花楸花粉超低温保存及再生

于琦; 杨雨荷; 刘欢; 郑昕雨; 孔繁秋; 张海莹; 宋红

doi:10.19303/j.issn.1008-0384.2019.04.005

花楸花粉超低温保存及再生

doi: 10.19303/j.issn.1008-0384.2019.04.005

东北林业大学园林学院, 黑龙江哈尔滨 150040

基金项目:

国家级大学生创新创业基金项目 201810225191

详细信息

作者简介:
于琦(1998-), 女, 研究方向:植物种质资源保护(E-mail:1508931645@qq.com)

通讯作者:
宋红(1965-), 女, 硕士, 高级工程师, 研究方向:植物种质资源保护(E-mail:851100942@qq.com)

中图分类号: S686
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- 被引次数: 0
出版历程
- 收稿日期: 2019-03-12
- 修回日期: 2019-04-03
- 刊出日期: 2019-04-28

Cryopreservation of Sorbus pohuashanensis Pollens

College of Landscape Architecture, Northeast Forestry University, Harbin, Heilongjiang 150040, China

摘要

摘要: 目的探讨花楸（Sorbus pohuashanensis）花粉超低温保存及再生的方法，以延长其花粉保存时间，为科研、生产提供理论和实践依据。方法分别设置不同培养基配方、不同光照、不同采集时间处理，进行花楸花粉离体萌发试验，测定各试验处理的花粉萌发率；设置不同冷冻方式和化冻方式处理，测定两种方式不同处理的花粉萌发率；制备不同含水率的花粉，分别测定其在超低温保存前、后的萌发率。结果（1）培养基为100 g·L^-1蔗糖+100 mg·L^-1硼酸的处理，花粉萌发率最高，达72.42%；（2）黑暗处理花粉萌发率为77.24%，比光照处理高8.96个百分点；（3）一天中，10：00和14：00采集的花粉，其活力显著高于其他时间采集的花粉；（4）-20℃预冷冻培养30 min后投入液氮保存24 h的处理效果最好，花粉萌发率达43.15%；（5）超低温保存后40℃水浴解冻5 min处理比其他解冻方式处理的花粉活力高，萌发率为35.70%；（6）花楸花粉超低温保存最适含水率为7.72%，此含水率下花粉超低温保存后萌发率最高，为31.53%。结论花楸花粉采用超低温技术保存24 h后仍具有活力，黑暗条件、花粉采集时间、花粉萌发培养基、花粉含水率、冷冻与化冻方式均会对花楸花粉超低温保存效果产生显著影响。
- 花楸 /
- 花粉 /
- 超低温保存 /
- 种质资源保护
Abstract: Objective Cryopreservation and regeneration of pollens of Sorbus pohuashanensis were investigatied. Method An in vitro method was developed with experiments on medium formulation, light exposure, and pollen collection for the pollen germination rate determination.The germination rates of S.pohuashanensis pollens treated by various freezing and thawing procedures and different moisture contents before and after the treatments were compared. Result (1) The medium for optimum pollen germination was found to consist of sucrose 100 g·L^-1 and boric acid 100 mg·L^-1 that delivered a rate of 72.42%. (2) Incubation in darkness yielded a germination rate of 77.24%, which was 8.96 percentage point higher than that under light exposure. (3) The vitality of pollens collected at 10 a.m. or 2 p.m. was significantly higher than other time in a day. (4) Among all cryopreservation methods tested, pre-freezing pollens at -20℃ for 30 min prior to submerging in liquid nitrogen for 24 h provided the greatest germination rate of 43.15% after thawing. (5) For thawing, soaking the frozen pollens in a 40℃ water bath for 5 min rendered the highest germination rate of 35.70%. (6) Moisture content of the pollens for the cryopreservation was determined to be 7.72% to result in the best germination rate of 31.53% after thawing. Conclusion The cryopreservation conditions for S. pohuashanensis pollens as established would result in high viability and germination rate after storage.
- Sorbus pohuashanensis /
- pollen /
- cryopreservation /
- germplasm preservation

HTML全文

图 1 不同培养基对花楸花粉萌发力的影响

注：不同大写字母表示相同的硼酸质量浓度条件下，不同蔗糖质量浓度处理间花粉萌发率的差异显著性，无相同大写字母者表示差异达显著水平(P＜0.05)；不同小写字母表示相同的蔗糖质量浓度条件下，不同硼酸质量浓度处理间花粉萌发率的差异显著性，无相同小写字母者表示差异达显著水平(P＜0.05)。

Figure 1. Effect of medium on germination of S. pohuashanensis pollens

Note: Different capital letters indicate significant differences in pollen germination rates under different concentrations of sucrose when the concentration of boric acid is the same.Different lowercases indicate significant differences in pollen germination rates under different concentrations of boric acid when the concentration of sucrose is the same.

下载: 全尺寸图片幻灯片

图 2 不同光照条件对花楸花粉的萌发率的影响

注：不同小写字母表示不同处理间花粉萌发率差异达显著水平(P＜0.05)。图 3~5同。

Figure 2. Effect of light exposure on germination rate of S. pohuashanensis pollens

Note: Different lowercases indicate significant differences at 0.05 level(P<0.05).The same as Fig. 3-5.

下载: 全尺寸图片幻灯片

图 3 不同采集时间对花楸花粉萌发率的影响

Figure 3. Effect of collection time on germination rate of S. pohuashanensis pollens

下载: 全尺寸图片幻灯片

图 4 不同预冷冻方式对花楸花粉萌发率的影响

注：A为逐级变温处理，CK为无预冷冻的对照处理。

Figure 4. Effect of pre-freezing on germination rate of S. pohuashanensis pollens after treatments

Note:A:Lower the temperature stage by stage, CK:Without prefreezing.

下载: 全尺寸图片幻灯片

图 5 不同化冻方式对花楸花粉萌发率的影响

注：CK为自来水，A为室温，B为40℃水浴。

Figure 5. Effect of thawing on germination rate of S. pohuashanensis pollens after treatments

Note:CK:Rinse underwater, A:At room temperature, B:Water bath at 40℃.

下载: 全尺寸图片幻灯片

图 6 不同含水率对花楸花粉冷冻前后萌发率的影响

注：不同大写字母表示冷冻前、后不同含水率的花粉萌发率差异显著(P＜0.05)；不同小写字母表示含水率相同条件下，冷冻前后花粉萌发率差异显著(P＜0.05)。

Figure 6. Effect of moisture content on germination rate of S. pohuashanensis pollen before and after treatments

Note: Different capital letters indicate significant differences in germination rate of pollen with different water content between different freezing treatments(P < 0.05). Different lowercases indicate rates of pollen germination are markedly different before and after freezing when the water content is the same (P < 0.05).

下载: 全尺寸图片幻灯片

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