文心兰花粉离体萌发培养基的筛选及萌发率的测定

罗远华; 陈燕; 方能炎; 林榕燕; 钟淮钦; 黄敏玲

doi:10.19303/j.issn.1008-0384.2023.11.005

文心兰花粉离体萌发培养基的筛选及萌发率的测定

doi: 10.19303/j.issn.1008-0384.2023.11.005

福建省农业科学院作物研究所/福建省农业科学院花卉研究中心/　福建省特色花卉工程技术研究中心，福建　福州　350013

基金项目: 福建省农业科学院对外合作项目（DWHZ-2022-09）；福建省农业科学院科技创新团队建设项目（CXTD2021010-2）；福建省农业高质量发展超越“5511”协同创新工程项目（XTCXGC2021016）

详细信息

作者简介:
罗远华（1981 —），男，硕士，助理研究员，主要从事花卉育种与栽培研究，E-mail：luoyh426@163.com

通讯作者:
黄敏玲（1960 —），女，研究员，主要从事花卉育种与生物技术研究，E-mail：huangml168@163.com

中图分类号: S682.31
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出版历程
- 收稿日期: 2023-08-21
- 修回日期: 2023-10-13
- 网络出版日期: 2023-12-21
- 刊出日期: 2023-11-28

Optimized Medium and Culture Conditions for Germination of Oncidium Pollens

Crop Research Institute/Flower Research Center/Fujian Engineering Research Center for Characteristic Floriculture, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian　350013, China

摘要

摘要: 目的研究不同培养基组分、培养温度及培养时间对文心兰花粉离体萌发的影响，以期筛选出文心兰花粉离体萌发的适宜培养基及培养条件，为文心兰种质资源创新利用提供科学依据。方法以盛花期当天开放的花朵为试验材料，采用液体培养基，比较蔗糖、H₃BO₃、Ca(NO₃) ·4H₂O、MgSO₄·7H₂O及KNO₃对文心兰花粉离体萌发的影响，正交试验筛选文心兰花粉离体萌发培养基的最佳组合及培养条件；在此基础上，测定收集保存的68份文心兰种质资源的花粉离体萌发率。结果一定质量浓度的蔗糖、H₃BO₃、Ca(NO₃) ·4H₂O、MgSO₄·7H₂O及KNO₃对文心兰花粉的离体萌发具有一定的促进作用，各因素的最佳作用效果从高到低依次为H₃BO₃、Ca(NO₃)·4H₂O、蔗糖、MgSO₄·7H₂O及KNO₃。正交试验表明适宜的液体培养基组合为100 g·L⁻¹蔗糖+10 mg·L⁻¹H₃BO₃+50 mg·L⁻¹Ca(NO₃)·4H₂O+20 mg·L⁻¹MgSO₄·7H₂O+30 mg·L⁻¹KNO₃，各成分的质量浓度与单因素试验的结果一致；培养条件为25 ℃、黑暗培养48 h。文心兰花粉离体萌发测定结果表明，60.29%的种质资源的花粉能离体萌发，萌发率普遍较低，为0.74%～49.65%。结论筛选出文心兰花粉离体萌发适宜的液体培养基组合及培养条件，不同文心兰种质资源的花粉离体萌发率存在明显差异，本研究结果可为后续文心兰种质资源的创新利用及他人研究等提供参考。
- 文心兰 /
- 花粉 /
- 离体萌发 /
- 液体培养基 /
- 萌发率
Abstract: Objective Medium formulation and culture conditions were optimized for in vitro pollen germination of Oncidium spp. Methods On a liquid culture medium, in vitro germination rates of the pollens collected from the flowers of various Oncidium germplasms on the day of full bloom were determined. Based on the effects of sucrose, H₃BO₃, Ca(NO3)·4H2O, MgSO4·7H2O, and KNO3 on the germination rate, an orthogonal experiment was applied to optimize the medium formulation and culture conditions. Subsequently, on the optimized medium under selected conditions, in vitro pollen germination rates of 68 collected Oncidium germplasms were determined. Results The degrees of importance of the medium ingredients to the pollen germination were found to be H₃BO₃＞Ca(NO3)·4H2O＞sucrose＞MgSO4·7H2O＞KNO3. The optimized formulation consisted of 100 g·L⁻¹ of sucrose, 10 mg·L⁻¹ of H₃BO₃, 50 mg·L⁻¹ of Ca(NO3)·4H2O, 20 mg·L⁻¹ of MgSO4·7H2O, and 30 mg·L⁻¹ of KNO3. The pollen culture was conducted at 25 ℃ for 48h in darkness to deliver a generally low germination rate of 0.74% to 49.65% on 60.29% of the Oncidium germplasms. Conclusion The liquid medium and culture conditions for an in vitro pollen germination of Oncidium were optimized. The germination rates differed significantly among the species.
- Oncidium spp. /
- pollen /
- in vitro germination /
- liquid medium /
- germination rate

