掌叶覆盆子组培快繁体系中生长调节剂与矿质元素的优化

doi:10.3969/j.issn.1004-1524.2022.07.10

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (7): 1431-1438.DOI: 10.3969/j.issn.1004-1524.2022.07.10

掌叶覆盆子组培快繁体系中生长调节剂与矿质元素的优化

汪秀媛¹(), 邹奕巧², 刘玲玲¹, 陈珍³^,^*(), 江景勇⁴

1.台州市林业技术推广总站,浙江台州 318000
2.台州市国土空间整治与生态修复中心,浙江台州 318000
3.台州学院生命科学学院,浙江台州 318000
4.台州市农业科学研究院园艺研究所,浙江临海 317000

收稿日期:2021-10-08 出版日期:2022-07-25 发布日期:2022-07-26
通讯作者: 陈珍
作者简介:* 陈珍,E-mail: chenzh@tzc.edu.cn
汪秀媛(1972—),女,浙江临海人,工程师,主要从事森林培育和生态保护等研究工作。E-mail: TZLYWXY@126.com
基金资助:
浙江省基础公益研究计划(公益项目)(LGN22C020001);台州市科技计划(21nya14)

Optimizing tissue culture media for Rubus chingii Hu multiplication: effects of plant growth regulators and mineral elements

WANG Xiuyuan¹(), ZOU Yiqiao², LIU Lingling¹, CHEN Zhen³^,^*(), JIANG Jingyong⁴

1. Taizhou Forestry Technology Extension Station, Taizhou 318000, Zhejiang, China
2. Taizhou Land and Space Remediation and Ecological Restoration Center, Taizhou 318000, Zhejiang, China
3. College of Life Sciences, Taizhou University, Taizhou 318000, Zhejiang, China
4. Institute of Horticulture, Taizhou Academy of Agricultural Sciences, Linhai 317000, Zhejiang, China

Received:2021-10-08 Online:2022-07-25 Published:2022-07-26
Contact: CHEN Zhen

摘要/Abstract

摘要：

掌叶覆盆子(Rubus chingii Hu)药食同源,营养保健价值凸显,经济效益显著。为加快掌叶覆盆子良种繁育,以药用成分含量较高的优质株系L14茎段为外植体,采用调整植物生长调节剂的组合、质量浓度,以及矿质元素含量的方式,优化组培快繁体系。结果表明,添加0.1 mg·L^-1萘乙酸(NAA)时,5.0~30.0 mg·L^-1 6-糠氨基腺嘌呤/激动素(KT)促进掌叶覆盆子组培苗增殖的效果与0.5~3.0 mg·L^-1 6-苄氨基腺嘌呤(6-BA)相当,可见,6-BA的作用强度约为KT的10倍。0.5~3.0 mgL^-1的6-BA会引起试管苗的玻璃化,0.1~1.0 mg·L^-1 KT的添加有助于减轻玻璃化,其中,0.5 mg·L^-1 6-BA+1.0 mg·L^-1 KT的细胞分裂素组合下掌叶覆盆子组培苗增殖系数最大,达10.33,且苗生长健壮。调整培养基中具体矿质元素的含量,可进一步促进试管苗的生长。因此,优化后的掌叶覆盆子增殖培养基配方为:改良的MS培养基(2倍浓度的Ca、Cu和Mn,1/4 Mg和1/2 Zn)+ 6-BA 0.5 mg·L^-1 + KT 1.0 mg·L^-1 + NAA 0.1 mg·L^-1,试管苗增殖良好,生长健壮,能有效抑制玻璃化的产生。

关键词: 掌叶覆盆子, 6-苄氨基腺嘌呤, 激动素, 矿质元素, 玻璃化

Abstract:

