Optimization of Hairy Roots Culture System of Broccoli Aiming at the Yield of Secondary Metabolites

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

Abstract

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

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.

Figures/Tables 12

References 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