10个菊花品种花粉生活力测定和贮藏方法研究

李亚军; 黄河; 牛雅静; 李霞

doi:10.19303/j.issn.1008-0384.2020.03.008

10个菊花品种花粉生活力测定和贮藏方法研究

doi: 10.19303/j.issn.1008-0384.2020.03.008

李亚军^1,,
黄河¹,
牛雅静²,
李霞^3, ,

1.
北京林业大学园林学院，北京　100083
2.
北京植物园，北京　100093
3.
北京城市学院城市建设学部，北京　100083

基金项目: 大北农青年教师科研基金项目（15ZK008）

详细信息

作者简介:
李亚军（1996−），女，硕士，研究方向：花卉分子生物学（E-mail：yajunli2018@bjfu.edu.cn）

通讯作者:
李霞（1985−），女，博士，讲师，研究方向：园林植物遗传育种（E-mail：lixia5966@163.com）

中图分类号: S 682.1+1
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出版历程
- 收稿日期: 2019-12-04
- 修回日期: 2020-02-17
- 刊出日期: 2020-03-01

Viability of Stored Chrysanthemum morifolium Pollens

LI Ya-jun^1
,,
HUANG He¹,
NIU Ya-jing²,
LI Xia^{3
, ,}

1.
School of Landscape Architecture, Beijing Forestry University, Beijing　100083, China
2.
Beijing Botanical Garden, Beijing　100093, China
3.
Urban Construction School, Beijing City University, Beijing　100083, China

摘要

摘要: 目的筛选菊花花粉离体培养生活力测定的最佳培养基配方，探究花粉贮藏的最适温度和耐贮藏时间，并挑选出较耐贮藏的菊花品种，为菊花新品种选育和种质资源保存提供参考。方法以10个菊花品种盛花期花粉为试验材料，通过离体萌发测定法筛选菊花花粉萌发最适培养基，以该培养基测定花粉在4℃、−20℃和−80℃ 3种温度下进行60 d或120 d贮藏周期的生活力变化趋势观测，每隔15 d取样检测1次；对贮藏120 d的花粉进行授粉，统计结实率。结果培养基为 ME₃+200 g·L⁻¹PEG 4000+6 g·L⁻¹琼脂的处理，10种花粉都能萌发，且萌发率较其他培养基有显著差异；花粉贮藏60 d后，所有花粉生活力均下降，其中‘5’、‘52’、‘64’和‘85’4个品种花粉生活力下降较缓慢；4种花粉贮藏120 d后，在−80℃贮藏的花粉生活力较高，花粉授粉后均能够结实，且品种‘5’花粉授粉结实率最高，平均每个授粉花序结实8粒。结论花粉萌发培养基、植物品种基因型、贮藏温度和贮藏时间都会影响菊花花粉生活力。本文研究表明，菊花花粉最适贮藏温度为−80℃，品种‘5’花粉可贮藏120 d保持较高的授粉结实率，在杂交育种中可优先考虑。
- 菊花 /
- 花粉生活力 /
- 贮藏 /
- 授粉
Abstract: Objective Viability of stored Chrysanthemum× morifolium pollens were determined for germplasm preservation. Method Pollen viability of 10 chrysanthemum cultivars were determined by an in vitro germination method for medium selection. Viability of pollens stored in the selected medium at 4℃, −20℃ or −80℃ for 60 d and 120 d was monitored every 15 d. After a 120 d storage, hybridization pollination with the pollens was performed for fructification evaluation. Result The medium for optimum pollen germination was ME₃ with the additions of 200 g·L⁻¹ PEG 4 000 and 6 g·L⁻¹ agar. All pollens stored in the chosen medium germinated with a significantly superior rate. The pollen viability decreased after 60d storage. Among the 10 chrysanthemum, Variety ‘5’, ‘52’, ‘64’, and ‘85’ had slower declining rates on the pollen viability but higher viability after the 120 d storage at −80℃ with full fructification upon pollination than the others. Of the 4 varieties, Variety ‘5’ had the highest pollen fructifying rate of 8 seeds per inflorescence on average. Conclusion The medium for pollen germination and genotype of cultivar as well as the temperature and duration of storage could all affect the viability of the chrysanthemum pollens. On the optimally formulated medium at −80℃ for 120 d, the pollens from Variety ‘5’ maintained a high viability with full fructification at pollination for satisfactory chrysanthemum cross breeding.
- Chrysanthemum morifolium /
- pollen viability /
- pollen storage /
- pollination

