Rhizome Proliferation and Bud differentiation of Cymbiduim sinense ‘Wuzicui’ × C. sinense ‘Honghua’ Hybrid

doi:10.3969/j.issn.1000-2561.2020.05.019

Abstract

Abstract:

Cymbiduim sinense is also called Bao Sui Lan (Heralding New Year Orchid), which belongs to Cymbiduim (Orchidaceae). It is loved by Chinese people for its robust plant type and flowering period during the Spring Festival. In view of the low proliferation rate and the difficulty of bud differentiation of the rhizome of C. sinense. In this study, the hybrid rhizomes of C. sinense were used as materials, through the combination of auxin, mitogen and organic additives in different concentrations, in order to find a suitable medium formula for the proliferation and differentiation of the rhizomes of hybrid C. sinense. The results showed that the most suitable medium for rhizomes proliferation was MS+ 8 mg/L 6-BA+0.1 mg/L NAA+0.8 mg/L TDZ+1.5 g/L AC+35.0 g/L Su+6.0 g/L Ag, with a proliferation coefficient 5.35 and the growth rate 3.44. The most suitable medium for leaf bud differentiation was MS+10.0 mg/L 6-BA+ 0.7 mg/L NAA+0.2 mg/L TDZ+1.5 g/L AC+ 35.0 g/L Su+6.0 g/L Ag, with a leaf bud differentiation rate 52.5%. The most suitable medium for leaf bud differentiation was MS+8.0 mg/L 6-BA+0.1 mg/L NAA+7.0 g/L peptone+100.0 mg/L CW+1.0 g/L AC, with a leaf bud differentiation rate 173.30%. This study explored the effects of hormones and additives on the proliferation and differentiation of the rhizomes, which could achieve large-scale reproduction of the tissue culture seedlings.

Key words: Cymbiduim sinense, rhizome, proliferation, differentiation, induction

CLC Number:

S682.31

XIE Juan,LI Long,XING Yue,WENG Qingshi,FAN Wanlin,PENG Donghui. Rhizome Proliferation and Bud differentiation of Cymbiduim sinense ‘Wuzicui’ × C. sinense ‘Honghua’ Hybrid[J]. Chinese Journal of Tropical Crops, 2020, 41(5): 985-993.

Figures/Tables 9

References 23

[1]	陈丽, 潘瑞炽, 陈汝民. 墨兰原球茎生长的研究[J]. 热带亚热带植物学报, 1999(1):59-64.
[2]	施福军, 莫昭展, 韦江萍, 等. 墨兰的无菌播种及根状茎的增殖研究[J]. 安徽农业科学, 2008,36(32):13968-13969, 13992.
[3]	左利娟, 汤久杨, 李志强. 墨兰杂交及F1代离体培养研究[J]. 北京农业职业学院学报, 2017,31(3):22-25.
[4]	丁雪珍, 韩磊, 张文静. 墨兰增殖培养基的筛选研究[J]. 北方园艺, 2009(8):208-209.
[5]	陈兰芬, 王晶, 田亦平, 等. 墨兰组织培养根状茎分化技术研究[J]. 河北林果研究, 2011,26(1):22-24.
[6]	鲁雪华, 郭文杰, 林勇. 墨兰的无菌播种和植株再生[J]. 亚热带植物科学, 1999(1):34-37.
[7]	傅雪琳, 张志胜, 何平, 等. 墨兰根状茎绿芽分化的研究[J]. 华南农业大学学报, 2000(3):53-55.
[8]	朱根发, 叶秀斌, 陈明莉, 等. 培养基不同成分对墨兰根状茎分化成苗的影响[J]. 中南林学院学报, 2003(5):42-44, 58.
[9]	许申平, 袁秀云, 王默霏, 等. 墨兰(Cymbiduim sinense)组培快繁技术体系研究[J]. 热带作物学报, 2018,39(5):926-930.
[10]	Yan X, Feng-an Y U, Zhen-hua P. Tissue culture of Cymbidium sinensis[J]. Forest Research, 2003,16(4):434-438.
[11]	Chen Y Y, Zhang Y, Zhang C, et al. A study on aseptic seed germination of interspecific hybrid between Cymbidium hybrida × C. sinense and C. faberi[J]. Acta Horticulturae Sinica, 2009,36(3):441-446.
[12]	Gao R, Wu S Q, Piao X C, et al. Micropropagation of Cymbidium sinense using continuous and temporary airlift bioreactor systems[J]. Acta Physiologiae Plantarum, 2014,36(1):117-124.
[13]	Chang C, Chang W. Effect of thidiazuron on bud development of Cymbidium sinense Willd in vitro[J]. Plant Growth Regulation, 2000,30(2):171-175.
[14]	徐程, 詹忠根, 张铭. 中国兰的组织培养[J]. 植物生理学通讯, 2002(2):171-174.
[15]	郑艳艳. 墨兰组培体系的优化及根状茎在生物反应器培养的研究[D]. 延吉: 延边大学, 2011.
[16]	刘昳雯, 许春梅, 王丹, 等. 莲瓣兰“滇梅”根状茎的增殖与分化技术研究[J]. 安徽农业科学, 2014,42(10):2890-2892.
[17]	Duan J X, Yazawa S. Floral induction and development in Phalaenopsis in vitro[J]. Plant Cell Tissue and Organ Culture, 1995,43(1):71-74. DOI URL
[18]	Duan J X, Yazawa S. In vitro floral development in × Doriella Tiny (Doritis pulcherrima × Kingiella philippinensis)[J]. Scientia Horticulturae, 1994,59(3-4):253-264. DOI URL
[19]	Duan J X, Yazawa S. In vitro flowering of Doriella, Phalaenopsis and Dendrobium[J]. Proceedings of NIOC, 1994: 87-96.
[20]	陈云喜, 何丹丹, 廖浩如, 等. 影响墨兰×兔耳兰根状茎芽分化的因素[J]. 中国农学通报, 2010,26(9):65-69.
[21]	Zeng S, Wu K, da Silva J A T, et al. Asymbiotic seed germination, seedling development and reintroduction of Paphiopedilum wardii Sumerh., an endangered terrestrial orchid[J]. Scientia Horticulturae, 2012,138:198-209.
[22]	罗虹. 活性炭对墨兰根状茎生长的影响[J]. 广东农业科学, 1998(1):30-31.
[23]	翁锦周, 林加耕, 林江波. 不同浓度活性炭对墨兰离体培养的影响[J]. 亚热带植物科学, 2006(3):37-38, 36.

