毛建草愈伤组织诱导及植株再生

doi:10.11983/CBB23178

摘要/Abstract

摘要： 毛建草(Dracocephalum rupestre)是一种重要的药用植物。然而, 其叶片外植体再生系统尚未建立。以毛建草大田叶片和组培苗叶片为外植体, 探讨植物生长调节剂对愈伤组织诱导和分化、不定芽增殖及生根的影响, 建立了叶片离体再生体系。结果表明, 大田叶片愈伤组织诱导的最适培养基为MS+1.0 mg·L⁻¹6-BA+0.1 mg·L⁻¹2,4-D+1.0 mg·L⁻¹IAA, 诱导率达84.51%, 不定芽分化最佳培养基为MS+3.0 mg·L⁻¹6-BA+0.5 mg·L⁻¹TDZ+0.5 mg·L⁻¹IAA, 分化率为66.37%; 组培苗叶片愈伤组织诱导的最适培养基为MS+2.0 mg·L⁻¹6-BA+0.1 mg·L⁻¹2,4-D+0.5 mg·L⁻¹IAA, 诱导率达86.73%, 不定芽分化最佳培养基为MS+2.0 mg·L⁻¹6-BA+2.0 mg·L⁻¹TDZ+0.05 mg·L⁻¹IAA, 分化率为53.48%。不定芽增殖适宜培养基为MS+2.0 mg·L⁻¹6-BA+0.05 mg·L⁻¹NAA, 增殖率为83.57%, 最适生根培养基为1/2MS+0.1 mg·L⁻¹NAA+0.1 mg·L⁻¹IBA, 生根率为86.97%; 在草炭:蛭石=1:1 (v/v)的混合基质中组培苗长势最好。该研究建立了毛建草叶片离体培养再生体系, 为毛建草种质资源保存和种苗快繁提供了技术支持。

关键词: 毛建草, 愈伤组织诱导, 分化, 增殖, 生根

Abstract: Dracocephalum rupestre is an important medicinal plant. However, its leaf explant regeneration system has yet to be established. Here we investigated the impact of plant growth regulators on callus induction, differentiation, adventitious bud proliferation, and rooting using field planted leaves and tissue-cultured seedling leaves of D. rupestre as explants. We found that the optimal medium for inducing callus from field planted leaves was MS+1.0 mg·L⁻¹ 6-BA+ 0.1 mg·L⁻¹ 2,4-D+1.0 mg·L⁻¹ IAA, achieving an induction rate of 84.51%. For adventitious bud differentiation, the preferred medium comprised MS+3.0 mg·L⁻¹ 6-BA+0.5 mg·L⁻¹TDZ+0.5 mg·L⁻¹IAA, resulting in a differentiation rate of 66.37%. Similarly, for inducing callus from tissue-cultured seedling leaves, the optimal medium included MS+2.0 mg·L⁻¹ 6-BA+0.1 mg·L⁻¹2,4-D+0.5 mg·L⁻¹ IAA, with an induction rate of 86.73%. The medium conductive to adventitious bud differentiation consisted of MS+2.0 mg·L⁻¹6-BA+2.0 mg·L⁻¹TDZ+0.05 mg·L⁻¹ IAA, yielding a differentiation rate of 53.48%. Furthermore, the appropriate medium for adventitious bud proliferation was MS+2.0 mg·L⁻¹ 6-BA+0.05 mg·L⁻¹ NAA, achieving a proliferation rate of 83.57%. The rooting medium was 1/2MS+0.1 mg·L⁻¹ NAA+0.1 mg·L⁻¹ IBA, resulting in a rooting rate of 86.97%. Tissue-cultured seedlings exhibited optimal growth in a mixed substrate of peat and vermiculite at a ratio of 1:1 (v/v). This study successfully established a leaf explant regeneration system for D. rupestre, providing valuable technical support for the conservation and rapid propagation of its germplasm resources.

Key words: Dracocephalum rupestre, callus induction, differentiation, proliferation, rooting

田旭平, 岳康杰, 王佳丽, 刘慧欣, 史子尹, 亢红伟. 毛建草愈伤组织诱导及植株再生. 植物学报, 2024, 59(4): 613-625.

