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西番莲种质资源花粉活力鉴定方法探究

项艳艳 黄伟杰 赖江龙 汤礼梅 林挺兴 李亮 魏秀清 许家辉

项艳艳,黄伟杰,赖江龙,等. 西番莲种质资源花粉活力鉴定方法探究 [J]. 福建农业学报,2024,39(3):302−309 doi: 10.19303/j.issn.1008-0384.2024.03.007
引用本文: 项艳艳,黄伟杰,赖江龙,等. 西番莲种质资源花粉活力鉴定方法探究 [J]. 福建农业学报,2024,39(3):302−309 doi: 10.19303/j.issn.1008-0384.2024.03.007
XIANG Y Y, HUANG W J, LAI J L, et al. Determination of Viability of Passion Fruit Pollens [J]. Fujian Journal of Agricultural Sciences,2024,39(3):302−309 doi: 10.19303/j.issn.1008-0384.2024.03.007
Citation: XIANG Y Y, HUANG W J, LAI J L, et al. Determination of Viability of Passion Fruit Pollens [J]. Fujian Journal of Agricultural Sciences,2024,39(3):302−309 doi: 10.19303/j.issn.1008-0384.2024.03.007

西番莲种质资源花粉活力鉴定方法探究

doi: 10.19303/j.issn.1008-0384.2024.03.007
基金项目: 福建省高质量发展超越“5511”协同创新工程项目(XTCXGC2021006);福建省种业创新与产业化工程项目(zycxny2021010-1);福建省农业科学院科技创新团队建设项目(CXTD2021008-2);福建省科技计划公益类专项(2021R1028002)
详细信息
    作者简介:

    项艳艳(1997 — ),女,硕士研究生,主要从事果树育种研究,E-mail:2711768629@qq.com

    通讯作者:

    许家辉(1971 — ),男,研究员,主要从事果树育种研究,E-mail:xjhui577@163.com

  • 中图分类号: S667.9

Determination of Viability of Passion Fruit Pollens

  • 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。
  • 图  1  花粉离体培养时间对花粉萌发率的影响

    A:不同时期花粉萌发率;B:a、b、c、d分别代表1、2、3、4 h等花粉萌发观测结果。

    Figure  1.  Effect of culture time on pollen germination rate

    A: pollen germination rate at different times. B: a, b, c, and d represent observed results on pollen germination at 1, 2, 3, and 4 h, respectively.

    图  2  不同方法检测西番莲花粉活力镜检效果。

    A:蓝冠;B:白花;C:香蜜;D:雅蜜;E:钦蜜9号;F:台农。

    Figure  2.  Microscopic examinations on passion fruit pollen viability detected by different methods

    A: Blue Crown; B: White Flowers; C: Xiangmi; D: Yami; E: Qinmi 9; F: Tainong.

    图  3  钦蜜9号不同贮藏温度下花粉萌发率的变化

    不同小写字母表示在0.05水平差异显著。

    Figure  3.  Germination rates of Qinmi 9 pollens stored under different temperatures

    Data with different lowercase letters indicate significant differences at 0.05 level.

    表  1  正交因素及水平

    Table  1.   Orthogonal factors and levers

    水平
    Level
    A.蔗糖
    Sucrose /(g·L−1
    B.H3BO3/
    (g·L−1
    C.Ca(NO3)2/
    (g·L−1
    D.PEG-4000/
    (g·L−1
    1 0 0 0 0
    2 50 0.01 0.01 50
    3 100 0.02 0.02 100
    4 150 0.03 0.03 150
    5 200 0.04 0.04 200
    下载: 导出CSV

    表  2  钦蜜9号花粉离体培养基筛选的L25(54)正交试验设计

    Table  2.   L25(54) orthogonal experimental design for culture medium of passion fruit pollens

