绒毛白蜡体胚诱导和植株再生

doi:10.11983/CBB15222

植物学报 ›› 2016, Vol. 51 ›› Issue (6): 807-816.DOI: 10.11983/CBB15222

绒毛白蜡体胚诱导和植株再生

燕丽萍^1,^2,³, 李丽^1,², 刘翠兰^1,², 吴德军^1,^2,,A;^*(), 王因花¹, 任飞¹, 赵梁军³

¹山东省林业科学研究院, 济南 250014
²山东省林木遗传改良重点实验室, 济南 250014
³中国农业大学观赏园艺与园林系, 北京 100193

收稿日期:2015-12-23 接受日期:2016-05-11 出版日期:2016-11-01 发布日期:2016-12-02
通讯作者: 吴德军
作者简介:
# 共同第一作者
基金资助:
“十二五”科技支撑计划(No.2013BAD01B06-6)和山东省科技发展计划(No.2014GNC111006)

Somatic Embryo Induction and Plantlet Regeneration of Fraxinus velutina

Liping Yan^1,2,3, Li Li^1,2, Cuilan Liu^1,2, Dejun Wu^1,2*, Yinhua Wang¹, Fei Ren¹, Liangjun Zhao³

¹Shandong Provincial Academy of Forestry, Jinan 250014, China
²Shandong Provincial Key Laboratory of Forest Tree Genetic Improvement, Jinan 250014, China
³Department of Ornamental Horticulture and Landscape Architecture, China Agricultural University, Beijing 100193, China

Received:2015-12-23 Accepted:2016-05-11 Online:2016-11-01 Published:2016-12-02
Contact: Wu Dejun
About author:
# Co-first authors

摘要/Abstract

摘要： 该文探讨了基本培养基、外植体、培养条件以及植物生长调节剂配比对绒毛白蜡(Fraxinus velutina)体胚诱导的影响。结果表明, 胚根是诱导体胚发生的最佳外植体; 体胚诱导的最适培养基为改良MS+2 mg·L^-1 6-BA+0.1 mg·L^-1 NAA、30 g·L^-1蔗糖、5.0 g·L^-1琼脂; 暗培养20天后进行光照培养(14小时光照/10小时黑暗), 光密度为100-120 μmol·m^-2·s^-1, 昼温度(25±2)°C, 夜温度(18±2)°C; 成功诱导出体细胞胚并获得再生植株, 体胚诱导率可达59.8%, 体胚萌发率达81.2%。壮苗最适培养基为改良WPM+0.5 mg·L^-1 6-BA+0.2 mg·L^-1ZT+0.01 mg·L^-1 NAA。生根最适培养基为改良1/2MS+1.0 mg·L^-1 IBA+0.05 mg·L^-1NAA+20 g·L^-1蔗糖, 生根率高达97.3%, 试管苗移栽成活率达97.8%。

Abstract: Using explants of Fraxinus velutina, we investigated the effects of basic medium, plant growth regulators, and concentrations of antioxidant medium, and culturing conditions on the induction of somatic embryogenesis in Fraxinus velutina. The best regeneration medium was Murashige and Skoog (MS) basal medium with Gamborg B5 vitamin supplementation, with 2.0 mg·L^-1 6-benzyladenine (6-BA), 0.1 mg·L^-1 naphthaleneacetic acid (NAA) at 20-day dark culture followed by light intensity of 100-120 μmol·m^-2·s^-1 irradiation for 14 h·d^-1, and the culture temperature was (25±2)°C at daytime and (18±2)°C at night. More than 81.2% radicle segments produced adventitious shoots, with a mean number of adventitious shoots per explant of 9.3. The optimal growing medium was modified WPM medium supplemented with 0.5 mg·L^-1 6-BA, 0.2 mg·L^-1 ZT and 0.01 mg·L^-1 NAA. More than 97.3% regeneration plantlets were able to root after transfer to modified half-strength MS medium supplemented with 1.0 mg·L^-1 indole-3-butyric acid, 0.05 mg·L^-1 NAA and 20 g·L^-1 sucrose; 97.8% of the plantlets survived.

燕丽萍, 李丽, 刘翠兰, 吴德军, 王因花, 任飞, 赵梁军. 绒毛白蜡体胚诱导和植株再生. 植物学报, 2016, 51(6): 807-816.

Liping Yan, Li Li, Cuilan Liu, Dejun Wu, Yinhua Wang, Fei Ren, Liangjun Zhao. Somatic Embryo Induction and Plantlet Regeneration of Fraxinus velutina. Chinese Bulletin of Botany, 2016, 51(6): 807-816.

