Establishment of Tissue Culture and Virus Eliminating Technology for Shallot

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

Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (6): 1426-1434.doi: 10.16420/j.issn.0513-353x.2023-0351

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Establishment of Tissue Culture and Virus Eliminating Technology for Shallot

ZHANG Chunyu, WANG Chen, LIU Jianqing, ZHAN Xiaolei, LI Xiaoyu, WANG Yongzhi^**()

Institute of Plant Protection，Jilin Academy of Agricultural Sciences，Gongzhuling，Jilin 136100，China

Received:2024-03-15 Revised:2024-04-20 Online:2024-12-18 Published:2024-06-22
Contact: WANG Yongzhi

Abstract

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

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.

Figures/Tables 6

References 24

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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

Establishment of Tissue Culture and Virus Eliminating Technology for Shallot

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NAA/（mg · L^-1）	6-BA/（mg · L^-1）	愈伤组织生芽率/% Germination rate of callus	愈伤组织平均分化苗数 The average number of callus differentiated plantlets
0.2	0.5	24.5	3.75
	1.0	42.2	5.00
	1.5	37.7	4.83
0.4	0.5	48.9	5.71
	1.0	82.2	9.23
	1.5	77.8	8.27
0.6	1.0	55.6	3.67
	1.5	28.9	2.40

培养基 Medium	NAA /（mg · L^-1）	生根率/% Rooting rate	根系生长状态 Growth state
MS	0	80.0	生长慢，较细Slow growth，thin roots
	0.5	73.3	生长慢，较细Slow growth，thin roots
1/2MS	0	100.0	生长健壮，较多Strong growth，more roots
	0.5	93.3	生长较慢Slow growth