草莓茎尖组织培养和快繁体系的建立

doi:10.16035/j.issn.1001-7283.2016.04.011

Abstract

Abstract:

Taking one main strawberry variety of Guinugan as sample,experiments were carried out to study the effects of 9 factors (HgCl₂,basal medium,white granulated sugar,etc.) on tip-shoot tissue culture of strawberry,and then to screen out a tip-shoot tissue culture technology for strawberry,which could produce better quality seedlings,and eventually to establish a relatively complete tip-shoot tissue culture and rapid propagation system in strawberry. The results showed that 9 factors had great effects on strawberry tip-shoot tissue culture.The best sterilization approach for explants was to treat 8-11min by 0.20% HgCl₂,the suitable primary culture medium was MS+6-BA 0.5-1.0mg/L+NAA 0.01mg/L+white granulated sugar 30g/L,the suitable proliferation medium was MS+6-BA 0.5-1.0mg/L+NAA 0.01-0.05mg/L+white granulated sugar 20-30g/L,and the suitable rooting medium was MS+NAA 0-0.01mg/L+IBA 0-0.1mg/L+IAA 0-0.1mg/L+ white granulated sugar 30g/L.

Key words: Strawberry, Tip-shoot, Tissue culture, Rapid propagation, Medium, Orthogonal experiment

Xiaoliang Li,Junyun Zhang,Zhong Zhang,Cuiping Zhang,Shixian Yang,Wenzhi Wang,Jiankang Zhang,Yonghua Duan. Tissue Culture and Rapid Propagation of Strawberry[J].Crops, 2016, 32(4): 68-74.

Figures/Tables 6

Table 1

Table 2

Table 3

Table 4

Table 5

Table 6

Effects of treatments on strawberry rhizogenesis and plant growth vigour in rooting culture"

处理 Treatments	根发生的部位Rhizogenesis site	植株生长势Plant growth vigour
1	基部（无愈伤组织）	较好：健壮、叶片较大、深绿、无玻璃化、无黄化
2	基部（无或少许愈伤组织）	较好：健壮、叶片较大、深绿、无玻璃化、无黄化
3	基部（无愈伤组织）或基部愈伤组织（中度）	较好：健壮、叶片较大、深绿、无玻璃化、无黄化
4	基部（无愈伤组织）或基部愈伤组织（少许或少量或中度）	一般：茎叶硬度一般、叶片大小一般、叶绿、无玻璃化、无黄化
5	基部（无愈伤组织）或基部愈伤组织（中度）	一般：茎叶硬度一般、叶片大小一般、叶绿、少许植株略玻璃化、无黄化
6	基部（无愈伤组织）或基部愈伤组织（偏多）	一般：茎叶硬度一般、叶片大小一般、叶绿、无玻璃化、无黄化
7	基部（无愈伤组织）或基部愈伤组织（少许或中度或偏多）	较差：茎叶较柔软、叶片较小、无玻璃化、略偏黄
8	基部（无愈伤组织）或基部愈伤组织（中度或偏多）	较差：茎叶较柔软、叶片较小、略玻璃化、略偏黄
9	基部（无愈伤组织）或基部愈伤组织（偏多或大量）	较差：茎叶较柔软、叶片较小、略玻璃化、略偏黄
10	基部（无愈伤组织）或叶柄处（或茎部）的绒毛状愈伤组织（少许）	一般：茎叶硬度一般、叶片大小一般、叶绿、无玻璃化、略偏黄
11	基部（无或少许愈伤组织）或叶柄处（或茎部）的绒毛状愈伤组织（少许）	一般：茎叶硬度一般、叶片大小一般、叶绿、略玻璃化或黄化
12	基部（无或少许愈伤组织）或叶柄处的绒毛状愈伤组织（少许）	一般：茎叶硬度一般、叶片大小一般、无玻璃化、略偏黄
13	基部（无或少许愈伤组织）或叶柄处的绒毛状愈伤组织（少许）	一般：茎叶硬度一般、叶片大小一般、略玻璃化或黄化
14	基部（无或少许愈伤组织）或叶柄处（或茎部）的绒毛状愈伤组织（少许）	一般：茎叶硬度一般、叶片大小一般、无玻璃化、略偏黄
15	基部（无或少许愈伤组织）或叶柄处（或茎部）的绒毛状愈伤组织（少许）	一般：茎叶硬度一般、叶片大小一般、无玻璃化、略偏黄
16	基部（无或少许愈伤组织）或叶柄处的绒毛状愈伤组织（少许）	较差：茎叶特别柔软、叶片较小、略玻璃化、偏黄
17	基部（无或少许愈伤组织）或叶柄处的绒毛状愈伤组织（少许）	较差：茎叶特别柔软、叶片较小、略玻璃化、偏黄
18	基部（无或少许愈伤组织）或叶柄处（或茎部）的绒毛状愈伤组织（中度）	较差：茎叶特别柔软、叶片较小、略玻璃化、偏黄

