Optimization of the tissue culture technology system of Dicranopteris pedata

doi:10.12302/j.issn.1000-2006.202110011

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2023, Vol. 47 ›› Issue (2): 107-114.doi: 10.12302/j.issn.1000-2006.202110011

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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

Abstract

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

CLC Number:

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, 2023, 47(2): 107-114.

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References 26

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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

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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