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

doi:10.3969/j.issn.1004-1524.2022.07.10

Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (7): 1431-1438.DOI: 10.3969/j.issn.1004-1524.2022.07.10

• Horticultural Science • Previous Articles Next Articles

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

Abstract

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

CLC Number:

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.

Figures/Tables 4

References 28

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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

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

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