Establishment of Plant Regeneration System from Hairy Roots of Aconitum coreanum

doi:10.11983/CBB14174

Abstract

Abstract: The hairy roots of Aconitum coreanum were used as explants to induce callus via differentiation in 1/2MS medium with 0.5-5 mg·L^-16-BA and 0-0.5 mg·L^-1NAA, then regeneration plantlets were induced from the callus. A whole transgenetic plantlet was obtained. The optimal medium for inducing callus and adventitious buds was 1/2MS+2.0 mg·L^-16-BA+0.2 mg·L^-1NAA+3% sugar. The optimal medium for inducing roots was 1/2MS+0.5 mg·L^-16-BA+0.1 mg·L^-1NAA+5 mg·L^-1GA+3% sugar.

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Lili Liu, Shubo Lu, Jiaping Xu, Qingtian Zhang, Changyu Li. Establishment of Plant Regeneration System from Hairy Roots of Aconitum coreanum[J]. Chinese Bulletin of Botany, 2015, 50(5): 623-627.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB14174

https://www.chinbullbotany.com/EN/Y2015/V50/I5/623

Figures/Tables 4

Figure 1 Different stages of plantlet regeneration from Aconitum coreanum hairy roots(A) Hairy root; (B) Base enlargement hairy roots; (C) Callus; (D) Buds; (E) A whole plantlet; (F) An albino seedling; (G) Induced roots. Bar=5 mm

Table 1 Effects of different plant growth regulators on inducing callus and adventitious buds of Aconitum coreanum

No. of medi- um	Phytohormone composition (mg·L^-1)		Color change of hairy roots	Callus induced	Adventitious buds induced
No. of medi- um	NAA	6-BA	Color change of hairy roots	Callus induced	Adventitious buds induced
M1	0	0.5	10% turned green	-	-
M2	0	1.0	30% turned green	-	-
M3	0	2.0	10% could see green spots	-	-
M4	0	4.0	5% could see green spots	-	-
M5	0	5.0	5% could see green spots	-	-
M6	0.1	0.5	50% turned green	-	3%
M7	0.2	0.5	60% turned yellow-green	20%	Few adventitious buds were induced
M8	0.5	0.5	40% turned yellow-green	20%	-
M9	0.1	1.0	90% turned yellow-green	-	10%
M10	0.2	1.0	40% turned yellow-green, 20% turned green	25%	Few adventitious buds were induced
M11	0.5	1.0	50% turned yellow-green	30%	Few adventitious buds were induced
M12	0.1	2.0	30% turned yellow-grown	30%	30%
M13	0.2	2.0	40% turned yellow-grown, 20% turned green	-	-
M14	0.5	2.0	40% turned yellow-green	Few callus were obtained	-
M15	0.1	4.0	30% turned green	-	-
M16	0.2	4.0	30% turned yellow-green	20%	40%
M17	0.5	4.0	40% turned yellow-green	20%	-
M18	0.1	5.0	20% turned green	-	-
M19	0.2	5.0	50% turned yellow-green, 50% turned green	50%	Few adventitious buds were induced
M20	0.5	5.0	50% turned yellow-green	20%	20%

Table 1 Effects of different plant growth regulators on inducing callus and adventitious buds of Aconitum coreanum

No. of medi- um	Phytohormone composition (mg·L^-1)		Color change of hairy roots	Callus induced	Adventitious buds induced
No. of medi- um	NAA	6-BA	Color change of hairy roots	Callus induced	Adventitious buds induced
M1	0	0.5	10% turned green	-	-
M2	0	1.0	30% turned green	-	-
M3	0	2.0	10% could see green spots	-	-
M4	0	4.0	5% could see green spots	-	-
M5	0	5.0	5% could see green spots	-	-
M6	0.1	0.5	50% turned green	-	3%
M7	0.2	0.5	60% turned yellow-green	20%	Few adventitious buds were induced
M8	0.5	0.5	40% turned yellow-green	20%	-
M9	0.1	1.0	90% turned yellow-green	-	10%
M10	0.2	1.0	40% turned yellow-green, 20% turned green	25%	Few adventitious buds were induced
M11	0.5	1.0	50% turned yellow-green	30%	Few adventitious buds were induced
M12	0.1	2.0	30% turned yellow-grown	30%	30%
M13	0.2	2.0	40% turned yellow-grown, 20% turned green	-	-
M14	0.5	2.0	40% turned yellow-green	Few callus were obtained	-
M15	0.1	4.0	30% turned green	-	-
M16	0.2	4.0	30% turned yellow-green	20%	40%
M17	0.5	4.0	40% turned yellow-green	20%	-
M18	0.1	5.0	20% turned green	-	-
M19	0.2	5.0	50% turned yellow-green, 50% turned green	50%	Few adventitious buds were induced
M20	0.5	5.0	50% turned yellow-green	20%	20%

