Establishment of Tissue Culture System for Axillary Bud Regeneration of Primula × pubescens

doi:10.11983/CBB21076

Abstract

Abstract: The effects of different concentrations of hormone on Primula × pubescens tissue culture were studied through callus induction and differentiation, adventitious bud proliferation culture and rooting. The optimal medium at different stages was selected to optimize the regeneration tissue culture system of Primula × pubescens. Our results showed that the best medium for callus induction and differentiation was MS+0.2 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA, with 84% induction rate and 67% budding rate; the optimal medium for adventitious bud proliferation was MS+0.5 mg∙L^-1 NAA+0.2 mg∙L^-1 6-BA, and the proliferation rate was 67%; MS+0.2 mg∙L^-1 NAA medium was the best for rooting and elongation of tissue culture seedlings. The average number of roots per plant was 9, and the rooting rate was 70%. This study established a highly effect tissue culture and regeneration system of Primula × pubescens, which laid a foundation for genetic research and germplasm innovation of other Primula plants.

Key words: Primula × pubescens, aseptic axillary bud, tissue culture, plant regeneration

Mengyue Li, Liu Liu, Yan Liu, Xiaoman Zhang. Establishment of Tissue Culture System for Axillary Bud Regeneration of Primula × pubescens[J]. Chinese Bulletin of Botany, 2021, 56(6): 732-739.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB21076

https://www.chinbullbotany.com/EN/Y2021/V56/I6/732

Figures/Tables 4

References 22

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No.	Culture medium	Inoculation number	Callus induction rate (%)	Callus growth situation	Callus differentiation rate (%)	Cluster bud growth situation
1	1/2MS+0.1 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	6.67±3.33 fg	+	0	-
2	1/2MS+0.1 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	15.67±2.96 efg	+	3.33±3.33 cd	+
3	1/2MS+0.1 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	6.67±6.67 fg	+	6.67±6.67 cd	+
4	1/2MS+0.2 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	63.33±8.81 abcd	++	13.33±3.33 bcd	+
5	1/2MS+0.2 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	43.33±12.02 bcde	+	20.00±11.55 bcd	+
6	1/2MS+0.2 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	76.33±8.21 ab	++	34.70±11.78 abcd	+
7	1/2MS+0.5 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	0	-	10.00±10.00 bcd	+
8	1/2MS+0.5 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	40.00±5.77 cdef	+	0	-
9	1/2MS+0.5 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	6.00±3.05 fg	+	67.00±11.36 a	++
10	1/2MS+1.0 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	6.67±3.33 fg	+	26.70±3.33 bcd	+
11	1/2MS+1.0 mg∙L^-1 NAA+1.0 mg∙L^-16-BA	30	0	-	23.33±3.33 bcd	+
12	1/2MS+1.0 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	0	-	30.00±30.00 abcd	++
13	MS+0.1 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	83.33±8.82 a	++++	40.00±11.55 abc	+++
14	MS+0.1 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	76.67±6.67 ab	++++	3.33±3.33 cd	+
15	MS+0.1 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	50.00±11.55 abcde	+++	16.70±3.33 bcd	+
16	MS+0.2 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	70.67±1.20 abc	++++	38.70±1.15 abc	++
17	MS+0.2 mg∙L^-1 NAA+1.0 mg∙L^-16-BA	30	84.00±11.37 a	++++	67.00±11.36 a	++++
18	MS+0.2 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	38.33±7.27 cdef	++	47.30±6.36 ab	+++
19	MS+0.5 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	70.00±5.77 abc	++++	0	-
20	MS+0.5 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	63.33±8.82 abcd	+++	0	-
21	MS+0.5 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	16.00±3.06 efg	+	6.67±6.67 cd	+
22	MS+1.0 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	32.00±2.00 defg	++	17.00±8.89 bcd	+
23	MS+1.0 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	0	-	6.67±6.67 cd	+
24	MS+1.0 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	0	-	13.33±8.82 bcd	+