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图 1 不同培养基成分对香水文心花粉离体萌发的影响

不同小写字母表示不同质量浓度间差异显著（P＜0.05）。图2同。

Figure 1. Effect of medium ingredients on in vitro pollen germination of Onc. Sharry Baby Sweet Fragrance

Data with different lowercase letters indicate significant differences between different mass concentrations at P＜0.05. Same for Fig.2.

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图 2 不同培养条件对香水文心花粉离体萌发的影响

Figure 2. Effect of culture conditions on in vitro pollen germination of Onc. Sharry Baby Sweet Fragrance

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表 1 单因素间香水文心花粉离体萌发率的比较

Table 1. In virto germination rates of Onc. Sharry Baby Sweet Fragrance by various single factors

因素 Factors	质量浓度 Mass concentration/ (mg·L⁻¹)	花粉萌发率 Pollen germination rate/%
蔗糖 Sucrose	50×10³	4.94±0.38 ef
	100×10³	9.73±0.57 c
	150×10³	9.46±0.32 c
H₃BO₃	5×10³	7.53±0.95 d
	10×10³	17.12±1.45 a
	20×10³	12.58±1.50 b
Ca(NO₃) ·4H₂O	40×10³	6.16±0.56 e
	50×10³	13.34±0.68 b
	60×10³	9.30±0.71 c
MgSO₄·7H₂O	20×10³	4.76±0.72 f
	30×10³	4.69±0.52 f
	40×10³	2.71±0.46 g
KNO₃	30×10³	3.01±0.20 g
	40×10³	1.75±0.55 gh
	50×10³	1.19±0.13 h
同列数据后不同小写字母表示在 0.05 水平差异显著。表2和表3同。 Data with different lowercase letters on same column indicate significant difference at 0.05 level. Same for Tables 2 and 3.

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表 2 文心兰花粉离体萌发的正交试验结果

Table 2. Pollen germination rates of Oncidium in orthogonal test

编号 No.	蔗糖 Sucrose/(g·L⁻¹)	H₃BO₃/ (mg·L⁻¹)	Ca(NO₃)·4H₂O/ (mg·L⁻¹)	MgSO₄·7H₂O/ (mg·L⁻¹)	KNO₃/ (mg·L⁻¹)	花粉萌发率 Pollen germination rate/%
编号 No.	蔗糖 Sucrose/(g·L⁻¹)	H₃BO₃/ (mg·L⁻¹)	Ca(NO₃)·4H₂O/ (mg·L⁻¹)	MgSO₄·7H₂O/ (mg·L⁻¹)	KNO₃/ (mg·L⁻¹)	香水文心 Onc.Sharry Baby ‘Sweet Fragrance’	蜜糖 Onc.Sweet Sugar
1	50	5	40	20	30	17.64±1.35 f	10.78±0.66 e
2	50	10	60	20	30	25.67±1.58 c	23.16±1.26 c
3	50	20	50	20	30	23.95±1.67 cd	22.43±0.64 c
4	100	5	60	20	30	21.89±1.65 de	16.48±1.57 d
5	100	10	50	20	30	43.25±1.43 a	32.91±1.37 a
6	100	20	40	20	30	34.08±2.06 b	28.69±1.23 b
7	150	5	50	20	30	23.87±1.75 cd	17.91±0.72 d
8	150	10	40	20	30	40.37±1.59 a	30.71±1.77 ab
9	150	20	60	20	30	36.02±1.20 b	24.69±1.78 c

