不同制样方法下6种耧斗菜花粉的形态特征

doi:10.3969/j.issn.1004-1524.20231181

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (1): 67-77.DOI: 10.3969/j.issn.1004-1524.20231181

不同制样方法下6种耧斗菜花粉的形态特征

廖珍凤¹(), 宋西娇¹, 沈梦梦¹, 肖彬¹, 周媛²^,^*()

1.浙江省农业科学院农产品质量安全危害因子与风险防控国家重点实验室,浙江杭州 310021
2.浙江省农业科学院园林植物与花卉研究所,浙江杭州 311251

收稿日期:2023-10-13 出版日期:2025-01-25 发布日期:2025-02-14
作者简介:廖珍凤(1991—),女,福建三明人,硕士,研究实习员,研究方向为电镜技术。E-mail:liaozhenfeng2@163.com
通讯作者: *周媛,E-mail:dazhouyuan19@163.com

Morphological characteristics of Aquilegia pollen grains from six species under different sample preparation methods

LIAO Zhenfeng¹(), SONG Xijiao¹, SHEN Mengmeng¹, XIAO Bin¹, ZHOU Yuan²^,^*()

1. State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
2. Institute of Garden Plants and Flowers, Zhejiang Academy of Agricultural Sciences, Hangzhou 311251, China

Received:2023-10-13 Online:2025-01-25 Published:2025-02-14

摘要/Abstract

摘要： 为了明确耧斗菜属不同品种间的花粉形态特征,并探究不同制样处理方式对花粉形态变化的影响,分别用冷冻扫描法、新鲜花粉干燥法、戊二醛固定法制备6种耧斗菜(Aquilegia)花粉样品。结果表明:在相同制样方法下,不同品种的花粉具有一定的共性和特异性;6种耧斗菜花粉均为等极的辐射对称型花粉,萌发沟为3沟型,覆盖层表面具刺状突起纹饰,说明不同品种耧斗菜之间具有高度的保守性;而极轴长度、赤道轴长度、极轴/赤道轴比值等方面存在显著差异,可作为耧斗菜品种分类鉴定的孢粉学依据。不同制样方法下,6种耧斗菜花粉大小排序基本一致,紫花耧斗菜的花粉最大,其次是无距耧斗菜,其余4种耧斗菜花粉大小差异不明显。冷冻扫描电镜法是研究耧斗菜孢粉学较理想的方法,最大程度地保持了花粉原本的超微结构特征。与冷冻扫描电镜法相比,新鲜花粉干燥法和戊二醛固定法制备的花粉极轴长度、P/E值和P×E值均存在不同程度的减小。该研究不仅成功建立了一套针对花粉超微结构观察的冷冻扫描技术体系,也为耧斗菜品种的分类鉴定提供了理论依据。

关键词: 耧斗菜, 花粉, 形态, 扫描电镜, 超微结构

Abstract:

To elucidate the pollen morphological characteristics of six Aquilegia species and to examine the impact of various sample preparation methods on alterations in pollen morphology, we prepared pollen samples using cryo-scanning electron microscopy, fresh pollen drying, and glutaraldehyde fixation techniques. The results revealed that under the same sample preparation method, different species of Aquilegia pollen exhibited certain commonness and specificity. All six species of Aquilegia pollen grains were characterized by isopolarity and radiate symmetry, with 3 germination furrow, covering layer surface ornamented with spinesis, indicating a high degree of conservatism among different species. Meanwhile, significant differences were observed in terms of polar axis length, equatorial axis length, polar axis/equatorial axis ratio, and other parameters, which could serve as palynological evidence for the classification and identification of Aquilegia species. Regardless of the sample preparation method, the size ranking of pollen grains among the six species of Aquilegia remained basically consistent. A. viridiflora var. Atropurpurear had the largest pollen grains, followed by A. ecalcarata, while the remaining four species showed no significant differences in pollen grain size. Cryo-scanning electron microscopy emerges was the preferred method for studying Aquilegia palynology due to its ability to preserve the ultrastructural characteristics of pollen grains. Compared with cryo-scanning electron microscopy, the data obtained through fresh pollen drying and glutaraldehyde fixation methods revealed varying degrees of reduction in polar axis length, P/E value, and P×E value. This research successfully established a cryo-scanning technique for ultrastructural observation of pollen grains and provided a theoretical foundation for the classification and identification of Aquilegia species in the field of palynology.

Key words: Aquilegia, pollen, morphology, scanning electron microscopy, ultrastructure

中图分类号:

S682.39

廖珍凤, 宋西娇, 沈梦梦, 肖彬, 周媛. 不同制样方法下6种耧斗菜花粉的形态特征[J]. 浙江农业学报, 2025, 37(1): 67-77.

