Optimization of Tissue Culture of Tip-shoot of Strawberry and SRAP Detection of Genetic Stability of Tissue Culture Seedling

doi:10.11924/j.issn.1000-6850.casb2022-1016

Abstract

Abstract:

The aim of this study was to optimize strawberry tip-shoot tissue culture breeding system based on the genetic stability and robustness of tissue culture seedlings, and to provide scientific and technical basis for strawberry factory cultivation. Taking two main strawberry varieties of ‘Fengguan’ and ‘Tianxianzui’ as samples, the concentration gradient of plant growth regulator was set, the basic formula of medium was adjusted, and the tip-shoot differentiation, the morphology of proliferating seedlings, the proliferation coefficient, the leaf and root of rooting seedlings were investigated. The differences of the amplified bands of SRAP markers in plants before and after tissue culture were compared. The medium formula suitable for primary culture was MS+6-BA 0.5 mg/L+ NAA 0.01 mg/L. The medium formula for proliferation culture was MS+ 6-BA 0.3 mg/L+ NAA 0.01 mg/L. And the medium formula for rooting culture was 1/2MS+ calcium nitrate 0.45 g/L. 19 pairs of SRAP primers were used for specific amplification of plant DNA before and after tissue culture, the obtained 72 gene locus bands did not change, and there was no difference in DNA fingerprints formed by the above bands. The tip-shoot tissue culture breeding system of strawberry obtained in this study had the characteristics of few plant growth regulators and low concentration, low proliferation coefficient, short tissue culture time, robust seedling and stable gene.

Key words: strawberry, tip-shoot, tissue culture, sequence-related amplified polymorphism, genetic stability

WANG Pei, LI Junjian, WANG Gaoqi, GENG Tengfei. Optimization of Tissue Culture of Tip-shoot of Strawberry and SRAP Detection of Genetic Stability of Tissue Culture Seedling[J]. Chinese Agricultural Science Bulletin, 2023, 39(34): 41-46.

Figures/Tables 8

References 27

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品种名称	采样点	采样时间
天仙醉	陕西省西安市长安区草莓种植户	2021年11月25日
凤冠	陕西省西安市高陵区东张寺村	2021年11月26日

品种名称	采样点	采样时间
天仙醉	陕西省西安市长安区草莓种植户	2021年11月25日
凤冠	陕西省西安市高陵区东张寺村	2021年11月26日

处理编号	6-BA浓度/(mg/L)	NAA浓度/(mg/L)
1	0.5	0.01
2	0.5	0.05
3	0.5	0.10
4	0.5	0.20
5	0.5	0.30
6	1.0	0.01
7	1.0	0.05
8	1.0	0.10
9	1.0	0.20
10	1.0	0.30
11	2.0	0.01
12	2.0	0.05
13	2.0	0.10
14	2.0	0.20
15	2.0	0.30

处理编号	6-BA浓度/(mg/L)	NAA浓度/(mg/L)
1	0.5	0.01
2	0.5	0.05
3	0.5	0.10
4	0.5	0.20
5	0.5	0.30
6	1.0	0.01
7	1.0	0.05
8	1.0	0.10
9	1.0	0.20
10	1.0	0.30
11	2.0	0.01
12	2.0	0.05
13	2.0	0.10
14	2.0	0.20
15	2.0	0.30

引物方向	引物名称	引物序列(5’-3’)	引物方向	引物名称	引物序列(5’-3’)
正向	me1	TGAGTCCAAACCGGATA	反向	em1	GACTGCGTACGAATTAAT
	me2	TGAGTCCAAACCGGAGC		em2	GACTGCGTACGAATTGAC
	me3	TGAGTCCAAACCGGAAT		em3	GACTGCGTACGAATTTGA
	me4	TGAGTCCAAACCGGACC		em4	GACTGCGTACGAATTAAC
	me5	TGAGTCCAAACCGGAAG		em5	GACTGCGTACGAATTGCA
	me6	TGAGTCCAAACCGGTAA		em6	GACTGCGTACGAATTCAA
	me7	TGAGTCCAAACCGGTCC		em7	GACTGCGTACGAATTCTG
	me8	TGAGTCCAAACCGGTGC		em8	GACTGCGTACGAATTCGA