A micropropagation system for Miscanthus×giganteus based on axillary buds and evaluation of its salt tolerance

doi:10.11686/cyxb2020229

Abstract

Abstract:

The aim of this study was to develop a micropropagation system for Miscanthus×giganteus and to evaluate its salt tolerance. We used nodal segments as explants and determined the optimum developmental stage to collect explants， the best culture medium for axillary bud induction， and the conditions for differentiation of callus and roots. Then， we established a regeneration system via tissue culture. The tissue culture system was based on the axillary shoot growth pathway. The best results were obtained by collecting nodal segments about 15 mm in length from plants at the 5-6 leaf stage， and sterilizing them by surface-spraying with 75% ethanol followed by immersion in 1.0% sodium hypochlorite for 20 min. Under these conditions， the pollution rate was zero and the adventitious bud induction rate was 85.2%. The most suitable medium for induction and subculturing of callus was Murashige and Skoog （MS） medium containing 5.0 mg?L^-1 6-benzylaminopurine and 0.25 mg?L^-1 1-naphthaleneacetic acid； the multiplication rate on this medium was 320.3%. For rooting and growth of cluster buds， the most suitable medium was MS medium containing 5.0 g?L^-1 activated carbon. The salt tolerance of Miscanthus×giganteus plantlets was evaluated under hydroponic culture conditions. Increasing concentrations of NaCl in the hydroponic culture medium led to decreased growth rate of the plantlets. The plantlets were able to survive normally in medium containing 0.8% NaCl in the hydroponic system， indicative of high salt tolerance. These results provide a theoretical and technical basis for the rapid propagation of Miscanthus×giganteus that can be further developed for genetic transformation and the production of Miscanthus×giganteus lines for cultivation in salinized areas.

Key words: Miscanthus×giganteus, nodal segments, micropropagation, salt tolerance

Ye WANG, Hui-ping CHEN, Run-zhi LI, Zhen PENG, Xi-feng FAN, Ju-ying WU, Liu-sheng DUAN. A micropropagation system for Miscanthus×giganteus based on axillary buds and evaluation of its salt tolerance[J]. Acta Prataculturae Sinica, 2021, 30(6): 214-220.

Figures/Tables 6

References 22

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消毒处理 Treatment	出芽率 Bud induction rate	污染率 Pollution rate
D₁	70.0±8.75b	0b
D₂	23.3±5.16c	0b
D₃	72.5±6.23b	12.5±2.57a
D₄	85.2±9.45a	0b

消毒处理 Treatment	出芽率 Bud induction rate	污染率 Pollution rate
D₁	70.0±8.75b	0b
D₂	23.3±5.16c	0b
D₃	72.5±6.23b	12.5±2.57a
D₄	85.2±9.45a	0b

6-BA浓度 6-BA concentration (mg·L^-1)	出芽率 Bud induction rate (%)	繁殖系数Multiplication coefficient (%)
6-BA浓度 6-BA concentration (mg·L^-1)	出芽率 Bud induction rate (%)	30 d	60 d	90 d
1.0	84.09±6.25a	145.83±15.69c	156.34±23.69c	126.16±25.34c
3.0	82.75±9.69a	187.50±21.35b	225.13±28.53b	243.78±31.25b
5.0	83.95±8.24a	320.29±20.45a	362.58±32.78a	342.13±29.14a
7.0	83.25±7.53a	184.69±24.62b	248.15±31.44b	276.24±30.25b

6-BA浓度 6-BA concentration (mg·L^-1)	出芽率 Bud induction rate (%)	繁殖系数Multiplication coefficient (%)
6-BA浓度 6-BA concentration (mg·L^-1)	出芽率 Bud induction rate (%)	30 d	60 d	90 d
1.0	84.09±6.25a	145.83±15.69c	156.34±23.69c	126.16±25.34c
3.0	82.75±9.69a	187.50±21.35b	225.13±28.53b	243.78±31.25b
5.0	83.95±8.24a	320.29±20.45a	362.58±32.78a	342.13±29.14a
7.0	83.25±7.53a	184.69±24.62b	248.15±31.44b	276.24±30.25b

NaCl浓度 NaCl concentration (%)	生长速率 Growth rate (cm·d^-1)		平均速率Average growth rate (cm·d^-1)	生物量Fresh weight (g·plant^-1)
NaCl浓度 NaCl concentration (%)	7 d	14 d	平均速率Average growth rate (cm·d^-1)	生物量Fresh weight (g·plant^-1)
0	0.15±0.030a	0.17±0.02a	0.160a	0.31±0.09a
0.2	0.12±0.010a	0.14±0.04b	0.130a	0.35±0.07a
0.4	0.02±0.004b	0.06±0.01c	0.040b	0.23±0.04b
0.6	0.02±0.009b	0.03±0.01c	0.025b	0.19±0.02b
0.8	0.01±0.007c	0c	0.005c	0.14±0.03c