万寿菊再生体系的建立及优化

doi:10.11983/CBB20062

植物学报 ›› 2020, Vol. 55 ›› Issue (6): 749-759.DOI: 10.11983/CBB20062

万寿菊再生体系的建立及优化

王亚琴¹^,², 韦陆丹¹, 王文静¹, 刘宝骏¹, 张春玲¹, 张俊卫¹, 何燕红¹^,^*()

¹华中农业大学园艺林学学院, 园艺植物生物学教育部重点实验室, 武汉 430070
²武汉市汉阳市政建设集团有限公司, 武汉 430050

收稿日期:2020-04-16 接受日期:2020-08-26 出版日期:2020-11-01 发布日期:2020-11-11
通讯作者: 何燕红
作者简介:*E-mail: hyh2010@mail.hzau.edu.cn
基金资助:
中央高校基本科研业务费专项(2662019PY072);国家自然科学基金(31672181)

The Establishment and Optimization of a Regeneration System for Marigold (Tagetes erecta)

Yaqin Wang¹^,², Ludan Wei¹, Wenjing Wang¹, Baojun Liu¹, Chunling Zhang¹, Junwei Zhang¹, Yanhong He¹^,^*()

¹Key Laboratory of Horticultural Plant Biology, Ministry of Education, College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan 430070, China
²Wuhan Hanyang Municipal Construction Group Co, Ltd, Wuhan 430050, China

Received:2020-04-16 Accepted:2020-08-26 Online:2020-11-01 Published:2020-11-11
Contact: Yanhong He

摘要/Abstract

摘要： 以40个不同基因型万寿菊(Tagetes erecta)叶片为外植体, 在相同条件下诱导不定芽分化, 获得最佳再生基因型; 然后分析不同激素组合、外植体切口方式、固化剂及蔗糖对万寿菊再生和玻璃化影响; 最后对不同类型的伸长培养基进行探索。结果表明, 最佳再生基因型为里程碑·黄色; 最佳再生培养基为MS+0.2 mg·L^‒1 TDZ+0.5 mg·L^‒1 IBA+8 g·L^‒1琼脂+40 g·L^‒1蔗糖, 再生率达70%, 玻璃化率降低至16%; 最适再生的小叶部位为全小叶; 最适伸长培养基为MS+8 g·L^‒1琼脂+30 g·L^‒1蔗糖, 伸长率达91.3%。该研究建立了高效稳定的万寿菊再生体系, 解决了万寿菊再生过程中严重的玻璃化问题, 可为万寿菊的遗传改良和基因功能研究奠定基础。

关键词: 万寿菊, 再生, 基因型, 玻璃化

Abstract: In this study, marigold (Tagetes erecta) leaves of 40 different genotypes were used as explants to induce adventitious buds formation to select the best regenerating genotype. In addition, main factors impacting the selected genotype’s regeneration and vitrification were analyzed, including hormone combinations, the way of explant incision, curing agent and sucrose, etc. Finally, different types of elongation medium were explored. The best regenerated genotype was marigold Milestone·Yellow. The best regeneration medium was MS media supplied with 0.2 mg·L^‒1 TDZ, 0.5 mg·L^‒1 IBA, 8 g·L^‒1 agar, and 40 g·L^‒1 sucrose, in which the regeneration rate was 70% and the vitrification rate was reduced to 16%. Whole leaflet was the most suitable explant among different leaflet segments; the best performing elongation medium was MS media supplied with 8 g·L^‒1 agar and 30 g·L^‒1 sucrose, and the elongation rate was 91.3%. This study establishes an efficient and stable regeneration system of marigold, and solved the serious vitrification problem commonly found during regeneration. It laid a foundation for marigold genetic improvement and gene function research.

Key words: marigold (Tagetes erecta), regeneration, genotype, vitrification

王亚琴, 韦陆丹, 王文静, 刘宝骏, 张春玲, 张俊卫, 何燕红. 万寿菊再生体系的建立及优化. 植物学报, 2020, 55(6): 749-759.