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https://www.chinbullbotany.com/CN/Y2016/V51/I6/807

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参考文献 32

[1]	陈之群, 刘志敏 (2006). 绒毛白蜡茎段的组织培养及植株再生. 安徽农业科学 34, 472-473.
[2]	洪源范, 沈雨佳, 洪青, 李顺鹏 (2006). 大叶白蜡的离体培养与快速繁殖. 植物生理学通讯 42, 1139-1141.
[3]	胡峰, 施琼, 黄烈健 (2014). 黑木相思愈伤组织诱导及植株再生. 植物学报 49, 603-610.
[4]	孔冬梅, 谭燕双, 沈海龙 (2003). 白蜡树属植物的组织培养和植株再生. 植物生理学通讯 39, 677-680.
[5]	孔冬梅, 沈海龙, 冯丹丹, 张莉杰 (2006). 水曲柳体细胞胚与合子胚发生的细胞学研究. 林业科学 42, 130-133.
[6]	李素华, 何松林 (2009). 基本培养基和激素对常青白蜡腋芽诱导的影响. 贵州农业科学 37, 175-176.
[7]	刘琳, 俞斌, 黄鹏燕, 赵华, 彭俊华, 陈鹏, 彭良才 (2013). 芒不同基因型愈伤组织诱导及分化的差异. 植物学报 48, 192-198.
[8]	孙桂君, 孔冬梅, 张丽杰, 沈海龙 (2010). 取材时期和母树来源对水曲柳体细胞胚诱导的影响. 东北林业大学学报 38, 28-30.
[9]	王彩云, 白吉刚, 杨玉萍, 赵路明, 季华 (1999). 对节白蜡的组织培养及植株再生. 植物生理学通讯 35, 299-300.
[10]	王磊, 李淑娟, 徐云刚, 詹亚光 (2008). 绒毛白蜡成熟胚和茎段培养繁殖体系的建立. 林业科技 33(3), 1-3.
[11]	杨玲, 单琳, 沈海龙, 祁永会 (2012). 花曲柳体胚发生和植株再生. 林业科学 48(4), 49-53.
[12]	张翠叶, 辛福梅, 杨小林, 洛桑曲杰 (2014). 川滇高山栎体胚诱导关键影响因素研究. 西北农林科技大学学报(自然科学版) 1, 51-56.
[13]	张存旭, 姚曾玉, 赵忠 (2005). 栓皮栎体胚诱导关键影响因素研究. 林业科学 41(2), 174-177.
[14]	Arnold S, Sabala I, Bozhkov1 P, Dyachok J, Filonova L (2002). Developmental pathways of somatic embryo- genesis.Plant Cell Tiss Org Cult 69, 233-249.
[15]	Bates S, Preece JE, Navarrete NE, Van WJ, Gaffney GR (1992). Thidiazuron stimulates shoot organogenesis and somatic embryogenesis in white ash (Fraxinus anericana L.). Plant Cell Tiss Org Cult 31, 21-29.
[16]	Capuana M, Petrini G, Marco AD, Giannini R (2007). Plant regeneration of common ash (Fraxinus excelsior L.) by somatic embryogenesis.In Vitro Cell Dev Biol-Plant 43, 101-110.
[17]	Gamborg OL, Miller RA, Ojima K (1968). Nutrient require- ments of suspension cultures of soybean rootcells.Exp Cell Res 50, 151-158.
[18]	Hammatt N (1994). Shoot initiation in the leaflet axils of compound leaves from micropropagated shoots of juvenile and mature common ash (Fraxinus excelsior L.).J Exp Bot 45, 871-875.
[19]	Hammatt N (1996). Fraxinus excelsior L. (common ash). In: Bajaj PS ed. Biotechnology in Agriculture and Forestry: Trees IV, Vol. 35. Berlin, Heidelberg: Springer-Verlin. pp. 172-193.
[20]	Kim MS, Schumann CM, Klopfenstein NB (1997). Effects of thidiazuron and benzyladenine on axillary shoot proliferation of three green ash (Fraxinus pennsylvanica Mar- sh.) clones.Plant Cell Tiss Org Cult 48, 45-52.
[21]	Lemmetyinen J, Keinonen-Mettala K, Lannenpaa M, Weissenberg K, Sopanen T (1998). Activity of the CaMV 35S promoter in various parts of transgenic early flower- ing birch clones.Plant Cell Rep 18, 243-248.
[22]	Manzanera JA, Astorrga R, Bueno MA (1993). Somatic embryo induction and germination in Quercus L.Silvae Genetica 42, 2-3.
[23]	McCown BH, Lloyd G (1981). Woody Plant Medium (WPM) —a mineral nutrient formulation for microculture of woody plant species.Hortscience 16, 453.
[24]	Murashige T, Skoog F (1962). A revised medium for rapid growth and bio assays with tohaoco tissue cultures.Physiol Plant 15, 473-497.
[25]	Oh MJ, Na HR, Choi HK, Liu JR, Kim SW (2010). High frequency plant regeneration system for Nymphoides coreana via somatic embryogenesis from zygotic embryo- derived embryogenic cell suspension cultures.Plant Biotech Rep 4, 125-128.
[26]	Preece JE, Bates S (1991). In vitro studies with white ash (Fraxinus americana) nodules. In: Ahuja MR ed. Woody Plant Biotechnology. New York: Plenum Press. pp. 37-44.
[27]	Silveira CE, Corrignies A (1994). Period of harvest spr- outing ability of cuttings and in vitro plant regeneration in Fraxinus excelsior.Can J Bot 72, 261-267.
[28]	Tabtett AM, Hammatt N (1992). Regeneration of shoots from embryo hypocotyls of common ash (Fraxinus excelsior).Cell Rep 11, 514-518.
[29]	Tonon G, Capuana M, Marco AD (2000). Plant regenera- tion of Fraxinus angustifolia by in vitro shoot organogenesis.Sci Hortic 87, 291-301.
[30]	Tonon G, Capuana M, Rossi C (2001). Somatic embryo- genesis and embryo encapsulation in Fraxinus angustifolia Vhal.J Hortic Sci BioTech 76, 753-757.
[31]	Wilhelm E (2000). Somatic embryogenesis in oak (Quercus spp.).In Vitro Cell Dev Biol Plant 36, 349-357.
[32]	Zhou HH, Li M, Zhao X, Fan XC, Guo AG Affiliated withZhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences (CAAS)(2010). Plant regeneration from in vitro leaves of the peach rootstock ‘Nemaguard’ (Prunus persica×P. davidiana).Plant Cell Tiss Organ Cult 101, 79-87.

