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

Abstract

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

CLC Number:

Q813.1

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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References 15

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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

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