Study on Influencing Factor of Bulblet Enlargement in vitro of Asiatic Lily

doi:10.16420/j.issn.0513-353x.2024-0368

Acta Horticulturae Sinica ›› 2025, Vol. 52 ›› Issue (6): 1644-1660.doi: 10.16420/j.issn.0513-353x.2024-0368

• Cultivation?Physiology & Biochemistry • Previous Articles Next Articles

Study on Influencing Factor of Bulblet Enlargement in vitro of Asiatic Lily

WU Xiaodan¹, KONG Xiangfeng¹, GAO Li², GONG Tiangeng¹, PIAO Meiling¹, YU Pengcheng¹, and JIA Guixia¹^,^*()

¹ Beijing Key Laboratory of Ornamental Plants Germplasm Innovation & Molecular Breeding，National Engineering Research Center for Floriculture，Beijing Laboratory of Urban and Rural Ecological Environment，Beijing Laboratory of Urban and Rural Ecological Environment，Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education，School of Landscape Architecture，Beijing Forestry University，Beijing 100083，China
² Beijing Green Garden Group Co，Ltd. Beijing 100067，China

Received:2024-09-01 Revised:2025-02-22 Online:2025-06-20 Published:2025-06-20
Contact: and JIA Guixia

Abstract

Abstract:

This study used tissue cultured bulblets of the new Asian lily cultivars‘Tianxin’and ‘Xiaohaixing’as materials to investigate the effects of 6-BA，NAA，and glyphosate on inducing adventitious buds on the scales. Based on the screening of suitable induction media for adventitious buds，the effects of plant growth regulators such as salicylic acid（SA），paclobutrazol（PP₃₃₃），and 2, 4-dichlorobrassinolide（2,4-EBR），sucrose，activated carbon（AC），NAA，6-BA，and basic media on the swelling of small bulbs were studied，and the expression changes of starch synthase genes SSS，SSS2，sugar efflux transporters genes SWEET14 and SWEET15 during the swelling development of small bulbs were analyzed. The results showed that‘Tianxin’and‘Xiaohaixing’germinated the most on MS + 0.5 mg · L^-1 6-BA + 0.2 mg · L^-1 NAA + 30 g · L^-1 sucrose medium，with differentiation coefficients of 2.78 and 2.03，respectively. The enlargement coefficients of‘Tianxin’in MS + 1.0 mg · L^-1 PP₃₃₃+ 0.05 mg · L^-1 2,4-EBR + 60 g · L^-1 sucrose，and MS + 0.5 mg · L^-1 6-BA + 0.1 mg · L^-1NAA + 60 g · L^-1 sucrose + 1.0 g · L^-1 activated carbon medium were 4.42，5.15，and 4.16，respectively. The suitable swelling medium for them was selected as MS + 1.0 mg · L^-1 NAA + 0.1-0.5 mg · L^-1 6-BA + 60-90 g · L^-1 sucrose + 1.0-2.0 g · L^-1 AC. The enlargement coefficients of‘Tianxin’and‘Xiaohaixing’in B5 medium were significantly higher than those in MS and WPM medium treatments，at 4.57 and 4.83，respectively. In B5 medium，the expression levels of soluble starch synthase genes SSS and SSS2，as well as sugar efflux transporters SWEET14 and SWEET15，gradually increase with the growth of the‘Tianxin’bulblet，reaching their highest levels during the rapid expansion phase of bulblet. In the leaves，the expression levels are highest at 36 days before entering the rapid expansion phase of bulblet.

Key words: Lilium, bulblet enlargement, tissue culture, carbohydrate metabolism

WU Xiaodan, KONG Xiangfeng, GAO Li, GONG Tiangeng, PIAO Meiling, YU Pengcheng, and JIA Guixia. Study on Influencing Factor of Bulblet Enlargement in vitro of Asiatic Lily[J]. Acta Horticulturae Sinica, 2025, 52(6): 1644-1660.

Figures/Tables 16

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基因名称 Gene name	正向引物（5′-3′） Forward primer sequence	反向引物（5′-3′） Reverse primer sequence
SSS（CL7226_C1）	AATGGTTGGAGGTGGAAATCT	GGCCTCCTTCGGAACATAAA
SSS2（CL3977_C2）	CACTGGTGGCCTTAGAGATAC	CCAGCATACTCTCCTTCGATAAA
SWEET14（CL255_C3）	TGACTCTTCTTGGTTGGATCTG	GGCATGAACTCGACACTCTT
SWEET15（CL7570_C1）	GGGTTCCAATCTCTTCCCTATG	TCCGATCGAGTTAATGGTGATG
Actin（内参基因 Reference gene）	CCCATTGAGCACGGCATTGTC	GGATTGAGAGGAGCTTCGGTGAGA

