珠葱组培脱毒技术研究

doi:10.16420/j.issn.0513-353x.2023-0351

摘要/Abstract

摘要：

以珠葱（Allium cepa var. aggregatum）鳞茎茎尖为外植体，以直接诱导丛生芽和愈伤组织诱导分化两种方式获得组培苗，对消毒处理方式、培养基植物生长调节剂配比、茎尖剥离大小等条件进行优化，采用ELISA检测方法对葱潜隐病毒（SLV）和洋葱黄矮病毒（OYDV）复合侵染的鳞茎组培苗的脱毒效果进行检测。结果表明：珠葱茎尖外植体采用75%乙醇浸泡30 s后用2%次氯酸钠溶液浸泡15 min消毒效果最佳，污染率为6.7%，成活率93.3%；确定最佳茎尖分生组织诱导培养基为MS + 0.5 mg · L^-1NAA + 1.0 mg · L^-16-BA，诱芽率达85.6%，平均分化芽数3.3个；最佳愈伤组织培养基为MS + 0.4 mg · L^-1 NAA + 0.5 mg · L^-1 2,4-D，愈伤率为71.7%；最佳生芽培养基为MS + 0.4 mg · L^-1NAA + 1.0 mg · L^-16-BA，生芽率达82.2%，平均分化苗数9.23个；在不加外源植物生长调节剂的1/2MS培养基中即可满足生根需求，生根率达100%；通过剥离0.3 ~ 0.4 mm的茎尖促愈伤获得珠葱脱毒种苗，利用ELISA检测组培苗中，脱毒率可达92%；对已完成脱毒且生长健壮的组培苗进行移栽，成活率为90%。

关键词: 珠葱, 茎尖诱导, 愈伤分化, 脱毒种苗

Abstract:

The bulb tip of shallot was used as explants to obtain tissue culture seedlings by direct induced clustered shoots and callus-induced differentiation. The disinfection treatment，plant growth regulator ratio of medium and the size of bulb tip were optimized，and the virus-free effect of shallot latent virus（SLV）and onion yellow dwarf virus（OYDV） in tissue culture seedlings was tested by ELISA. The results showed that the bulb tip explants were soaked in 75% ethanol for 30 s and 2% sodium hypochlorite solution for 15 min. The pollution rate was 6.7% and the survival rate was 93.3%. The optimal bulb tip induction medium was MS + 0.5 mg · L^-1NAA + 1.0 mg · L^-16-BA，with the bud induction rate of 85.6% and the average number of differentiated buds was 3.3. The optimal callus medium was MS + 0.4 mg · L^-1NAA + 0.5 mg · L^-12,4-D and the callus rate was 71.7%. The optimal bud-forming medium was MS + 0.4 mg · L^-1NAA + 1.0 mg · L^-16-BA，with the bud-forming rate was 82.2% and the average number of seedlings was 9.23. The rooting requirement could be satisfied in 1/2MS medium without plant growth regulator ratio，and the rooting rate could reach 100%. The shallot virus-free seedlings were obtained by stripping the bulb tips of 0.3-0.4 mm，with the virus-free rate of 92%. The survival rate of tissue culture seedlings was 90% after transplanting.

Key words: shallot, bulb tip induction, callus differentiation, virus-free seedling

张春雨, 王晨, 刘建青, 战笑蕾, 李小宇, 王永志. 珠葱组培脱毒技术研究[J]. 园艺学报, 2024, 51(6): 1426-1434.

ZHANG Chunyu, WANG Chen, LIU Jianqing, ZHAN Xiaolei, LI Xiaoyu, WANG Yongzhi. Establishment of Tissue Culture and Virus Eliminating Technology for Shallot[J]. Acta Horticulturae Sinica, 2024, 51(6): 1426-1434.

图/表 6

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处理浓度/% Concentrations of NaClO	处理时间/min Time of treatment	污染率/% Contamination rate	成活率/% Survival rate
2	5	83.3	16.7
	10	43.3	56.7
	15	6.7	93.3
	20	6.7	83.3
10	5	13.3	86.7
	10	3.3	73.3

处理浓度/% Concentrations of NaClO	处理时间/min Time of treatment	污染率/% Contamination rate	成活率/% Survival rate
2	5	83.3	16.7
	10	43.3	56.7
	15	6.7	93.3
	20	6.7	83.3
10	5	13.3	86.7
	10	3.3	73.3

NAA/（mg · L^-1）	6-BA/（mg · L^-1）	诱芽率/% Rate of induced buds	平均分化芽数 Average number of differentiated buds	株高/cm Plant height
0.2	0.5	24.4	1.0	3.30 e
	0.8	26.7	1.0	3.48 d
	1.0	41.1	1.0	3.79 c
	1.5	47.8	1.2	3.37 de
0.5	0.8	51.1	1.4	3.82 c
	1.0	85.6	3.3	4.43 a
	1.5	57.8	1.0	4.13 b
0.8	1.0	43.3	1.1	3.80 c
	1.5	46.7	1.0	3.70 c

NAA/（mg · L^-1）	6-BA/（mg · L^-1）	诱芽率/% Rate of induced buds	平均分化芽数 Average number of differentiated buds	株高/cm Plant height
0.2	0.5	24.4	1.0	3.30 e
	0.8	26.7	1.0	3.48 d
	1.0	41.1	1.0	3.79 c
	1.5	47.8	1.2	3.37 de
0.5	0.8	51.1	1.4	3.82 c
	1.0	85.6	3.3	4.43 a
	1.5	57.8	1.0	4.13 b
0.8	1.0	43.3	1.1	3.80 c
	1.5	46.7	1.0	3.70 c

NAA / （mg · L^-1）	2,4-D / （mg · L^-1）	愈伤率/% Callus rate	生长状态 Growth state
0.2	0.5	41.7	淡黄色或白色颗粒状，较疏松但小 Pale yellow or white grainy，relatively loose but small
	0.8	38.3	淡黄色颗粒状，较致密 Light yellow granular，relatively dense
	1.0	31.7	淡黄色或黄褐色颗粒状，较致密 Light yellow or yellowish brown granular，relatively dense
0.4	0.5	71.7	淡黄色和白色颗粒状，较疏松 Pale yellow or white grainy，relatively loose
	0.8	45.0	淡黄色或黄褐色，较疏松 Light yellow or yellowish brown，relatively loose
	1.0	36.7	黄色或黄褐色，堆积且致密 Yellow or yellowish brown，packed and dense
0.6	0.5	66.7	黄色，较疏松 Yellow，relatively loose
	0.8	56.7	黄褐色或较深黄色，较坚硬 Yellowish brown or darker yellow，harder