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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2018, Vol. 38 Issue (10): 48-54    DOI: 10.13523/j.cb.20181006
Orginal Article     
Establishment of High Frequency Regeneration via Leaf Explants of ‘Guichang’ Kiwifruit (Actinidia chinensis)
Xu-peng ZHAO,Xiao-peng ZHAO,Hao SHI,Xue-mei CHEN,Ting JIANG,Yan LIU()
Guiyang University, Guiyang 550005, China
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Abstract  

A high efficient in vitro regeneration system was developed from leaf explants of ‘Guichang’ kiwifruit (Actinidia chinensis) and the multiplication coefficient and rooting rate of adventitious buds were also optimized. Results revealed that the adventitious buds developing directly from leaf explants were noticed after 30d of culture. The maximum regeneration frequency of adventitious buds is 95.8% and 15.7 shoots was observed in each leaf explants when MS medium was supplemented with 4.0mg/L 6-BA+0.4mg/L NAA. The optimal culture medium for bud proliferation is MS+3.0mg/L 6-BA+0.3mg/L NAA+0.2mg/L GA3 and the proliferation coefficient reached 8.15 in every subculture during 1 to 6 subcultures. On the rooting medium with 1/2 MS+1.0mg/L IBA for 15d, the adventitious plantlets were successively transferred into agar power and matrix perlite supplied with 1/2 MS liquid medium for 15d and 95.83% of them rooted and the roots were also heavy and strong very much. 49 out of 50 plantlets (99%) survived acclimatization in the greenhouse after transplanting them to the Mixed matrix of pearlite and soil(1∶4)for two weeks. In conclusion, a highly efficient regeneration protocol via leaf explants of ‘Guichang’ kiwifruit was successfully established and it may provide theoretical and technical guidance for micropropagation and genetic transformation studies of ‘Guichang’ kiwifruit.

Key words： ‘Guichang’ kiwifruit (Actinidia chinensis);      Leaves      Adventitious buds      Plant regeneration     
Received: 01 July 2018      Published: 09 November 2018
ZTFLH:  Q813.1+2  
Corresponding Authors: Yan LIU     E-mail: gyly68@sina.com
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Xu-peng ZHAO
Xiao-peng ZHAO
Hao SHI
Xue-mei CHEN
Ting JIANG
Yan LIU
Cite this article:

Xu-peng ZHAO,Xiao-peng ZHAO,Hao SHI,Xue-mei CHEN,Ting JIANG,Yan LIU. Establishment of High Frequency Regeneration via Leaf Explants of ‘Guichang’ Kiwifruit (Actinidia chinensis). China Biotechnology, 2018, 38(10): 48-54.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20181006     OR     https://manu60.magtech.com.cn/biotech/Y2018/V38/I10/48

编号
No.		 植物生长调节剂/mg/L
Plant growth regulators		 接种外植体数
No. of leaves		 出芽叶片数/个
Explants No.of
buds formation		 出芽率/%
Induction rate of
adventitious buds		 平均芽长/cm
Average No.of buds
A1 6-BA 3.0+NAA 0.2 24 9.3±0.88d 37.5±4.17f 3.7±0.33d
A2 6-BA 4.0+NAA 0.2 24 10.7±1.20d 45.8±6.36ef 7.3±0.67c
A3 6-BA 5.0+NAA 0.2 24 15.0±2.08c 62.5±8.67de 4.3±0.67d
A4 6-BA 3.0+NAA 0.4 24 16.0±1.73bc 66.7±7.22cd 10.7±1.45b
A5 6-BA 4.0+NAA 0.4 24 23.0±1.00a 95.8±4.17a 15.7±0.88a
A6 6-BA 5.0+NAA 0.4 24 19.7±1.20ab 83.3±4.17abc 10.3±0.88b
A7 6-BA 3.0+NAA 0.6 24 16.3±1.76bc 70.8±4.81bcd 11.0±1.25b
A8 6-BA 4.0+NAA 0.6 24 23.0±0.58a 95.8±2.41a 11.3±1.20b
A9 6-BA 5.0+NAA 0.6 24 21.0±1.53a 87.5±6.36ab 10.0±0.58bc
Table 1 Effects of plant growth regulators on dedifferentiation of buds from leaves of ‘Guichang’ kiwifruit
编号
No.		 植物生长调节剂/mg/L
Plant growth regulators		 增殖率/%
Multiplied
rate/%		 各代繁殖系数/倍 Propagation coefficients
in clonal generations /Times		 平均增殖倍数Average
multiplied times
1 2 3 4 5 6
B1 6-BA 2.0+NAA 0.3+GA3 0.2 73.6±3.67b 2.11 2.07 2.00 1.89 1.88 1.97 1.99±0.03e
B2 6-BA 3.0+NAA 0.3+GA3 0.2 100±0.00a 8.14 8.22 8.17 8.29 8.03 8.03 8.15±0.04a
B3 6-BA 4.0+NAA 0.3+GA3 0.2 100±0.00a 5.36 5.40 5.38 5.44 5.25 5.23 5.34±0.03d
B4 6-BA 2.0+NAA 0.6+GA3 0.2 61.1±3.67b 1.76 1.79 1.72 1.86 1.78 1.76 1.78±0.02f
B5 6-BA 3.0+NAA 0.6+GA3 0.2 95.8±2.41a 5.68 5.67 5.53 5.75 5.60 5.60 5.64±0.03c
B6 6-BA 4.0+NAA 0.6+GA3 0.2 100±0.00a 6.21 6.22 6.06 6.19 5.97 6.14 6.13±0.04b
Table 2 Effects of plant growth regulators on multiplication of adventitious buds of ‘Guichang’ kiwifruit
培养基
编号
Media No.		 吲哚丁酸
IBA/mg-1		 接种不定芽数
No. of adventitious
bud inoculated		 生根不定芽数
No. of adventitious
bud rooted		 生根率/%
Rate of rooting		 平均根数/条
Average No.
of root		 平均根长/cm
Average length
of root
C1 0 24 10.33±0.88e 43.06±3.67e 0.78±0.12d 0.44±0.08b
C2 0.4 24 12.00±2.31de 50.00±9.62de 1.22±0.22d 0.51±0.05b
C3 0.6 24 15.67±1.45cd 65.28±6.05cd 2.01±0.14c 0.85±0.01a
C4 0.8 24 20.33±1.20ab 84.72±5.01ab 3.47±0.26b 0.86±0.01a
C5 1.0 24 23.67±0.33a 98.61±1.39a 5.31±0.15a 0.81±0.01a
C6 1.2 24 19.33±0.67bc 80.56±2.78bc 3.53±0.10b 0.76±0.02a
Table 3 Effects of IBA concentration on rooting of adventitious buds of ‘Guichang’ Kiwifruit
Fig.1 Plant regeneration from leaf of ‘Guichang’ kiwifruit (a) Leaves inoculated two weeks later (b) Adventitious buds directly regenerated from leaves(The red part is leaf) (c) Adventitious buds cultured in light(The red part is leaf) (d) Adventitious buds proliferated in one subculture (e) Thick root system of test tube plantlet (f) Acclimatized plantlets after transplanting to matrix soil for two weeks
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