LIAO Zhenfeng, SONG Xijiao, SHEN Mengmeng, XIAO Bin, ZHOU Yuan. Morphological characteristics of Aquilegia pollen grains from six species under different sample preparation methods[J]. Acta Agriculturae Zhejiangensis, 2025, 37(1): 67-77.

图/表 4

图1 冷冻扫描制样法得到的6种耧斗菜花粉形态的扫描电镜照片 a1,无距耧斗菜;a2,无距耧斗菜极面观;a3,无距耧斗菜赤道面观;b1,扇形耧斗菜;b2,扇形耧斗菜极面观;b3,扇形耧斗菜赤道面观;c1,紫花耧斗菜;c2,紫花耧斗菜极面观;c3,紫花耧斗菜赤道面观;d1,尖萼耧斗菜;d2,尖萼耧斗菜极面观;d3,尖萼耧斗菜赤道面观;e1,黄花耧斗菜;e2,黄花耧斗菜极面观;e3,黄花耧斗菜赤道面观;f1,黄花尖萼耧斗菜;f2,黄花尖萼耧斗菜极面观;f3,黄花尖萼耧斗菜赤道面观。a1、b1、c1、d1、e1、f1的标尺为100 μm;a2、a3、b2、b3、c2、c3、d2、d3、e2、e3、f2、f3的标尺为10 μm。下同。

Fig.1 Scanning electron microscopy images of pollen grains of six Aquilegia species obtained by cryo-scanning electron microscopy method a1, Aquilegia ecalcarata; a2, Polar view of Aquilegia ecalcarata; a3, Equatorial view of Aquilegia ecalcarata; b1, Aquilegia flabellate; b2, Polar view of Aquilegia flabellate; b3, Equatorial view of Aquilegia flabellate; c1, Aquilegia viridiflora var. atropurpurea; c2, Polar view of Aquilegia viridiflora var. atropurpurea; c3, Equatorial view of Aquilegia viridiflora var. atropurpurea; d1, Aquilegia oxysepala; d2, Polar view of Aquilegia oxysepala; d3, Equatorial view of Aquilegia oxysepala; e1, Aquilegia chrysantha; e2, Polar view of Aquilegia chrysantha; e3, Equatorial view of Aquilegia chrysantha; f1, Aquilegia oxysepala var. oxysepala f. pallidiflora; f2, Polar view of Aquilegia oxysepala var. oxysepala f. pallidiflora; f3, Equatorial view of Aquilegia oxysepala var. oxysepala f. pallidiflora. The bars of figures a1, b1, c1, d1, e1 and f1 were 100 μm, the bars of figures a2, a3, b2, b3, c2, c3, d2, d3, e2, e3, f2, f3 were 10 μm.The same as below.

图2 新鲜花粉干燥法得到的6种耧斗菜花粉形态的扫描电镜照片

Fig.2 Scanning electron microscopy images of pollen grains of six Aquilegia species obtained by fresh pollen drying method

图3 戊二醛固定法得到的6种耧斗菜花粉形态的扫描电镜照片

Fig.3 Scanning electron microscopy images of pollen grains of six Aquilegia species obtained by glutaraldehyde fixation method

表1 三种方法得到的6种耧斗菜花粉的形态特征

Table 1 Morphological characteristics of pollen grains of six Aquilegia species obtained by three methods

方法Method	品种Species	P/μm	E/μm	P/E	P×E/(μm×μm)
冷冻扫描法	无距耧斗菜A. ecalcarata	30.55±2.32 b	15.92±1.01 c	1.92±0.13 a	487.72±58.32 b
Cryo-scanning	扇形耧斗菜A. flabellate	24.27±2.13 e	17.12±0.78 b	1.42±0.16 d	414.93±34.65 c
electron microscopy	紫花耧斗菜	34.45±2.00 a	21.55±1.91 a	1.61±0.13 c	744.01±91.59 a
	A. viridiflora var. atropurpurea
	尖萼耧斗菜A. oxysepala	27.19±1.81 c	15.5±0.62c	1.76±0.13 b	421.30±32.24 c
	黄花耧斗菜A. chrysantha	25.73±1.65 d	15.82±0.89 c	1.63±0.14 c	407.02±34.90 c
	黄花尖萼耧斗菜	27.20±1.20 c	14.74±0.91 d	1.85±0.13 a	401.01±32.36 c
	A. oxysepala var. oxysepala f. pallidiflora
新鲜花粉干燥法	无距耧斗菜A. ecalcarata	25.04±1.13 a	15.37±1.80 bc	1.65±0.18 a	385.17±49.36 a
Fresh pollen drying	扇形耧斗菜A. flabellate	24.45±1.44a	15.19±1.01 bc	1.62±0.13 ab	371.66±35.68 ab
	紫花耧斗菜	24.69±1.80 a	15.75±1.12 b	1.57±0.14 b	389.24±44.32 a
	A. viridiflora var. atropurpurea
	尖萼耧斗菜A. oxysepala	23.60±1.16 b	14.95±0.78 c	1.58±0.09 ab	353.04±30.05 b
	黄花耧斗菜A. chrysantha	22.36±1.02 c	16.69±1.15 a	1.34±0.09 c	373.52±35.08 ab
	黄花尖萼耧斗菜	24.60±0.95 a	15.05±0.71 c	1.64±0.10 ab	370.18±21.40 ab
	A. oxysepala var. oxysepala f. pallidiflora
戊二醛固定法	无距耧斗菜A. ecalcarata	23.91±1.24 a	15.13±0.86 d	1.58±0.10 a	362.19±31.95 b
Glutaraldehyde	扇形耧斗菜A. flabellate	19.06±1.49 c	15.86±1.50 cd	1.21±0.13 c	302.69±38.76 cd
fixation	紫花耧斗菜	23.20±1.09 b	18.10±2.59 a	1.32±0.27 b	419.39±61.36 a
	A. viridiflora var. atropurpurea
	尖萼耧斗菜A. oxysepala	17.85±1.03 e	16.34±1.02 bc	1.09±0.07 d	292.19±30.24 cd
	黄花耧斗菜A. chrysantha	18.33±0.71 de	16.86±1.02 b	1.09±0.05 d	309.52±27.30 c
	黄花尖萼耧斗菜	18.67±1.02 cd	15.14±1.25 d	1.24±0.09 c	283.20±33.77 d
	A. oxysepala var. oxysepala f. pallidiflora

