Chin Bull Bot ›› 2018, Vol. 53 ›› Issue (5): 686-692.doi: 10.11983/CBB17122

• TECHNIQUES AND METHODS • Previous Articles     Next Articles

Plant Regeneration by Inducing Axillary Buds of Sterile Seedlings of Primula denticulata

Zheng Yunfeng, Zhang Xiaoman*(), Liu Xiao   

  1. College of Landscape Architecture and Tourism, Hebei Agriculture University, Baoding 071000, China
  • Received:2017-06-28 Accepted:2017-10-07 Online:2018-11-29 Published:2018-09-10
  • Contact: Zhang Xiaoman E-mail:173610925@qq.com
  • About author:

    † These authors contributed equally to this paper

Abstract:

The axillary buds of sterile seedlings of Primula denticulata were used as explants. By disinfecting the explants, inducing adventitious buds, multiplication of adventitious buds, rooting culture and transplantation, the best medium of each stage was selected to establish the regeneration system of P. denticulata. The optimal medium of P. denticulata for inducing adventitious shoots from explants was MS+1 mg·L-1 NAA+0.1 mg·L-1 6-BA, and the induction rate was 73.33%. The optimal medium for inducing multiple shoots was MS+1 mg·L-1 NAA+0.5 mg·L-1 6-BA, and the proliferation rate was 85.19%. The optimal medium for rooting was 1/2MS+0.2 mg·L-1 NAA, and the rooting rate was 95.59%. Tissue cultured P. denticulata seedlings were transplanted into a mixture of peat and perlite, and the volume ratio was 3:1, with maximum survival rate 96.67%.

Key words: Primula denticulata, axillary bud of sterile seedling, multiple shoots, plant regeneration

Table 1

Effects of different plant growth regulator proportions on adventitious buds induction from the axillary bud of Primula denticulata in MS and 1/2MS medium (means±SE)"

Culture medium Vaccination
number
Adventitious buds
number
Rate of adventitious bud
induction (%)
Growth
condition
MS 45 0 0 -
MS+0.1 mg·L-1 NAA+0.1 mg·L-1 6-BA 45 13 28.89±4.44 a ++
MS+0.5 mg·L-1 NAA+0.1 mg·L-1 6-BA 45 32 71.11±4.44 b +++
MS+1 mg·L-1 NAA+0.1 mg·L-1 6-BA 45 33 73.33±3.85 b ++++
MS+0.1 mg·L-1 NAA+0.5 mg·L-1 6-BA 45 4 8.89±2.22 a +
MS+0.5 mg·L-1 NAA+0.5 mg·L-1 6-BA 45 7 15.56±2.22 a ++
MS+1 mg·L-1 NAA+0.5 mg·L-1 6-BA 45 9 20.00±7.70 a ++
MS+0.1 mg·L-1 NAA+1 mg·L-1 6-BA 45 4 8.89±2.22 a +
MS+0.5 mg·L-1 NAA+1 mg·L-1 6-BA 45 7 15.56±2.22 a +
MS+1 mg·L-1 NAA+1 mg·L-1 6-BA 45 10 22.22±5.88 a ++
MS+0.1 mg·L-1 NAA+2 mg·L-1 6-BA 45 3 6.67±3.85 a +
MS+0.5 mg·L-1 NAA+2 mg·L-1 6-BA 45 14 31.11±5.88 a +
MS+1 mg·L-1 NAA+2 mg·L-1 6-BA 45 16 35.56±2.22 a ++
1/2MS 45 0 0 -
1/2MS+0.1 mg·L-1 NAA+0.1 mg·L-1 6-BA 45 1 2.22±2.22 b +
1/2MS+0.5 mg·L-1 NAA+0.1 mg·L-1 6-BA 45 13 28.89±2.22 b +
1/2MS+1 mg·L-1 NAA+0.1 mg·L-1 6-BA 45 33 73.33±3.85 c ++
1/2MS+0.1 mg·L-1 NAA+0.5 mg·L-1 6-BA 45 3 6.67±3.85 a +
1/2MS+0.5 mg·L-1 NAA+0.5 mg·L-1 6-BA 45 3 6.67±3.85 a +
1/2MS+1 mg·L-1 NAA+0.5 mg·L-1 6-BA 45 31 68.89±2.22 c ++
1/2MS+0.1 mg·L-1 NAA+1 mg·L-1 6-BA 45 1 2.22±2.22 b +
1/2MS+0.5 mg·L-1 NAA+1 mg·L-1 6-BA 45 13 28.89±2.22 b +
1/2MS+1 mg·L-1 NAA+1 mg·L-1 6-BA 45 14 31.11±2.22 b +
1/2MS+0.1 mg·L-1 NAA+2 mg·L-1 6-BA 45 14 31.11±2.22 b +
1/2MS+0.5 mg·L-1 NAA+2 mg·L-1 6-BA 45 31 68.89±2.22 c ++
1/2MS+1 mg·L-1 NAA+2 mg·L-1 6-BA 45 43 95.56±2.22 d +++