No.	Culture medium	Inoculation number	Callus induction rate (%)	Callus growth situation	Callus differentiation rate (%)	Cluster bud growth situation
1	1/2MS+0.1 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	6.67±3.33 fg	+	0	-
2	1/2MS+0.1 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	15.67±2.96 efg	+	3.33±3.33 cd	+
3	1/2MS+0.1 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	6.67±6.67 fg	+	6.67±6.67 cd	+
4	1/2MS+0.2 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	63.33±8.81 abcd	++	13.33±3.33 bcd	+
5	1/2MS+0.2 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	43.33±12.02 bcde	+	20.00±11.55 bcd	+
6	1/2MS+0.2 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	76.33±8.21 ab	++	34.70±11.78 abcd	+
7	1/2MS+0.5 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	0	-	10.00±10.00 bcd	+
8	1/2MS+0.5 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	40.00±5.77 cdef	+	0	-
9	1/2MS+0.5 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	6.00±3.05 fg	+	67.00±11.36 a	++
10	1/2MS+1.0 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	6.67±3.33 fg	+	26.70±3.33 bcd	+
11	1/2MS+1.0 mg∙L^-1 NAA+1.0 mg∙L^-16-BA	30	0	-	23.33±3.33 bcd	+
12	1/2MS+1.0 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	0	-	30.00±30.00 abcd	++
13	MS+0.1 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	83.33±8.82 a	++++	40.00±11.55 abc	+++
14	MS+0.1 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	76.67±6.67 ab	++++	3.33±3.33 cd	+
15	MS+0.1 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	50.00±11.55 abcde	+++	16.70±3.33 bcd	+
16	MS+0.2 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	70.67±1.20 abc	++++	38.70±1.15 abc	++
17	MS+0.2 mg∙L^-1 NAA+1.0 mg∙L^-16-BA	30	84.00±11.37 a	++++	67.00±11.36 a	++++
18	MS+0.2 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	38.33±7.27 cdef	++	47.30±6.36 ab	+++
19	MS+0.5 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	70.00±5.77 abc	++++	0	-
20	MS+0.5 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	63.33±8.82 abcd	+++	0	-
21	MS+0.5 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	16.00±3.06 efg	+	6.67±6.67 cd	+
22	MS+1.0 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	32.00±2.00 defg	++	17.00±8.89 bcd	+
23	MS+1.0 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	0	-	6.67±6.67 cd	+
24	MS+1.0 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	0	-	13.33±8.82 bcd	+

No.	Culture medium	Inoculation number	Multiplication rate (%)	Growth situation
1	1/2MS+0.2 mg∙L^-1 NAA+0.2 mg∙L^-1 6-BA	30	0	-
2	1/2MS+0.2 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	3.33±3.33 cd	+
3	1/2MS+0.2 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	6.67±6.67 cd	+
4	1/2MS+0.2 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	13.33±3.33 bcd	+
5	1/2MS+0.5 mg∙L^-1 NAA+0.2 mg∙L^-1 6-BA	30	20.00±11.55 bcd	+
6	1/2MS+0.5 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	34.67±11.78 abcd	+
7	1/2MS+0.5 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	10.00±10.00 bcd	+
8	1/2MS+0.5 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	0	-
9	1/2MS+1.0 mg∙L^-1 NAA+0.2 mg∙L^-1 6-BA	30	67.00±11.36 a	++
10	1/2MS+1.0 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	26.67±3.33 bcd	+
11	1/2MS+1.0 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	23.33±3.33 bcd	+
12	1/2MS+1.0 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	30.00±0.00 abcd	+
13	MS+0.2 mg∙L^-1 NAA+0.2 mg∙L^-1 6-BA	30	40.00±11.55 abc	+++
14	MS+0.2 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	3.33±3.33 cd	+
15	MS+0.2 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	16.67±3.33 bcd	++
16	MS+0.2 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	38.00±1.15 abc	++
17	MS+0.5 mg∙L^-1 NAA+0.2 mg∙L^-1 6-BA	30	67.00±11.35 a	++++
18	MS+0.5 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	47.33±6.36 ab	+++
19	MS+0.5 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	0	-
20	MS+0.5 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	0	-
21	MS+1.0 mg∙L^-1 NAA+0.2 mg∙L^-1 6-BA	30	6.67±6.67 cd	+
22	MS+1.0 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	17.00±8.89 bcd	++
23	MS+1.0 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	6.67±6.67 cd	+
24	MS+1.0 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	13.33±8.82 bcd	++

