白蜡花粉最佳离体萌发培养基筛选

doi:10.12302/j.issn.1000-2006.202104026

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南京林业大学学报（自然科学版） ›› 2023, Vol. 47 ›› Issue (3): 70-76.doi: 10.12302/j.issn.1000-2006.202104026

白蜡花粉最佳离体萌发培养基筛选

刘蓉¹^,²(), 吴德军¹^,^*(), 王因花¹, 任飞¹, 李丽¹, 燕丽萍¹, 周晓锋²

1.山东省林业科学研究院,山东省林木遗传改良重点实验室,山东济南 250014
2.中国农业大学烟台研究院,山东烟台 264670

收稿日期:2021-04-26 接受日期:2021-08-24 出版日期:2023-05-30 发布日期:2023-05-25
通讯作者: 吴德军
基金资助:
山东省农业良种工程项目(2019LZGC009);国家重点研发计划项目(2021YFD22003030109)

Screening of optimal germination medium for in vitro Fraxinus

LIU Rong¹^,²(), WU Dejun¹^,^*(), WANG Yinhua¹, REN Fei¹, LI Li¹, YAN Liping¹, ZHOU Xiaofeng²

1. Shandong Provincial Academy of Forestry, Shandong Provincial Key Laboratory of Forest Tree Genetic Improvement, Jinan 250014,China
2. Yantai Institute of China Agricultural University,Yantai 264670,China

Received:2021-04-26 Accepted:2021-08-24 Online:2023-05-30 Published:2023-05-25
Contact: WU Dejun

摘要/Abstract

摘要：

【目的】筛选白蜡(Fraxinus sp.)花粉最适离体培养基,能够为后续的授粉受精和育种工作提供一定的参考依据。【方法】通过收集绒毛白蜡‘金箭’(F. velutina ‘Jinjian’)和美国红梣‘鲁蜡5号’(F. pennsylvanica ‘Lula 5’)的花粉,比较固体培养基和液体培养基上花粉的萌发情况,选用适宜的离体培养法观察培养时间对花粉萌发情况的影响,设计正交试验,筛选最佳离体萌发培养基。【结果】①固体培养基上‘金箭’的花粉萌发率均显著高于液体培养基,02和03号固体培养基上‘鲁蜡5号’的花粉萌发率显著高于液体培养基。②培养2.5 h左右的花粉适宜进行生活力的观测。③正交试验表明:‘金箭’花粉的最适萌发培养基为150 g/L蔗糖+20 mg/L硼酸+30 mg/L氯化钙,‘鲁蜡5号’花粉的最适萌发培养基为150 g/L蔗糖+40 mg/L硼酸+30 mg/L氯化钙。【结论】采用150 g/L蔗糖+20 mg/L硼酸+30 mg/L氯化钙的固体培养基培养2.5 h后适宜统计两种白蜡花粉的萌发情况,但不同种的花粉萌发率与花粉管长度的相关性有差异。

关键词: 白蜡, 花粉, 离体培养, 培养基

Abstract:

【Objective】Screening of the optimal medium in vitro for pollen can provide a valuable reference for subsequent pollination, fertilization and breeding of Fraxinus. 【Method】 A comparative analysis of pollen germination in Fraxinus velutina cv. Jinjian and F. pennsylvanica cv. Lula 5 was conducted on the solid and liquid media. An appropriate culture method in vitro was selected to observe the influence of culture time on pollen germination and screening for the optimal germination medium in vitro through orthogonal experiments. 【Result】The results showed that the pollen germination rate of ‘Jinjian’ on the solid medium was significantly higher than that on the liquid medium. The pollen germination rate of ‘Lula 5’ on the solid medium No. 02 and 03 was significantly higher than that on the liquid medium. Pollen cultured for approximately 2.5 h was suitable for the observation of viability. The orthogonal test showed that the optimum medium for pollen germination of ‘Jinjian’ was 150 g/L sucrose + 20 mg/L boric acid + 30 mg/L calcium chloride. Meanwhile, the optimum medium for pollen germination of ‘Lula 5’ was 150 g/L sucrose + 40 mg/L boric acid + 30 mg/L calcium chloride. 【Conclusion】 Pollen germination of ‘Jinjian’ and ‘Lula 5’ was suitable to calculate after 2.5 h of cultivation on the solid medium with 150 g/L sucrose + 20 mg/L boric acid + 30 mg/L calcium chloride. However, the correlation between the pollen germination rate and pollen tube length of different species was different.