表1 三种方法得到的6种耧斗菜花粉的形态特征

Table 1 Morphological characteristics of pollen grains of six Aquilegia species obtained by three methods

方法Method	品种Species	P/μm	E/μm	P/E	P×E/(μm×μm)
冷冻扫描法	无距耧斗菜A. ecalcarata	30.55±2.32 b	15.92±1.01 c	1.92±0.13 a	487.72±58.32 b
Cryo-scanning	扇形耧斗菜A. flabellate	24.27±2.13 e	17.12±0.78 b	1.42±0.16 d	414.93±34.65 c
electron microscopy	紫花耧斗菜	34.45±2.00 a	21.55±1.91 a	1.61±0.13 c	744.01±91.59 a
	A. viridiflora var. atropurpurea
	尖萼耧斗菜A. oxysepala	27.19±1.81 c	15.5±0.62c	1.76±0.13 b	421.30±32.24 c
	黄花耧斗菜A. chrysantha	25.73±1.65 d	15.82±0.89 c	1.63±0.14 c	407.02±34.90 c
	黄花尖萼耧斗菜	27.20±1.20 c	14.74±0.91 d	1.85±0.13 a	401.01±32.36 c
	A. oxysepala var. oxysepala f. pallidiflora
新鲜花粉干燥法	无距耧斗菜A. ecalcarata	25.04±1.13 a	15.37±1.80 bc	1.65±0.18 a	385.17±49.36 a
Fresh pollen drying	扇形耧斗菜A. flabellate	24.45±1.44a	15.19±1.01 bc	1.62±0.13 ab	371.66±35.68 ab
	紫花耧斗菜	24.69±1.80 a	15.75±1.12 b	1.57±0.14 b	389.24±44.32 a
	A. viridiflora var. atropurpurea
	尖萼耧斗菜A. oxysepala	23.60±1.16 b	14.95±0.78 c	1.58±0.09 ab	353.04±30.05 b
	黄花耧斗菜A. chrysantha	22.36±1.02 c	16.69±1.15 a	1.34±0.09 c	373.52±35.08 ab
	黄花尖萼耧斗菜	24.60±0.95 a	15.05±0.71 c	1.64±0.10 ab	370.18±21.40 ab
	A. oxysepala var. oxysepala f. pallidiflora
戊二醛固定法	无距耧斗菜A. ecalcarata	23.91±1.24 a	15.13±0.86 d	1.58±0.10 a	362.19±31.95 b
Glutaraldehyde	扇形耧斗菜A. flabellate	19.06±1.49 c	15.86±1.50 cd	1.21±0.13 c	302.69±38.76 cd
fixation	紫花耧斗菜	23.20±1.09 b	18.10±2.59 a	1.32±0.27 b	419.39±61.36 a
	A. viridiflora var. atropurpurea
	尖萼耧斗菜A. oxysepala	17.85±1.03 e	16.34±1.02 bc	1.09±0.07 d	292.19±30.24 cd
	黄花耧斗菜A. chrysantha	18.33±0.71 de	16.86±1.02 b	1.09±0.05 d	309.52±27.30 c
	黄花尖萼耧斗菜	18.67±1.02 cd	15.14±1.25 d	1.24±0.09 c	283.20±33.77 d
	A. oxysepala var. oxysepala f. pallidiflora

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不同制样方法下6种耧斗菜花粉的形态特征

Morphological characteristics of Aquilegia pollen grains from six species under different sample preparation methods

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