No.	Culture medium	Inoculation number	Multiplication rate (%)	Growth situation
1	1/2MS+0.2 mg∙L^-1 NAA+0.2 mg∙L^-1 6-BA	30	0	-
2	1/2MS+0.2 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	3.33±3.33 cd	+
3	1/2MS+0.2 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	6.67±6.67 cd	+
4	1/2MS+0.2 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	13.33±3.33 bcd	+
5	1/2MS+0.5 mg∙L^-1 NAA+0.2 mg∙L^-1 6-BA	30	20.00±11.55 bcd	+
6	1/2MS+0.5 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	34.67±11.78 abcd	+
7	1/2MS+0.5 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	10.00±10.00 bcd	+
8	1/2MS+0.5 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	0	-
9	1/2MS+1.0 mg∙L^-1 NAA+0.2 mg∙L^-1 6-BA	30	67.00±11.36 a	++
10	1/2MS+1.0 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	26.67±3.33 bcd	+
11	1/2MS+1.0 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	23.33±3.33 bcd	+
12	1/2MS+1.0 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	30.00±0.00 abcd	+
13	MS+0.2 mg∙L^-1 NAA+0.2 mg∙L^-1 6-BA	30	40.00±11.55 abc	+++
14	MS+0.2 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	3.33±3.33 cd	+
15	MS+0.2 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	16.67±3.33 bcd	++
16	MS+0.2 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	38.00±1.15 abc	++
17	MS+0.5 mg∙L^-1 NAA+0.2 mg∙L^-1 6-BA	30	67.00±11.35 a	++++
18	MS+0.5 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	47.33±6.36 ab	+++
19	MS+0.5 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	0	-
20	MS+0.5 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	0	-
21	MS+1.0 mg∙L^-1 NAA+0.2 mg∙L^-1 6-BA	30	6.67±6.67 cd	+
22	MS+1.0 mg∙L^-1 NAA+0.5 mg∙L^-1 6-BA	30	17.00±8.89 bcd	++
23	MS+1.0 mg∙L^-1 NAA+1.0 mg∙L^-1 6-BA	30	6.67±6.67 cd	+
24	MS+1.0 mg∙L^-1 NAA+2.0 mg∙L^-1 6-BA	30	13.33±8.82 bcd	++

No.	Culture medium	Inoculation number	Rooting number	Rooting rate (%)	Average number of bars	Growth situation
1	MS	20	13	65.00±5.00 a	4	+
2	MS+0.2 mg∙L^-1 IBA	20	10	50.00±5.77 ab	2	+
3	MS+0.3 mg∙L^-1 IBA	20	12	60.00±8.17 ab	5	+
4	MS+0.1 mg∙L^-1 NAA	20	2	10.00±5.77 c	1	+
5	MS+0.1 mg∙L^-1 NAA+0.2 mg∙L^-1 IBA	20	1	5.00±5.00 c	2	+
6	MS+0.1 mg∙L^-1 NAA+0.3 mg∙L^-1 IBA	20	5	25.00±9.57 bc	4	+
7	MS+0.2 mg∙L^-1 NAA	20	14	70.00±5.77 a	9	++++
8	MS+0.2 mg∙L^-1 NAA+0.2 mg∙L^-1IBA	20	10	50.00±5.77 ab	3	++
9	MS+0.2 mg∙L^-1 NAA+0.3 mg∙L^-1 IBA	20	11	55.00±12.58 ab	10	+++