处理 Treatments	A:NAA浓度 NAA concentration /(mg·L^-1)	B:6-BA浓度 6-BA concentration /(mg·L^-1)	C:TDZ浓度 TDZ concentration /(mg·L^-1)		生长速度 Growth speed		增殖系数 Multiplication coefficient
1	0.10	2.00	0.20		3.45±0.53^bc		5.35±0.10^a
2	0.10	5.00	0.50		3.34±0.19^c		4.08±0.30^bc
3	0.10	8.00	0.80		4.46±0.02^a		5.05±1.05^ab
4	0.30	2.00	0.50		3.34±0.36^c		3.05±0.70^c
5	0.30	5.00	0.80		3.75±0.46^bc		4.13±1.13^bc
6	0.30	8.00	0.20		3.37±0.50^c		3.07±0.36^c
7	0.50	2.00	0.80		2.55±0.32^d		3.00±0.20^c
8	0.50	5.00	0.20		4.04±0.03^ab		4.08±0.08^bc
9	0.50	8.00	0.50	3.37±0.31^c		3.58±0.08^c
Ka1	3.75	3.11	3.62	Ra值:B>A>C 最优水平:A1B3C1
Ka2	3.49	3.71	3.35
Ka3	3.32	3.73	3.59
Ra	0.43	0.62	0.27
Kb1	4.83	3.80	4.16	Rb值:A>C>B 最优水平:A1B2C1
Kb2	3.41	4.10	3.57
Kb3	3.56	3.90	4.06
Rb	1.42	0.30	0.59

处理 Treatments	A:NAA浓度 NAA concentration /(mg·L^-1)	B:6-BA浓度 6-BA concentration /(mg·L^-1)	C:TDZ浓度 TDZ concentration /(mg·L^-1)		生长速度 Growth speed		增殖系数 Multiplication coefficient
1	0.10	2.00	0.20		3.45±0.53^bc		5.35±0.10^a
2	0.10	5.00	0.50		3.34±0.19^c		4.08±0.30^bc
3	0.10	8.00	0.80		4.46±0.02^a		5.05±1.05^ab
4	0.30	2.00	0.50		3.34±0.36^c		3.05±0.70^c
5	0.30	5.00	0.80		3.75±0.46^bc		4.13±1.13^bc
6	0.30	8.00	0.20		3.37±0.50^c		3.07±0.36^c
7	0.50	2.00	0.80		2.55±0.32^d		3.00±0.20^c
8	0.50	5.00	0.20		4.04±0.03^ab		4.08±0.08^bc
9	0.50	8.00	0.50	3.37±0.31^c		3.58±0.08^c
Ka1	3.75	3.11	3.62	Ra值:B>A>C 最优水平:A1B3C1
Ka2	3.49	3.71	3.35
Ka3	3.32	3.73	3.59
Ra	0.43	0.62	0.27
Kb1	4.83	3.80	4.16	Rb值:A>C>B 最优水平:A1B2C1
Kb2	3.41	4.10	3.57
Kb3	3.56	3.90	4.06
Rb	1.42	0.30	0.59