Xuping Tian, Kangjie Yue, Jiali Wang, Huixin Liu, Ziyin Shi, Hongwei Kang. Callus Induction and Plant Regeneration of Dracocephalum rupestre. Chinese Bulletin of Botany, 2024, 59(4): 613-625.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB23178

https://www.chinbullbotany.com/CN/Y2024/V59/I4/613

图/表 11

参考文献 24

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Type	6-BA (mg∙L^-1)	TDZ (mg∙L^-1)	KT (mg∙L^-1)	Occurrence time (d)	Induction rate (%)	Callus growth state
Leaves of field planted individual	0.5	-	-	10.25±4.65 a	40.36±3.17 b	Yellow with a firm and dense texture
	1.0	-	-	7.58±1.29 b	65.21±9.94 a	Pale yellow-green, loose, and a softer texture
	2.0	-	-	6.92±1.17 b	67.07±9.05 a	Pale yellow, loose and watery in appearance, with a soft texture
	-	0.5	-	16.25±3.39 a	37.98±5.73 b	Green, loose, and soft in texture, with a loose consistency
	-	1.0	-	15.83±3.58 a	40.17±7.53 b	Green, loose, with white spots, a soft texture
	-	2.0	-	11.50±2.88 b	62.53±6.66 a	Green, loose, with white spots, a soft texture
	-	-	0.5	9.50±2.60 a	35.19±6.73 b	Yellow, compact, with a somewhat hard texture
	-	-	1.0	8.17±2.44 ab	57.06±9.06 a	Brownish-yellow, compact, with a soft texture
	-	-	2.0	6.67±1.45 b	42.53±5.84 b	Brown, loose, resembling water stains, with a soft texture
Leaves of tissue-cultured seedling	0.5	-	-	11.08±2.69 a	47.33±8.53 b	Yellow with a firm and dense texture
	1.0	-	-	10.33±1.48 a	55.76±7.85 ab	Yellow-green, loose, with a relatively soft texture
	2.0	-	-	8.17±1.27 b	68.01±6.46 a	Yellow-green, loose, with a relatively soft texture
	-	0.5	-	14.17±2.99 a	33.63±5.79 b	Green, loose, with a soft texture
	-	1.0	-	12.33±1.03 ab	45.36±9.64 ab	Green, compact, with a hard texture
	-	2.0	-	10.33±2.63 b	55.61±9.07 a	Green, loose, with a relatively soft texture
	-	-	0.5	12.92±3.62 a	40.75±8.32 b	Pale yellow, loose, with a hard texture
	-	-	1.0	11.25±2.65 a	63.59±7.07 a	Pale yellow, loose, with a softer texture
	-	-	2.0	8.83±1.58 b	52.29±9.41 ab	Pale yellow, loose, and loosely resembling water stains in texture

Type	6-BA (mg∙L^-1)	TDZ (mg∙L^-1)	KT (mg∙L^-1)	Occurrence time (d)	Induction rate (%)	Callus growth state
Leaves of field planted individual	0.5	-	-	10.25±4.65 a	40.36±3.17 b	Yellow with a firm and dense texture
	1.0	-	-	7.58±1.29 b	65.21±9.94 a	Pale yellow-green, loose, and a softer texture
	2.0	-	-	6.92±1.17 b	67.07±9.05 a	Pale yellow, loose and watery in appearance, with a soft texture
	-	0.5	-	16.25±3.39 a	37.98±5.73 b	Green, loose, and soft in texture, with a loose consistency
	-	1.0	-	15.83±3.58 a	40.17±7.53 b	Green, loose, with white spots, a soft texture
	-	2.0	-	11.50±2.88 b	62.53±6.66 a	Green, loose, with white spots, a soft texture
	-	-	0.5	9.50±2.60 a	35.19±6.73 b	Yellow, compact, with a somewhat hard texture
	-	-	1.0	8.17±2.44 ab	57.06±9.06 a	Brownish-yellow, compact, with a soft texture
	-	-	2.0	6.67±1.45 b	42.53±5.84 b	Brown, loose, resembling water stains, with a soft texture
Leaves of tissue-cultured seedling	0.5	-	-	11.08±2.69 a	47.33±8.53 b	Yellow with a firm and dense texture
	1.0	-	-	10.33±1.48 a	55.76±7.85 ab	Yellow-green, loose, with a relatively soft texture
	2.0	-	-	8.17±1.27 b	68.01±6.46 a	Yellow-green, loose, with a relatively soft texture
	-	0.5	-	14.17±2.99 a	33.63±5.79 b	Green, loose, with a soft texture
	-	1.0	-	12.33±1.03 ab	45.36±9.64 ab	Green, compact, with a hard texture
	-	2.0	-	10.33±2.63 b	55.61±9.07 a	Green, loose, with a relatively soft texture
	-	-	0.5	12.92±3.62 a	40.75±8.32 b	Pale yellow, loose, with a hard texture
	-	-	1.0	11.25±2.65 a	63.59±7.07 a	Pale yellow, loose, with a softer texture
	-	-	2.0	8.83±1.58 b	52.29±9.41 ab	Pale yellow, loose, and loosely resembling water stains in texture

