竹叶兰种子无菌萌发与组培快繁研究

夏春英; 谢小敏; 刘江枫; 郑诚乐

doi:10.19303/j.issn.1008-0384.2020.11.006

竹叶兰种子无菌萌发与组培快繁研究

doi: 10.19303/j.issn.1008-0384.2020.11.006

1.
福建农林大学园艺学院，福建　福州　350002
2.
福州市于山风景区管理处，福建　福州　350001

基金项目: 福州市园林局科研项目（201404）

详细信息

作者简介:
夏春英（1996−），女，硕士，研究方向：花卉与景观园艺（E-mail：1144179784@qq.com）

通讯作者:
郑诚乐（1963−），男，博士，教授，研究方向：花卉与景观园艺、果树栽培生理与生化（E-mail：zcl2003@126.com）

中图分类号: S 682.31
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出版历程
- 收稿日期: 2020-04-18
- 修回日期: 2020-08-06
- 网络出版日期: 2020-11-24
- 刊出日期: 2020-11-30

Optimization of Media for Seed Germination and Propagation of Arundina graminifolia

1.
College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China
2.
Management Office of Yu Shan Scenic Area, Fuzhou, Fujian　350001, China

摘要

摘要: 目的筛选出竹叶兰各生长阶段适宜的培养基配方，建立竹叶兰组培快繁体系。方法以竹叶兰种子为材料，采用1/2 MS固体培养基，设计正交试验比较不同种类植物生长调节剂和添加物的浓度配比对竹叶兰种子无菌萌发、芽增殖生根等关键环节的影响。结果（1）竹叶兰种子萌发的最适培养基为1/2 MS+琼脂7 g·L⁻¹+蔗糖30.0 g·L⁻¹+活性炭1 g·L⁻¹+椰子乳100 mL·L⁻¹+NAA 1.0 mg·L⁻¹+6-BA 2.0 mg·L⁻¹+花宝1号0.5 g·L⁻¹，萌发率达83.79%；（2）芽增殖的最适培养基为1/2 MS+琼脂7 g·L⁻¹+蔗糖30 g·L⁻¹+活性炭1 g·L⁻¹+IBA 1 mg·L⁻¹+6-BA 3 mg·L⁻¹+NAA 0.5 mg·L⁻¹+100 g·L⁻¹土豆泥，增殖率1.36；（3）芽生根培养的最适培养基为1/2 MS+琼脂7 g·L⁻¹+蔗糖30 g·L⁻¹+活性炭1 g·L⁻¹+IBA 0.2 mg·L⁻¹+6-BA 2 mg·L⁻¹+NAA （0.2～0.5） mg·L⁻¹+100 g·L⁻¹香蕉泥；（4）炼苗移栽的最适培养基质为珍珠岩 ∶ 泥炭土=1 ∶ 1，成活率高达98%，移栽效果良好。结论适宜的椰子乳有利于竹叶兰种子无菌萌发，通过正交试验所建立的竹叶兰组培快繁体系，可为竹叶兰野生种质资源的保护和开发利用提供保障。
- 竹叶兰 /
- 无菌萌发 /
- 炼苗移栽 /
- 正交设计 /
- 组织培养
Abstract: Objective Formulation of culture media was optimized to facilitate the seed germination, tissue propagation, and various developmental stages of Arundina graminifolia. Method An orthogonal experiment was conducted to optimize the media for aseptic germination of the seeds and development of the seedlings of A. graminifolia using 1/2 MS as the base. Application of growth regulators and additives in the solid medium formulation was evaluated in accordance to the seed germination, seedling growth, root development and transplanting efficiency. Result The optimum media for various developmental stages of A. graminifolia were determined to include (1) 1/2 MS + 7 g agar·L⁻¹ + 30.0 g sucrose·L⁻¹ + 1 g activated carbon·L⁻¹ + 100 mL coconut milk·L⁻¹ + 1.0 mg NAA·L⁻¹ + 2.0 mg 6-BA·L⁻¹ + 0.5g Huabao No. 1·L⁻¹ to reach a seed germination rate of 83.79% . (2) 1/2 MS + 7 g agar·L⁻¹ + 3 0g sucrose·L⁻¹ + 1 g activated carbon·L⁻¹ + 1 mg IBA·L⁻¹ + 3 mg 6-BA·L⁻¹ + 0.5 mg NAA·L⁻¹ + 100 g mashed potato·L⁻¹ to achieve an average bud proliferation rate of 1.36. (3) 1/2 MS + 7 g agar·L⁻¹ + 30 g sucrose·L⁻¹ + 1 g activated carbon·L⁻¹ + 0.2 mg IBA·L⁻¹ + 2 mg 6-BA·L⁻¹ + 0.2-0.5 mg NAA·L⁻¹ + 100 g banana puree·L⁻¹ for an optimal root development. (4) Perlite:peat soil mixture at 1 ∶ 1 ratio to yield a seedling survival rate of 98% with desirable transplanting and acclimatization results. Conclusion Coconut milk in culture medium was found to significantly benefit the seed germination. The individually formulated media for various growth stages of A. graminifolia seedlings would help ensure the success in preserving the wild germplasms.
- Arundina graminifolia (D. Don) Hochr /
- aseptic germination /
- acclimatization and transplant /
- orthogonal design /
- tissue culture