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图 1 10个菊花品种头状花序

Figure 1. Inflorescence of 10 cultivars of C. morifolium

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表 1 10个菊花品种头状花序相关性状

Table 1. Inflorescence traits of 10 cultivars of C. morifolium

品种 Cultivar	花序性状 Inflorescence traits
品种 Cultivar	花色 Color	瓣型 Petal shape	重瓣性 Double trait/ whorl	始花期 First flowering/(M-D)	盛花期 Peak flowering/(M-D)	末花期 Last flowering/(M-D)
‘5’	紫 Purple	平瓣 Flat	1～2	10-25	11-07	11-16
‘16’	红 Red	平瓣 Flat	3～5	10-02	11-05	11-22
‘22’	复 Multiple	平瓣 Flat	1～2	10-02	11-07	11-15
‘30’	红 Red	管瓣 Tubular	1～2	10-26	11-01	11-23
‘45’	黄 Yellow	平瓣 Flat	2～3	10-02	11-05	11-15
‘52’	复 Multiple	匙瓣 Spoon	1～2	10-22	11-01	11-17
‘64’	红 Red	平瓣 Flat	1～2	10-29	11-12	11-18
‘85’	粉 Pink	匙瓣 Spoon	3～5	11-02	11-19	11-25
‘132’	粉 Pink	平瓣 Flat	2～3	10-22	11-15	11-02
‘317’	紫 Purple	匙瓣 Spoon	1	10-15	10-31	11-12

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表 2 花粉在7种培养基中的萌发率

Table 2. Germination rates of pollens stored on 7 different media

编号 No.	培养基成分 Media composition					花粉萌发率 Pollen germination rate/%
编号 No.	蔗糖 Sucrose/ （g·L⁻¹）	硼酸 Boric acid/ （g·L⁻¹）	ME₃	PEG4000/ （g·L⁻¹）	琼脂 Agar/ （g·L⁻¹）	‘5’	‘16’	‘22’	‘30’	‘45’	‘52’	‘64’	‘85’	‘132’	‘317’
1	50	0.6	-	-	-	0	0	0	0	0	0	0	0	0	0
2	100	0.6	-	-	-	0	0	0	0	0	0	0	0	0	0
3	150	0.6	-	-	-	3.30±b	3.21±0.27 a	2.56±0.13 b	2.01±0.20 b	0.86±0.05 d	2.67±0.17 b	3.01±0.36 b	2.33±0.21 c	2.12c	2.67±0.15 b
4	200	0.6	-	-	-	4.12b	4.22±0.28 a	3.53±0.15 a	2.67±0.22 a	1.14±0.13 b	3.56±0.16 a	3.91 a	3.43 a	2.45 b	3.24±0.22 a
5	-	-	√		6	0	0	0	0	0	0	0	0	0	0
6	200	-	√		6	3.98 b	4.23±0.22 a	3.45±0.15 a	2.67 a	1.09 c	3.44±0.12 a	3.86 a	2.98 b	1.98±0.13 d	3.32±0.20 a
7	-	-	√	200	6	5.52 a	4.13±0.15 a	3.63±0.18 a	2.72 a	1.24 a	3.67±0.13 a	3.83 a	3.51 a	2.67±0.18 a	3.32±0.19 a
注：表内同列数据后不同小写字母表示同一品种在不同培养基上的花粉萌发率差异显著（P＜0.05）。 Note: Different letters on same column indicate significant differences in pollen germination rates of same cultivar on different media（P＜0.05）.