Yaqin Wang, Ludan Wei, Wenjing Wang, Baojun Liu, Chunling Zhang, Junwei Zhang, Yanhong He. The Establishment and Optimization of a Regeneration System for Marigold (Tagetes erecta). Chinese Bulletin of Botany, 2020, 55(6): 749-759.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB20062

https://www.chinbullbotany.com/CN/Y2020/V55/I6/749

图/表 9

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No.	Genotype	Phenotype		Regeneration rate (%)		Source	Regeneration type
1	Perfect·Golden	Short-height plant, yellow-oran- ge color, double petal		54.21±12.11 a		Chunhe Gardening (F₁)	Easy
2	Milestone·Yellow	Short-height plant, yellow color, double petal		51.44±8.29 ab		Sainuo Gardening (F₁)	Easy
3	Proud·Orange-yellow	Short-height plant, yellow-oran- ge color, double petal		41.47±6.12 abc		Dannuo Gardening (F₁)	Moderate
4	Chrysanthemum marigold·Orange	Short-height plant, orange color, double petal		36.23±11.95 abcd		Huaxianzi Gardening (F₁)	Moderate
5	Xunlei·Yellow	Short-height plant, yellow color, double petal		33.48±13.73 abcde		Chunhe Gardening (F₁)	Moderate
6	Galore·Yellow	Medium-height plant, yellow co- lor, double petal		31.95±18.22 abcdef		Hongyue Gardening (F₁)	Moderate
7	Proud·Golden	Short-height plant, yellow-oran- ge color, double petal		30.50±10.33 abcdef		Dannuo Gardening (F₁)	Moderate
8	Double marigold· Light beige	Medium-height plant, yellow color, double petal		22.01±14.56 bcdefg		Jinqiu Gardening (F₁)	Difficult
9	DC	Short-height plant, orange color, double petal		21.33±13.66 cdefgh		Self breeding (inbred line) *	Difficult
10	Youyi·Yellow	Short-height plant, yellow color, double petal		18.00±1.83 cdefgh		Chunhe Gardening (F₁)	Difficult
11	Miracle·Yellow	Short-height plant, yellow color, double petal		13.29±9.62 cdefghi		Huaxianzi Gardening (F₁)	Difficult
12	Old Inka·Orange	Medium-height plant, orange co- lor, double petal		13.24±10.50 defghij		Hongyue Gardening (F₁)	Difficult
13	Liberty·Yellow	Short-height plant, yellow color, double petal		12.95±8.82 defghij		Huaxianzi Gardening (F₁)	Difficult
14	6210	Medium-height plant, yellow-oran- ge color, double petal		10.00±11.08 fghij		Self breeding (inbred line) *	Difficult
15	F8	Medium-height plant, orange co- lor, simple petal		9.33±5.96 efghij		Self breeding (inbred line) *	Difficult
No.	Genotype		Phenotype		Regeneration rate (%)	Source	Regeneration type
16	Fengsheng·Orange		Short-height plant, yellow-orange color, double petal		7.11±7.70 ghij	Shijie seed (F₁)	Difficult
17	AJ		Short-height plant, yellow-orange color, double petal		7.02±13.41 ghij	Self breeding (inbred line) *	Difficult
