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中国农学通报 ›› 2019, Vol. 35 ›› Issue (32): 53-56.doi: 10.11924/j.issn.1000-6850.casb18070095

• 林学 园艺 园林 • 上一篇    下一篇

酸枣花粉原生质体的分离条件研究

牛瑜菲, 彭建营   

  1. 河北农业大学 园艺学院
  • 收稿日期:2018-07-21 修回日期:2018-09-29 接受日期:2018-10-24 出版日期:2019-11-19 发布日期:2019-11-19
  • 通讯作者: 彭建营
  • 基金资助:
    国家自然科学基金资助项目“中国枣属植物的分子系统学及枣的品种分类学研究”(30270927);河北省自然科学基金项目“枣2n 花粉形成 机制和三倍体赞皇大枣起源的研究”(C2011204122),“‘无核小枣’木质素合成关键酶基因的克隆和表达研究”(C2016204144)。

Isolation Conditions of Pollen Protoplasts in Wild Jujube

  • Received:2018-07-21 Revised:2018-09-29 Accepted:2018-10-24 Online:2019-11-19 Published:2019-11-19

摘要: 旨为酸枣倍性育种、细胞融合提供材料,为枣品种杂交培育、品种更新开辟新径。以酸枣四分体时期的花粉为材料,采用酶解法分离原生质体,血球计数器计算原生质体产量以及FDA测定原生质体的活力,探讨花粉原生质体的分离条件。试验结果表明,甘露醇作为渗透压调节剂,在1%蜗牛酶+0.5%纤维素酶的混合酶液中,静止酶解4 h,四分体时期的花粉原生质体分离率最高,达到4.8×105个/mL,且通过0.01%的FDA活力检测,四分体花粉原生质体的活力为21.1%;不同浓度的甘露醇影响花粉原生质体的分离,试验得出,在0.3mol/L的甘露醇中,四分体时期的花粉原生质体分离率达到3.8×105个/mL,且活力为27.3%。四分体时期花粉原生质体的成功分离为后续三倍体育种提供了材料,为酸枣花粉其它时期的原生质体分离提供了实践基础。

关键词: 兰州市, 兰州市, 城市绿地, 观赏价值, 层次分析法

Abstract: The aim is to provide materials for ploidy breeding and cell fusion of wild jujube, and to open up a new path for hybrid breeding and variety renewal of jujube. The protoplast was separated by enzymatic hydrolysis from the pollen of wild jujube tetrad. The protoplast yield was calculated by hemocyte counter and the protoplast viability was determined by FDA. The results showed that when mannitol was used as osmotic regulator in the mixed enzyme solution of 1% snail enzyme and 0.5% cellulase, after 4 hours of enzymatic hydrolysis,the isolation rate of Pollen Protoplasts was the highest at the tetrad stage, reaching 4.8×105A/mL, by 0.01% FDA activity, the vitality of Pollen Protoplasts the tetrad stage was 21.1%. Degree of mannitol affected the isolation of pollen protoplasts. In 0.3 mol/L mannitol, the isolation rate of Pollen Protoplasts at tetrad stage was 3.8 ×105A/mL , and the vitality was 27.3%.isolation of Pollen Protoplasts in tetrad stage provides materials for subsequent triploid breeding, and provides practical basis for protoplast isolation in other pollen stages of wild jujube.

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