光培养条件对东北红豆杉外植体组培效果的影响

doi:10.13229/j.cnki.jdxbgxb20200563

吉林大学学报(工学版) ›› 2021, Vol. 51 ›› Issue (6): 2313-2318.doi: 10.13229/j.cnki.jdxbgxb20200563

• 农业工程·仿生工程 • 上一篇

光培养条件对东北红豆杉外植体组培效果的影响

王淑杰¹(),赵子瑞¹,孟慧雯¹,李晓卉¹,张雅静¹,周亚军²()

^1.吉林大学生物与农业工程学院，长春 130022
^2.吉林大学食品科学与工程学院，长春 130022

收稿日期:2020-07-27 出版日期:2021-11-01 发布日期:2021-11-15
通讯作者: 周亚军 E-mail:shujie@jlu.edu.cn;zhouyajun109@yahoo.com
作者简介:王淑杰（1965-），女，教授，博士. 研究方向：资源环境可持续发展理论与技术. E-mail：shujie@jlu.edu.cn
基金资助:
科技部“十三五”国家科技计划发展项目(3G016W112418);吉林省科技发展计划项目(20180201009NY);长春市科技发展计划项目(NK15SS22)

Effect of light culture condition on explants of Taxus cuspidate

Shu-jie WANG¹(),Zi-rui ZHAO¹,Hui-wen MENG¹,Xiao-hui LI¹,Ya-jing ZHANG¹,Ya-jun ZHOU²()

^1.College of Biological and Agricultural Engineering，Jilin University，Changchun 130022，China
^2.College of Food Science and Engineering，Jilin University，Changchun 130022，China

Received:2020-07-27 Online:2021-11-01 Published:2021-11-15
Contact: Ya-jun ZHOU E-mail:shujie@jlu.edu.cn;zhouyajun109@yahoo.com

摘要/Abstract

摘要：

为了研究不同光培养条件对东北红豆杉外植体生长状况的影响，本文以带芽幼茎为试验材料，探讨了在光培养条件下，培养基中蔗糖的最佳用量，并采用正交试验设计分析得到适合东北红豆杉外植体生长的最佳光培养条件。试验结果表明：在光质为LED红蓝光（色比R∶B=3∶1），CO₂体积浓度为1500 mg/L，光照强度为12.5 μmol·m^-2·s^-1，光周期为10 h/d的环境下，培养基中蔗糖的最佳用量为10 g/L。此时，外植体芽萌动率和出愈率最高，芽萌动时间和初愈时间短，褐化率最低。正交试验得到适合东北红豆杉外植体生长的最佳光培养条件组合为：CO₂体积浓度为3000 mg/L+光照强度为37.5 μmol·m^-2·s^-1+光周期为14 h/d。验证试验表明：该环境下外植体与对照组（无光）对比，染菌率降低了4.33%，芽萌动时间和初愈时间分别降低了2.33、3.75 d，芽萌动率和初愈率分别提高了16%、6.33%，愈伤组织鲜重增长倍数提高了0.35倍，紫杉醇含量提高了0.09 mg/L，为东北红豆杉在光培养环境条件下组织培养提供了理论基础和技术支持。

关键词: 组织培养, 光培养条件, 东北红豆杉, 外植体, 愈伤组织, 蔗糖

Abstract:

The effect of light culture technology on the explants growth of Taxus cuspidate was studied. The young stems of Taxus cuspidate were used as the explants. The optimal dosage of sucrose in the culture medium were researched. The orthogonal test was applied to study the optimal light culture conditions (CO₂ concentration, light intensity, light cycle) for explants of Taxus cuspidate. The results show that the optimal concentration of sucrose was 10 g/L, under the 3∶1 of red to blue light ratio, 1500 mg/L of CO₂ concentration, 12.5 mol·m^-2·s^-1 of light intensity, and 10 h/d of photoperiod. Under this condition, the explants have the highest bud germination rate and healing rate, short bud germination time and initial healing time, and the lowest browning rate. The optimal combination of factors in the light culture on explants of Taxus cuspidate was as follows: CO₂ concentration was 3000 mg/L, the light intensity was 37.5 μmol·m^-2·s^-1, and the photoperiod was 14 h/d. The verification test showed that compared with the control group (without light), the bacteria rate, bud germination time and initial healing time of explants under light culture are reduced by 4.33%, 2.33 d, and 3.75 d, respectively. Bud germination rate, healing rate, fresh weight growth rate of callus and paclitaxel content are increased by 16%, 6.33%, 0.35 times, 0.09 mg/L, respectively, which meant that the optimization is obvious. This study suggests that the light culture provides theoretical basis and technical support for tissue culture of Taxus cuspidate under light environment.

Key words: tissue culture, light culture, Taxus cuspidate, explant, callus, sucrose

中图分类号:

Q813.1

王淑杰,赵子瑞,孟慧雯,李晓卉,张雅静,周亚军. 光培养条件对东北红豆杉外植体组培效果的影响[J]. 吉林大学学报(工学版), 2021, 51(6): 2313-2318.

Shu-jie WANG,Zi-rui ZHAO,Hui-wen MENG,Xiao-hui LI,Ya-jing ZHANG,Ya-jun ZHOU. Effect of light culture condition on explants of Taxus cuspidate[J]. Journal of Jilin University(Engineering and Technology Edition), 2021, 51(6): 2313-2318.