基因名称 Gene name	正向引物（5′-3′） Forward primer sequence	反向引物（5′-3′） Reverse primer sequence
SSS（CL7226_C1）	AATGGTTGGAGGTGGAAATCT	GGCCTCCTTCGGAACATAAA
SSS2（CL3977_C2）	CACTGGTGGCCTTAGAGATAC	CCAGCATACTCTCCTTCGATAAA
SWEET14（CL255_C3）	TGACTCTTCTTGGTTGGATCTG	GGCATGAACTCGACACTCTT
SWEET15（CL7570_C1）	GGGTTCCAATCTCTTCCCTATG	TCCGATCGAGTTAATGGTGATG
Actin（内参基因 Reference gene）	CCCATTGAGCACGGCATTGTC	GGATTGAGAGGAGCTTCGGTGAGA

处理 Treatment	生长调节剂/（mg · L^-1） Growth regulator			甜心 Tianxin			小海星 Xiaohaixing
处理 Treatment	6-BA	NAA	6-BA/ NAA	出芽时间/d Budding time	诱导率 Inductivity	分化系数 Coefficient of differentiation	出芽时间/d Budding time	诱导率 Inductivity	分化系数 Coefficient of differentiation
1	0.5	0.1	5.0	8 ~ 24	1.00 ± 0 a	2.44 ± 0.25 abc	13 ~ 30	0.90 ± 0.11 ab	1.92 ± 0.29 a
2	0.5	0.2	2.2	8 ~ 24	0.90 ± 0.09 ab	2.78 ± 0.69 a	13 ~ 30	0.89 ± 0.01 ab	2.03 ± 0.18 a
3	0.5	1.0	0.5	14 ~ 30	0.82 ± 0.08 abc	2.14 ± 0.29 abcd	24 ~ 45	0.37 ± 0.10 de	1.25 ± 0.45 bcd
4	1.0	0.1	10.0	14 ~ 30	0.96 ± 0.06 a	2.52 ± 0.63 ab	18 ~ 35	0.84 ± 0.05 abc	2.01 ± 0.16 a
5	1.0	0.2	5.0	14 ~ 30	0.89 ± 0.12 ab	2.12 ± 0.51 abcd	18 ~ 35	0.83 ± 0.13 abc	1.72 ± 0.54 ab
6	1.0	1.0	1.0	20 ~ 35	0.74 ± 0.18 bcd	1.55 ± 0.16 d	30 ~ 55	0.25 ± 0.15 ef	1.22 ± 0.40 bcd
7	1.5	0.1	15.0	18 ~ 35	0.92 ± 0.07 ab	2.73 ± 0.25 a	24 ~ 45	0.92 ± 0.06 a	1.81 ± 0.45 ab
8	1.5	0.2	7.5	18 ~ 35	0.89 ± 0.09 ab	2.09 ± 0.19 abcd	24 ~ 45	0.75 ± 0.13 bc	1.94 ± 0.65 a
9	1.5	1.0	1.5	24 ~ 45	0.63 ± 0.07 d	1.60 ± 0.40 d	30 ~ 50	0.19 ± 0.17 fg	1.01 ± 0.58 cd
10	2.0	0.1	20.0	24 ~ 45	0.83 ± 0.16 abc	2.46 ± 0.64 abc	24 ~ 45	0.71 ± 0.14 c	1.64 ± 0.38 abc
11	2.0	0.2	10.0	24 ~ 45	0.74 ± 0.12 bcd	1.82 ± 0.17 cd	24 ~ 45	0.47 ± 0.04 d	1.55 ± 0.41 abc
12	2.0	1.0	2.0	24 ~ 45	0.66 ± 0.22 cd	1.89 ± 0.32 bcd	40 ~ 65	0.08 ± 0.06 g	0.67 ± 0.52 d