Key words: Fraxinus plants, pollen, germination in vitro, medium

中图分类号:

S722

刘蓉,吴德军,王因花,等. 白蜡花粉最佳离体萌发培养基筛选[J]. 南京林业大学学报（自然科学版）, 2023, 47(3): 70-76.

LIU Rong, WU Dejun, WANG Yinhua, REN Fei, LI Li, YAN Liping, ZHOU Xiaofeng. Screening of optimal germination medium for in vitro Fraxinus[J].Journal of Nanjing Forestry University (Natural Science Edition), 2023, 47(3): 70-76.DOI: 10.12302/j.issn.1000-2006.202104026.

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水平 level	萌发率/% germination rate			花粉管长度/μm pollen tube length
水平 level	蔗糖(A) sucrose	H₃BO₃ (B)	CaCl₂ (C)	蔗糖(A) sucrose	H₃BO₃ (B)	CaCl₂ (C)
1	18.423 b	40.393 a	35.523 a	116.27 a	134.92 a	105.74 a
2	48.147 a	29.430 a	40.737 a	144.92 a	110.83 a	145.36 a
3	42.513 ab	39.260 a	32.823 a	106.79 a	122.22 a	116.88 a

水平 level	萌发率/% germination rate			花粉管长度/μm pollen tube length
水平 level	蔗糖(A) sucrose	H₃BO₃ (B)	CaCl₂ (C)	蔗糖(A) sucrose	H₃BO₃ (B)	CaCl₂ (C)
1	42.637 a	52.350 ab	40.647 a	149.18 a	167.43 a	146.56 a
2	56.410 a	30.887 b	53.553 a	170.41 a	125.56 a	168.59 a
3	43.260 a	59.070 a	48.107 a	131.94 a	158.52 a	136.37 a

白蜡花粉最佳离体萌发培养基筛选

Screening of optimal germination medium for in vitro Fraxinus

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水平 level	因素factor
水平 level	蔗糖/(g·L^-1) sucrose	硼酸/(mg·L^-1) H₃BO₃	氯化钙/(mg·L^-1) CaCl₂
1	100	20	20
2	150	30	30
3	200	40	40

培养基号 medium No.	蔗糖(A) /(g·L^-1) sucrose	硼酸(B) /(mg·L^-1) H₃BO₃	氯化钙(C) /(mg·L^-1) CaCl₂	萌发率/% germination rate	花粉管长度/μm pollen tube length
1	100	20	20	21.89±3.54 c	107.68±29.76 b
2	100	30	30	11.29±1.64 c	113.46±9.84 b
3	100	40	40	22.09±3.98 c	127.66±14.05 b
4	150	20	30	60.86±3.35 a	197.49±13.60 a
5	150	30	40	37.95±6.64 b	123.37±35.51 b
6	150	40	20	45.63±9.89 ab	113.89±40.19 b
7	200	20	40	38.43±3.95 b	99.60±67.72 b
8	200	30	20	39.05±5.07 b	95.66±33.80 b
9	200	40	30	50.06±4.04 ab	125.12±49.14 b

培养基号 medium No.	蔗糖(A)/ (g·L^-1) sucrose	硼酸(B)/ (mg·L^-1) H₃BO₃	氯化钙(C)/ (mg·L^-1) CaCl₂	萌发率/% germination rate	花粉管长度/μm pollen tube length
1	100	20	20	44.12±4.06 c	152.68±23.41 ab
2	100	30	30	27.17±4.03 d	135.96±68.81 b
3	100	40	40	56.62±4.92 b	158.89±40.20 ab
4	150	20	30	69.12±2.29 a	221.25±51.01 a
5	150	30	40	43.89±3.23 c	121.84±41.49 b
6	150	40	20	56.22±3.48 b	168.13±14.79 ab
7	200	20	40	43.81±6.32 c	128.39±41.15 b
8	200	30	20	21.60±2.80 d	118.88±32.28 b
9	200	40	30	64.37±1.63 ab	148.55±52.19 ab

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