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表 3 10个菊花品种花粉原始生活力和贮藏60 d后生活力比较

Table 3. Initial and after 60d storage viability of pollens from 10 cultivars of C. morifolium

品种 Cultivar	原始萌发率 Pollen germination rate before storage/%			贮藏60 d萌发率 Pollen germination rate after storage for 60 d/%			萌发率下降值 Decline value
品种 Cultivar	4℃	−20℃	−80℃	4℃	−20℃	−80℃	4℃	−20℃	−80℃
‘5’	5.62±0.97 a	5.69±0.84 a	5.73±0.84 a	3.51±0.23 a	3.61±0.38 a	3.86±0.06 a	2.11 c	2.08 b	1.87 b
‘16’	4.13±0.24 b	4.11±0.22 b	4.15±0.29 b	1.70±0.27 b	2.01±0.10 b	2.12±0.31 b	2.43 b	2.10 b	2.03 b
‘22’	3.30±0.23 cd	3.33±0.07 cd	3.30±0.11 cde	1.66±0.11 b	1.83±0.14 bc	1.96±0.06 b	1.64 e	1.50 c	1.34 d
‘30’	2.70±0.23 d	2.77±0.09 de	2.80±0.13 def	0.79±0.04 def	0.83±0.13 ef	1.02±0.17 de	1.91 cd	1.94 b	1.78 b
‘45’	1.20±0.19 e	1.24±0.27 f	1.22±0.29 f	0.69±0.47 ef	1.01±0.19 de	1.13±0.12 de	0.51 f	0.23 e	0.09 f
‘52’	3.67±0.20 bc	3.66±0.20 bc	3.68±0.19 bc	1.52±0.17 bc	1.63±0.17 c	1.82±0.14 bc	2.15 c	2.03 b	1.86 b
‘64’	2.67±0.37 d	2.62±0.42 e	2.59±0.37 f	1.01± 0.19 de	1.02±0.17 de	1.23±0.31 d	1.66 de	1.60 c	1.36 c
‘85’	3.20±0.22 cd	3.16±0.28 cde	3.20±0.30 cdef	1.79±0.27 b	1.98±0.17 b	2.18±0.52 b	1.41 e	1.18 d	1.02 e
‘132’	2.67±0.39 d	2.70±0.36 de	2.68±0.39 ef	1.21±0.12 cd	1.23±0.11 d	1.48±0.34 cd	1.46 e	1.47 c	1.20 de
‘317’	3.30±0.14 cd	3.37±0.26 cd	3.41±0.24 cd	0.45±0.31 f	0.56±0.17 f	0.73±0.26 e	2.85 a	2.81 a	2.68 a
注：表内同列数据后不同小写字母表示相同贮藏温度下不同品种之间花粉萌发率差异显著（P＜0.05）。 Note: Different letters on same column indicate significant difference in pollen germination rate /decline of different cultivars at same storage temperature（P＜0.05）.

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表 4 不同贮藏温度及天数对4个菊花品种花粉生活力的影响

Table 4. Effects of storage temperature and duration on pollen viability of 4 cultivars of C. morifolium