Type	6-BA (mg∙L^-1)	IAA (mg∙L^-1)	Occurrence time (d)	Induction rate (%)	Callus growth state
Leaves of field planted individual	0.5	0	10.25±4.60 b	40.37±6.89 b	Yellow, compact, and with a hard texture
	0.5	0.1	15.25±3.72 c	39.09±8.97 bc	Green, loose, with a softer texture
	0.5	0.5	9.58±1.40 ab	33.28±7.29 bc	Pale yellow, loose, with a harder texture
	0.5	1.0	8.08±1.96 a	26.72±5.74 c	Yellow, resembling water stains, soft texture
	1.0	0	7.58±2.29 a	65.33±6.56 a	Pale yellow, loose, softer texture
	1.0	0.1	14.67±3.08 c	55.46±5.64 ab	Green, compact, harder texture
	1.0	0.5	10.83±3.33 b	47.51±6.74 b	Pale yellow, loose, soft texture
	1.0	1.0	9.58±2.39 ab	33.72±9.32 bc	Green loose, soft texture
	2.0	0	6.92±2.17 a	67.55±6.54 a	Pale yellow, loose, watery appearance, soft texture
	2.0	0.1	11.67±1.20 b	63.29±7.96 a	Pale green, loose, soft texture
	2.0	0.5	12.16±2.46 bc	54.29±8.57 ab	Yellow-green, loose, softer texture
	2.0	1.0	14.00±3.43 c	41.31±9.03 b	Green, loose, softer texture
Leaves of tissue- cultured seedling	0.5	0	11.08±2.69 b	47.81±5.75 b	Yellow, compact, hard texture
	0.5	0.1	12.00±3.38 bc	34.52±6.34 bc	Yellow-green, loose, soft texture
	0.5	0.5	13.00±3.43 bc	31.96±4.75 bc	Yellow-green, compact, softer texture
	0.5	1.0	18.08±4.17 c	24.07±7.22 c	Yellow-green, loose, soft texture
	1.0	0	10.33±2.48 b	55.67±4.21 ab	Yellow-green, loose, softer texture
	1.0	0.1	10.33±2.58 b	51.34±1.99 ab	Yellow-green, loose, soft texture
	1.0	0.5	11.33±3.42 b	46.21±11.23 b	Yellow-green, loose, softer texture
	1.0	1.0	15.42±3.22 c	39.14±9.41 bc	Yellow-green, loose, soft texture
	2.0	0	8.17±1.27 a	68.73±13.47 a	Yellow-green, loose, softer texture
	2.0	0.1	9.67±2.30 ab	55.24±6.91 ab	Yellow-green, loose, soft texture
	2.0	0.5	9.88±2.65 ab	51.33±13.54 ab	Green, loose, softer texture
	2.0	1.0	10.33±2.79 b	46.61±14.26 b	Yellow-green, loose, softer texture