18	Inka·Orange-yellow		Short-height plant, yellow-orange color, double petal		5.11±7.87 ghij	Chunhe Gardening (F₁)	Difficult
19	S3		Medium-height plant, yellow color, simple petal		4.92±2.75 ghij	Self breeding (inbred line) *	Difficult
20	S12		Short-height plant, yellow color, simple petal		4.67±10.43 hij	Self breeding (inbred line) *	Difficult
21	S5		Short-height plant, yellow-orange color, simple petal		4.67±5.06 ghij	Self breeding (inbred line) *	Difficult
22	SW2		Medium-height plant, yellow color, double petal		4.67±10.43 hij	Self breeding (inbred line) *	Difficult
23	Hawaii		Medium-height plant, yellow-oran- ge color, double petal		4.22±5.87 ghij	Thailand (F₁)	Difficult
24	6170		Medium-height plant, yellow-oran- ge color, double petal		2.89±1.62 ghij	Self breeding (inbred line) *	Difficult
25	Jindun·Yellow		Short-height plant, yellow color, double petal		2.71±4.36 hij	Sainuo Gardening (F₁)	Difficult
26	GFR		Medium-height plant, orange color, double petal		1.92±1.75 hij	Self breeding (inbred line) *	Difficult
27	DH		Short-height plant, yellow color, double petal		1.33±1.83 ij	Self breeding (inbred line) *	Difficult
28	Vanilla		Medium-height plant, yellow-w- hite color, double petal		0.91±2.03 ij	Thailand (F₁)	Unable
29	Double Chrysanthe- mum marigold·Orange		Short-height plant, yellow-oran- ge color, double petal		0.71±1.60 ij	Jinqiu Gardening (F₁)	Unable
30	6155		Medium-height plant, yellow color, double petal		0.00±0.00 j	Self breeding (inbred line) *	Unable
31	9905		Medium-height plant, yellow-oran- ge color, double petal		0.00±0.00 j	Self breeding (inbred line) *	Unable
32	Antigua·Orange		Short-height plant, yellow-orange color, double petal		0.00±0.00 j	Huaxianzi Gardening (F₁)	Unable
33	Discovery·Orange		Short-height plant, orange color, double petal		0.00±0.00 j	Sainuo Gardening (F₁)	Unable
34	Double cut flower marigold·Lemon		Tall-height plant, yellow color, double petal		0.00±0.00 j	Jinqiu Gardening (F₁)	Unable
35	Double marigold·Golden		Medium-height plant, yellow color, double petal		0.00±0.00 j	Jinqiu Gardening (F₁)	Unable
36	Marigold·Deep orange		Tall-height plant, orange color, double petal		0.00±0.00 j	Jinqiu Gardening (F₁)	Unable
37	MB		Short-height plant, yellow color, double petal		0.00±0.00 j	Self breeding (inbred line) *	Unable
38	New Fengsheng·Yellow		Short-height plant, yellow color, double petal		0.00±0.00 j	Sainuo Gardening (F₁)	Unable
39	S13		Medium-height plant, yellow color, double petal		0.00±0.00 j	Self breeding (inbred line) *	Unable
40	Yingtai·Golden		Medium-height plant, yellow color, double petal		0.00±0.00 j	Shijie seed (F₁)	Unable