源 Source	因变量 Dependent variable		III类平方和 Class III square sum	自由度 Degree of freedom	均方 Mean square	F	P
修正模型	生长速度		3.430^a	6	0.572	2.073	0.103
	增殖系数		13.127^b	6	2.188	4.062	0.008
截距	生长速度		334.432	1	334.432	1212.398	0
	增殖系数		417.563	1	417.563	775.349	0
A:NAA浓度/(mg·L^-1)	生长速度		0.821	2	0.411	1.489	0.25
		增殖系数	10.908	2	5.454	10.127	0.001
B:6-BA浓度/(mg·L^-1)		生长速度	2.223	2	1.112	4.030	0.034
		增殖系数	0.413	2	0.207	0.384	0.686
C:TDZ浓度/(mg·L^-1)		生长速度	0.386	2	0.193	0.699	0.509
		增殖系数	1.805	2	0.903	1.676	0.212
误差		生长速度	5.517	20	0.276	—	—
		增殖系数	10.771	20	0.539	—	—
总计		生长速度	343.379	27	—	—	—
		增殖系数	441.460	27	—	—	—
修正后总计		生长速度	8.947	26	—	—	—
		增殖系数	23.898	26	—	—	—

源 Source	因变量 Dependent variable		III类平方和 Class III square sum	自由度 Degree of freedom	均方 Mean square	F	P
修正模型	生长速度		3.430^a	6	0.572	2.073	0.103
	增殖系数		13.127^b	6	2.188	4.062	0.008
截距	生长速度		334.432	1	334.432	1212.398	0
	增殖系数		417.563	1	417.563	775.349	0
A:NAA浓度/(mg·L^-1)	生长速度		0.821	2	0.411	1.489	0.25
		增殖系数	10.908	2	5.454	10.127	0.001
B:6-BA浓度/(mg·L^-1)		生长速度	2.223	2	1.112	4.030	0.034
		增殖系数	0.413	2	0.207	0.384	0.686
C:TDZ浓度/(mg·L^-1)		生长速度	0.386	2	0.193	0.699	0.509
		增殖系数	1.805	2	0.903	1.676	0.212
误差		生长速度	5.517	20	0.276	—	—
		增殖系数	10.771	20	0.539	—	—
总计		生长速度	343.379	27	—	—	—
		增殖系数	441.460	27	—	—	—
修正后总计		生长速度	8.947	26	—	—	—
		增殖系数	23.898	26	—	—	—

处理 Treatments	A:NAA浓度 NAA concentration/(mg·L^-1)	B:6-BA浓度 6-BA concentration/(mg·L^-1)	C:TDZ浓度 TDZ concentration/(mg·L^-1)	叶芽分化率 Leaf bud differentiation rate/%		株高 Plant height/cm
1	0.30	8.00	0.20	37.50±10.61^ab		2.79±0.06^bc
2	0.30	10.00	0.50	30.00±21.21^abc		4.04±0.77^ab
3	0.30	12.00	0.80	22.50±10.61^bc		1.35±0.92^c
4	0.50	8.00	0.50	5.00±0.00^c		1.35±0.11^c
5	0.50	10.00	0.80	40.00±7.07^ab		4.11±0.98^ab
6	0.50	12.00	0.20	27.50±3.54^abc		2.71±0.98^bc
7	0.70	8.00	0.80	22.50±7.50^bc		4.98±0.31^a
8	0.70	10.00	0.20	52.50±10.61^a		5.36±1.31^a
9	0.70	12.00	0.50	30.00±14.14^abc		4.46±0.72^ab
Ka1	0.30	0.22	0.39		Ra值:B>C>A 最优水平:A3B2C1
Ka2	0.24	0.41	0.22
Ka3	0.35	0.27	0.28
Ra	0.11	0.19	0.18
Kb1	2.73	3.04	3.62		Rb值:A>B>C 最优水平:A3B2C1
Kb2	2.72	4.50	3.28
Kb3	4.93	2.84	3.48
Rb	2.21	1.66	0.34