Rubus chingii Hu is a typical dual-function medicine and edible food with prominent nutraceutical, health and economic values. In order to accelerate propagation of R. chingii with high quality, the stems of line 14 with higher contents of medicinal ingredients were used as explants. The effects of combinations and concentrations of plant growth regulators and mineral elements on multiplication and growth of R. chingii plantlets were investigated in this paper, and optimized media was obtained. The results showed that when the concentration of naphthylacetic acid (NAA) was 0.1 mg·L^-1, the effect of 5.0-30.0 mg·L^-1 kinetin (KT) on growth and multiplication of seedlings was equivalent with 0.5-3.0 mg·L^-1 6-benzylamine adenine (6-BA). Therefore, the action concentration of KT was 10 times that of 6-BA. 0.5-3.0 mg·L^-1 6-BA could caused vitrification of tissue culture plantlets and the supplement of 0.1-1.0 mg·L^-1 KT to the media alleviated the vitrification. The multiplication coefficient reached a maximum level of 10.33 under the combination of the two cytokinin, 0.5 mg·L^-1 6-BA and 1.0 mg·L^-1 KT, and the seedlings grew well. Optimization of mineral elements contents in media improved the growth of seedlings. All together, the best media for R. chingii multiplication were: modified MS medium (2-fold concentrations of Ca、Cu and Mn with 1/4 Mg and 1/2 Zn) added with 6-BA 0.5 mg·L^-1, KT 1.0 mg·L^-1 and NAA 0.1 mg·L^-1. Thus, the tissue culture seedlings multiplied and grew well without vitrification.

Key words: Rubus chingii Hu, N⁶-benzylaminopurine, kinetin, nutrient element, vitrification

中图分类号:

汪秀媛, 邹奕巧, 刘玲玲, 陈珍, 江景勇. 掌叶覆盆子组培快繁体系中生长调节剂与矿质元素的优化[J]. 浙江农业学报, 2022, 34(7): 1431-1438.

WANG Xiuyuan, ZOU Yiqiao, LIU Lingling, CHEN Zhen, JIANG Jingyong. Optimizing tissue culture media for Rubus chingii Hu multiplication: effects of plant growth regulators and mineral elements[J]. Acta Agriculturae Zhejiangensis, 2022, 34(7): 1431-1438.

图/表 4

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6-BA/ (mg·L^-1)	KT/ (mg·L^-1)	增殖系数 Multiplication coefficient	苗生长状况 Growing situation of seedlings
0	0	0 e	黄化Etiolated seedlings
0	0.05	0 e	黄化Etiolated seedlings
0	0.10	0.67±0.61 de	生长缓慢,浅绿Slow growing and light green seedlings
0	0.50	0 e	苗有一定生长,绿色Certain growing and green seedlings
0	1.00	0 e	苗较健壮,深绿Relatively well and dark green seedlings
0.05	0	0.33±0.33 e	苗细小,浅绿Weak and light green seedlings
0.05	0.05	0.50±0.19 de	部分黄化Certain etiolated seedlings
0.05	0.10	0.78±0.51 de	浅绿Light green seedlings
0.05	0.50	0 e	深绿Dark green seedlings
0.05	1.00	0.83±0.43 de	苗生长较好,深绿Relatively well and dark green seedlings
0.10	0	0.58±0.57 e	黄化、玻璃化Etiolated and vitrified seedlings
0.10	0.05	0.67±0.67 e	苗细小,浅绿Weak and light green seedlings
0.10	0.10	1.17±0.24 de	苗细小,浅绿Weak and light green seedlings
0.10	0.50	2.11±0.51 d	苗生长较好,深绿Relatively well and dark green seedlings
0.10	1.00	1.44±0.19 de	苗生长较好,深绿Relatively well and dark green seedlings
0.50	0	5.33±0.47 c	轻微玻璃化Slightly vitrified seedlings
0.50	0.05	8.08±1.23 b	轻微玻璃化Slightly vitrified seedlings
0.50	0.10	7.78 ±0.77 b	苗较健壮,深绿Relatively well and dark green seedlings
0.50	0.50	5.67±0.54 c	苗较健壮,深绿
0.50	1.00	10.33±0.67 a	苗健壮,深绿Seedlings grow well with dark green leaves
1.00	0	7.89±1.02 b	严重玻璃化Severe vitrified seedlings
1.00	0.05	8.42±2.42 b	轻微玻璃化Slightly vitrified seedlings
1.00	0.10	7.67±2.26 b	轻微玻璃化Slightly vitrified seedlings
1.00	0.50	8.25±1.60 b	苗较健壮,深绿Relatively well and dark green seedlings
1.00	1.00	7.50±1.04 b	苗健壮,深绿Relatively well and dark green seedlings

