorescence positions and mannitol concentrations on its isolation were determined.[Result]The results indicated that the rate of protoplast isolation increased with elongation of low temperature（4℃） duration,the highest rate was recorded as 19.40% after treating pollens 7 days with 0.6 M mannitol,followed by 18.94% after treating pollens for 5 days.The rate of protoplast isolation from anthers of top inflorescence was the highest（19.11%） ,and that of protoplasts from anther of middle and lower inflorescence was lower.Compared to 1.2 M mannitol,the rate of protoplast isolation was higher in 0.6 M mannitol.[Conclusion]The low temperature duration,pollen maturity and mannitol concentration have significant effects on isolation of protoplast from pollen.The highest protoplast may be obtained from pollens of middle and upper part of anther and treating them for 5-7 days under 0-4℃ low temperature with 0.6 M mannitol.

关 键 词：酿酒葡萄 桂萄一号 花粉 原生质体 分离条件 

分 类 号：S663.101[农业科学—果树学]