芒萁组培技术体系的优化

doi:10.12302/j.issn.1000-2006.202110011

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南京林业大学学报（自然科学版） ›› 2023, Vol. 47 ›› Issue (2): 107-114.doi: 10.12302/j.issn.1000-2006.202110011

芒萁组培技术体系的优化

张港隆¹^,²(), 朱恩永³, 刘丽婷¹^,^*(), 杨嘉麒², 汪雁楠¹, 莫晓勇²

1.江西省林业科学院,江西南昌 330032
2.华南农业大学,广东广州 510642
3.信丰县林业发展中心,江西赣州 341600

收稿日期:2021-10-08 修回日期:2022-04-27 出版日期:2023-03-30 发布日期:2023-03-28
通讯作者: * 刘丽婷(39191393@qq.com),副研究员。
基金资助:
江西省重点研发项目(20181ACG70020);江西省林业科技创新项目(202214)

Optimization of the tissue culture technology system of Dicranopteris pedata

ZHANG Ganglong¹^,²(), ZHU Enyong³, LIU Liting¹^,^*(), YANG Jiaqi², WANG Yannan¹, MO Xiaoyong²

1. Jiangxi Academy of Forestry, Nanchang 330032, China
2. South China Agricultural University, Guangzhou 510642, China
3. Xinfeng County Forestry Development Center, Ganzhou 341600, China

Received:2021-10-08 Revised:2022-04-27 Online:2023-03-30 Published:2023-03-28

摘要/Abstract

摘要：

【目的】芒萁(Dicranopteris pedate)是尾矿及酸性边坡修复的优良植物。优化芒萁组培技术,提高组培效率,可为困难立地修复提供大量芒萁孢子体。【方法】以芒萁孢子为外植体,研究外植体收集时间、消毒时间与孢子萌发率的关系;通过改变培养基中无机盐浓度、蔗糖浓度及植物生长调节剂类型及浓度,研究孢子萌发、原叶体增殖、孢子体生成3个过程的最佳培养基配方。【结果】春季2—4月采集的芒萁孢子萌发活性最强;孢子诱导萌发最佳处理为DKW +5%(质量分数) NaClO消毒15 s,萌发率97.15%,20 d原叶体可见;原叶体最佳增殖培养基为1/2 MS+蔗糖5 g/L + 6-BA 0.5 mg/L + KT 0.5 mg/L,30 d质量增殖倍数为7.28;孢子体诱导生成最佳培养基为1/2 MS+蔗糖30 g/L + NAA 1.0 mg/L,第8天孢子体即可产生,生成率53.33%。【结论】芒萁孢子活性与生长区域、生成月份存在关联性,广州地区3月的孢子活性最强;在幼孢子体的瓶苗生成率较低的情况下,研发原叶体绿色球状体(prothallus green globular body, PGGB)高效增殖为核心的芒萁苗繁育技术,有助于实现芒萁苗大量、快速生产。

关键词: 芒萁, 孢子收集时间, 孢子活性, 原叶体绿色球状体(PGGB), 生态修复

Abstract:

【Objective】 Optimizing tissue culture techniques of Dicranopteris pedata is needed to improve the success rate and productivity of tissue culture.【Method】 Dicranopteris pedata spores were used as explants to study the relationship between the explant collection time, disinfection time and spore germination rate. By using the different concentration of inorganic salt and sucrose, along with the type and concentration of the plant growth regulator in the medium, the optimal medium formula for the spore germination, prothallus proliferation and sporophyte formation were studied.【Result】 March was the optimal time for collecting spores; the optimal treatment for inducing spore germination was DKW + NaClO (5%) disinfection for 15 s, with which the germination rate was 97.15%. Prothallus appeared after 20 days and the optimal medium for prothallus proliferation was 1/2 MS + sucrose 5 g/L + 6-BA 0.5 mg/L + KT 0.5 mg/L. The mass proliferation multiple was 7.28 after 30 days; the optimal medium for sporophyte induction was 1/2 MS + sucrose 30 g/L + NAA 1.0 mg/L; sporophytes appeared after 8 days, and the formation rate was 53.33%.【Conclusion】 Spore activity of Dicranopteris pedata is related to the growth area and formation month, and the activity of spores is strongest in Guangzhou in March. In the case of low bottle seedling rates of young sporophytes, the seedling breeding technology with efficient proliferation of prothallus green globular body as the core has been developed, which will contribute to the realization of large-scale and rapid production of Dicranopteris pedata seedlings.

Key words: Dicranopteris pedata, spore collection time, spore activity, prothallus green globular body (PGGB), ecological restoration

中图分类号:

张港隆,朱恩永,刘丽婷,等. 芒萁组培技术体系的优化[J]. 南京林业大学学报（自然科学版）, 2023, 47(2): 107-114.

