Tissue Culture and Plant Regeneration of Leaves of Primulina guigangensis

doi:10.3969/j.issn.1000-2561.2019.01.015

Abstract

Abstract:

The effects of different media on the primary induction, adventitious buds proliferation, callus differentiation and rooting culture were investigated using the leaves of P. guigangensis as the explants. The results showed that the rip cutting was the suitable method; the suitable medium for adventitious buds regeneration was MS+6-BA 4.0 mg/L+IAA 1.5 mg/L; the suitable medium for callus induction was MS+6-BA 3.0-5.0 mg/L+2,4-D 0.5-1.5 mg/L; both rate of adventitious buds induction and rate of callus induction were 100.00%; the suitable medium for callus differentiation was MS+KT 1.0 mg/L+NAA 0.2 mg/L+potato 30 g/L+banana 30 g/L+apple 20 g/L+coconut juice 100 mL/L, and the differentiation coefficient was 12.64; the suitable medium for adventitious buds proliferation was MS+ZT 1.0 mg/L+NAA 0.10 mg/L+potato 30 g/L+banana 30 g/L+apple 20 g/L+coconut juice 100 mL/L, and the multiplication coefficient was 8.55; the suitable medium for rooting culture was 3/4 MS+NAA 0.01-0.05 mg/L+active carbon 1.0-3.0 g/L and the rooting rate was 100%. It was concluded that the leaves rip cutting could be used to establish the tissue culture system of P. guigangensis by the regeneration methods of adventitious buds and callus which had high induction and proliferation rates of adventitious buds and good rooting growth.

Key words: Primulina guigangensis, tissue culture, regeneration system, natural extracts

YAN Haixia,HUANG Changyan,ZHANG Zibin,CUI Xueqiang,DENG Jieling,GUAN Shikai,BU Zhaoyang. Tissue Culture and Plant Regeneration of Leaves of Primulina guigangensis[J]. Chinese Journal of Tropical Crops, 2019, 40(1): 98-106.

Figures/Tables 10

References 21

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处理 Treatment	切割方式 Method	培养基 Medium	接种数 Number of inoculated	不定芽诱导率 Rate of adventitious bud induction/%	愈伤诱导率 Rate of callus induction/%	不定芽 Number of adventitious bud	诱导时间 Time of induction/d
C₁	纵切	MS+6-BA 3.0 mg/L+IAA 1.0 mg/L	100	100.00^a	0.00^d	116.33^cd	29.33^a
C₂	纵切	MS+6-BA 4.0 mg/L+IAA 1.5 mg/L	100	100.00^a	0.00^d	156.00^a	24.67^a
C₃	纵切	MS+6-BA 5.0 mg/L+IAA 2.0 mg/L	100	100.00^a	0.00^d	138.67^b	24.00^a
C₄	纵切	MS+6-BA 3.0mg/L+2,4-D 0.5mg/L	100	0.00^c	100.00^a	0.00^f	30.00^a
C₅	纵切	MS+6-BA 4.0mg/L+2,4-D 1.0mg/L	100	0.00^c	100.00^a	0.00^f	29.33^a
C₆	纵切	MS+6-BA 5.0mg/L+2,4-D 1.5mg/L	100	0.00^c	100.00^a	0.00^f	25.33^a
C₇	横切	MS+6-BA 3.0 mg/L+IAA 1.0 mg/L	100	94.33^b	0.00^d	107.00^e	29.33^a
C₈	横切	MS+6-BA 4.0 mg/L+IAA 1.5 mg/L	100	95.00^b	0.00^d	114.00^d	26.00^a
C₉	横切	MS+6-BA 5.0 mg/L+IAA 2.0 mg/L	100	96.00^b	0.00^d	121.33^c	26.00^a
C₁₀	横切	MS+6-BA 3.0mg/L+2,4-D 0.5mg/L	100	0.00^c	90.00^c	0.00^f	27.00^a
C₁₁	横切	MS+6-BA 4.0mg/L+2,4-D 1.0mg/L	100	0.00^c	92.33^b	0.00^f	26.00^a
C₁₂	横切	MS+6-BA 5.0mg/L+2,4-D 1.5mg/L	100	0.00^c	93.00^b	0.00^f	25.00^a

处理 Treatment	切割方式 Method	培养基 Medium	接种数 Number of inoculated	不定芽诱导率 Rate of adventitious bud induction/%	愈伤诱导率 Rate of callus induction/%	不定芽 Number of adventitious bud	诱导时间 Time of induction/d
C₁	纵切	MS+6-BA 3.0 mg/L+IAA 1.0 mg/L	100	100.00^a	0.00^d	116.33^cd	29.33^a
C₂	纵切	MS+6-BA 4.0 mg/L+IAA 1.5 mg/L	100	100.00^a	0.00^d	156.00^a	24.67^a
C₃	纵切	MS+6-BA 5.0 mg/L+IAA 2.0 mg/L	100	100.00^a	0.00^d	138.67^b	24.00^a
C₄	纵切	MS+6-BA 3.0mg/L+2,4-D 0.5mg/L	100	0.00^c	100.00^a	0.00^f	30.00^a
C₅	纵切	MS+6-BA 4.0mg/L+2,4-D 1.0mg/L	100	0.00^c	100.00^a	0.00^f	29.33^a
C₆	纵切	MS+6-BA 5.0mg/L+2,4-D 1.5mg/L	100	0.00^c	100.00^a	0.00^f	25.33^a
C₇	横切	MS+6-BA 3.0 mg/L+IAA 1.0 mg/L	100	94.33^b	0.00^d	107.00^e	29.33^a
C₈	横切	MS+6-BA 4.0 mg/L+IAA 1.5 mg/L	100	95.00^b	0.00^d	114.00^d	26.00^a
C₉	横切	MS+6-BA 5.0 mg/L+IAA 2.0 mg/L	100	96.00^b	0.00^d	121.33^c	26.00^a
C₁₀	横切	MS+6-BA 3.0mg/L+2,4-D 0.5mg/L	100	0.00^c	90.00^c	0.00^f	27.00^a
C₁₁	横切	MS+6-BA 4.0mg/L+2,4-D 1.0mg/L	100	0.00^c	92.33^b	0.00^f	26.00^a
C₁₂	横切	MS+6-BA 5.0mg/L+2,4-D 1.5mg/L	100	0.00^c	93.00^b	0.00^f	25.00^a

