以次生代谢物产量为目标的西兰花毛状根培养技术体系优化

doi:10.13560/j.cnki.biotech.bull.1985.2021-0143

摘要/Abstract

摘要：

本研究旨在探究不同单因素条件下西兰花毛状根培养体系中萝卜硫苷（glucoraphanin,GRA）和萝卜硫素（sulforaphane,SF）释放的关系。通过10 mmol/L茉莉酸甲酯（methyl jasmonate,MeJA）处理,探究了在不同接种量、培养基体积、pH、培养温度及摇床转速下对西兰花毛状根中GRA和SF释放的影响;在单因素实验基础上,选取培养温度、毛状根接种量及pH进行响应面实验。结果表明：当培养温度为25.44℃,毛状根接种量为0.15 g,pH 5.56,西兰花毛状根中GRA和SF总产量为2.08 mg/flask,与回归模型预测理论总产量2.44 mg/flask相近;不同培养条件下,西兰花毛状根培养体系中GRA和SF释放至培养基中的量随之不同。

关键词: 西兰花毛状根, 萝卜硫苷, 萝卜硫素, 响应面法, 释放机理

Abstract:

This study focuses on the relationship between the release of glucoraphanin（GRA）and sulforaphane（SF）in broccoli hairy root culture system under different single factors. Through 10 mmol/L methyl jasmonate（MeJA）treatment,the effects of different inoculation amount,medium volume,pH,culture temperature and rotating speed on the release of GRA and SF in hairy roots were investigated. On the basis of single factor experiments,the culture temperature,hairy root inoculation amount and pH were subjected to response surface experiments. The results showed that total GRA and SF yields in the broccoli hairy roots were 2.08 mg under culture temperature of 25.44℃,hairy root inoculation amount of 0.15 g and pH of 5.56,which was close to the theoretical total yield of 2.44 mg predicted by regression model. The results demonstrated that the amount of GRA and SF released into the medium varied under different culture conditions.

Key words: hairy roots of broccoli, glucoraphanin, sulforaphane, response surface, release mechanism

张秀民, 马绍英, 杨洁, 包金玉, 张晓玲, 田鹏, 路亚琦, 李胜. 以次生代谢物产量为目标的西兰花毛状根培养技术体系优化[J]. 生物技术通报, 2021, 37(8): 75-84.

ZHANG Xiu-min, MA Shao-ying, YANG Jie, BAO Jin-yu, ZHANG Xiao-ling, TIAN Peng, LU Ya-qi, LI Sheng. Optimization of Hairy Roots Culture System of Broccoli Aiming at the Yield of Secondary Metabolites[J]. Biotechnology Bulletin, 2021, 37(8): 75-84.

图/表 12

参考文献 32

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因素 Factor	因素Factor
因素 Factor	接种量 Inoculation amount/g	培养基体积 Medium volume/mL	转速 Rotating speed/（r·min^-1）	温度 Temperature/℃	pH
接种量Inoculation amount/g	0.05 0.1 0.15 0.2 0.25	100	110	25	6.0
培养基体积Medium volume/mL	0.15	80 90 100 110 120	110	25	6.0
转速Rotating speed/（r·min^-1）	0.15	100	90 100 110 120 130	25	6.0
温度Temperature/℃	0.15	100	110	21 23 25 27 29	6.0
pH	0.15	100	110	25	5.0 5.5 6.0 6.5 7.0

因素 Factor	因素Factor
因素 Factor	接种量 Inoculation amount/g	培养基体积 Medium volume/mL	转速 Rotating speed/（r·min^-1）	温度 Temperature/℃	pH
接种量Inoculation amount/g	0.05 0.1 0.15 0.2 0.25	100	110	25	6.0
培养基体积Medium volume/mL	0.15	80 90 100 110 120	110	25	6.0
转速Rotating speed/（r·min^-1）	0.15	100	90 100 110 120 130	25	6.0
温度Temperature/℃	0.15	100	110	21 23 25 27 29	6.0
pH	0.15	100	110	25	5.0 5.5 6.0 6.5 7.0

方差来源Source of variance	平方和Sum of squares	自由度df	均方Means square	F	P	显著性Significance
模型Model	2.73	9	0.30	7.78	0.0065	**
A-培养温度Culture temperature	0.00072	1	0.00072	0.18	0.6805
B-接种量Inoculation amount	0.18	1	0.18	4.69	0.0671
C-pH	0.028	1	0.028	0.71	0.4281
AB	0.068	1	0.068	1.73	0.2296
AC	0.022	1	0.022	0.58	0.4725
BC	0.055	1	0.055	1.41	0.2730
A²	0.99	1	0.99	25.35	0.0015	**
B²	0.58	1	0.58	14.95	0.0062	**
C²	0.55	1	0.55	14.16	0.0071	**
残差Residual	0.27	7	0.039
失拟项Lack of fit	0.14	3	0.046	1.38	0.3701
纯误差Pure error	0.13	4	0.034
总值Total value	3.01	16
R²=0.9091 R²_Adj=0.8629

方差来源Source of variance	平方和Sum of squares	自由度df	均方Means square	F	P	显著性Significance
模型Model	2.73	9	0.30	7.78	0.0065	**
A-培养温度Culture temperature	0.00072	1	0.00072	0.18	0.6805
B-接种量Inoculation amount	0.18	1	0.18	4.69	0.0671
C-pH	0.028	1	0.028	0.71	0.4281
AB	0.068	1	0.068	1.73	0.2296
AC	0.022	1	0.022	0.58	0.4725
BC	0.055	1	0.055	1.41	0.2730
A²	0.99	1	0.99	25.35	0.0015	**
B²	0.58	1	0.58	14.95	0.0062	**
C²	0.55	1	0.55	14.16	0.0071	**
残差Residual	0.27	7	0.039
失拟项Lack of fit	0.14	3	0.046	1.38	0.3701
纯误差Pure error	0.13	4	0.034
总值Total value	3.01	16
R²=0.9091 R²_Adj=0.8629