Table 6

References 14

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指标Index	0.15% HgCl₂					0.20% HgCl₂
指标Index	2~3min	4~5min	6~7min	8~9min	10~11min	2~3min	4~5min	6~7min	8~9min	10~11min
细菌污染率Bacteria contamination rates	80.13	55.00	48.75	38.12	25.00	72.50	52.54	43.61	36.84	23.24
死亡率Death rates	0	0	10.00	12.26	21.32	0	5.00	11.83	13.45	22.67

处理Treatments	植物形态 Plant morphology	芽的量(%) Percent of buds	幼苗的量(%) Percent of seedlings	增殖分化情况 Proliferation and differentiation	增殖系数 Proliferation coefficients
1	丛芽（芽块）或幼苗	6.69~31.25	68.75~93.31	生长势较好、健壮、无愈伤组织、深绿、无玻璃化、无黄化、幼苗个体形态较好	22.00±2.34b
2	丛芽（芽块）或幼苗	81.23~95.27	4.73~18.77	生长势较好、健壮、无愈伤组织、深绿、无玻璃化、无黄化、幼苗个体形态较差	25.70±1.54a
3	芽块（愈伤组织）或幼苗	61.32~78.47	21.53~38.68	生长势较好、健壮、深绿、无玻璃化、无黄化、愈伤组织偏多或大量	10.40±1.00c
4	芽块（愈伤组织）或幼苗	90.80~100.00	0~9.20	生长势一般、大部分绿色、少量黄化、玻璃化、愈伤组织中度	4.40±0.34d
5	芽块（愈伤组织）或幼苗	60.00~83.33	16.67~40.00	生长势一般、大部分绿色、少量黄化、玻璃化、愈伤组织偏多或大量	5.70±0.45d
6	丛芽（芽块）或幼苗	60.00~66.68	33.32~40.00	生长势一般、大部分绿色、少量黄化、少量玻璃化、无或少许愈伤组织	9.50±0.83cd
7	芽块（愈伤组织）或幼苗	81.82~100.00	0~18.18	生长势较差、大部分浅绿色、少量黄化、略玻璃化、愈伤组织大量	6.70±0.62d
8	丛芽（芽块）或幼苗	95.00~100.00	0~5.00	生长势较差、大部分黄色（黄化现象突出）、少量褐色、无玻璃化、无愈伤组织	11.30±1.63c
9	芽块（愈伤组织）	100.00	0	生长势较差、黄色或褐色、质量较差（小、紧密、硬）、玻璃化和黄化现象突出、死亡现象明显、愈伤组织偏多或大量	9.00±0.54cd