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表 3 68份文心兰种质资源花粉离体萌发率

Table 3. Statistical in vitro pollen germination rates of 68 Oncidium germplasms

序号 No.	种质名称 Name of material	花粉形态特征 Pollen morphology	花粉萌发率 Pollen germination rate/%	序号 No.	种质名称 Name of material	花粉形态特征 Pollen morphology	花粉萌发率 Pollen germination rate/%
1	香水235 Pi-Payk#235	形状规则，内含物充实	49.65±1.62 a	22	法国香水 French Fragrance	较多形状不规则，内含物皱缩、空泡化	6.22±0.95 jkl
2	香水文心 Sweet Fragrance	形状规则，内含物充实	43.25±1.43 b	23	香水254 Mon-Tho#254	形状规则，内含物充实	5.30±0.94 klm
3	蜜糖 Sweet Sugar	形状规则，内含物充实	32.91±1.37 c	24	XM-1	较多形状不规则，内含物皱缩、空泡化	5.10±0.74 klmn
4	小蜜蜂 Little Bee	形状规则，内含物充实	30.89±1.74 c	25	欧洲之星 Eurostar	较多形状不规则，内含物皱缩、空泡化	4.67±1.43 klmn
5	百万金币 Million Dollar	形状规则，内含物充实	20.88±3.07 d	26	粉星 Massai Pink	少量形状不规则，内含物皱缩	4.54±0.94 klmno
6	红樱桃 Red Cherry	少量形状不规则，内含物皱缩	20.42±1.89 d	27	红星 Massai Red	形状规则，内含物充实	4.50±1.33 klmnop
7	芒果文心 Mango	形状规则，内含物充实	20.27±1.59 d	28	紫薇 Ziwei	少量形状不规则，内含物皱缩	4.26±1.16 lmnop
8	红日 Red Sun	少量形状不规则，内含物皱缩	14.78±2.41 e	29	金香 Fragrant Gold	较多形状不规则，内含物皱缩、空泡化	4.09±0.36 lmnop
9	剑叶文心 Red Panther	形状规则，内含物充实	14.14±1.48 e	30	香香公主 Princess Fragrance	形状规则，内含物充实 Regular shape，inclusion sufficient	3.69±1.06 mnop
10	门神 Everglades	形状规则，内含物充实	13.89±1.79 e	31	紫香兰 Dark Matter	少量形状不规则，内含物皱缩	3.65±0.82 mnop
11	香水305 See-Daa#305	形状规则，内含物充实	11.36±1.60 f	32	黄色精灵 Golden Spirit	较多形状不规则，内含物皱缩、空泡化	3.24±0.67 mnopq
12	香水252 Mon-Tho#252	形状规则，内含物充实	10.91±1.98 fg	33	蕙心兰 Dawn	较多形状不规则，内含物皱缩、空泡化	3.13±0.56 mnopq
13	黄金天使 Golden Angel	少量形状不规则，内含物皱缩	9.76±1.38 fgh	34	黄花斑点 Alpine	较多形状不规则，内含物皱缩、空泡化	2.71±0.68 nopqr
14	香水304 Jairak Catt #304	形状规则，内含物充实	9.71±0.88 fgh	35	美少女 Beauty Lady	形状规则，内含物充实	2.56±0.60 nopqr
15	红鸟 Red bird	形状规则，内含物充实	8.69±0.76 ghi	36	香水美人 Kathrin Zoch	较多形状不规则，内含物皱缩、空泡化	2.09±0.48 opqr
16	三色香水 Tricolor	少量形状不规则，内含物皱缩	8.34±1.55 hij	37	花精灵 Flower Fairy	少量形状不规则，内含物皱缩，细胞质空泡化	2.02±0.60 pqr
17	香水607 Jairak Catt #607	形状规则，内含物充实	7.74±1.04 hij	38	铁板文心 Beauty Gold	少量形状不规则，内含物皱缩	1.91±0.51 pqr
18	香水599 Jairak Catt#599	形状规则，内含物充实	6.93±0.70 ijk	39	黄金午后 Rich Yellow	少量形状不规则，内含物皱缩，细胞质空泡化	1.12±0.19 qr
19	花猫 Carmela	少量形状不规则，内含物皱缩、空泡化	6.77±1.38 ijk	40	香水564 Jairak Fragrance#564	少量形状不规则，内含物皱缩	1.08±0.19 qr
20	情人 Lover	少量形状不规则，内含物皱缩	6.43±0.65 ijkl	41	光华 Chian-Tzy Deilight	较多形状不规则，内含物皱缩、空泡化	0.74±0.09 r
21	香水307 Pra-Lak#307	少量形状不规则，内含物皱缩	6.29±1.45 ijkl

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