ZHANG Ganglong, ZHU Enyong, LIU Liting, YANG Jiaqi, WANG Yannan, MO Xiaoyong. Optimization of the tissue culture technology system of Dicranopteris pedata[J].Journal of Nanjing Forestry University (Natural Science Edition), 2023, 47(2): 107-114.DOI: 10.12302/j.issn.1000-2006.202110011.

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变异来源(因素) source of variation(foctor)	平方和 sum of squares (SS)	自由度 degree of freedom (df)	均方 mean square (MS)	H统计量 H statistic	P
基本培养基(A) basic culture media	55 229.804	3	18 409.935	72.60	0.000
消毒时间(B) disinfection time	83.861	3	27.954	0.11	0.991
基本培养基×消毒时间(A×B) basic culture media× disinfection time	2 184.546	9	242.727	2.87	0.969
误差error	14 012.125	79	177.369
总计total	71 510.336	94	760.748

处理号 treatment No.	因素水平 factor level		评价指标 evaluating indicator
处理号 treatment No.	A	B	萌发率/% germination rate	最早萌发时间/d earliest germination time	染菌瓶数 number of infected bottles
1	1	1	19.08±11.26 c	27	0
2	1	2	42.39±16.81 bc	26	0
3	1	3	47.32±13.43 bc	26	0
4	1	4	19.99±12.43 c	27	1
5	2	1	70.08±14.24 abc	24	0
6	2	2	87.43±3.01 abc	24	0
7	2	3	76.40±1.91 abc	25	0
8	2	4	82.44±3.86 abc	24	0
9	3	1	94.16±0.80 abc	22	0
10	3	2	79.58±15.42 abc	24	0
11	3	3	92.89±1.28 abc	22	0
12	3	4	94.74±0.56 abc	22	0
13	4	1	97.15±0.69 a	21	0
14	4	2	95.44±0.93 ab	21	0
15	4	3	96.54±0.95 a	20	0
16	4	4	97.08±0.68 a	21	0
K1	32.20	70.12
K2	79.09	76.27
K3	90.34	78.29
K4	96.55	73.56
R	64.35	8.17

变异来源(因素) fact or(source of variation)	平方和 SS	df	均方 MS	H统计量 H statistic	P
基本培养基(C) basic culture media	6 242.709	2	3 121.354	8.086	0.018
蔗糖(D)sucrose	8 800.858	2	4 400.429	11.399	0.003
6-BA(E)	23 703.507	2	11 851.753	30.702	0.000
KT(F)	2 312.997	2	1 156.499	2.996	0.224
误差error	37 688.977	94	400.947
总计total	78 749.048	102	772.049

处理号 treatment No.	因素水平 factor level				评价指标 evaluating indicator
处理号 treatment No.	C	D	E	F	30天后质量增殖倍数 mass proliferation times in 30 days
1	1	1	1	1	2.60±0.22 abc
2	2	1	2	2	7.28±0.60 a
3	3	1	3	3	2.03±0.08 cd
4	1	2	2	3	3.06±0.34 ab
5	2	2	3	1	3.29±0.81 abcd
6	3	2	1	2	2.16±0.13 bcd
7	1	3	3	2	1.66±0.07 d
8	2	3	1	3	2.13±0.06 bcd
9	3	3	2	1	2.57±0.13 abc
K1	2.44	3.97	2.29	2.82
K2	4.23	2.84	4.30	3.70
K3	2.25	2.12	2.32	2.41
R	1.98	1.85	2.01	1.29

变异来源(因素) source of variation	平方和 SS	df	均方 MS	F	P
基本培养基(G) basic culture media	0.173	2	0.086	3.736	0.045
蔗糖sucrose(H)	0.031	2	0.015	0.663	0.528
6-BA(I)	0.240	2	0.120	5.177	0.018
NAA(J)	0.122	2	0.061	2.627	0.101
误差error	0.393	17	0.23

芒萁组培技术体系的优化

Optimization of the tissue culture technology system of Dicranopteris pedata

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处理号 treatment No.	因素水平 factor level				评价指标 evaluating indicator
处理号 treatment No.	G	H	I	J	孢子体生成率/% sporophyte formation rate	最早生成时间/d the earliest formation time
1	1	1	1	1	10.00±10.00 bc	15
2	2	1	2	2	33.33±6.67 abc	12
3	3	1	3	3	20.00±11.55 bc	14
4	1	2	2	3	20.00±0.00 bc	15
5	2	2	3	1	6.70±6.67 c	16
6	3	2	1	2	40.00±11.55 ab	10
7	1	3	3	2	6.70±6.67 c	16
8	2	3	1	3	53.33±6.67 a	8
9	3	3	2	1	26.67±13.33 abc	13
K1	12.2	21.1	34.4	14.5
K2	31.1	22.2	26.7	26.7
K3	28.9	28.9	11.1	31.1
R	18.9	7.8	23.3	16.6

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