    编号
    No.
    因素
    Factors
    萌发率
    Germination rate/%
    花粉管长度
    Pollen tube length/μm
    A B C D
    1 1 1 1 1 0 j
    2 1 2 2 2 13.81±1.88 i 158.5±4.03 ab
    3 1 3 3 3 48.25±3.12 g 152.92±2.73 ab
    4 1 4 4 4 64.50±1.68 f 126.24±5.96 e
    5 1 5 5 5 75.53±3.07 bcde 108.29±10.29 f
    6 2 1 2 3 69.11±11.13 def 147.43±4.46 bc
    7 2 2 3 4 79.85±1.03 abc 132.59±8.86 de
    8 2 3 4 5 63.65±4.68 f 109.89±5.34 f
    9 2 4 5 1 12.99±2.23 i 150.65±13.17 b
    10 2 5 1 2 72.26±1.83 cdef 167.15±4.46 a
    11 3 1 3 5 45.95±0.61 g 143.76±7.77 bcd
    12 3 2 4 1 0 j
    13 3 3 5 2 76.99±5.37 abcde 126.75±5.77 e
    14 3 4 1 3 67.23±7.08 f 134.33±12.72 cde
    15 3 5 2 4 85.56±1.82 a 157.36±4.01 ab
    16 4 1 4 2 6.5±3.04 ij
    17 4 2 5 3 28.73±17.87 h 124.43±12.31 e
    18 4 3 1 4 83.08±3.5 ab 120.2±3.76 ef
    19 4 4 2 5 0.27±0.47 j
    20 4 5 3 1 0.18±0.17 j
    21 5 1 5 4 0.54±0.47 j
    22 5 2 1 5 1.03±0.4 j
    23 5 3 2 1 0 j
    24 5 4 3 2 1.66±2.88 j
    25 5 5 4 3 25.94±3.2 h 158.54±9.45 ab
    k1 40.42 24.42 44.72 2.63
    k2 59.57 24.68 33.75 34.24
    k3 41.64 54.40 34.53 47.85
    k4 23.75 29.33 32.12 62.71
    k5 5.83 51.89 38.96 37.29
    R 53.74 29.98 12.60 60.08
    数据表示为平均值±标准差。同列数据用不同小写字母标识表示在0.05水平差异显著。
    Data are represented as mean ± standard deviation; those with different lowercase letters on same column indicate significant differences at P<0.05.
    下载: 导出CSV

    表  3  钦蜜9号离体萌发率方差分析结果

    Table  3.   Variance analysis on in vitro germination rate of Qinmi 9

    变异来源
    Source of variation
    III 类平方和
    Class III sum of squares
    自由度
    Degree of freedom
    均方
    Mean square
    F 显著性
    Significance(P
    蔗糖
    Sources
    9986.344 4 2496.586 8.365 0.006**
    H3BO3 4465.681 4 1116.420 3.740 0.53
    Ca(NO3)2 505.625 4 126.406 0.424 0.788
    PEG-4000 9836.241 4 2459.06 8.239 0.006**
    误差
    Error
    2387.782 8 298.473
    **表示在0.01水平差异极显著。
    **indicates extremely significant difference at 0.01 level.
    下载: 导出CSV

    表  4  不同方法检测的西番莲花粉活力

    Table  4.   Passion fruit pollen viability detected by different methods

    种质
    Germplasm
    染色率
    Staining rate/%
    离体萌发率
    In vitro
    germination
    rate/%
    I2-IK TTC 亚历山大
    Alexandria
    台农
    Tainong
    83.83±3.76 a 67.69±5.29 a 81.19±2.78 a 74.13±2.89 a
    钦蜜9号
    Qinmi 9
    81.79±7.75 b 70.29±4.47 b 97.95±1.14 a 79.76±7.80 b
    雅蜜
    Yami
    87.85±3.22 b 69.97±5.92 c 95.42±2.16 a 63.71±2.84 c
    白花
    White Flowers
    84.32±7.73 b 77.41±6.33bc 97.22±1.89 a 63.7±1.00 c
    蓝冠
    Blue Crown
    80.41±3.26 a 64.85±4.41 b 78.50±5.28 a 60.79±4.61 b
    香蜜
    Xiangmi
    84.32±7.70 b 63.78±1.11 c 98.75±1.88 a 71.34±6.97 c
    数据表示为平均值±标准差,同行数据后不同小写字母表示在0.05水平差异显著。
    Data are mean±SD; those with different lowercase letters on same row indicate significant difference at 0.05 level.
    下载: 导出CSV
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  • 收稿日期:  2024-02-06
  • 修回日期:  2024-03-01
  • 网络出版日期:  2024-03-28
  • 刊出日期:  2024-03-28

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