6-BA/ (mg·L^-1)	KT/ (mg·L^-1)	增殖系数 Multiplication coefficient	苗生长状况 Growing situation of seedlings
0	0	0 e	黄化Etiolated seedlings
0	0.05	0 e	黄化Etiolated seedlings
0	0.10	0.67±0.61 de	生长缓慢,浅绿Slow growing and light green seedlings
0	0.50	0 e	苗有一定生长,绿色Certain growing and green seedlings
0	1.00	0 e	苗较健壮,深绿Relatively well and dark green seedlings
0.05	0	0.33±0.33 e	苗细小,浅绿Weak and light green seedlings
0.05	0.05	0.50±0.19 de	部分黄化Certain etiolated seedlings
0.05	0.10	0.78±0.51 de	浅绿Light green seedlings
0.05	0.50	0 e	深绿Dark green seedlings
0.05	1.00	0.83±0.43 de	苗生长较好,深绿Relatively well and dark green seedlings
0.10	0	0.58±0.57 e	黄化、玻璃化Etiolated and vitrified seedlings
0.10	0.05	0.67±0.67 e	苗细小,浅绿Weak and light green seedlings
0.10	0.10	1.17±0.24 de	苗细小,浅绿Weak and light green seedlings
0.10	0.50	2.11±0.51 d	苗生长较好,深绿Relatively well and dark green seedlings
0.10	1.00	1.44±0.19 de	苗生长较好,深绿Relatively well and dark green seedlings
0.50	0	5.33±0.47 c	轻微玻璃化Slightly vitrified seedlings
0.50	0.05	8.08±1.23 b	轻微玻璃化Slightly vitrified seedlings
0.50	0.10	7.78 ±0.77 b	苗较健壮,深绿Relatively well and dark green seedlings
0.50	0.50	5.67±0.54 c	苗较健壮,深绿
0.50	1.00	10.33±0.67 a	苗健壮,深绿Seedlings grow well with dark green leaves
1.00	0	7.89±1.02 b	严重玻璃化Severe vitrified seedlings
1.00	0.05	8.42±2.42 b	轻微玻璃化Slightly vitrified seedlings
1.00	0.10	7.67±2.26 b	轻微玻璃化Slightly vitrified seedlings
1.00	0.50	8.25±1.60 b	苗较健壮,深绿Relatively well and dark green seedlings
1.00	1.00	7.50±1.04 b	苗健壮,深绿Relatively well and dark green seedlings

植物生长调节剂 Plant growth regulator/(mg·L^-1)		增殖系数 Multiplication coefficient	苗生长状况 Growing situation of seedlings
KT	0.10	0.17±0.29 e	苗细小Weak seedlings
	0.50	0.25±0.01 e	苗有一定生长Certain growing seedlings
	1.00	0.44±0.19 e	苗较健壮,深绿Relatively well and dark green seedlings
	2.00	0.75±0.25 e	苗较健壮,深绿Relatively well and dark green seedlings
	5.00	4.33±0.63 c	苗健壮,深绿Seedlings grow well with dark green leaves
	10.00	5.33±0.77 ab	苗健壮,深绿Seedlings grow well with dark green leaves
	20.00	4.89±0.38 bc	玻璃化,浅绿Vitrified and light green seedlings
	30.00	6.11±0.19 a	玻璃化,浅绿Vitrified and light green seedlings
6-BA	0.01	0.11±0.33 e	苗细小Weak seedlings
	0.05	0.67±0.33 e	苗细小Weak seedlings
	0.10	0.67±0.29 e	苗小,浅绿Small and light green seedlings
	0.20	2.89±0.69 d	苗较健壮,浅绿Relatively well and light green seedlings
	0.50	5.00±0.88 bc	轻微玻璃化Slightly vitrified seedlings
	1.00	5.67±0.58 ab	轻微玻璃化Slightly vitrified seedlings
	2.00	6.00±0.01 a	玻璃化Vitrified seedlings
	3.00	5.89±0.51 a	玻璃化Vitrified seedlings