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图 1 竹叶兰种子萌发

注：A.种胚变绿；B.种胚突破种皮形成带芽原球茎；C.未生根实生苗

Figure 1. Seed germination of A. graminifolia

Note: A: Embryo turned green in color; B: Embryo broke through seed coat and formed protocorm; C: Seedlings prior to rooting.

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表 1 不同培养基组分对竹叶兰种子萌发的影响

Table 1. Effect of formulated media on A. graminifolia seed germination

处理 Treatment	椰子乳含量 Content of Coconut milk/ （mL·L⁻¹）	NAA含量 Content of NAA/ （mg·L⁻¹）	6-BA 含量 6-BA/（Content of 6-BA/（mg·L⁻¹）	花宝1号含量 Content of Hua-bao No.1/（g·L⁻¹）	萌发率 Germination rate/%
1	50	0.1	1.5	0.5	72.68±1.48 ab
2	50	0.5	2	1	73.72±8.28 ab
3	50	1	2.5	1.5	75.44±4.76 a
4	100	0.1	2.5	1	73.84±0.42 ab
5	100	0.5	1.5	1.5	76.60±4.07 a
6	100	1	2	0.5	83.79±5.06 a
7	150	0.1	2	1.5	60.36±6.58 bc
8	150	0.5	2.5	0.5	52.01±7.75 c
9	150	1	1.5	1	59.97±8.55 bc
K₁	221.84	206.88	209.26	208.48	T=628.41
K₂	234.23	202.32	217.87	207.52
K₃	172.34	219.21	201.29	212.41
k₁	73.95 a	68.96 b	69.75 b	69.49 b
k₂	78.08 a	67.44 b	72.62 a	69.17 b
k₃	57.45 b	73.07 a	67.10 b	70.80 a
R	20.63	5.63	5.53	0.32
注：数据后不同小写字母表示差异达0.05显著水平。表2～5同。 Note: Data with different lowercase letters indicate significant difference at 0.05 level. Same for Table 2-5.

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表 2 不同培养基组分对芽增殖的影响

Table 2. Effect of formulated media on A. graminifolia bud proliferation

处理 Treatment	IBA含量 Content of IBA/ （mg·L⁻¹）	6-BA含量 Content of 6-BA/ （mg·L⁻¹）	NAA含量 Content of NAA/ （mg·L⁻¹）	添加物 Additives	增殖系数 Proliferation rate
1	0.2	1	0.2	1	1.160±0.652 a
2	0.2	2	0.5	2	1.153±1.058 a
3	0.2	3	1	3	1.247±0.997 a
4	0.5	1	0.5	3	1.020±0.905 a
5	0.5	2	1	1	0.853±0.545 a
6	0.5	3	0.2	2	1.007±0.901 a
7	1	1	1	2	1.233±1.253 a
8	1	2	0.2	1	1.120±0.687 a
9	1	3	0.5	3	1.360±0.823 a
K₁	3.560	3.413	3.287	3.133	T=10.153
K₂	2.880	3.127	3.533	3.393
K₃	3.713	3.613	3.333	3.627
k₁	1.187 a	1.138 a	1.096 a	1.044 a
k₂	0.960 b	1.042 a	1.178 a	1.131 a
k₃	1.238 a	1.204 a	1.111 a	1.209 a
R	0.278	0.162	0.082	0.164
注：添加物1为100 g·L⁻¹香蕉泥；添加物2为50 g·L⁻¹香蕉泥+50 g·L⁻¹土豆泥；添加物3为100 g·L⁻¹土豆泥。表3同。 Note: Additive 1: 100 g banana puree·L⁻¹; Additive 2: 50 g banana puree·L⁻¹ + 50 g mashed potatoes·L⁻¹; Additive 3: 100 g mashed potatoes·L⁻¹. Same for Table 3.