编号 No.	培养基 Medium	分化系数 Differentiation coefficient	生长情况 Growing situation
Y₁	MS+KT 0.5 mg/L+NAA 0.1 mg/L	5.29^g	芽点多,极少玻璃化,不定芽长势一般
Y₂	MS+KT 0.5 mg/L+NAA 0.2 mg/L	5.72^e	芽点多,极少玻璃化,不定芽长势良
Y₃	MS+KT 0.5 mg/L+NAA 0.3 mg/L	5.45^f	芽点多,部分玻璃化,不定芽长势差
Y₄	MS+KT 1.0 mg/L+NAA 0.1 mg/L	5.81^de	芽点很多,极少玻璃化,不定芽长势良
Y₅	MS+KT 1.0 mg/L+NAA 0.2 mg/L	6.43^a	芽点很多,极少玻璃化,不定芽长势优
Y₆	MS+KT 1.0 mg/L+NAA 0.3 mg/L	6.13^b	芽点很多,玻璃化严重,不定芽长势差
Y₇	MS+KT 1.5 mg/L+NAA 0.1 mg/L	5.93^cd	芽点很多,部分玻璃化,不定芽长势良
Y₈	MS+KT 1.5 mg/L+NAA 0.2 mg/L	6.16^b	芽点很多,部分玻璃化,不定芽长势一般
Y₉	MS+KT 1.5 mg/L+NAA 0.3 mg/L	6.03^bc	芽点很多,玻璃化严重,不定芽长势差

编号 No.	培养基 Medium	分化系数 Differentiation coefficient	生长情况 Growing situation
Y₁	MS+KT 0.5 mg/L+NAA 0.1 mg/L	5.29^g	芽点多,极少玻璃化,不定芽长势一般
Y₂	MS+KT 0.5 mg/L+NAA 0.2 mg/L	5.72^e	芽点多,极少玻璃化,不定芽长势良
Y₃	MS+KT 0.5 mg/L+NAA 0.3 mg/L	5.45^f	芽点多,部分玻璃化,不定芽长势差
Y₄	MS+KT 1.0 mg/L+NAA 0.1 mg/L	5.81^de	芽点很多,极少玻璃化,不定芽长势良
Y₅	MS+KT 1.0 mg/L+NAA 0.2 mg/L	6.43^a	芽点很多,极少玻璃化,不定芽长势优
Y₆	MS+KT 1.0 mg/L+NAA 0.3 mg/L	6.13^b	芽点很多,玻璃化严重,不定芽长势差
Y₇	MS+KT 1.5 mg/L+NAA 0.1 mg/L	5.93^cd	芽点很多,部分玻璃化,不定芽长势良
Y₈	MS+KT 1.5 mg/L+NAA 0.2 mg/L	6.16^b	芽点很多,部分玻璃化,不定芽长势一般
Y₉	MS+KT 1.5 mg/L+NAA 0.3 mg/L	6.03^bc	芽点很多,玻璃化严重,不定芽长势差

编号No.	培养基Medium	分化系数Differentiation coefficient	生长情况Growing situation
CK	Y₅	6.43^j	芽点多,极少玻璃化,不定芽长势优
W₁	Y₅+土豆10 g/L	6.89^hi	芽点多,无玻璃化,不定芽长势优
W₂	Y₅+土豆20 g/L	7.18^defg	芽点较多,无玻璃化,不定芽长势优
W₃	Y₅+土豆30 g/L	7.67^b	芽点极多,无玻璃化,不定芽长势优
W₄	Y₅+土豆40 g/L	7.20^defg	芽点极多,无玻璃化,不定芽长势优
W₅	Y₅+香蕉10 g/L	6.48^j	芽点多,无玻璃化,不定芽长势优
W₆	Y₅+香蕉20 g/L	7.05^fgh	芽点较多,无玻璃化,不定芽长势优
W₇	Y₅+香蕉30 g/L	7.29^def	芽点极多,无玻璃化,不定芽长势优
W₈	Y₅+香蕉40 g/L	6.98^gh	芽点极多,无玻璃化,不定芽长势优
W₉	Y₅+苹果10 g/L	6.11^k	芽点多,无玻璃化,不定芽长势优
W₁₀	Y₅+苹果20 g/L	7.43^bcd	芽点极多,无玻璃化,不定芽长势优
W₁₁	Y₅+苹果30 g/L	6.87^hi	芽点较多,无玻璃化,不定芽长势优
W₁₂	Y₅+苹果40 g/L	6.68^ij	芽点极多,无玻璃化,不定芽长势优
W₁₃	Y₅+椰汁50 mL/L	7.34^cde	芽点多,无玻璃化,不定芽长势优
W₁₄	Y₅+椰汁100 mL/L	8.17^a	芽点极多,无玻璃化,不定芽长势优
W₁₅	Y₅+椰汁150 mL/L	7.56^bc	芽点较多,无玻璃化,不定芽长势优
W₁₆	Y₅+椰汁200 mL/L	7.12^efgh	芽点极多,无玻璃化,不定芽长势优