Table 2 Grow phenomena of the materials subcultured at the same inducing medium

No. of medium	Phytohormone composition (mg·L^-1)				Description of hairy root growing and differentiation status
No. of medium	NAA		6-BA
M6	0.1		0.5		The material turned albino and died
M7	0.2		0.5		30% of the material turned brown, 10% turned green, 30% turned yellow slim hairy roots, 10% produced cluster buds
M8		0.5		0.5		80% turned yellow slim hairy roots, the base section of hairy roots became enlargement and 10% base section turned green
M9	0.1		1.0		The material turned albino and died
M10	0.2		1.0		The material turned albino and died
M11		0.5		1.0		30% base section of hairy roots turned green, 20% new produced hairy roots grew long and thin, others were short and thick
M12		0.1		2.0		30% of the material turned green, 30% kept yellow-green and enlargement
M13		0.2		2.0		The material turned albino and died
M14		0.5		2.0		50% of the material turned green and was going to be buds, hairy roots grew thick, 40% hairy roots were becoming brown
M15		0.1		4.0		40% of the material turned green and some were going to be buds, hairy roots were enlargement and strong, 50% were becoming brown
M16		0.2		4.0		The material turned albino and died
M17		0.5		4.0		The material turned albino and died
M18		0.1		5.0		The material turned albino and died
M19		0.2		5.0		40% of the material turned green, 20% kept yellow-green and grew strongly, 20% turned brown
M20	0.5		5.0		50% of the material turned green and grew strongly, 10% became cluster plantlets, 20% turned brown

Table 2 Grow phenomena of the materials subcultured at the same inducing medium

No. of medium	Phytohormone composition (mg·L^-1)				Description of hairy root growing and differentiation status
No. of medium	NAA		6-BA
M6	0.1		0.5		The material turned albino and died
M7	0.2		0.5		30% of the material turned brown, 10% turned green, 30% turned yellow slim hairy roots, 10% produced cluster buds
M8		0.5		0.5		80% turned yellow slim hairy roots, the base section of hairy roots became enlargement and 10% base section turned green
M9	0.1		1.0		The material turned albino and died
M10	0.2		1.0		The material turned albino and died
M11		0.5		1.0		30% base section of hairy roots turned green, 20% new produced hairy roots grew long and thin, others were short and thick
M12		0.1		2.0		30% of the material turned green, 30% kept yellow-green and enlargement
M13		0.2		2.0		The material turned albino and died
M14		0.5		2.0		50% of the material turned green and was going to be buds, hairy roots grew thick, 40% hairy roots were becoming brown
M15		0.1		4.0		40% of the material turned green and some were going to be buds, hairy roots were enlargement and strong, 50% were becoming brown
M16		0.2		4.0		The material turned albino and died
M17		0.5		4.0		The material turned albino and died
M18		0.1		5.0		The material turned albino and died
M19		0.2		5.0		40% of the material turned green, 20% kept yellow-green and grew strongly, 20% turned brown
M20	0.5		5.0		50% of the material turned green and grew strongly, 10% became cluster plantlets, 20% turned brown

Table 3 Media selection of rooting from regenerated plantlet in Aconitum coreanum

Phytohormone composition (mg·L^-1)	No. of root
Phytohormone composition (mg·L^-1)	0.1	0.5	1.0	1.5
IBA	1	2	6	6
NAA	5	3	1	0
IAA	0	1	3	4
0.5 mg·L^-16-BA+NAA	4	3	2	1
0.5 mg·L^-16-BA +0.5 mg·L^-1GA+ NAA	8	3	1	0

Table 3 Media selection of rooting from regenerated plantlet in Aconitum coreanum

Phytohormone composition (mg·L^-1)	No. of root
Phytohormone composition (mg·L^-1)	0.1	0.5	1.0	1.5
IBA	1	2	6	6
NAA	5	3	1	0
IAA	0	1	3	4
0.5 mg·L^-16-BA+NAA	4	3	2	1
0.5 mg·L^-16-BA +0.5 mg·L^-1GA+ NAA	8	3	1	0

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