珍稀濒危物种堇叶紫金牛高效快繁体系的建立

doi:10.11983/CBB16233

植物学报 ›› 2017, Vol. 52 ›› Issue (6): 764-773.DOI: 10.11983/CBB16233

珍稀濒危物种堇叶紫金牛高效快繁体系的建立

孙英坤¹, 胡绍庆², 庞基良³, 高凯¹, 刘华红¹, 陈焕伟⁴, 姚涛¹, 陈林敬¹, 沈柏春^1,^*()

¹杭州市园林绿化股份有限公司, 杭州 310000
²浙江理工大学建筑工程学院, 杭州 310018
³杭州师范大学生命与环境科学学院, 杭州 310036
⁴龙泉市林业科学研究院, 龙泉 323700

收稿日期:2016-11-30 接受日期:2017-05-04 出版日期:2017-11-01 发布日期:2018-02-22
通讯作者: 沈柏春
基金资助:
浙江省花卉新品种选育重大科技专项(No.2012C12909-9)和杭州市科技计划(No.20150932H01)

Establishment of a Tissue Culture and Propagation System for Ardisia violacea, a Rare and Endangered Species

Sun Yingkun¹, Hu Shaoqing², Pang Jiliang³, Gao Kai¹, Liu Huahong¹, Chen Huanwei⁴, Yao Tao¹, Chen Linjing¹, Shen Bochun^1,^*()

¹Hangzhou Landscaping Incorporated, Hangzhou 310000, China
²College of Civil Engineering and Architecture, Zhejiang Sci-Tech University, Hangzhou 310018, China
³College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 310036, China
⁴Longquan Forestry Research Institute, Longquan 323700, China

Received:2016-11-30 Accepted:2017-05-04 Online:2017-11-01 Published:2018-02-22
Contact: Shen Bochun

摘要/Abstract

摘要： 筛选堇叶紫金牛(Ardisia violacea)野生优株, 以其当年新发带休眠腋芽茎段为外植体, 通过启动培养、丛生芽诱导增殖、壮苗培养、生根培养和炼苗移栽等过程建立其组培快繁技术体系。研究结果表明, 最佳启动培养基为MS+0.80 mg·L^-1 KT+0.10 mg·L^-1 NAA+0.10 mg·L^-1 IBA, 腋芽萌发率达92.60%; 最佳丛生芽诱导增殖培养基为MS1+0.50 mg·L^-1 TDZ+0.10 mg·L^-1 NAA, 平均增殖系数达8.60; 最佳壮苗培养基为MS+1.00 mg·L^-1 KT+0.50 mg·L^-1NAA; 最佳生根培养基为1/2MS+2.00 mg·L^-1 IBA+1.00 mg·L^-1 NAA+1.00 mg·L^-1 AC, 平均生根率达98.70%; 采用松鳞和泥炭(2:1, v/v)作为炼苗基质, 炼苗成活率可达85.30%。实验成功建立了堇叶紫金牛高效组培快繁技术体系, 经验证该体系能够满足规模化生产的需求。

关键词: 堇叶紫金牛, 启动培养, 丛生芽

Abstract: Shoot segments with axillary buds from young branches of wild Ardisia violacea with excellent characters were cultured in vitro. Through bud induction, multiple shoots proliferation, subculture, rooting and transplantation, a regeneration system was established. The optimal bud induction medium was MS+0.80 mg?L^-1 KT+0.10 mg?L^-1 NAA+0.10 mg?L^-1 IBA, and the germination frequency of axillary buds reached 92.60%; the optimal multiple shoot proliferation medium was MS1+0.50 mg?L^-1 TDZ+0.10 mg?L^-1 NAA, and the average proliferation coefficient reached 8.60; the optimal subculture medium was MS+1.00 mg?L^-1 KT+0.50 mg?L^-1 NAA; the optimal rooting medium was 1/2MS+2.00 mg?L^-1 IBA+1.00 mg?L^-1 NAA+1.00 mg?L^-1 AC, and the average frequency of shoot rooting reached 98.70%. Using pine bark:peat (2:1, v/v) as a matrix, the transplant survival rate could reach 85.30%. We established a high-frequency regeneration system for A. violacea that could meet the needs of large-scale production.

Key words: Ardisia violacea, bud induction, multiple shoots

孙英坤, 胡绍庆, 庞基良, 高凯, 刘华红, 陈焕伟, 姚涛, 陈林敬, 沈柏春. 珍稀濒危物种堇叶紫金牛高效快繁体系的建立. 植物学报, 2017, 52(6): 764-773.

