春兰‘黄梅’×‘黄荷’杂交F<sub>1</sub>根状茎的组培快繁研究

漆子钰; 周育真; 艾叶; 彭东辉

doi:10.19303/j.issn.1008-0384.2019.09.006

春兰‘黄梅’×‘黄荷’杂交F₁根状茎的组培快繁研究

doi: 10.19303/j.issn.1008-0384.2019.09.006

漆子钰^{1, 2,},
周育真¹,
艾叶¹,
彭东辉^1, ,

1.
福建农林大学园林学院，福建　福州　350002
2.
江西省宜春市园林局，江西　宜春　336000

基金项目: 福建省种业创新与产业化工程林业项目（ZYCX-LY-2017005）

详细信息

作者简介:
漆子钰（1992−），女，硕士，助理工程师，研究方向：园林植物与应用研究（E-mail：qiziyu2013@126.com）

通讯作者:
彭东辉（1971−），男，博士，教授，研究方向：园林植物与应用研究（E-mail：fjpdh@126.com）

中图分类号: S 682.31
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出版历程
- 收稿日期: 2019-05-15
- 修回日期: 2019-07-18
- 刊出日期: 2019-09-01

Tissue Culture for Rapid Propagation of A Cymbidium goeringii Hybrid

QI Zi-yu^{1, 2
,},
ZHOU Yu-zheng¹,
AI Ye¹,
PENG Dong-hui^{1
, ,}

1.
College of Landscape Architecture, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China
2.
Yichun Garden Bureau, Yichun, Jiangxi　336000, China

摘要

摘要: 目的筛选春兰组培各阶段的最佳培养条件，建立优良的春兰快繁体系，为春兰大规模生产开发提供科学依据。方法以春兰‘黄梅’×‘黄荷’F₁无菌播种形成的根状茎为试验材料，通过基本培养基（1/2MS、改良1/2MS、HyponexⅡ）和植物生长调节剂（6-BA、NAA、TDZ、IBA）等因子不同组合，围绕增殖、分化、生根等阶段建立其再生体系，并对培育出的组培苗进行移栽练苗。结果增殖最适培养基为3.0 g·L⁻¹ HyponexⅡ+0.5 mg·L⁻¹ 6-BA+3.0 mg·L⁻¹ NAA+1.0 g·L⁻¹ 活性炭（AC）+30.0 g·L⁻¹ 白砂糖（Su）+7.0 g·L⁻¹ 琼脂（Ag），生长速度和增殖系数分别为7.31和6.02；分化最适培养基为2.0 mg·L⁻¹ 6-BA+0.3 mg·L⁻¹ NAA+30.0 g·L⁻¹ Su+7.0 g·L⁻¹ Ag，分化率、芽诱导个数和株高分别为90.00%、5.44个和2.53 cm；生根最适培养基为1.5 mg·L⁻¹ IBA+2.0 g·L⁻¹蛋白胨+1.0 g·L⁻¹ AC+25.0 g·L⁻¹ Su+7.0 g·L⁻¹ Ag，生根率、生根数和平均根长分别为100.00%、8.03条和3.00 cm；春兰组培苗练苗移栽60 d后存活率达96.53%。结论新型基本培养基HyponexⅡ（3.0 g·L⁻¹）对春兰增殖培养有高效促进作用，高水平的IBA（1.5 mg·L⁻¹）有利于春兰组培苗生根壮苗。
- 春兰 /
- 增殖培养 /
- 分化培养 /
- 生根培养
Abstract: Objective Optimum conditions for an efficient, rapid propagation from rhizomes of a Cymbidium goeringii hybrid were investigated. Method Rhizomes of the aseptically seeded F₁ hybrid between two C. goeringii cultivars, Huang-mei and Huang-he, were cultivated on various media (i.e., 1/2MS、modified 1/2MS, and HyponexⅡ) with added plant growth regulators (i.e., 6-BA, NAA, TDZ, and/or IBA) to monitor the plant growth throughout propagation, differentiation, and rooting of the regeneration process. The seedlings were subsequently transplanted outdoor to determine their survival rate. Result In the propagation stage, the optimal medium was 3.0 g HyponexⅡ·L⁻¹ + 0.5 mg 6-BA·L⁻¹ + 3.0 mg NAA·L⁻¹ + 1.0 g activated carbon (AC)·L⁻¹ + 30.0 g sugar (Su)·L⁻¹ + 7.0 g agar (Ag)·L⁻¹ to achieve a growing rate of 7.31 and a multiplication coefficient of 6.02. The optimal medium for differentiation was 2.0 mg 6-BA·L⁻¹ + 0.3 mg NAA·L⁻¹ + 30.0 g Su·L⁻¹ + 7.0 g Ag·L⁻¹ to deliver an averaged 90.00% differentiation, 5.44 buds per rhizome segment, and 2.53 cm height of seedling. For optimum rooting, the medium was 1.5 mg IBA·L⁻¹ + 2.0 g peptone·L⁻¹ + 1.0 g AC·L⁻¹ + 25.0 g Su·L⁻¹ + 7.0 g Ag·L⁻¹ to result in 100.00% root-generation with an averaged 8.03 roots at 3.00 cm long per plant. The survival rate of plantlets 60 d after transplantation was up to 96.53%. Conclusion HyponexⅡ applied at 3.0 g·L⁻¹ was found highly efficient for the C. goeringii rhizome regeneration; and, the addition of 1.5 mg·L⁻¹ of IBA in the medium significantly enhanced the rooting for the seedlings.
- Cymbidium goeringii /
- propagation culture /
- differentiation culture /
- rooting culture

