Regeneration System of Codonopsis pilosula Resources: Seed Sterilization Methods

doi:10.11924/j.issn.1000-6850.casb2023-0822

Abstract

Abstract:

As an authentic medicinal material in Shanxi Province, Codonopsis pilosula (Franch.) Nannf. is well-known throughout the country for its quality and curative effect. In order to effectively protect the high-quality germplasm resources of Codonopsis pilosula, an efficient regeneration system of Codonopsis pilosula was constructed. In this experiment, Codonopsis pilosula seeds were used as test materials, using 4 different treatment methods, after isolation and wrapping, two different sterilizers were used to treat mercury and sodium hypochlorite, respectively, and the damage to the seeds by different treatments was observed to determine the best sterilization method.The results showed that under different sterilization treatments, the seed contamination rate was 0-92.7%, and the survival rate was 0.8%-91.3%. Based on the combination of pollution rate and survival rate, treatment 29 was selected as the best result and followed by treatments 30. After wrapping the seeds in method 4, the best effect was achieved after 60-120 s of mercury sterilization, and the survival rate of the seeds was 82.00%-91.3%, while the contamination rate was 2.17%-3.83%. This experiment establishes a seed sterilization method suitable for the regeneration system of Codonopsis pilosula and lays the foundation for biotechnological breeding of Codonopsis pilosula.

Key words: Codonopsis pilosula (Franch.) Nannf., seed, contamination rate, survival rate, callus tissue, regeneration system, sterilization method

WANG Qiubao, MA Hongbin, WANG Yaoqin, TIAN Hongling, LIU Yuexian, PEI Shuaishuai, ZUO Xianqiang. Regeneration System of Codonopsis pilosula Resources: Seed Sterilization Methods[J]. Chinese Agricultural Science Bulletin, 2024, 40(27): 40-45.

Figures/Tables 5

References 30

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包裹方法、灭菌剂	灭菌用时	处理	污染率/%	死亡率/%	成活率/%
A1	30 s	1	71.5	0	28.5
	60 s	2	41.2	4.8	54.0
	120 s	3	14.0	36.8	49.2
	180 s	4	9.7	65.7	25.3
	240 s	5	5.7	88.3	6.3
A2	5 min	6	79.0	0	21.0
	10 min	7	33.5	24	40.5
	15 min	8	11.0	34.8	50.8
	20 min	9	7.0	65.7	26.5
B1	30 s	10	89.7	0	10.3
	60 s	11	67.2	3.5	29.3
	120 s	12	51.5	22.8	25.7
	180 s	13	31.5	62.8	5.7
	240 s	14	6.3	90.8	2.7
B2	5 min	15	92.7	0	7.3
	10 min	16	37.8	24.5	37.7
	15 min	17	24.5	42.7	31.2
	20 min	18	13.5	49.5	35.8
C1	30 s	19	68.5	0	31.5
	60 s	20	36.7	5.2	57.3
	120 s	21	11.8	37.3	51.2
	180 s	22	7.3	68.7	25.7
	240 s	23	3.2	91.7	4.8
C2	5 min	24	75.2	0	24.8
	10 min	25	29.2	29	41.5
	15 min	26	10.5	31.8	55.3
	20 min	27	5.7	55.8	41.2
D1	30 s	28	39.0	0	60.8
	60 s	29	3.8	5.2	91.3
	120 s	30	2.2	15.5	82.0
	180 s	31	0.0	95.7	4.3
	240 s	32	0.0	99.2	0.8
D2	5 min	33	68.2	0	31.5
	10 min	34	7.3	32.8	59.7
	15 min	35	4.3	38.3	56.5
	20 min	36	2.7	62	33.7

包裹方法、灭菌剂	灭菌用时	处理	污染率/%	死亡率/%	成活率/%
A1	30 s	1	71.5	0	28.5
	60 s	2	41.2	4.8	54.0
	120 s	3	14.0	36.8	49.2
	180 s	4	9.7	65.7	25.3
	240 s	5	5.7	88.3	6.3
A2	5 min	6	79.0	0	21.0
	10 min	7	33.5	24	40.5
	15 min	8	11.0	34.8	50.8
	20 min	9	7.0	65.7	26.5
B1	30 s	10	89.7	0	10.3
	60 s	11	67.2	3.5	29.3
	120 s	12	51.5	22.8	25.7
	180 s	13	31.5	62.8	5.7
	240 s	14	6.3	90.8	2.7
B2	5 min	15	92.7	0	7.3
	10 min	16	37.8	24.5	37.7
	15 min	17	24.5	42.7	31.2
	20 min	18	13.5	49.5	35.8
C1	30 s	19	68.5	0	31.5
	60 s	20	36.7	5.2	57.3
	120 s	21	11.8	37.3	51.2
	180 s	22	7.3	68.7	25.7
	240 s	23	3.2	91.7	4.8
C2	5 min	24	75.2	0	24.8
	10 min	25	29.2	29	41.5
	15 min	26	10.5	31.8	55.3
	20 min	27	5.7	55.8	41.2
D1	30 s	28	39.0	0	60.8
	60 s	29	3.8	5.2	91.3
	120 s	30	2.2	15.5	82.0
	180 s	31	0.0	95.7	4.3
	240 s	32	0.0	99.2	0.8
D2	5 min	33	68.2	0	31.5
	10 min	34	7.3	32.8	59.7
	15 min	35	4.3	38.3	56.5
	20 min	36	2.7	62	33.7

处理	灭菌用时	处理	污染率/%	成活率/%
D1	180 s	31	0.00±0.00^aA	4.33±0.29^fE
D1	240 s	32	0.00±0.00^aA	0.83±0.76^gE
D1	120 s	30	2.17±1.89^abAB	82.00±0.87^bB
D2	20 min	36	2.67±2.36^abAB	33.67±3.88^eD
C1	240 s	23	3.17±0.76^bAB	4.83±0.76^fE
D1	60 s	29	3.83±2.08^bB	91.33±2.75^aA
D2	15 min	35	4.33±0.76^bB	56.50±2.00^dC
D1	30 s	28	39.00±1.32^cC	60.83±1.26^cC

处理	灭菌用时	处理	污染率/%	成活率/%
D1	180 s	31	0.00±0.00^aA	4.33±0.29^fE
D1	240 s	32	0.00±0.00^aA	0.83±0.76^gE
D1	120 s	30	2.17±1.89^abAB	82.00±0.87^bB
D2	20 min	36	2.67±2.36^abAB	33.67±3.88^eD
C1	240 s	23	3.17±0.76^bAB	4.83±0.76^fE
D1	60 s	29	3.83±2.08^bB	91.33±2.75^aA
D2	15 min	35	4.33±0.76^bB	56.50±2.00^dC
D1	30 s	28	39.00±1.32^cC	60.83±1.26^cC

处理	外植体	接种数	出愈数	出愈率/%
29	叶	195	167	85.6
29	茎段	135	128	94.8
30	叶	195	141	72.3
30	茎段	135	110	81.5