Basal medium	6-BA (mg·L^-1)	NAA (mg·L^-1)	Callus induction rate (%)	Induction rate of somatic embryo (%)	Somatic embryo shoot regeneration rate (%)
MS	2.0	0.1	79.4±10.7 b	34.6±5.3 b	61.2±12.8 b
Modified MS	2.0	0.1	89.3±12.2 a	59.8±4.5 a	81.2±14.5 a
WPM	2.0	0.1	71.2±8.4 c	25.4±5.3 c	41.2±9.3 c
Modified WPM	2.0	0.1	75.5±9.8 c	25.9±4.7 c	45.7±11.4 c

Basal medium	6-BA (mg·L^-1)	NAA (mg·L^-1)	Callus induction rate (%)	Induction rate of somatic embryo (%)	Somatic embryo shoot regeneration rate (%)
MS	2.0	0.1	79.4±10.7 b	34.6±5.3 b	61.2±12.8 b
Modified MS	2.0	0.1	89.3±12.2 a	59.8±4.5 a	81.2±14.5 a
WPM	2.0	0.1	71.2±8.4 c	25.4±5.3 c	41.2±9.3 c
Modified WPM	2.0	0.1	75.5±9.8 c	25.9±4.7 c	45.7±11.4 c

Explant type	Callus induction rate (%)	Induction rate of somatic embryo (%)	Somatic embryo shoot regeneration rate (%)
Cotyledon	57.5±11.6 b	23.8±5.9 b	21.7±8.8 b
Epicotyl	29.4±10.7 c	10.8±5.5 bc	4.7±2.9 c
Hypocotyl	35.7±9.3 c	11.4±5.3 c	5.1±2.3 c
Radicle	85.5±11.8 a	65.9±12.7 a	75.9±10.5 a

Explant type	Callus induction rate (%)	Induction rate of somatic embryo (%)	Somatic embryo shoot regeneration rate (%)
Cotyledon	57.5±11.6 b	23.8±5.9 b	21.7±8.8 b
Epicotyl	29.4±10.7 c	10.8±5.5 bc	4.7±2.9 c
Hypocotyl	35.7±9.3 c	11.4±5.3 c	5.1±2.3 c
Radicle	85.5±11.8 a	65.9±12.7 a	75.9±10.5 a

Darkness (days)	6-BA (mg·L^-1)	NAA (mg·L^-1)	Induction rate of somatic embryo (%)	Somatic embryo shoot regeneration rate (%)
0	2.0	0.1	10.8±3.7 c	0 c
10	2.0	0.1	30.8±5.5 b	41.2±9.3 b
20	2.0	0.1	59.7±7.1 a	79.2±14.5 a
30	2.0	0.1	25.9±4.8 a	45.7±11.4 c