Sun Yingkun, Hu Shaoqing, Pang Jiliang, Gao Kai, Liu Huahong, Chen Huanwei, Yao Tao, Chen Linjing, Shen Bochun. Establishment of a Tissue Culture and Propagation System for Ardisia violacea, a Rare and Endangered Species. Chinese Bulletin of Botany, 2017, 52(6): 764-773.

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https://www.chinbullbotany.com/CN/Y2017/V52/I6/764

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参考文献 23

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Treatment	Basic medium	TDZ (mg?L^-1)	NAA (mg?L^-1)	The average prolix- feration coefficient	Multiple shoots growth condition
1	MS	0.80	1.00	7.26±0.316 dE	Poor growth; small buds; leaves almost undifferentiated
2	MS	0.50	0.50	8.54±0.314 bBC	Poor growth; small buds; leaves almost undifferentiated
3	MS	0.10	0.10	8.47±0.155 bBC	Poor growth; small buds; leaves almost undifferentiated
4	MS1	0.80	0.50	8.42±0.053 bcBC	Vigorous growth; bud clusters; bud clusters could be sep- arated into several independent and complete shoots; leav- es almost differentiated
5	MS1	0.50	0.10	8.60±0.3126 bB	Vigorous growth; bud clusters; bud clusters could be sep- arated into several independent and complete shoots; leav- es almost differentiated
6	MS1	0.10	1.00	7.65±0.132 dDE	Vigorous growth; bud clusters; bud clusters could be sep- arated into several independent and complete shoots; leav- es almost differentiated
7	MS2	0.80	0.10	9.20±0.223 aA	Vigorous growth; bud clusters; bud clusters couldn’t be sep- arated into several independent and complete shoots; leav- es almost undifferentiated
8	MS2	0.50	1.00	9.48±0.164 aA	Vigorous growth; bud clusters; bud clusters couldn’t be sep- arated into several independent and complete shoots; leav- es almost undifferentiated
9	MS2	0.10	0.50	8.05±0.239 cCD	Vigorous growth; bud clusters; bud clusters couldn’t be sep- arated into several independent and complete shoots; leav- es almost undifferentiated
K1	8.0900	8.2933	8.1300
K2 K3 R	8.2233 8.9100 0.8200	8.8733 8.0567 0.8166	8.3367 8.7567 0.6267

Treatment	Basic medium	TDZ (mg?L^-1)	NAA (mg?L^-1)	The average prolix- feration coefficient	Multiple shoots growth condition
1	MS	0.80	1.00	7.26±0.316 dE	Poor growth; small buds; leaves almost undifferentiated
2	MS	0.50	0.50	8.54±0.314 bBC	Poor growth; small buds; leaves almost undifferentiated
3	MS	0.10	0.10	8.47±0.155 bBC	Poor growth; small buds; leaves almost undifferentiated
4	MS1	0.80	0.50	8.42±0.053 bcBC	Vigorous growth; bud clusters; bud clusters could be sep- arated into several independent and complete shoots; leav- es almost differentiated
5	MS1	0.50	0.10	8.60±0.3126 bB	Vigorous growth; bud clusters; bud clusters could be sep- arated into several independent and complete shoots; leav- es almost differentiated
6	MS1	0.10	1.00	7.65±0.132 dDE	Vigorous growth; bud clusters; bud clusters could be sep- arated into several independent and complete shoots; leav- es almost differentiated
7	MS2	0.80	0.10	9.20±0.223 aA	Vigorous growth; bud clusters; bud clusters couldn’t be sep- arated into several independent and complete shoots; leav- es almost undifferentiated
8	MS2	0.50	1.00	9.48±0.164 aA	Vigorous growth; bud clusters; bud clusters couldn’t be sep- arated into several independent and complete shoots; leav- es almost undifferentiated
9	MS2	0.10	0.50	8.05±0.239 cCD	Vigorous growth; bud clusters; bud clusters couldn’t be sep- arated into several independent and complete shoots; leav- es almost undifferentiated
K1	8.0900	8.2933	8.1300
K2 K3 R	8.2233 8.9100 0.8200	8.8733 8.0567 0.8166	8.3367 8.7567 0.6267