处理 Treatment	生长调节剂/（mg · L^-1） Growth regulator			甜心 Tianxin			小海星 Xiaohaixing
处理 Treatment	6-BA	NAA	6-BA/ NAA	出芽时间/d Budding time	诱导率 Inductivity	分化系数 Coefficient of differentiation	出芽时间/d Budding time	诱导率 Inductivity	分化系数 Coefficient of differentiation
1	0.5	0.1	5.0	8 ~ 24	1.00 ± 0 a	2.44 ± 0.25 abc	13 ~ 30	0.90 ± 0.11 ab	1.92 ± 0.29 a
2	0.5	0.2	2.2	8 ~ 24	0.90 ± 0.09 ab	2.78 ± 0.69 a	13 ~ 30	0.89 ± 0.01 ab	2.03 ± 0.18 a
3	0.5	1.0	0.5	14 ~ 30	0.82 ± 0.08 abc	2.14 ± 0.29 abcd	24 ~ 45	0.37 ± 0.10 de	1.25 ± 0.45 bcd
4	1.0	0.1	10.0	14 ~ 30	0.96 ± 0.06 a	2.52 ± 0.63 ab	18 ~ 35	0.84 ± 0.05 abc	2.01 ± 0.16 a
5	1.0	0.2	5.0	14 ~ 30	0.89 ± 0.12 ab	2.12 ± 0.51 abcd	18 ~ 35	0.83 ± 0.13 abc	1.72 ± 0.54 ab
6	1.0	1.0	1.0	20 ~ 35	0.74 ± 0.18 bcd	1.55 ± 0.16 d	30 ~ 55	0.25 ± 0.15 ef	1.22 ± 0.40 bcd
7	1.5	0.1	15.0	18 ~ 35	0.92 ± 0.07 ab	2.73 ± 0.25 a	24 ~ 45	0.92 ± 0.06 a	1.81 ± 0.45 ab
8	1.5	0.2	7.5	18 ~ 35	0.89 ± 0.09 ab	2.09 ± 0.19 abcd	24 ~ 45	0.75 ± 0.13 bc	1.94 ± 0.65 a
9	1.5	1.0	1.5	24 ~ 45	0.63 ± 0.07 d	1.60 ± 0.40 d	30 ~ 50	0.19 ± 0.17 fg	1.01 ± 0.58 cd
10	2.0	0.1	20.0	24 ~ 45	0.83 ± 0.16 abc	2.46 ± 0.64 abc	24 ~ 45	0.71 ± 0.14 c	1.64 ± 0.38 abc
11	2.0	0.2	10.0	24 ~ 45	0.74 ± 0.12 bcd	1.82 ± 0.17 cd	24 ~ 45	0.47 ± 0.04 d	1.55 ± 0.41 abc
12	2.0	1.0	2.0	24 ~ 45	0.66 ± 0.22 cd	1.89 ± 0.32 bcd	40 ~ 65	0.08 ± 0.06 g	0.67 ± 0.52 d

	因子Factor	平方和SS	自由度df	均方MS	F	P
甜心 Tianxin	6-BA	1.301	3	0.434	2.413	0.081
	NAA	4.832	2	2.416	13.443	0.000^**
	6-BA × NAA	1.774	6	0.296	1.645	0.161
	误差Error	7.009	39	0.180
	总计Total	254.532	51
小海星Xiaohaixing	6-BA	1.709	3	0.570	2.774	0.051
	NAA	8.709	2	4.354	21.204	0.000^**
	6-BA × NAA	0.456	6	0.076	0.370	0.894
	误差Error	9.857	48	0.205
	总计Total	158.494	60

Study on Influencing Factor of Bulblet Enlargement in vitro of Asiatic Lily

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草甘膦/mg · L^-1 Glyphosate	‘甜心’‘Tianxin’		‘小海星’‘Xiaohaixing’
草甘膦/mg · L^-1 Glyphosate	诱导率 Inductivity	分化系数 Coefficient of differentiation	诱导率 Inductivity	分化系数 Coefficient of differentiation
0（对照 Control）	0.89 ± 0 ab	2.26 ± 0.39 ab	0.89 ± 0.11 ab	2.59 ± 0.82
1.0	0.93 ± 0.13 a	2.70 ± 0.93 a	0.96 ± 0.06 a	3.56 ± 1.34
2.0	0.71 ± 0.06 bc	1.44 ± 0.12 b	0.78 ± 0.11 ab	2.51 ± 0.56
3.0	0.56 ± 0.12 c	1.30 ± 0.50 b	0.74 ± 0.06 b	2.33 ± 0.51

处理 Treatments	NAA/（mg · L^-1）	蔗糖/（g · L^-1） Sucrose	AC/（g · L^-1）	6-BA/（mg · L^-1）	膨大系数Expansion coefficient
处理 Treatments	NAA/（mg · L^-1）	蔗糖/（g · L^-1） Sucrose	AC/（g · L^-1）	6-BA/（mg · L^-1）	甜心Tianxin	小海星Xiaohaixing
1	0.1	30	0.5	0.1	2.67 ± 0.73 de	3.05 ± 1.26 de
2	0.1	60	1.0	0.5	4.16 ± 1.34 a	4.12 ± 1.47 abc
3	0.1	90	2.0	1.0	3.27 ± 1.54 bcd	3.85 ± 0.91 abcd
4	0.5	30	1.0	1.0	2.40 ± 0.60 e	2.95 ± 0.78 e
5	0.5	60	2.0	0.1	3.63 ± 1.24 abc	4.25 ± 1.28 abc
6	0.5	90	0.5	0.5	3.91 ± 1.64 ab	3.55 ± 0.69 bcde
7	1.0	30	2.0	0.5	2.96 ± 0.79 cde	4.35 ± 1.17 ab
8	1.0	60	0.5	1.0	2.79 ± 0.91 de	3.43 ± 0.88 cde
9	1.0	90	1.0	0.1	3.13 ± 1.11 bcde	4.43 ± 1.65 a

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