Screening of optimal germination medium for in vitro Fraxinus

doi:10.12302/j.issn.1000-2006.202104026

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2023, Vol. 47 ›› Issue (3): 70-76.doi: 10.12302/j.issn.1000-2006.202104026

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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 E-mail:492721695@qq.com;sdlky412@163.com

Abstract

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

CLC Number:

S722

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, 2023, 47(3): 70-76.

Figures/Tables 11

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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

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