Plant Regeneration System from Cotyledon of Camellia japonica (Naidong)

doi:10.7525/j.issn.1673-5102.2021.03.001

Abstract

Abstract:

As the floral emblem of Qingdao，Camellia japonica（Naidong） is a precious ornamental species during the winter of the north， but the quantity of C. japonica（Naidong） cultivars cannot meet requirements for application in landscape at present， so it is necessary to enlarge propagation by the means of tissue culture. In order to establish the regeneration system of C. japonica（Naidong）， different factors affected on callus induction and plantlet regeneration were studied from mature cotyledons of C. japonica （Naidong）. The optimum timing of sterilization in C. japonica（Naidong） seeds was soaked for 8 min in the 2% NaClo， callus induction rate was 100%. The optimal hormone combination of cotyledon callus subculture was 0.5 mg·L^-1 NAA+1.5 mg·L^-1 6-BA， the multiplication rate was 4.06. After 2-time subcultured， the callus was transferred to MS+1.0mg·L^-1 NAA+10.0 mg·L^-1 6-BA for differentiation of adventitious buds， the rate of adventitious bud differentiation was 10.7%. The shoot base was immersed in 500 mg·L^-1 IBA solution for 70 min， then transferred to 1/2MS， the rate of rooting could reach 40%. Regeneration system from mature cotyledons of C.japonica（Naidong） by inducing callus， adventitiousshoots and roots was established， which would provide technical support for the vegetative production and innovation of its germplasm resources.

Key words: Camellia japonica（Naidong）, cotyledon, callus, plantlet regeneration

CLC Number:

S685.14

Hong-Mei TIAN, Xiao GUO, Kui-Ling WANG, Ying-Kun SUN. Plant Regeneration System from Cotyledon of Camellia japonica (Naidong)[J]. Bulletin of Botanical Research, 2021, 41(3): 321-328.

Figures/Tables 8

Fig.1

Fig.2

Fig.3

Table 2

Effects of different hormone combinations on cotyledon callus growth

实验编号 Test No.	激素配比 Hormone combination		增殖倍数 Multiplication rate	生长状态 Growing status
实验编号 Test No.	NAA（mg·L^-1）	6-BA（mg·L^-1）	增殖倍数 Multiplication rate	生长状态 Growing status
1	0.1	2.0	2.31	黄绿色，微泛红，颗粒细小紧致 Yellow-green and pink,creamy and compact
2	0.1	1.5	2.77	浅黄绿色，少数微红，比较水润 Pale greenish-yellow,a little callus was pink,moist
3	0.1	1.0	1.15	30 %的愈伤组织为黄绿色，其余为灰褐色 Thirty percent callus was yellow-green,the rest was beige
4	0.1	0.5	2.42	表面干燥，颗粒状明显，生长量大 The surface was dry,granular,growth increment larger
5	0.2	2.0	2.08	浅黄绿色，比较水润Pale greenish-yellow,moist
6	0.2	1.5	1.77	黄绿色，比较水润Yellow-green,moist
7	0.2	1.0	1.67	黄绿色，表面比较干燥Yellow-green,the surface was dry
8	0.2	0.5	1.27	黄绿色，表面干燥，愈伤边缘略发黑 Yellow-green , the surface was dry,callus of peripheral was black
9	0.3	2.0	1.85	微泛红，表面干燥，颗粒细小紧致 Pink,the surface was dry,creamy,creamy and compact
10	0.3	1.5	2.67	黄绿色，表面稍微干燥Yellow-green,the surface was a little dry
11	0.3	1.0	1.86	黄绿色，比较水润Yellow-green,moist
12	0.3	0.5	1.91	黄绿色，比较水润Yellow-green,moist
13	0.4	2.0	1.89	浅黄绿色，比较水润Pale greenish-yellow,moist
14	0.4	1.5	3.29	愈伤组织为浅红色和黄绿色，6.67%的愈伤出现不均质颗粒 Yellow-green and lighter red,nodular of 6.67 % callus
15	0.4	1.0	3.16	黄绿色，少数微微泛红，表面干燥 Yellow-green,a little callus was pink,the surface was dry
16	0.4	0.5	1.17	干燥，愈伤表面呈黑褐色Dry,the surface of callus was dark brown
17	0.5	2.0	2.38	黄绿色，比较水润Yellow-green,moist
18	0.5	1.5	4.06	黄绿色，微微泛红，20%的愈伤出现不均质颗粒 Yellow-green and pink,nodular of 20% callus
19	0.5	1.0	2.35	黄绿色，表面稍微干燥yellow-green,the surface was a little dry
20	0.5	0.5	1.83	愈伤微微泛红，较水润Pink and moist

Table 2

Fig.4

Table3

Fig.5

Table 4

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编号 Test No.	不同激素配比 Different hormone combination(mg·L^-1)	丛生芽诱导率 The rate of differentiation(%)	愈伤组织生长状态 Growing status of callus
1	0.25IBA+106-BA	0	愈伤组织呈灰褐色Beige
2	0.5IBA+106-BA	0	与培养基接触的愈伤组织呈黑褐色，质地松散 The callus contacted to the medium was dark brown and loose structure
3	0.25NAA+106-BA	1.50	黄绿色，质地紧实，比较水润Yellow-green, compact, moist
4	0.5NAA+106-BA	0.89	浅黄绿色，较干燥，质地紧实Pale greenish-yellow,dry,compact
5	0.75NAA+106-BA	3.90	黄绿色，质地紧实，比较水润The callus was yellow-green,moist,compact
6	1NAA+106-BA	10.70	愈伤组织黄绿色，质地紧实且水润有光泽Yellow-green,compact and moist

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