绒毛白蜡体胚诱导和植株再生

Somatic Embryo Induction and Plantlet Regeneration of Fraxinus velutina

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Plant growth regulator (mg·L^-1)		Induction rate of somatic embryo (%)	Somatic embryo shoot regeneration rate (%)	Plant growth regulator (mg·L^-1)		Induction rate of somatic embryo (%)	Somatic embryo shoot regeneration rate (%)
6-BA	NAA	Induction rate of somatic embryo (%)	Somatic embryo shoot regeneration rate (%)	TDZ	NAA	Induction rate of somatic embryo (%)	Somatic embryo shoot regeneration rate (%)
1.0	0	0±0.0 d	0±0.0 d	1.0	0	0±0.0 d	0±0.0 d
	0.1	50.3±11.7 bc	34.2±11.3 c		0.1	40.4±10.3 bc	24.8±8.7 c
	0.25	40.7±9.5 c	44.3±13.0 bc		0.25	38.7±9.7 bc	31.5±12.3 bc
1.5	0	0±0.0 d	0±0.0 d	1.5	0	0±0.0	0±0.0 d
	0.1	61.8±10.3 ab	46.4±21.9 bc		0.1	50.9±9.8 b	57.2±16.9 a
	0.25	58.8±11.8 b	57.8±19.7 b		0.25	48.5±12.3 b	44.8±14.5 b
2.0	0	0±0.0 d	0±0.0 d	2.0	0	0±0.0 d	0±0.0 d
	0.1	68.9±12.5 a	78.9±24.8 a		0.1	56.5±11.7 a	45.2±14.5 b
	0.25	57.6±10.3 b	63.2±17.6 ab		0.25	59.4±12.4 a	43.9±20.0 b

Treatment		Plant growth regulator			Average plantlet (length·cm^-1)
Treatment		6-BA (mg·L^-1)	ZT (mg·L^-1)	NAA (mg·L^-1)	Average plantlet (length·cm^-1)
1		0.10	0.10	0.01	2.1
2		0.10	0.20	0.05	4.3
3		0.10	0.50	0.10	3.1
4		0.20	0.10	0.05	3.5
5		0.20	0.20	0.50	3.1
6		0.20	0.50	0.01	4.2
7		0.50	0.10	0.10	5.7
8		0.50	0.20	0.01	6.9
9		0.50	0.50	0.05	3.6
Average plantlet (length· cm^-1)	T1	3.6	4.0	4.9
	T2	4.0	4.9	3.9
	T3	5.2	3.8	4.0
	R	1.6	1.1	1.0

Treatment	Plant growth regulator		1/2MS		Modified 1/2MS
Treatment	IBA (mg·L^-1)	NAA (mg·L^-1)	Root rate (%)	The average number of root	Root rate (%)	The average number of root
1	0.1	-	0.0±0.0 e	0.0±0.0 d	0.0±0.0 e	0.0±0.0 d
2	0.5	-	11.6±1.3 d	2.7±0.5 c	21.3±2.1 d	3.5±0.3 c
3	1.0	-	24.9±2.4 c	3.3±0.8 c	31.5±1.8 cd	4.1±0.5 b
4	1.5	-	17.5±3.1 cd	3.1±0.3 c	26.6±3.2 d	3.6±0.3 c
5	-	0.01	8.6±2.8 d	3.8±0.3 b	14.9±1.9 de	3.3±0.5 c
6	-	0.05	13.4±3.7 cd	3.4±0.6 c	18.8±1.7 d	4.1±0.6 b
7	-	0.1	9.6±1.9 d	4.1±0.8 b	12.3±2.2 de	4.2±0.8 b
8	-	0.2	3.9±1.1 de	4.4±0.7 b	9.4±1.9 de	3.9±0.8 b
9	0.5	0.05	31.8±4.8 bc	5.7±0.6 a	67.6±3.8 bc	4.9±0.7 a
10	1.0	0.05	69.4±3.9 a	5.0±0.9 a	97.3±5.1 a	5.1±0.9 a
11	0.5	0.1	53.7±5.2 ab	5.4±0.9 a	77.6±4.3 b	5.3±0.6 a
12	1.0	0.1	67.8±4.4 a	5.7±0.5 a	76.8±4.7 b	6.4±0.8 a
13	0.5	0.2	32.5±4.9 bc	5.0±0.7 a	44.9±3.3 c	6.3±1.1 a
14	1.0	0.2	46.9±5.3 b	5.3±0.8 a	56.4±4.4 bc	5.6±0.7 a

Treatment	Sand:soil:medium (v:v:v)	Survival rate (%)
1	4:1:1	97.8 a
2	4:1:2	87.6 b
3	2:2:1	73.4 c

Treatment	Sand:soil:medium (v:v:v)	Survival rate (%)
1	4:1:1	97.8 a
2	4:1:2	87.6 b
3	2:2:1	73.4 c