品种 Cultivar	贮藏温度 Temperature	花粉萌发率 Pollen germination rate/%
品种 Cultivar	贮藏温度 Temperature	1 d	15 d	30 d	45 d	60 d	75 d	90 d	105 d	120 d
‘5’	4℃	5.62±0.97 Ca	4.86±1.00 Cb	4.72±0.14 Cb	3.51±0.14 Cc	3.51±0.23 Cc	2.59±0.56 Cd	2.33±0.13 Cd	1.98±0.29 Ce	1.21±0.16 Cf
	−20℃	5.69±0.84 Ba	5.02±0.39 Bb	5.02±0.19 Bb	3.89±0.19 Bc	3.61±0.38 Bcd	3.08±0.17 Bd	2.89±0.25 Be	2.15±0.17 Bf	1.52±0.12 Bg
	−80℃	5.73±0.84 Aa	5.43±0.41 Aab	5.21±0.2 Ab	4.35±0.34 Ac	3.86±0.06 Ad	3.41±0.26 Ae	3.14±0.2 Aef	2.45±0.26 Af	1.63±0.25 Ag
‘52’	4℃	3.67±0.20 ABa	3.09±0.38 Cb	2.24±0.17 Cc	1.73±0.31 Cd	1.52±0.17 Be	1.29±0.08 Be	1.02±0.13 Cf	0.92±0.17 Cg	0.79±0.27 Ch
	−20℃	3.66±0.20 Ba	3.26±0.30 Bb	2.51±0.11 Bc	2.09±0.18 Bd	1.63±0.17 Be	1.35±0.23 Bf	1.23±0.22 Bf	1.19±0.20 Bf	0.98±0.31 Bg
	−80℃	3.68±0.19 Aa	3.49±0.22 Ab	2.91±0.50 Ac	2.21±0.09 Ad	1.82±0.14 Ae	1.63±0.04 Ae	1.45±0.21 Af	1.41±0.12 Ag	1.33±0.18 Ah
‘64’	4℃	2.67±0.37 Aa	1.34±0.24 Cb	1.08±0.26 Cc	1.01±0.46 Cd	1.01±0.19 Ce	0.90±0.21 Ce	0.85±0.23 Cf	0.78±0.26 Cg	0.57±0.11 Ch
	−20℃	2.62±0.42 Ba	1.79±0.19 Bb	1.25±0.16 Bc	1.17±0.17 Bd	1.02±0.17 Be	1.03±0.10 Bf	1.02±0.27 Bf	1.02±0.16 Bf	0.69±0.16 Bg
	−80℃	2.59±0.37 Ca	2.12±0.19 Ab	1.58±0.12 Ac	1.45±0.19 Ad	1.23±0.31 Ae	1.22±0.14 Ae	1.19±0.08 Af	1.16±0.19 Ag	0.89±0.29 Ah
‘85’	4℃	3.21±0.22 Aa	2.31±0.69 Cb	2.19±0.16 Cb	1.84±0.30 Cc	1.79±0.27 Cc	1.71±0.47 Cc	1.70±0.76Cc	1.21±0.18 Cd	1.17±0.14 Cd
	−20℃	3.16±0.28 Ba	2.61±0.18 Bb	2.46±0.26 Bb	2.01±0.19 Bc	1.98±0.17 Bc	1.83±0.26 Bc	1.78±0.59 Bc	1.43±0.18 Bd	1.21±0.11 Bd
	−80℃	3.20±0.30 Aa	2.93±0.23 Aab	2.72±0.56 Ab	2.38±0.19 Ac	2.18±0.52 Acd	1.93±0.33 Ad	1.91±0.43 Ad	1.62±0.14 Ae	1.35±0.30 Ae
注：不同大写字母表示相同贮藏时间内不同贮藏温度下的花粉萌发率差异显著（P＜0.05）；不同小写字母表示相同贮藏温度下不同贮藏时间内的花粉萌发率差异显著（P＜0.05）。 Note: Different capital letters indicate significant difference in pollen germination rate at different storage temperatures for same storage duration（P＜0.05）; different lowercase letters indicate significant difference in pollen germination rate stored at same temperature for different durations（P＜0.05）.

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表 5 当季花粉做父本的杂交结果

Table 5. Hybridization pollination using fresh pollens as male parents

杂交组合 Hybrid combination	父本 Male	母本 Female	授粉花序数 Number of pollination inflorescences/个	种子数 Number of seeds/粒	平均每个花序种子数 Average number of seeds per inflorescence/粒
1	‘5’	‘45’	9	90	10a
2	‘52’	‘45’	7	49	7ab
3	‘64’	‘45’	8	32	4bc
4	‘85’	‘45’	10	30	3c
注：不同小写字母表示不同杂交组合间授粉结实率差异显著（P＜0.05）。 Note: Different letters indicate significant differences in fructification between cross combinations（P＜0.05）.

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表 6 120 d贮藏周期花粉做父本的杂交结果

Table 6. Hybridization pollination using pollens stored for 120 d as male parents

花粉 Pollen	贮藏温度 Storage temperature /℃	母本 Female	授粉花序数/个 Number of pollination inflorescences	种子数 Number of seeds/粒	平均每个花序种子数 Average number of seeds per inflorescence/粒
‘5’	4	‘45’	6	0	0
	−20		8	16	2b
	−80		7	42	6a
‘52’	4	‘45’	6	0	0
	−20		7	0	0
	−80		9	18	2b
‘64’	4	‘45’	8	0	0
	−20		5	0	0
	−80		8	16	2b
‘85’	4	‘45’	7	0	0
	−20		6	0	0
	−80		6	6	1b
注：不同小写字母表示不同杂交组合间授粉结实率差异显著（P＜0.05）。 Note: Different letters indicate significant differences in fructification between cross combinations（P＜0.05）.

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