植物生长调节剂 Plant growth regulator/(mg·L^-1)		增殖系数 Multiplication coefficient	苗生长状况 Growing situation of seedlings
KT	0.10	0.17±0.29 e	苗细小Weak seedlings
	0.50	0.25±0.01 e	苗有一定生长Certain growing seedlings
	1.00	0.44±0.19 e	苗较健壮,深绿Relatively well and dark green seedlings
	2.00	0.75±0.25 e	苗较健壮,深绿Relatively well and dark green seedlings
	5.00	4.33±0.63 c	苗健壮,深绿Seedlings grow well with dark green leaves
	10.00	5.33±0.77 ab	苗健壮,深绿Seedlings grow well with dark green leaves
	20.00	4.89±0.38 bc	玻璃化,浅绿Vitrified and light green seedlings
	30.00	6.11±0.19 a	玻璃化,浅绿Vitrified and light green seedlings
6-BA	0.01	0.11±0.33 e	苗细小Weak seedlings
	0.05	0.67±0.33 e	苗细小Weak seedlings
	0.10	0.67±0.29 e	苗小,浅绿Small and light green seedlings
	0.20	2.89±0.69 d	苗较健壮,浅绿Relatively well and light green seedlings
	0.50	5.00±0.88 bc	轻微玻璃化Slightly vitrified seedlings
	1.00	5.67±0.58 ab	轻微玻璃化Slightly vitrified seedlings
	2.00	6.00±0.01 a	玻璃化Vitrified seedlings
	3.00	5.89±0.51 a	玻璃化Vitrified seedlings

元素 Element		增殖系数 Multiplication coefficient	苗生长状况 Growing situation of seedlings
Ca²⁺	1/4 MS	4.08±1.07 a	部分玻璃化Certain vitrified seedlings
	1/2 MS	5.75±1.10 a	黄化,玻璃化Etiolated and vitrified seedlings
	1×MS	5.25±0.69 a	苗较健壮Relative well seedlings
	2×MS	4.58±1.17 a	苗健壮,深绿Seedlings grow well with dark green leaves
	4×MS	5.44±1.39 a	玻璃化Vitrified seedlings
Mg²⁺	1/4 MS	7.67±1.33 a	苗健壮,深绿Seedlings grow well with dark green leaves
	1/2 MS	4.58±1.62 b	苗健壮,深绿Seedlings grow well with dark green leaves
	1×MS	5.25±0.69 b	苗较健壮Relative well seedlings
	2×MS	4.22±1.54 b	苗矮小Weak seedlings
	4×MS	3.50±0.43 b	苗矮小,玻璃化Weak and vitrified seedlings
Cu²⁺	1/4 MS	5.17±1.11 ab	黄化Etiolated seedlings
	1/2 MS	6.17±1.69 a	黄化Etiolated seedlings
	1×MS	5.25±0.69 ab	苗较健壮Relative well seedlings
	2×MS	6.08±1.07 a	苗健壮Seedlings grow well
	4×MS	4.00±0.58 b	苗健壮,深绿Seedlings grow well with dark green leaves
Mn²⁺	1/4 MS	5.83±1.29 ab	黄化,玻璃化Etiolated and vitrified seedlings
	1/2 MS	5.42±0.63 ab	玻璃化Vitrified seedlings
	1×MS	5.25±0.69 b	苗较健壮Relative well seedlings
	2×MS	5.92±0.83 ab	苗健壮,深绿Seedlings grow well with dark green leaves
	4×MS	6.83±1.14 a	苗较健壮Relative well seedlings
Zn²⁺	1/4 MS	5.75±1.10 a	部分黄化Certain Etiolated seedlings
	1/2 MS	5.83±1.23 a	苗健壮,深绿Seedlings grow well with dark green leaves
	1×MS	5.25±0.69 a	苗较健壮Relative well seedlings
	2×MS	5.42±0.88 a	苗较健壮,浅绿Relative well and light green seedlings
	4×MS	5.45±0.79 a	轻微玻璃化Certain vitrified seedlings

掌叶覆盆子组培快繁体系中生长调节剂与矿质元素的优化

Optimizing tissue culture media for Rubus chingii Hu multiplication: effects of plant growth regulators and mineral elements

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