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蔗糖体积浓度/（g·L^-1）	染菌率/%	芽萌动率/%	芽萌动时间/d	初愈时间/d	出愈率/%	褐化率/%	外植体生长状态（针叶形态、颜色）
0	13.00^d	24.75^d	20.25^a	28.50^a	8.25^d	66.67^a	针叶枯萎、色褐
5	18.75^c	50.67^c	17.25^b	23.25^b	33.25^cd	25.00^c	针叶萎蔫、色浅褐
10	19.25^c	91.75^a	14.50^c	20.25^c	83.50^ab	8.33^e	针叶茂盛、色绿
15	32.25^b	72.25^b	16.00^bc	21.25^bc	71.00^ab	12.50^d	针叶茂盛、色黄绿
20	40.00^a	55.67^c	16.67^b	22.00^bc	44.33^cb	33.33^b	针叶萎蔫、色黄

试验号	染菌率/%	芽萌动率/%	芽萌动时间/d	出愈率/%	初愈时间/d	愈伤组织鲜重/倍	紫杉醇产量/（mg·L^-1）
1	66.67^a	16.67^hi	15.00^c	58.33^cd	17.50^cd	2.18^cd	0.41^fgh
2	46.67^abc	25.00^gh	14.40^d	50.00^d	18.00^cd	1.57^fg	0.39^ghi
3	53.33^abc	75.00^c	14.60^cd	33.33^f	19.50^c	1.65^fg	0.28^j
4	40.00^abc	66.67^d	15.00^c	25.00^g	24.25^b	1.28^h	0.35ⁱ
5	60.00^ab	58.33^e	14.67^cd	41.67^e	18.67^cd	1.56^fgh	0.36^hi
6	40.00^abc	83.33^b	15.75^b	66.67^c	18.75^cd	2.11^cde	0.41^fgh
7	33.33^abc	41.67^f	15.00^c	75.00^bc	17.00^d	1.85^ef	0.98^b
8	26.67^abc	91.67^a	14.40^d	91.67^ab	16.75^d	2.55^b	0.82^c
9	46.67^abc	16.67^hi	14.80^cd	66.67^c	18.50^cd	1.59^fg	0.42^fg
10	20.00^abc	25.00^gh	14.25^d	41.67^e	19.00^c	2.04^de	0.61^e
11	53.33^abc	33.33^g	14.20^d	83.33^b	16.50^d	2.96^a	0.47^f
12	33.33^abc	75.00^c	13.60^f	100.00^a	16.25^d	2.37^bc	0.95^b
13	13.33^bc	16.67^hi	14.25^d	50.00^d	17.00^d	1.67^fg	0.41^fg
14	6.67^c	41.67^f	15.00^c	41.67^e	17.75^cd	1.50^gh	1.15^a
15	20.00^abc	50.00^ef	14.75^cd	25.00^g	18.25^cd	2.18^cd	0.94^b
16	26.67^abc	8.33ⁱ	18.00^a	8.33^h	27.50^a	1.26^h	0.70^d

试验号	A/（mg·L^-1）	B/（μmol·m^-2·s^-1）	C/（h·d^-1）
Y_j₁	5.68	6.02	5.98
Y_j₂	7.91	7.24	7.43
Y_j₃	7.68	7.43	7.64
Y_j₄	6.74	7.30	6.95
R_j	2.23	1.42	1.66
因素主次	A、C、B
最优组合	A₂B₃C₃

光培养条件对东北红豆杉外植体组培效果的影响

Effect of light culture condition on explants of Taxus cuspidate

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水平	因素
水平	A/（mg·L^-1）	B（μmol·m^-2·s^-1）	C/（h·d^-1）
1	1500	12.5	10
2	3000	25	12
3	4500	37.5	14
4	6000	50	16

试验号	A/（mg·L^-1）	B（μmol·m^-2·s^-1）	C/（h·d^-1）
1	1	1	1
2	1	2	2
3	1	3	3
4	1	4	4
5	2	1	2
6	2	2	1
7	2	3	4
8	2	4	3
9	3	1	3
10	3	2	4
11	3	3	1
12	3	4	2
13	4	1	4
14	4	2	3
15	4	3	2
16	4	4	1

试验号	A/（mg·L^-1）	B（μmol·m^-2·s^-1）	C/（h·d^-1）
1	1	1	1
2	1	2	2
3	1	3	3
4	1	4	4
5	2	1	2
6	2	2	1
7	2	3	4
8	2	4	3
9	3	1	3
10	3	2	4
11	3	3	1
12	3	4	2
13	4	1	4
14	4	2	3
15	4	3	2
16	4	4	1

试验号	A/（mg·L^-1）	B（μmol·m^-2·s^-1）	C/（h·d^-1）
1	1	1	1
2	1	2	2
3	1	3	3
4	1	4	4
5	2	1	2
6	2	2	1
7	2	3	4
8	2	4	3
9	3	1	3
10	3	2	4
11	3	3	1
12	3	4	2
13	4	1	4
14	4	2	3
15	4	3	2
16	4	4	1