• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

西番莲种质资源花粉活力鉴定方法探究

Determination of Viability of Passion Fruit Pollens

    摘要
  • 摘要:
      目的  明确西番莲属植物花粉最佳离体培养基、离体培养最适观测时间、花粉的最适染色法和花粉贮藏条件，快速精准鉴定西番莲属种质资源的花粉活力，为其种质资源利用与新品种选育提供科学依据。
      方法  以6份西番莲属种质资源的花粉为试验材料，设计正交试验，探究不同浓度的蔗糖、H3BO3、Ca(NO3)2和聚乙二醇-4000（PEG-4000）对花粉离体萌发率的影响；利用最佳花粉离体培养基，筛选花粉离体培养最适观测时间；采用I2-KI染色法、TTC染色法与亚历山大染色法，探究最适的西番莲花粉染色法；通过不同贮藏温度与时间处理，筛选花粉的最佳贮藏条件。
      结果  正交试验筛选出最佳培养基配方为100 g·L−1 蔗糖+0.04 g·L−1 H3BO3+0.01 g·L−1 Ca(NO3)2+150 g·L−1 PEG-4000+200 mg·L−1 MgSO4·7H2O+100 mg·L−1 KNO3，pH值5.5；花粉离体培养1 h为最适观测时间；TTC染色法染色效果好且结果与离体萌发率无显著差异，可有效检测西番莲花粉活力；花粉贮藏24 h以内，25 ℃与4 ℃的贮藏温度下花粉活力分别为30.48%、26.69%，花粉贮藏时间在1～7 d，4 ℃下花粉活力仍保持在26.54%～27.05%。
      结论  筛选出西番莲属植物花粉离体萌发的最佳培养基组合、离体培养最适观测时间、花粉最适染色法与花粉贮藏最佳条件，为西番莲属植物花粉活力的快速筛选和品种改良提供理论依据。

     

    Abstract:
      Objective   Method for determining the viability of passion fruit pollens was explored to facilitate the utilization of existing germplasms and breeding new varieties.
      Methods  Medium and conditions for in vitro culture of Passiflora pollens was optimized. Appropriate staining methods to determine and storage conditions to maximize the pollen viability were investigated. Contents of sucrose, H3BO3, Ca(NO3)2, and polyethylene glycol-4000 (PEG-4000) of the medium were optimized in an orthogonal experiment on pollens of 6 germplasms. Pollen germination on the finalized medium for varied culture times was observed. The staining of I2-KI, TTC, and Alexander methods and storage for different durations at different temperatures were compared.
      Results   The optimum medium was formulated with 100 g·L−1 sucrose, 0.02 g·L−1 H3BO3, 0.04 g·L−1 Ca(NO3)2, 150 g·L−1 PEG-4000, 200 g·L−1 MgSO4·7H2O, and 100 g·L−1 KNO3, then adjusted to pH 5.5. The pollen germination was examined in 1 h of the in vitro culture. TTC staining showed satisfactory effect with no significant deviations from the in vitro observation indicating it an applicable indicator for pollen viability. In 24 h of storage at 25 ℃ the pollen viability was 30.48%; and at 4 ℃, it was 26.69% and remained 26.54%–29.05% for 7 d.
      Conclusion  The medium formulation and in vitro culture time as well as the staining method and storage conditions of passion fruit pollens were determined making a rapid and reliable procedure to maximize the pollen germination available.

     

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