| 马铃薯成熟花粉原生质体分离 |
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| 引用本文： | 张峰,王蒂,王玉萍.马铃薯成熟花粉原生质体分离[J].中国农业科学,2004,37(9):1358-1362. |
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| 作者姓名： | 张峰 王蒂 王玉萍 |
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| 作者单位： | 1. 甘肃农业大学生命科学技术学院,兰州,730070
2. 甘肃农业大学农学院,兰州,730070 |
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| 基金项目： | 国家教委高等学校优秀青年教师教学和科研奖励基金资助项目(9830596),国家“863”计划资助项目(2001AA241132) |
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| 摘 要： | 初步研究了马铃薯成熟花粉原生质体的分离条件,首次采用"低温-萌发-酶解"三步法从马铃薯成熟花粉中分离出原生质体。其分离途径是：经6℃低温处理的成熟花粉在萌发液中萌发出短花粉管时转入酶液。在酶液作用下,花粉发生质壁分离,花粉管脱落,随后原生质体缓慢地从花粉管断裂处挤出。在萌发30 min和1.0 mol·L-1甘露醇渗透压下,原生质体分离效果最佳,分离率最高达19.40%。用0.1%荧光增白剂检测脱壁完全,FDA荧光检测表明花粉原生质体具有生活力。
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| 关 键 词： | 马铃薯 花粉原生质体 分离 |
| 收稿时间： | 2003-9-22 |
Isolation of Pollen Protoplasts in Solanum tuberosum |
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| ZHANG Feng,WANG Di,WANG Yu-ping.Isolation of Pollen Protoplasts in Solanum tuberosum[J].Scientia Agricultura Sinica,2004,37(9):1358-1362. |
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| Authors: | ZHANG Feng WANG Di WANG Yu-ping |
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| Institution: | ZHANG Feng1,WANG Di1,WANG Yu-ping2 |
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| Abstract: | Mature pollen protoplasts of Solanum tuberosum were isolated by means of "cold pretreatment- germination-enzy-
matic treatment". The buds were first incubated at 6℃, then mature pollens were germinated. When most of the pollens grains had just produced short pollen tubes, they were transferred to an enzymatic solution. The process of pollen protoplasts isolation was as follows: firstly, plasm membrance seperated from the cell wall, then the protoplasts inside pollen wall were partially extruded through the germinal hole after the action of enzyme solution, and finally the spherical protoplasts were detached from the exine of pollens. Pollen protoplasts yield was greatly influenced by the germination duration time and mannitol concentrations in enzymatic solution, and 30 min germination and 1 mol·L-1 mannitol were optimal. The enzymatic solution consists of 1% cellulase (R-10), 0.5% hemicellulase (Sigma), 1% pectinlase (Serva), 0.5% potassium dextran sulfate, 1% PVP, 0.01% CH and K3 medium. The highest pollen protoplasts yield was 19.40%. Wall degraded pollens were identified with 0.1% calcoflower white test and its vitality was tested by fluorochromatic reaction with fluorescein diacetate (FDA). |
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| Keywords: | Solanum tuberosum L Pollen protoplasts Isolation |
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