Treatment	Basic medium	KT (mg?L^-1)	NAA (mg?L^-1)	The average height of plantlet (cm)	The thickness and strength of the plantlet stem
1	MS	1.00	0.50	4.62±0.20 cB	+ + + + +
2	MS	0.80	0.10	3.50±0.74 C	+ + + +
3	MS	0.50	0.05	4.82±0.11 bcAB	+ + +
4	MS1	1.00	0.10	5.60±0.11 aA	+
5	MS1	0.80	0.05	5.18±0.27 abcAB	+ +
6	MS1	0.50	0.50	5.42±0.05 abAB	+
7	MS2	1.00	0.05	4.90±0.11 bcAB	+ +
8	MS2	0.80	0.50	4.87±0.25 bcAB	+ + +
9	MS2	0.50	0.10	3.64±0.82 dC	+ + + +
K1	4.3133	5.0400	4.9700
K2 K3 R	5.4000 4.4700 1.0867	4.5167 4.6267 0.5233	4.2467 4.9667 0.7233

Treatment	Basic medium	KT (mg?L^-1)	NAA (mg?L^-1)	The average height of plantlet (cm)	The thickness and strength of the plantlet stem
1	MS	1.00	0.50	4.62±0.20 cB	+ + + + +
2	MS	0.80	0.10	3.50±0.74 C	+ + + +
3	MS	0.50	0.05	4.82±0.11 bcAB	+ + +
4	MS1	1.00	0.10	5.60±0.11 aA	+
5	MS1	0.80	0.05	5.18±0.27 abcAB	+ +
6	MS1	0.50	0.50	5.42±0.05 abAB	+
7	MS2	1.00	0.05	4.90±0.11 bcAB	+ +
8	MS2	0.80	0.50	4.87±0.25 bcAB	+ + +
9	MS2	0.50	0.10	3.64±0.82 dC	+ + + +
K1	4.3133	5.0400	4.9700
K2 K3 R	5.4000 4.4700 1.0867	4.5167 4.6267 0.5233	4.2467 4.9667 0.7233

Treatment	Basic medium	IBA (mg?L^-1)	NAA (mg?L^-1)	Frequency of shoots rooting	The average root number of per plantlet	The average length of the roots of per plantlet (cm)
1	MS	2.00	1.50	93.30%±0.021 dC	3.60±0.6 abcABC	2.96±0.10 eC
2	MS	1.50	1.00	95.30%±0.031 cdBC	2.80±0.36 cdeBCDE	4.22±0.11 cB
3	MS	1.00	0.50	93.80%±0.013 dC	2.50±053 deCDE	4.60±0.28 bcB
4	1/2MS	2.00	1.00	98.70%±0.006 abAB	4.60±0.26 Aa	3.18±0.08 dC
5	1/2MS	1.50	0.50	99.10%±0.008 abAB	2.00±0.95 eDE	4.74±0.07 bAB
6	1/2MS	1.00	1.50	97.20%±0.017 abcABC	3.10±0.62 cdBCDE	3.35±0.13 deC
7	1/4MS	2.00	0.50	96.80%±0.026 bcABC	3.40±0.26 bcdABCD	3.48±0.23 dC
8	1/4MS	1.50	1.50	100.00%±0.000 aA	1.90±0.89 eE	5.26±0.16 aA
9	1/4MS	1.00	1.00	98.20%±0.003 abAB	4.20±0.44 abAB	3.50±0.27 dC
K1	0.9408	0.9627	0.9683		Frequency of shoots rooting
K2 K3 R	0.9833 0.9833 0.0425	0.9808 0.9640 0.0181	0.9734 0.9657 0.0077		Frequency of shoots rooting
K1	2.9667	3.8667	2.8667
K2 K3 R	3.2333 3.1667 0.2666	2.2333 3.2667 1.6334	3.8667 2.6333 1.2334		The average root number of per plantlet
K1	3.9267	3.2856	3.8567
K2 K3 R	3.8356 4.0800 0.2444	4.7400 3.8167 1.4544	3.7122 4.2733 0.5611		The average length of the roots of per plantlet

珍稀濒危物种堇叶紫金牛高效快繁体系的建立

Establishment of a Tissue Culture and Propagation System for Ardisia violacea, a Rare and Endangered Species

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Batch	Multiple shoots (bottle)	Regenerated plantlets (bottle)	Rooting plantlets (bottle)
1	526	4860	4773
2 3	508 620	4816 5282	4623 5176
4	582	5122	5081

Batch	Multiple shoots (bottle)	Regenerated plantlets (bottle)	Rooting plantlets (bottle)
1	526	4860	4773
2 3	508 620	4816 5282	4623 5176
4	582	5122	5081