No.	Genotype	Phenotype		Regeneration rate (%)		Source	Regeneration type
1	Perfect·Golden	Short-height plant, yellow-oran- ge color, double petal		54.21±12.11 a		Chunhe Gardening (F₁)	Easy
2	Milestone·Yellow	Short-height plant, yellow color, double petal		51.44±8.29 ab		Sainuo Gardening (F₁)	Easy
3	Proud·Orange-yellow	Short-height plant, yellow-oran- ge color, double petal		41.47±6.12 abc		Dannuo Gardening (F₁)	Moderate
4	Chrysanthemum marigold·Orange	Short-height plant, orange color, double petal		36.23±11.95 abcd		Huaxianzi Gardening (F₁)	Moderate
5	Xunlei·Yellow	Short-height plant, yellow color, double petal		33.48±13.73 abcde		Chunhe Gardening (F₁)	Moderate
6	Galore·Yellow	Medium-height plant, yellow co- lor, double petal		31.95±18.22 abcdef		Hongyue Gardening (F₁)	Moderate
7	Proud·Golden	Short-height plant, yellow-oran- ge color, double petal		30.50±10.33 abcdef		Dannuo Gardening (F₁)	Moderate
8	Double marigold· Light beige	Medium-height plant, yellow color, double petal		22.01±14.56 bcdefg		Jinqiu Gardening (F₁)	Difficult
9	DC	Short-height plant, orange color, double petal		21.33±13.66 cdefgh		Self breeding (inbred line) *	Difficult
10	Youyi·Yellow	Short-height plant, yellow color, double petal		18.00±1.83 cdefgh		Chunhe Gardening (F₁)	Difficult
11	Miracle·Yellow	Short-height plant, yellow color, double petal		13.29±9.62 cdefghi		Huaxianzi Gardening (F₁)	Difficult
12	Old Inka·Orange	Medium-height plant, orange co- lor, double petal		13.24±10.50 defghij		Hongyue Gardening (F₁)	Difficult
13	Liberty·Yellow	Short-height plant, yellow color, double petal		12.95±8.82 defghij		Huaxianzi Gardening (F₁)	Difficult
14	6210	Medium-height plant, yellow-oran- ge color, double petal		10.00±11.08 fghij		Self breeding (inbred line) *	Difficult
15	F8	Medium-height plant, orange co- lor, simple petal		9.33±5.96 efghij		Self breeding (inbred line) *	Difficult
No.	Genotype		Phenotype		Regeneration rate (%)	Source	Regeneration type
16	Fengsheng·Orange		Short-height plant, yellow-orange color, double petal		7.11±7.70 ghij	Shijie seed (F₁)	Difficult
17	AJ		Short-height plant, yellow-orange color, double petal		7.02±13.41 ghij	Self breeding (inbred line) *	Difficult
18	Inka·Orange-yellow		Short-height plant, yellow-orange color, double petal		5.11±7.87 ghij	Chunhe Gardening (F₁)	Difficult
19	S3		Medium-height plant, yellow color, simple petal		4.92±2.75 ghij	Self breeding (inbred line) *	Difficult
20	S12		Short-height plant, yellow color, simple petal		4.67±10.43 hij	Self breeding (inbred line) *	Difficult
21	S5		Short-height plant, yellow-orange color, simple petal		4.67±5.06 ghij	Self breeding (inbred line) *	Difficult
22	SW2		Medium-height plant, yellow color, double petal		4.67±10.43 hij	Self breeding (inbred line) *	Difficult
23	Hawaii		Medium-height plant, yellow-oran- ge color, double petal		4.22±5.87 ghij	Thailand (F₁)	Difficult
24	6170		Medium-height plant, yellow-oran- ge color, double petal		2.89±1.62 ghij	Self breeding (inbred line) *	Difficult
25	Jindun·Yellow		Short-height plant, yellow color, double petal		2.71±4.36 hij	Sainuo Gardening (F₁)	Difficult
26	GFR		Medium-height plant, orange color, double petal		1.92±1.75 hij	Self breeding (inbred line) *	Difficult
27	DH		Short-height plant, yellow color, double petal		1.33±1.83 ij	Self breeding (inbred line) *	Difficult
28	Vanilla		Medium-height plant, yellow-w- hite color, double petal		0.91±2.03 ij	Thailand (F₁)	Unable
29	Double Chrysanthe- mum marigold·Orange		Short-height plant, yellow-oran- ge color, double petal		0.71±1.60 ij	Jinqiu Gardening (F₁)	Unable
30	6155		Medium-height plant, yellow color, double petal		0.00±0.00 j	Self breeding (inbred line) *	Unable
31	9905		Medium-height plant, yellow-oran- ge color, double petal		0.00±0.00 j	Self breeding (inbred line) *	Unable
32	Antigua·Orange		Short-height plant, yellow-orange color, double petal		0.00±0.00 j	Huaxianzi Gardening (F₁)	Unable
33	Discovery·Orange		Short-height plant, orange color, double petal		0.00±0.00 j	Sainuo Gardening (F₁)	Unable
34	Double cut flower marigold·Lemon		Tall-height plant, yellow color, double petal		0.00±0.00 j	Jinqiu Gardening (F₁)	Unable
35	Double marigold·Golden		Medium-height plant, yellow color, double petal		0.00±0.00 j	Jinqiu Gardening (F₁)	Unable
36	Marigold·Deep orange		Tall-height plant, orange color, double petal		0.00±0.00 j	Jinqiu Gardening (F₁)	Unable
37	MB		Short-height plant, yellow color, double petal		0.00±0.00 j	Self breeding (inbred line) *	Unable
38	New Fengsheng·Yellow		Short-height plant, yellow color, double petal		0.00±0.00 j	Sainuo Gardening (F₁)	Unable
39	S13		Medium-height plant, yellow color, double petal		0.00±0.00 j	Self breeding (inbred line) *	Unable
40	Yingtai·Golden		Medium-height plant, yellow color, double petal		0.00±0.00 j	Shijie seed (F₁)	Unable