处理Treatments	株高Plant height (cm)	叶片数Leaf number	根长Root length (cm)	根粗Root diameter (mm)	生根量Root number
1	2.34±0.13b	6.90±0.59bc	2.75±0.22a	0.19±0.02a	6.80±0.95bc
2	2.38±0.14ab	7.90±0.62ab	2.47±0.23ab	0.17±0.03ab	7.40±0.22bc
3	1.53±0.13c	5.30±0.47c	1.14±0.09d	0.14±0.01b	6.30±0.60bc
4	2.04±0.13bc	6.30±0.21c	1.67±0.11c	0.12±0.01bc	7.40±0.78bc
5	2.28±0.17b	6.80±0.33bc	1.55±0.13cd	0.10±0.01c	9.60±0.75a
6	2.48±0.23ab	7.10±0.28bc	1.84±0.17bc	0.11±0.01bc	9.50±0.96a
7	2.63±0.27ab	7.40±0.40bc	1.90±0.16bc	0.12±0.01bc	10.70±0.60a
8	2.70±0.29ab	6.50±0.34bc	2.75±0.33a	0.17±0.03ab	7.40±0.85bc
9	2.86±0.19a	6.40±0.31c	2.25±0.19ab	0.14±0.02b	7.80±0.53b
10	1.66±0.16c	7.90±0.38ab	0.95±0.10d	0.07±0.01c	5.90±0.43c
11	2.16±0.19bc	9.00±0.58a	2.01±0.30bc	0.10±0.01c	3.80±0.59d
12	2.16±0.13bc	7.70±0.54b	1.14±0.20d	0.08±0.01c	4.60±0.40cd
13	2.19±0.23b	8.90±0.60ab	1.87±0.18bc	0.09±0.01c	4.30±0.30cd
14	2.75±0.20ab	8.10±0.64ab	2.21±0.14b	0.09±0.01c	6.30±0.54bc
15	2.66±0.14ab	8.60±0.58ab	2.11±0.11bc	0.09±0.01c	3.60±0.52d
16	2.84±0.21ab	8.10±0.35ab	2.62±0.15ab	0.09±0.02c	4.00±0.33d
17	2.75±0.09ab	8.10±0.41ab	2.18±0.30bc	0.08±0.01c	4.60±0.70cd
18	2.70±0.24ab	8.20±0.51ab	2.46±0.12ab	0.10±0.01c	4.90±0.46cd

Tissue Culture and Rapid Propagation of Strawberry

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处理 Treatments	基础培养基 Basal medium	6-BA (mg/L)	NAA (mg/L)	白砂糖(g/L) White granulated sugar
1	MS	0.5	0.01	20
2	MS	1.0	0.05	25
3	MS	2.0	0.10	30
4	1/2 MS	0.5	0.05	30
5	1/2 MS	1.0	0.10	20
6	1/2 MS	2.0	0.01	25
7	1/4 MS	0.5	0.10	25
8	1/4 MS	1.0	0.01	30
9	1/4 MS	2.0	0.05	20

试验类型Experimental types	处理Treatments	基础培养基Basal medium	NAA(mg/L)	吲哚丁酸IBA(mg/L)	吲哚乙酸IAA(mg/L)	AC(%)
正交试验L₉(3⁴)	1	MS	0	0	0	—
Orthogonal experiments	2	MS	0.01	0.1	0.1	—
	3	MS	0.05	0.2	0.2	—
	4	1/2 MS	0	0.1	0.2	—
	5	1/2 MS	0.01	0.2	0	—
	6	1/2 MS	0.05	0	0.1	—
	7	1/4 MS	0	0.2	0.1	—
	8	1/4 MS	0.01	0	0.2	—
	9	1/4 MS	0.05	0.1	0	—
对比试验	10	MS	0	0	0	0.1
Comparative experiments	11	MS	0.01	0.1	0.1	0.1
	12	MS	0.05	0.2	0.2	0.1
	13	1/2 MS	0	0.1	0.2	0.1
	14	1/2 MS	0.01	0.2	0	0.1
	15	1/2 MS	0.05	0	0.1	0.1
	16	1/4 MS	0	0.2	0.1	0.1
	17	1/4 MS	0.01	0	0.2	0.1
	18	1/4 MS	0.05	0.1	0	0.1

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