Optimization of Tissue Culture and Plant Regeneration System of Mature Embryo of Leymus chinensis

doi:10.11983/CBB19223

Abstract

Abstract: Leymus chinensis is a heterotetraploid grass. It is hard to use genetic transformation method for improving breeding since the efficiency of regeneration using callus induced from mature embryo is very low. In this study, we optimized the protocol of culturing mature embryo with L. chinensis as explants by screening optimum plant hormone concentrations, light conditions and culturing temperatures at different culturing stages including callus induction, differentiation, rooting and transplanting. Our results showed at the callus induction stage, the induction rate could reach up to 74.1% under 2.0 mg·L ^-12,4-D, variable temperature and dark culture. The differentiation rate could reach up to 57.1% with optimum condition of 1.0 mg·L ^-16-BA and 1.0 mg·L ^-1NAA. In the rooting stage, the survival rate was 100% when cultured with 0.25 mg·L ^-1NAA.

Key words: mature embryo, Leymus chinensis, callus, plant hormone, plant regeneration

Yan Xiao,Zhenxing Wang,Dongming Li,Yanhua Qi, Enhebayaer. Optimization of Tissue Culture and Plant Regeneration System of Mature Embryo of Leymus chinensis[J]. Chinese Bulletin of Botany, 2020, 55(2): 192-198.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB19223

https://www.chinbullbotany.com/EN/Y2020/V55/I2/192

Figures/Tables 4

References 7

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6-BA/NAA (mg·L^-1)	Callus number	The number of adventitious buds in proliferation	The number of roots in reproduction	Differentiation efficiency (%)
0.5/0.25	21	0	0	0
1.0/0.25	21	1A	4+	4.8±4.8 ab
2.0/0.25	21	0	0	0
3.0/0.25	21	3AB	0	14.3±8.3 abcd
4.0/0.25	21	8AB	0	38.1±4.8 efgh
5.0/0.25	21	4A	0	19.1±4.8 abcde
0.5/0.5	21	0	0	0
1.0/0.5	21	7BC	6+	33.3±9.5 defg
2.0/0.5	21	10BC	5+	47.6±9.5 gh
3.0/0.5	21	7AB	4	33.3±4.8 defg
4.0/0.5	21	10AB	0	47.6±9.5 gh
5.0/0.5	21	3BC	0	14.3±0 abcd
0.5/1.0	21	2A	8+	9.5±4.8 abc
1.0/1.0	21	12AB	4	57.1±4.8 h
2.0/1.0	21	9A	0	42.9±8.3 fgh
3.0/1.0	21	5AC	3	23.8±8.3 bcdef
4.0/1.0	21	4A	1+	19.1±4.8 abcde
5.0/1.0	21	5A	0	23.8±12.6 bcdef
0.5/2.0	21	5ABC	9+	23.8±4.8 bcdef
1.0/2.0	21	1A	5	4.8±4.8 ab
2.0/2.0	21	0	0	0
3.0/2.0	21	2A	1+	9.5±4.8 abc
4.0/2.0	21	2AB	4	9.5±4.8 abc
5.0/2.0	21	0	0	0
0.5/3.0	21	1A	3	4.8±4.8 ab
1.0/3.0	21	4A	3	19.1±4.8 abcde
2.0/3.0	21	7AB	3	33.3±4.8 defg
3.0/3.0	21	10ABC	1	47.6±9.5 gh
4.0/3.0	21	0	0	0
5.0/3.0	21	0	0	0
0.5/4.0	21	6A	3	28.6±8.2 cdefg
1.0/4.0	21	0	0	0
2.0/4.0	21	8AB	2	38.1±4.8 efgh
3.0/4.0	21	0	0	0
4.0/4.0	21	3AB	5	14.3±8.2 abcd
5.0/4.0	21	6AB	0	28.6±8.2 cdefg

6-BA/NAA (mg·L^-1)	Callus number	The number of adventitious buds in proliferation	The number of roots in reproduction	Differentiation efficiency (%)
0.5/0.25	21	0	0	0
1.0/0.25	21	1A	4+	4.8±4.8 ab
2.0/0.25	21	0	0	0
3.0/0.25	21	3AB	0	14.3±8.3 abcd
4.0/0.25	21	8AB	0	38.1±4.8 efgh
5.0/0.25	21	4A	0	19.1±4.8 abcde
0.5/0.5	21	0	0	0
1.0/0.5	21	7BC	6+	33.3±9.5 defg
2.0/0.5	21	10BC	5+	47.6±9.5 gh
3.0/0.5	21	7AB	4	33.3±4.8 defg
4.0/0.5	21	10AB	0	47.6±9.5 gh
5.0/0.5	21	3BC	0	14.3±0 abcd
0.5/1.0	21	2A	8+	9.5±4.8 abc
1.0/1.0	21	12AB	4	57.1±4.8 h
2.0/1.0	21	9A	0	42.9±8.3 fgh
3.0/1.0	21	5AC	3	23.8±8.3 bcdef
4.0/1.0	21	4A	1+	19.1±4.8 abcde
5.0/1.0	21	5A	0	23.8±12.6 bcdef
0.5/2.0	21	5ABC	9+	23.8±4.8 bcdef
1.0/2.0	21	1A	5	4.8±4.8 ab
2.0/2.0	21	0	0	0
3.0/2.0	21	2A	1+	9.5±4.8 abc
4.0/2.0	21	2AB	4	9.5±4.8 abc
5.0/2.0	21	0	0	0
0.5/3.0	21	1A	3	4.8±4.8 ab
1.0/3.0	21	4A	3	19.1±4.8 abcde
2.0/3.0	21	7AB	3	33.3±4.8 defg
3.0/3.0	21	10ABC	1	47.6±9.5 gh
4.0/3.0	21	0	0	0
5.0/3.0	21	0	0	0
0.5/4.0	21	6A	3	28.6±8.2 cdefg
1.0/4.0	21	0	0	0
2.0/4.0	21	8AB	2	38.1±4.8 efgh
3.0/4.0	21	0	0	0
4.0/4.0	21	3AB	5	14.3±8.2 abcd
5.0/4.0	21	6AB	0	28.6±8.2 cdefg