Crops ›› 2017, Vol. 33 ›› Issue (6): 60-64.doi: 10.16035/j.issn.1001-7283.2017.06.011

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Establisment of Highly Effective in Vitro Regeneration System of Dioscorea alata L.

Xu Yan,Zhang Shaolong,Chen Yuhua,Li Hui   

  1. Shantou Agricultural Science Institute,Shantou 515021,Guangdong,China
  • Received:2017-07-11 Revised:2017-08-24 Online:2017-12-15 Published:2018-08-26
  • Contact: Shaolong Zhang

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Abstract:

In order to establish an efficient in vitro regeneration system of Dioscorea alata L., the branch nodes were as explants and the key factors affecting plant regeneration in vitro were studied. As results, the best sterilization method for explants was 75% alcohol used for 30 seconds and 0.1% HgCl2 adding a few drops of Tween-20 for 11 minutes. The optimum medium for inducing buds from explants was MS+0.8mg/L 6-BA+0.1mg/L NAA, resulting in the induction rate 80.0%. The best multiplication medium was MS+1.2mg/L 6-BA, the multiplication coefficient was 2.15. The optimum rooting medium was 1/2 MS+0.5mg/L NAA+0.2mg/L IBA+0.3g/L AC, with rooting rate 84.4% after 30 days and 98.9% after 50 days. The surviving rate of plantlets was above 80.0%, after transplanting into the mixture of 1/2 pearlite and 1/2 coconut shell powder. The plantlets grew well and produced tubers. It was established an efficient in vitro regeneration system of Dioscorea alata L., with the branch nodes as explants and axillary bud proliferation method, which laid a foundation for rapid propagation, germplasm resources conservation and cultivar improvement of Dioscorea alata L..

Key words: Dioscorea alata L., Tissue culture, In vitro regeneration

Table 1

Effects of different sterilization methods on explants of Dioscorea alata L."

0.1% HgCl2消毒时间(min)
Sterilization time of 0.1% HgCl2		 污染率(%)
Contamination rate		 诱导率(%)
Induction rate
9 10.0 55.0
11 5.0 52.5
13 5.0 50.0

Table 2

Effects of different hormone combinations on explants of Dioscorea alata L."

编号
Code		 处理 [6-BA(mg/L)+NAA(mg/L)]
Treatment		 诱导率(%)
Induction rate		 芽异常率(%)
Abnormality rate		 长势
Growth potential		 芽高(cm)
Bud height
对照（CK） 0.0+0.0 35.0±7.6d 0.0±0.0 一般 0.1~0.5
T1 0.5+0.05 40.0±5.0cd 3.0±3.0 好 0.1~7.0
T2 0.8+0.05 48.3±4.4bcd 5.9±0.2 较好 0.1~5.0
T3 1.2+0.05 70.0±5.0ab 17.2±2.4 较好 0.1~5.5
T4 1.5+0.05 61.7±7.3abc 21.0±5.2 较好 0.1~4.0
T5 0.5+0.10 38.3±8.8cd 5.8±3.4 较好 0.1~5.0
T6 0.8+0.10 80.0±5.8a 7.5±0.4 好 0.1~6.0
T7 1.2+0.10 68.3±6.0ab 15.2±6.4 好 0.1~5.5
T8 1.5+0.10 65.0±14.4ab 19.8±7.3 较好 0.1~5.0

Table 3

Effects of different hormone combinations on subculture multiplication"

编号
Code		 处理 [6-BA(mg/L)+NAA(mg/L)]
Treatment		 增殖系数
Coefficient of multiplication		 芽异常率(%)
Abnormality rate		 长势
Growth potential		 芽高(cm)
Bud height
ST1 0.5+0.0 1.65±0.15b 2.0±1.0d 较好 0.1~4.5
ST2 0.8+0.0 1.73±0.16b 4.5±2.3cd 好 0.1~5.0
ST3 1.2+0.0 2.15±0.03a 6.2±0.8cd 好 0.1~5.0
ST4 1.5+0.0 1.77±0.12ab 8.4±1.2bcd 好 0.1~6.0
ST5 2.0+0.0 1.85±0.09ab 14.6±3.0ab 较好 0.1~4.0
ST6 0.5+0.1 1.53±0.17b 3.0±3.0d 好 0.1~5.5
ST7 0.8+0.1 1.58±0.09b 9.5±1.7abcd 好 0.1~5.0
ST8 1.2+0.1 1.67±0.16b 11.6±2.6abc 好 0.1~6.0
ST9 1.5+0.1 1.72±0.07b 14.4±4.2ab 好 0.1~5.0
ST10 2.0+0.1 1.73±0.13b 17.2±2.9a 较好 0.1~4.0

Table 4

Effects of mediums on roots formation"

处理Treatment
[NAA (mg/L)+IBA(mg/L)+AC(g/L)]		 生根率(%)
Rooting rate		 平均根数
Average root number		 基部生根率(%)
Rooting rate of base		 长势
Growth potential		 根长(cm)
Root length
30d 50d 30d 50d 50d 50d 50d
0.5+0.2+0.3 84.4±11.0a 98.9±1.1a 2.76±0.10 3.60±0.22a 30.0±8.8a 良好 0.1~6.5
0.5+1.0+0.3 72.2±7.3a 92.2±4.0a 3.34±0.04 3.86±0.30a 16.7±7.0a 良好 0.1~7.0
0.5+1.0+0.5 66.7±5.1a 98.9±1.1a 3.13±0.25 3.64±0.22a 20.0±8.4a 良好 0.1~6.5

Table 5

Effects of mediums on plantlet transplant surviving rate %"

基质
Medium		 成活率Surviving rate
重复ⅠRepeatⅠ 重复ⅡRepeat Ⅱ
园土∶椰糠=1∶1 70.0 40.0
珍珠岩∶椰糠=1∶1 90.0 80.0

Table 6

Survival rate of plantlets transplanted to field"

移植日期
Transplant date		 移植苗数
Transplant number		 成活率(%)
Surviving rate
4月15日 50 84.0
5月6日 80 81.3
5月16日 81 77.8
5月29日 102 82.4
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