贵长猕猴桃茎段愈伤组织诱导条件优化研究

doi:10.16178/j.issn.0528-9017.20230743

摘要/Abstract

摘要：

以贵长猕猴桃茎段为材料,在基本培养基(MS)上诱导其产生愈伤组织,采用响应面法优化贵长猕猴桃愈伤组织诱导条件。在单因素分析的基础上,以6-苄基腺嘌呤(6-BA)质量浓度、2, 4-二氯苯氧乙酸(2,4-D)质量浓度和0.1%氯化汞(HgCl₂)处理外植体时间为自变量,以贵长猕猴桃茎段愈伤组织的诱导率为评价指标,设计三因素三水平响应面法优化实验。结果表明,贵长猕猴桃最优愈伤组织诱导条件为:6-BA质量浓度为0.4 mg·L^-1、2, 4-D质量浓度为2.0 mg·L^-1、0.1%氯化汞处理时间为4.0 min,该诱导条件下贵长猕猴桃茎段愈伤组织诱导率为85.71%,与预测值的相对误差较小,实验值与预测值较吻合,贵长猕猴桃茎段愈伤组织诱导条件经优化后诱导率得到有效提高,可为贵长猕猴桃组织培养的进一步研究提供依据。

关键词: 贵长猕猴桃, 愈伤组织, 茎段, 组织培养, 响应面法

Abstract:

Using the stem segments of kiwifruit as materials, callus induction was induced on MS medium, and the response surface methodology was used to optimize the induction conditions of kiwifruit callus. On the basis of single factor analysis, a three factor three-level response surface methodology was designed to optimize the experiment, with 6-BA mass concentration, 2, 4-D mass concentration, and 0.1% mercuric chloride treatment time as independent variables, and the induction rate of callus tissue in the stem segment of kiwifruit as the evaluation index. The results showed that the optimal conditions for inducing callus tissue in kiwifruit were 6-BA concentration of 0.4 mg·L^-1, 2, 4-D concentration of 2.0 mg·L^-1, and 0.1% mercuric chloride treatment time of 4.0 minutes. Under these induction conditions, the induction rate of callus tissue in kiwifruit stem segments was 85.71%, which had a relatively small relative error with the predicted value. The experimental value was in good agreement with the predicted value. After optimizing the induction conditions, the induction rate of callus tissue in kiwifruit stem segments was effectively improved, providing a basis for further research on kiwifruit tissue culture.

Key words: Guichang kiwifruit, callus, stem segments, tissue culture, response surface methodology

中图分类号:

S663.4

杨椿玉, 辛佳乐, 潘小舆, 班虎检, 韩世明, 王月霞. 贵长猕猴桃茎段愈伤组织诱导条件优化研究[J]. 浙江农业科学, 2024, 65(11): 2601-2606.

YANG Chunyu, XIN Jiale, PAN Xiaoyu, BAN Hujian, HAN Shiming, WANG Yuexia. Study on optimization of callus induction conditions in stem segment of Guichang kiwifruit[J]. Journal of Zhejiang Agricultural Sciences, 2024, 65(11): 2601-2606.

图/表 10

表1 单因素实验设计

Table 1 Single factor experimental design

0.1%HgCl₂/min	6-BA/(mg·L^-1)	2, 4-D/(mg·L^-1)
1.0	0.1	0.5
2.0	0.2	1.0
3.0	0.3	1.5
4.0	0.4	2.0
5.0	0.5	2.5
6.0	0.6	3.0

表2 响应面因素水平设计

Table 2 Response surface factor level design

水平	因素
水平	A,6-BA/ (mg·L^-1)	B,2, 4-D/ (mg·L^-1)	C,0.1%HgCl₂/ min
-1	0.2	1.0	2.0
0	0.4	2.0	4.0
1	0.6	3.0	6.0

图1 0.1% HgCl2处理时间对贵长猕猴桃茎段愈伤组织诱导的影响

Fig.1 Effect of 0.1% mercuric chloride treatment time on callus induction in stem segments of kiwifruit

图2 6-BA质量浓度对贵长猕猴桃茎段愈伤组织诱导的影响

Fig.2 Effect of 6-BA mass concentration on callus induction in stem segments of kiwifruit

图3 2, 4-D质量浓度对贵长猕猴桃茎段愈伤组织诱导的影响

Fig.3 Effect of 2, 4-D mass concentration on callus induction in stem segments of kiwifruit

表3 响应面实验设计及结果

Table 3 Response surface experiment design and results

实验号	因素			诱导率/ %
实验号	A,6-BA/ (mg·L^-1)	B,2, 4-D/ (mg·L^-1)	C,0.1%HgCl₂/ min	诱导率/ %
1	0.2	1.0	4.0	57.14
2	0.6	1.0	4.0	71.43
3	0.2	3.0	4.0	71.43
4	0.6	3.0	4.0	78.57
5	0.2	2.0	2.0	57.14
6	0.6	2.0	2.0	67.86
7	0.2	2.0	6.0	50.00
8	0.6	2.0	6.0	60.71
9	0.4	1.0	2.0	64.29
10	0.4	3.0	2.0	78.57
11	0.4	1.0	6.0	57.14
12	0.4	3.0	6.0	78.57
13	0.4	2.0	4.0	85.71
14	0.4	2.0	4.0	89.29
15	0.4	2.0	4.0	89.29
16	0.4	2.0	4.0	85.71
17	0.4	2.0	4.0	89.29

表4 回归模型方差分析

Table 4 Variance analysis of regression model

方差来源	平方和	自由度	均方	F值	P	显著程度
模型	2 702.27	9	300.25	44.49	<0.000 1	^***
A	229.62	1	229.62	34.02	0.000 6	^**
B	408.12	1	408.12	60.47	0.000 1	^**
C	57.46	1	57.46	8.51	0.022 4	^*
AB	12.78	1	12.78	1.89	0.211 2
AC	2.50×10^-5	1	2.50×10^-5	3.70×10^-6	0.998 5
BC	12.78	1	12.78	1.89	0.211 2
A²	881.03	1	881.03	130.54	<0.000 1	^***
B²	59.22	1	59.22	8.77	0.021 0	^*
C²	881.03	1	881.03	130.54	<0.000 1	^***
残差	47.24	7	6.75
失拟误差	31.86	3	10.62	2.76	0.175 6
纯误差	15.38	4	3.84
总和	2 749.51	16

图4 6-BA和2, 4-D质量浓度对愈伤组织诱导率影响的等高线图及响应面

Fig.4 Contour plot and response surface of the effects of 6-BA and 2, 4-D mass concentration on callus induction rate

图5 6-BA质量浓度和0.1% HgCl2处理时间对愈伤组织诱导率影响的等高线图及响应面

Fig.5 Contour plot and response surface of the effects of 6-BA mass concentration and 0.1% mercuric chloride treatment time on callus induction rate

图6 2, 4-D质量浓度和0.1% HgCl2处理时间对愈伤组织诱导率影响的等高线图及响应面

Fig.6 Contour plot and response surface of the effects of 2, 4-D mass concentration and 0.1% mercuric chloride treatment time on callus induction rate

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