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图 1 离体培养各个阶段

注：A为增殖培养第1 d；B为增殖培养第15 d；C为增殖培养120 d；D为分化培养第1 d；E为分化培养第15 d；F为分化培养120 d；G为生根培养120 d；H为组培苗练苗移栽60 d。

Figure 1. Tissue cultures at various development stages

Note: A is multiplication culture on 1^st day; B, 15^th day; and, C, 120^th day. D is differentiation culture on 1^st day; E, 15^th day; and, F, 120^th day. G is rooting culture on 120^th day. H indicates plantlets on 60^th day after transplantation.

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表 1 根状茎增殖培养配方正交设计L₉（3⁴）

Table 1. Orthogonal design L₉(3⁴) on culture media for rhizome propagation

水平 Level	因素 Factor
水平 Level	基本培养基 Basic medium	6-BA/(mg·L⁻¹)	NAA/(mg·L⁻¹)
1	1/2MS	0.1	2.0
2	Revised 1/2MS	0.3	3.0
3	Hyponex Ⅱ3.0 g·L⁻¹	0.5	5.0
注：改良1/2MS为不含有微量元素的1/2MS培养基。 Note: Modified ½ MS was regular ½ MS without added microelements.

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表 2 根状茎分化培养试验设计

Table 2. Experimental design on culture media for rhizome differentiation

处理号 Number	NAA/(mg·L⁻¹)	6-BA/(mg·L⁻¹)	TDZ/(mg·L⁻¹)
1	0.3	1.0	0.0
2	0.3	2.0	0.0
3	0.3	3.0	0.0
4	0.3	0.0	0.5
5	0.3	0.0	1.0
6	0.3	0.0	1.5
7	0.5	1.0	0.0
8	0.5	2.0	0.0
9	0.5	3.0	0.0
10	0.5	0.0	0.5
11	0.5	0.0	1.0
12	0.5	0.0	1.5

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表 3 组培苗生根培养全因素设计

Table 3. All factor design on culture media for plantlet rooting

处理号 Number	IBA/(mg·L⁻¹)	蛋白胨 Peptone/(g·L⁻¹)
1	0.5	1.0
2	0.5	2.0
3	0.5	3.0
4	1.0	1.0
5	1.0	2.0
6	1.0	3.0
7	1.5	1.0
8	1.5	2.0
9	1.5	3.0

