Establishment of an Efficient Regeneration System in Iris lacteal var. chinensis with Immature Embryo

doi:10.7525/j.issn.1673-5102.2023.03.002

Abstract

Abstract:

The effect of sampling time and plant growth regulators on the induction of callus， somatic embryo and adventitious bud were investigated with immature embryos of Iris lacteal var. chinensis as explants. The results showed that the most feasible sampling time for immature embryos was 35-45 d after pollination， the young embryos were fully developed， but the endosperm was not completely hardened， and the young embryo was easy to dissect. The optimum medium for callus induction was MS+1.0 mg·L^-1 2，4-D+0.2 mg·L^-1 6-BA with the induction rate of 83.33%. The appropriate medium for somatic cell induction was MS+0.25 mg·L^-1 2，4-D+0.05 mg·L^-1 6-BA with the induction rate of 71.67%. The most suitable medium for adventitious bud induction and proliferation was MS+0.2 mg·L^-1 TDZ+0.05 mg·L^-1 NAA with the induction rate of 78.33% and proliferation coefficient 3.65 in this experiment. The rooting rate was more than 95% after four weeks cultivation on MS medium supplemented with 0.1 mg·L^-1 NAA. Then， the medium was washed off and explants were transplanted to sterilized vermiculite， and the survival rate was more than 95%. This study has established an efficient regeneration system of I. lacteal var. chinensis embryos， which can lay a foundation for genetic transformation and gene function analysis of I. lacteal var. chinensis.

Key words: Iris lacteal var. chinensis, immature embryo, callus induction, in vitro, regeneration

CLC Number:

S682.1+9

Wenjie AN, Yinjie WANG, Qingquan LIU, Yongheng YANG, Ting ZHANG, Shaoru FAN, Yongxia ZHANG, Haiyan YUAN. Establishment of an Efficient Regeneration System in Iris lacteal var. chinensis with Immature Embryo[J]. Bulletin of Botanical Research, 2023, 43(3): 333-339.

Figures/Tables 5

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Table 1

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处理 Treatment	2，4-D /（mg·L^-1）	6-BA /（mg·L^-1）	愈伤组织出现时间 Time of callus formation /d	愈伤组织诱导率 Callus induction rate /%	愈伤组织状态 Callus status
C1	0.5	0.1	20	53.33±8.82c	浅黄色，结构紧密，颗粒状 Light yellow，compact strcture， granular
C2	1.0	0.2	12	83.33±3.34ab	浅黄色，结构紧密，颗粒状 Light yellow，compact strcture， granular
C3	1.0	0.5	12	76.67±3.34b	黄色，大颗粒 Yellow， big granular
C4	1.5	0.2	10	81.11±8.39ab	淡黄色和透明白色，致密颗粒和松软组织 Light yellow and white，compact and soft strcture
C5	1.5	0.5	11	77.78±3.85ab	黄色和白色，大颗粒和松软组织 Yellow and white，big granular and soft strcture
C6	2.0	0.2	10	87.78±5.09a	淡黄色和透明白色，致密颗粒和松软组织 Light yellow and white，compact and soft strcture
C7	2.0	0.5	10	83.33±3.34ab	黄色和白色，大颗粒和松软组织 Yellow and white， big granular and soft strcture

处理 Treatment	TDZ /（mg·L^-1）	6-BA /（mg·L^-1）	NAA /（mg·L^-1）	不定芽诱导率 Induction rate /%	增殖系数 Proliferation coefficient
S1	0.5	0	0.1	55.00±5.00b	1.80±0.25c
S2	1.0	0	0.2	78.33±5.77a	3.65±0.26a
S3	1.5	0	0.2	68.33±2.89a	3.45±0.40ab
S4	0	0.5	0.1	33.33±7.64c	1.30±0.23d
S5	0	1.0	0.2	51.67±2.89b	2.23±0.10c
S6	0	1.5	0.2	45.00±8.66b	3.07±0.25b

处理 Treatment	NAA /（mg·L^-1）	生根率 Induction rate /%	根的状态 Roots status
R1	0	100	细弱，数量少 Weak，small quantity
R2	0.1	100	柔软、健壮，数量多 Soft and strong，large quantity
R3	0.2	100	柔软、健壮，数量多 Soft and strong，large quantity
R4	0.5	100	粗短、质硬、易断，数量多 Short，hard，easy to break，large quantity

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