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表 3 不同培养基组分对芽生根的影响

Table 3. Effect of formulated media on rooting of A. graminifolia

处理 Treatment	IBA Content of IBA/ （mg·L⁻¹）	6-BA含量 Content of 6-BA/ （mg·L⁻¹）	NAA含量 Content of NAA/ （mg·L⁻¹）	添加物 Additives	生根数 Number of rootingoting	根长 Root length/ cmth/cm
1	0.2	1	0.2	1	3.273±1.668 a	2.221±1.362 a
2	0.2	2	0.5	2	2.600±1.268 ab	2.617±1.473 a
3	0.2	3	1	3	1.120±1.263 c	1.286±1.523 b
4	0.5	1	0.5	3	0.813±0.830 c	0.719±0.790 bc
5	0.5	2	1	1	0.847±0.954 c	0.341±0.369 c
6	0.5	3	0.2	2	0.693±0.834 c	0.452±0.581 c
7	1	1	1	2	0.947±0.887 c	0.588±0.523 bc
8	1	2	0.2	1	2.073±1.641 b	0.950±0.702 bc
9	1	3	0.5	3	0.553±0.716 c	0.520±0.680 bc
K_a1	6.993	5.033	6.040	6.193	T_a=12.920
K_a2	2.353	5.520	3.967	4.240
K_a3	3.573	2.367	2.913	2.487
k_a1	2.331 a	1.678 a	2.013 a	2.064 a
k_a2	0.784 b	1.840 a	1.322 b	1.413 b
k_a3	1.191 b	0.789 b	0.971 b	0.829 c
R_a	1.547	1.051	1.042	1.236
K_b1	6.123	3.528	3.622	3.512	T_b=9.693
K_b2	1.512	3.908	3.856	3.656
K_b3	2.057	2.258	2.215	2.525
k_b1	2.041 a	1.176 a	1.207 a	1.171 a
k_b2	0.504 b	1.303 a	1.285 a	1.219 a
k_b3	0.686 b	0.753 b	0.738 b	0.842 b
R_b	1.537	0.550	0.547	0.377

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表 4 不同基质移栽成活率

Table 4. Survival rates of transplanted A. graminifolia seedlings on different substrates

处理 Treatment	基质种类及配比 Matrix types and proportions	成活率 Rate of survival/%
1	水苔（Sphaghnum）	34.00±1.00 e
2	蛭石（Vermiculite）	88.00±2.00 b
3	泥炭土（Peat soil）	86.00±1.00 b
4	珍珠岩（Perlite）∶泥炭土（Peat soil）=1∶1	98.00±2.65 a
5	珍珠岩（Perlite）∶蛭石（Vermiculite）=1∶1	78.00±2.65 c
6	珍珠岩（Perlite）∶泥炭土（Peat soil）∶蛭石（Vermiculite）=1∶1∶1	80.00±3.61 c
7	珍珠岩（Perlite）∶蛭石（Vermiculite）∶草木灰（Plant ash）=1∶1∶1	60.00±4.00 d

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表 5 不同基质移栽苗生长情况

Table 5. Growth of transplanted A. graminifolia seedlings on different substrates

处理 Treatment	株高 Plant height/ cm	叶数 Leaf number	中间叶长 Length of middle leaf/cm	中间叶宽 Width of middle leaf/cm	茎粗 Stem diameter/ mm	根数 Root number	最长根长 The longest root length/ mm	最长根粗 The longest root width/ mm	鲜重 Fresh weight/g
1	3.18±0.82 ab	4.89±1.05 cd	4.64±0.30 a	0.37±0.04 ab	1.64±0.10 b	5.67±1.53 a	38.79±4.56 ab	0.40±0.14 d	0.11±0.01 d
2	2.96±0.70 b	5.10±0.76 bcd	3.73±1.05 bc	0.34±0.06 abc	2.12±0.39 ab	6.97±2.04 a	53.64±19.52 a	1.03±0.15 a	0.23±0.11 ab
3	3.78±1.06 a	5.69±0.93 ab	3.89±0.84 b	0.38±0.05 a	2.37±0.38 a	7.45±2.40 a	38.54±19.29 ab	0.77±0.14 bc	0.24±0.11 a
4	3.34±1.25 ab	5.81±1.05 a	3.21±1.24 bcd	0.34±0.06 abc	2.28±0.55 a	6.62±1.76 a	36.51±12.65 ab	0.80±0.19 bc	0.19±0.11 abcd
5	2.76±0.56 b	5.95±1.08 a	3.02±0.86 cd	0.32±0.06 bcd	2.24±0.59 a	6.80±2.33 a	43.43±14.21 ab	0.91±0.20 ab	0.21±0.11 abc
6	2.81±0.70 b	5.52±0.64 abc	2.69±0.43 d	0.30±0.04 cd	1.78±0.35 b	5.52±1.16 a	37.94±14.78 ab	0.70±0.13 c	0.14±0.05 bcd
7	2.65±0.65 b	4.43±0.53 d	2.56±0.70 d	0.28±0.04 d	1.67±0.48 b	6.00±2.00 a	34.77±10.36 b	0.66±0.11 c	0.12±0.08 cd

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