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表 4 不同处理对根状茎增殖培养的影响

Table 4. Effects of medium composition on rhizome propagation

处理号 Number	A：基本培养基 Basic medium	B：6-BA/(mg·L⁻¹)	C：NAA/(mg·L⁻¹)	生长速度 Speed of growth	增殖系数 Proliferation coefficient
1	1	1	1	2.53±0.35 Bd	7.40±0.42 Aa
2	1	2	2	4.19±0.14 Bbcdd	6.69±0.0 5 Aab
3	1	3	3	4.76±0.31 ABbc	7.46±0.66 Aa
4	2	1	2	2.91±0.18 Bcd	6.27±0.31 Aab
5	2	2	3	3.46±0.15 Bbcd	7.20±0.30 Aa
6	2	3	1	4.19±0.47 Bbcd	5.85±0.07 Aab
7	3	1	3	5.21±0.41 ABb	5.41±0.54 Ab
8	3	2	1	5.10±0.53 ABbc	5.93±0.10 Aab
9	3	3	2	7.31±0.27 Aa	6.02±0.82 Aab
ka1	3.83	3.55	3.94	Ra值排序：A＞B＞C 最优水平：A₃B₃C₂ Rb值排序：A＞C＞B 最优水平：A₁B₂C₃
ka2	3.52	4.25	4.80
ka3	5.87	5.42	4.48
Ra	2.35	1.87	0.87
kb1	7.18	6.36	6.39
kb2	6.44	6.61	6.33
kb3	5.79	6.45	6.69
Rb	1.39	0.25	0.37
注：ka表示生长速度；Ra表示生长速度的极差；kb表示增殖系数；Rb表示增殖系数的极差。大写字母表示在0.01水平下的差异，小写字母表示0.05水平下的差异，下同。 Note: ka stands for speed of growth; Ra, variance of ka; kb, multiplication coefficient; and, Rb, variance of kb. Capital letters indicate significantly different at P＜0.01, lowercase significantly different at P＜0.05. Same for Table 5&6.

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表 5 NAA、6-BA和TDZ对根状茎芽分化的影响

Table 5. Effects of varied concentrations of NAA, 6-BA, and TDZ on rhizome differentiation

处理号 Number	分化率 Differentiation rate /%	芽诱导个数 Number of Induced bud /个	株高 Plant height/cm
1	98.00±6.32 Aa	4.07±0.53 BCDEbcde	2.70±0.81 Aa
2	90.00±10.44 ABabcd	5.44±0.78 Aa	2.53±0.75 Aa
3	94.55±12.93 ABab	4.90±0.52 ABab	1.51±0.42 DEFcd
4	78.08±19.10 Bcd	4.48±0.71 ABCDbc	1.70±0.36 CDEc
5	91.67±13.37 ABabc	4.78±0.63 ABCab	1.26±0.27 EFde
6	77.14±15.41 Bd	4.07±0.50 BCDEbcde	1.06±0.16 Fe
7	94.00±13.50 ABab	3.55±0.36 CDEcde	2.77±0.90 Aa
8	87.27±13.48 ABabcd	3.31±0.33 DEde	2.49±0.79 ABa
9	80.00±16.33 ABcd	3.09±0.33 Ee	2.07±0.55 BCb
10	46.00±9.66 Ce	3.15±0.48 Ee	1.77±0.47 CDbc
11	88.33±10.30 ABabcd	4.18±0.47 BCDEbcd	1.51±0.30 DEFcd
12	81.43±11.43 ABbcd	3.80±0.45 BCDEcde	1.47±0.33 DEFcd

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表 6 不同IBA和蛋白胨对组培苗生根的影响

Table 6. Effects of varied concentrations of IBA and peptone on rooting of seedlings

处理号 Number	生根率 Rooting rate/%	生根数 Number of roots/条	平均根长 Average root length /cm
1	72.73±16.61 Aa	5.25±0.76 ABb	2.65±1.19 Bc
2	82.86±12.87 Aa	5.89±1.16 ABab	2.80±1.20 ABbc
3	83.33±18.25 Aa	4.53±0.61 Bb	2.77±1.13 Bbc
4	89.47±15.29 Aa	5.25±1.00 ABb	2.75±1.30 Bbc
5	88.24±15.94 Aa	5.29±1.79 ABb	2.91±1.27 ABabc
6	77.42±12.62 Aa	6.87±1.09 ABab	3.18±1.12 Aa
7	97.14±9.24 Aa	6.46±1.17 ABab	3.19±1.20 Aa
8	100.00±0 Aa	8.03±1.38 Aa	3.00±1.16 ABab
9	96.43±7.91 Aa	6.31±1.37 ABab	2.75±1.08 Bbc

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