2种溲疏叶片不定芽诱导的组织培养体系构建

doi:10.11707/j.1001-7488.LYKX20230500

Abstract

Abstract:

Objective: Seeds of Deutzia are small, and their survival rate is low. Because the branches of Deutzia are hollow, branch propagation is difficult. This study aims to establish a complete tissue culture system through the induction of adventitious buds from the leaves of plants in the genus Deutzia, and lay the foundation for the genetic transformation system and molecular breeding. It also provides theoretical basis and practical guidance for the innovation of de novo domestication of wild Deutzia. Method: In this study, branches with leaves of D. parviflora and D. baroniana, two common species of Deutzia in the Qinling Mountains of Shaanxi Province, were used as explants. The explants were used for primary culture, subculture, rooting culture, and then, the leaves of the cultured plantlets were used to induce callus and adventitious buds. Result: The optimal disinfection method of explants was to disinfect the surface with 75% alcohol for 10s and 0.1% mercuric chloride for 3 min. The survival rate was 100% and the contamination rate was 0. The optimal medium for D. parviflora subculture was MS + 2 mg?L^?1 6-BA, and the proliferation coefficient was 2.8. The optimal medium for D. baroniana subculture was MS + 5 mg?L^?1 6-BA^?1, and the proliferation coefficient was 4.53. The optimal medium for rooting culture was 1/2 MS + 0.8 mg?L^?1 IBA, with the proliferation coefficient of 1.74 for D. parviflora and 7.86 for D. baroniana. The explants for callus induction were the leaves on the 2–4 nodes from top to bottom of 30 day old root seedlings. The optimal medium for D. parviflora leaves to induce callus was WPM + 1 mg?L^?1 2,4-D, with the induction rate of 63%. The optimal medium for D. baroniana leaves to induce callus was WPM + 0.1 mg?L^?1 2,4-D, and the induction rate was 70%. Conclusion: This study has successfully established a regeneration system for adventitious shoot induction and plant formation using the leaves of D. parviflora and D. baroniana as explants. The optimal medium for D. parviflora adventitious bud differentiation from leaves was MS + 2 mg?L^?1 6-BA + 0.3 mg?L^?1 IBA. The optimal medium for the adventitious bud differentiation from D. baroniana leaves was MS + 5 mg?L^?1 6-BA+ 0.05 mg?L^?1 NAA.

Key words: Deutzia, adventitious bud, shade-tolerant, shrubs, de novo domestication

CLC Number:

S722.3⁺7

Yaxin Wang,Xin Shang,Yingnan Hao,Xiaoyi Ning,Yue Ma,Sinan Gan,Jinyu Li,Xin Zhang. Establishment of Tissue Culture Systems for Inducing Adventitious Buds from Leaves of Two Deutzia Species[J]. Scientia Silvae Sinicae, 2024, 60(10): 164-174.

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消毒处理 Disinfection treatment		成活率 Survival rate (%)	污染率 Contamination rate (%)
75%乙醇 75% Ethanol	0.1%升汞 0.1% Mercuric chloride	成活率 Survival rate (%)	污染率 Contamination rate (%)
30 s	9 min	20.0±10.7c	0.0±0.0c
30 s	6 min	53.3±13.3b	0.0±0.0c
30 s	3 min	86.7±9.1a	0.0±0.0c
30 s	1 min	93.3±6.7a	26.7±11.8b
10 s	9 min	86.7±9.1a	0.0±0.0c
10 s	6 min	100.0±0.0a	0.0±0.0c
10 s	3 min	100.0±0.0a	0.0±0.0c
10 s	1 min	100.0±0.0a	46.7±13.3a

培养基 Medium	6-BA/(mg?L^?1)	NAA/(mg?L^?1)	增殖系数 Proliferation coefficient
MS	1	0.1	0.20±0.03d
MS	2	0.1	0.33±0.03cd
MS	3	0.1	0.13±0.02d
MS	1	0	1.20±0.05b
MS	2	0	2.00±0.09a
MS	3	0	0.80±0.05bc

培养基 Medium	6-BA/(mg?L^?1)	NAA/(mg?L^?1)	增殖系数 Proliferation coefficient
MS	1	0	1.67±0.98b
MS	2	0	1.87±0.96b
MS	3	0	1.93±0.73b
MS	4	0	3.13±2.53ab
MS	5	0	4.53±3.56a

培养基 Medium	6-BA/ (mg?L^?1)	NAA/ (mg?L^?1)	IBA/ (mg?L^?1)	生根系数 Rooting coefficient	生根状态 State of rooting
MS	1	0.1	0	1.74±0.23a	粗且多 Thick and numerous
1/2MS	0	0	0.8	0.45±0.09b	细且少 Thin and few

培养基 Medium	IBA/(mg?L^?1)	生根系数 Rooting coefficient	生根状态 State of rooting
1/2MS	0.4	3.43±1.57b	细且少 Thin and few
1/2MS	0.6	4.00±1.29ab	细且少 Thin and few
1/2MS	0.8	7.86±5.98a	粗且多 Thick and numerous
1/2MS	1.0	5.00±1.00ab	细且少 Thin and few
1/2MS	1.2	4.00±1.28ab	细且少 Thin and few

Establishment of Tissue Culture Systems for Inducing Adventitious Buds from Leaves of Two Deutzia Species

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培养基 Medium	愈伤组织诱导率 Induction rate of callus (%)
培养基 Medium	第1片 First leaf	第2片 Second leaf	第3片 Third leaf	第4片 Fourth leaf	第5片 Fifth leaf
WPM+2,4-D1	32.00±12.34ab	60.00±12.47a	54.00±12.13a	43.50±12.0a	5.50±2.93b

培养基Medium	2,4-D/(mg?L^?1)	愈伤组织诱导率 Induction rate of callus (%)		愈伤状态 State of callus
培养基Medium	2,4-D/(mg?L^?1)	小花溲疏 D. parviflora	钩齿溲疏 D. baroniana	小花溲疏 D. parviflora	钩齿溲疏 D. baroniana
WPM	0.01	13.33±9.09b	26.67±11.82b	白色紧实 White and tight	白色疏松 White and loose
WPM	0.1	26.67±11.82b	73.33±11.82a	白色紧实 White and tight	白色疏松 White and loose
WPM	1	66.67±12.60a	13.33±9.09b	黄色疏松 Yellow and loose	白色疏松 White and loose

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