Type	6-BA (mg∙L^-1)	IAA (mg∙L^-1)	Occurrence time (d)	Induction rate (%)	Callus growth state
Leaves of field planted individual	0.5	0	10.25±4.60 b	40.37±6.89 b	Yellow, compact, and with a hard texture
	0.5	0.1	15.25±3.72 c	39.09±8.97 bc	Green, loose, with a softer texture
	0.5	0.5	9.58±1.40 ab	33.28±7.29 bc	Pale yellow, loose, with a harder texture
	0.5	1.0	8.08±1.96 a	26.72±5.74 c	Yellow, resembling water stains, soft texture
	1.0	0	7.58±2.29 a	65.33±6.56 a	Pale yellow, loose, softer texture
	1.0	0.1	14.67±3.08 c	55.46±5.64 ab	Green, compact, harder texture
	1.0	0.5	10.83±3.33 b	47.51±6.74 b	Pale yellow, loose, soft texture
	1.0	1.0	9.58±2.39 ab	33.72±9.32 bc	Green loose, soft texture
	2.0	0	6.92±2.17 a	67.55±6.54 a	Pale yellow, loose, watery appearance, soft texture
	2.0	0.1	11.67±1.20 b	63.29±7.96 a	Pale green, loose, soft texture
	2.0	0.5	12.16±2.46 bc	54.29±8.57 ab	Yellow-green, loose, softer texture
	2.0	1.0	14.00±3.43 c	41.31±9.03 b	Green, loose, softer texture
Leaves of tissue- cultured seedling	0.5	0	11.08±2.69 b	47.81±5.75 b	Yellow, compact, hard texture
	0.5	0.1	12.00±3.38 bc	34.52±6.34 bc	Yellow-green, loose, soft texture
	0.5	0.5	13.00±3.43 bc	31.96±4.75 bc	Yellow-green, compact, softer texture
	0.5	1.0	18.08±4.17 c	24.07±7.22 c	Yellow-green, loose, soft texture
	1.0	0	10.33±2.48 b	55.67±4.21 ab	Yellow-green, loose, softer texture
	1.0	0.1	10.33±2.58 b	51.34±1.99 ab	Yellow-green, loose, soft texture
	1.0	0.5	11.33±3.42 b	46.21±11.23 b	Yellow-green, loose, softer texture
	1.0	1.0	15.42±3.22 c	39.14±9.41 bc	Yellow-green, loose, soft texture
	2.0	0	8.17±1.27 a	68.73±13.47 a	Yellow-green, loose, softer texture
	2.0	0.1	9.67±2.30 ab	55.24±6.91 ab	Yellow-green, loose, soft texture
	2.0	0.5	9.88±2.65 ab	51.33±13.54 ab	Green, loose, softer texture
	2.0	1.0	10.33±2.79 b	46.61±14.26 b	Yellow-green, loose, softer texture

Type	6-BA (mg∙L^-1)	2,4-D (mg∙L^-1)	Occurrence time (d)	Induction rate (%)	Callus growth state
Leaves of field planted individual	0.5	0	10.25±2.46 b	40.38±8.73 b	Yellow, compact, hard texture
	0.5	0.1	11.42±1.79 bc	25.77±6.33 c	Pale yellow, loose, soft texture
	0.5	0.5	10.67±2.65 b	41.83±6.86 bc	Brownish-yellow, compact, somewhat hard texture
	0.5	1.0	15.00±4.66 c	44.08±9.91 bc	Brownish-yellow, resembling water stains, soft texture
	1.0	0	7.58±2.29 a	65.38±13.16 a	Pale yellow, loose, softer texture
	1.0	0.1	10.08±3.20 b	43.33±12.02 b	Pale yellow, dense, softer texture
	1.0	0.5	12.92±3.71 bc	51.16±15.02 ab	Pale yellow, loose, softer texture
	1.0	1.0	12.00±2.76 bc	56.32±9.31 ab	Yellow, compact, somewhat hard texture
	2.0	0	6.92±2.17 a	67.52±13.25 a	Pale yellow, loose, resembling water stains, soft texture
	2.0	0.1	7.91±2.22 a	60.83±11.02 a	Yellow, resembling water stains, soft texture
	2.0	0.5	10.33±1.45 b	57.12±13.37 ab	Yellow, compact, softer texture
	2.0	1.0	13.67±2.05 bc	48.53±8.04 b	Pale yellow, loose, softer texture
Leaves of tissue-cultured seedling	0.5	0	11.08±3.69 b	47.51±11.05 b	Yellow, compact, hard texture
	0.5	0.1	16.42±3.08 c	44.17±9.31 b	Pale yellow, loose, soft texture
	0.5	0.5	11.08±2.44 b	39.98±8.74 bc	Pale yellow, loose, hard texture
	0.5	1.0	16.33±5.30 c	19.03±6.33 c	Yellow, loose, soft texture
	1.0	0	10.33±3.48 ab	55.62±10.15 ab	Yellow-green, loose, softer texture
	1.0	0.1	15.17±3.17 c	49.71±10.63 b	Green loose, softer texture
	1.0	0.5	10.42±3.44 ab	44.54±9.63 b	Yellow, compact, hard texture
	1.0	1.0	10.92±2.51 ab	38.05±8.94 bc	Pale yellow, loose, softer texture
	2.0	0	8.17±2.27 a	68.23±14.06 a	Yellow-green, loose, softer texture
	2.0	0.1	11.67±2.54 b	66.11±11.82 a	Green, loose, softer texture
	2.0	0.5	9.42±3.33 a	53.42±11.44 ab	Pale yellow, loose, softer texture
	2.0	1.0	9.83±3.44 a	50.76±14.13 b	Pale yellow, loose, softer texture