Figure 1

Adventitious bud induction, proliferation, rooting and transplanting of Primula denticulata(A) Adventitious buds induced from axillary buds; (B) Adventitious buds multiplication; (C) Rooting; (D) Transplanting the regeneration of Primula denticulata in peat:perlite=3:1 (v/v). Bars=2 cm"

Table 2

Effects of different plant growth regulator proportions on adventitious buds multiplication of Primula denticulata (means±SE)"

Culture medium Vaccination
number
The number of adventitious
buds in proliferation
Shoot multiplication
rate (%)
Growth
condition
MS 27 1 3.70±3.70 a +
MS+0.2 mg·L-1 NAA+0.2 mg·L-1 6-BA 27 2 7.40±7.40 a +
MS+0.2 mg·L-1 NAA+0.5 mg·L-1 6-BA 27 2 7.40±0.00 a +
MS+0.2 mg·L-1 NAA+1 mg·L-1 6-BA 27 2 7.40±3.70 a +
MS+0.2 mg·L-1 NAA+2 mg·L-1 6-BA 27 3 11.11±0.00 ab +
MS+0.6 mg·L-1 NAA+0.2 mg·L-1 6-BA 27 7 25.93±3.70 abc ++
MS+0.6 mg·L-1 NAA+0.5 mg·L-1 6-BA 27 20 74.07±13.35 d +++
MS+0.6 mg·L-1 NAA+1 mg·L-1 6-BA 27 13 48.15±3.70 bcd +++
MS+0.6 mg·L-1 NAA+2 mg·L-1 6-BA 27 9 33.33±0.00 abc ++
MS+1 mg·L-1 NAA+0.2 mg·L-1 6-BA 27 17 62.96±3.70 cd ++
MS+1 mg·L-1 NAA+0.5 mg·L-1 6-BA 27 23 85.19±3.70 d ++++
MS+1 mg·L-1 NAA+1 mg·L-1 6-BA 27 21 77.78±6.42 d +++
MS+1 mg·L-1 NAA+2 mg·L-1 6-BA 27 21 77.78±0.00 d +++

Table 3

Effects of different plant growth regulator proportions on rooting of Primula denticulata adventitious buds (means±SE)"

Culture medium Vaccination
number
Rooting
number
Rooting rate
(%)
Average rooting
number
Growth
condition
MS 27 23 85.19±3.70 bc 5 +++
1/2MS 27 24 88.89±0.00 c 3 +++
MS+0.1 mg·L-1 NAA 27 3 11.11±6.42 a 2 +
MS+0.2 mg·L-1 NAA 27 19 70.37±9.80 abc 3 ++
MS+0.2 mg·L-1 NAA+0.3 mg·L-1 IBA 27 24 88.89±6.42 c 5 +++
MS+0.3 mg·L-1 IBA 27 11 40.74±13.35 abc 2 +
1/2MS+0.1 mg·L-1 IBA 27 18 66.67±11.11 abc 3 +
1/2MS+0.3 mg·L-1 IBA 27 17 62.96±13.35 abc 3 +
1/2MS+1 mg·L-1 IBA 27 7 25.93±9.80 ab 2 +
1/2MS+0.2 mg·L-1 NAA 27 25 95.59±3.70 c 6 ++++
1/2MS+0.2 mg·L-1 NAA+0.3 mg·L-1 IBA 27 5 18.52±9.78 a 2 +

Table 4

Effects of different substrate combinations on tissue culture seedling transplanting of Primula denticulata (means±SE)"

Treatment Transplanting
number
Transplanting survival
number
Transplanting survival
rate (%)
Growth
condition
Peat:perlite=1:1 30 23 76.67±3.33 a ++
Peat:perlite=2:1 30 26 86.67±3.33 a +++
Peat:perlite=3:1 30 29 96.67±3.33 b ++++
Peat:perlite:vermiculite=1:1:1 30 28 93.33±3.33 b +++
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