No.	6-BA (mg?L^-1)	IAA (mg?L^-1)	Regeneration rate (%)
1	3	1.0	37.50±15.00 b
2	3	1.5	40.00±14.14 b
3	3	1.8	35.00±5.77 b
4	3	2.0	52.50±5.00 ab
5	3	2.5	47.50±17.08 ab
6	3	3.0	65.00±12.91 a

No.	6-BA (mg?L^-1)	IAA (mg?L^-1)	Regeneration rate (%)
1	3	1.0	37.50±15.00 b
2	3	1.5	40.00±14.14 b
3	3	1.8	35.00±5.77 b
4	3	2.0	52.50±5.00 ab
5	3	2.5	47.50±17.08 ab
6	3	3.0	65.00±12.91 a

No.	ZT (mg?L^-1)	IAA (mg?L^-1)	Regeneration rate (%)
1	0.3	0.5	37.50±9.57 bc
2	0.3	1.0	35.00±10.00 bc
3	0.3	2.0	22.50±12.58 c
4	0.6	0.5	67.50±9.57 a
5	0.6	1.0	40.00±8.16 bc
6	0.6	2.0	55.00±10.00 ab
7	1.8	0.5	52.50±15.00 ab
8	1.8	1.0	55.00±19.15 ab
9	1.8	2.0	27.50±22.17 c

万寿菊再生体系的建立及优化

The Establishment and Optimization of a Regeneration System for Marigold (Tagetes erecta)

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相关文章 15

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Metrics

本文评价

No.	TDZ (mg?L^-1)	IBA (mg?L^-1)	Regeneration rate (%)
1	0.5	0.3	62.50±12.58 ab
2	0.5	0.5	55.00±17.32 ab
3	0.5	1.0	67.50±15.00 a
4	1.0	0.3	40.00±14.14 bcd
5	1.0	0.5	57.50±20.62 ab
6	1.0	1.0	47.50±9.57 abcd
7	1.5	0.3	32.50±12.58 cd
8	1.5	0.5	52.50±9.57 abc
9	1.5	1.0	27.50±9.57 d

No.	TDZ (mg?L^-1)	IBA (mg?L^-1)	Regeneration rate (%)
1	0.1	0.25	22.50±15.00 c
2	0.1	0.50	40.00±8.16 ab
3	0.2	0.25	35.00±12.91 ab
4	0.2	0.50	62.50±12.58 a
5	0.3	0.25	35.00±12.91 ab
6	0.3	0.50	47.50±12.58 ab
7	0.4	0.25	35.00±12.91 ab
8	0.4	0.50	32.50±17.08 ab

Code	Treatment	Explant No.	Elongation rate (%)	Status
MS1	No hormone	46	91.30	Strong; elongate 4-5 cm
MS2	0.1 mg·L^?1 TDZ; chang to 0.01 mg·L^?1 TDZ after 15 days	60	65.00	Healthy; elongate 3-4 cm
MS3	0.3 mg·L^?1 6-BA+1.0 mg·L^?1 NAA	53	30.19	Weak; elongate 0-2 cm
MS4	0.6 mg·L^?1 6-BA+1.0 mg·L^?1 NAA	55	18.18	Weak; elongate 0-1 cm
MS5	0.9 mg·L^?1 6-BA+1.0 mg·L^?1 NAA	57	12.28	Weak; elongate 0-1 cm
MS6	0.2 mg·L^?1 6-BA+0.6 mg·L^?1 NAA	45	33.33	Weak; elongate 0-2 cm

Leaflet segment	Regeneration rate (%)	Status
Whole leaflet	66.67±11.55 a	Callus and buds differentiated at the incision
Basal segment	63.49±8.64 a	Callus were differentiated at both end incisions, but only adventitious buds were differentiated in basal wound
Top segment	29.84±11.75 b	Callus and buds differentiated at the incision

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