Elimination Technology of Sugarcane Streak Mosaic Virus in Chewing Cane

doi:10.13560/j.cnki.biotech.bull.1985.2025-0268

Abstract

Abstract:

Objective This study is aimed to investigate efficient virus elimination technology for Sugarcane Streak Mosaic Virus (SCSMV) in chewing cane ‘Badila’, providing technical support for improving sugarcane yield. Method Using SCSMV-infected ‘Badila’ as experimental material, six hot water treatment groups were established: Untreated control, 50 ℃ for 2 h, 50 ℃ for 3 h, 52 ℃ for 3 h, 54 ℃ for 3 h, and 58 ℃ for 1 h. Virus-free plantlets were produced through shoot-tip culture and bud-tip culture. The virus-free plantlets were subjected to molecular detection of SCSMV, and their agronomic traits were investigated after pot cultivation to comprehensively analyze the virus elimination effects. Result Hot water treatment significantly reduced bud germination rates, with temperature and treatment duration showing a negative correlation with germination rate. No sprouting was observed at 54 ℃ and 58 ℃, making it impossible to obtain shoot-tip explants for tissue culture. However, the treatment had minimal impact on the survival rate of bud-tip culture. Even at 54 ℃ and 58 ℃, the bud apical meristems still maintained relatively high survival rates (83.3% and 76.9%, respectively). In terms of virus elimination, hot water treatment at 50 ℃ for 2 h, 50 ℃ for 3 h, and 52 ℃ for 3 h combined with shoot-tip culture only reduced the SCSMV detection rate to 80%. Under the same conditions, bud-tip culture reduced the rates to 70%, 60%, and 30%, respectively. Notably, under 58 ℃ treatment for 1 h, bud-tip culture achieved complete virus elimination (100%). Field trials demonstrated that virus-free sugarcane plants exhibited significantly better growth and agronomic traits compared to the control group. Plant height, internode length, single stem weight, brix and sugar content increased by 43.5%, 53.5%, 58.3%, 12.4% and 30.0%, respectively. Conclusion This study successfully established a virus elimination system combining hot water treatment and bud-tip culture, characterized by high plantlet regeneration and virus elimination efficiency.

Key words: chewing cane, SCSMV, heat treatment, bud-tip culture, virus elimination

HU Xin, LUO Zheng-ying, LIU Xin-long, WU Cai-wen, WU Zhuan-di. Elimination Technology of Sugarcane Streak Mosaic Virus in Chewing Cane[J]. Biotechnology Bulletin, 2025, 41(9): 62-70.

Figures/Tables 7

Fig. 1 Flowchart of the bud-tips culture technique of chewing caneA: Hot water treatment. B: Removing bud scales. C: Extracting bud tip. D: Inoculation of bud-tip onto MS medium. E: Differentiation of bud-tip into seedlings. F: Subculture of seedlings into clumping seedlings

Fig. 2 RNA extraction and molecular detection of SCSMV in chewing caneA: Extraction of total RNA from chewing cane leaves. B: PCR detection of SCSMV virus. M indicates the DNA molecular marker, N is the negative control using sterile water, P is the positive control carrying SCSMV virus, and samples are numbered 1‒20

Fig. 3 Homology analysis of PCR product sequences

Table 1 Effects of different hot water treatment conditions on the survival rates of stem buds

编号 No.	温水处理条件 Hot water treatment conditions					芽尖组织培养 Bud-tip culture
编号 No.	温水处理条件 Hot water treatment conditions	出芽率Germination rate (%)	茎尖成活率Survival rate of stem tips (%)	污染率 Contamination rate (%)	芽苗状态 Emergence of seedlings	芽尖成活率 Survival rate of bud-tips (%)	污染率 Contamination rate (%)	芽苗状态 Emergence of seedlings
A	未处理	86.3	81.8	15.7	丛生芽，长势良好	87.0	11.9	丛生芽，长势良好
B	50 ℃ 2 h	37.5	89.3	10.0	丛生芽，长势良好	95.2	5.0	丛生芽，长势良好
C	50 ℃ 3 h	23.8	85.7	13.2	丛生芽，长势良好	90.9	5.6	丛生芽，长势良好
D	52 ℃ 3 h	12.9	84.6	15.0	丛生芽，长势良好	90.9	3.1	丛生芽，长势良好
E	54 ℃ 3 h	0	-		-	83.3	2.3	丛生芽，长势良好
F	58 ℃ 1 h	0	-		-	76.9	0	丛生芽，长势良好

Fig. 4 SCSMV molecular detection in tissue-cultured seedlings of chewing cane under different treatment conditionsA: Non-hot water treatment group. B: The 50 ℃ 2 h hot water treatment group. C: The 50 ℃ 3 h hot water treatment group. D: The 52 ℃ 3 h hot water treatment group. E: The 54 ℃ 3 h treatment group. F: The 58 ℃ 1 h treatment group. A‒D: Samples 1‒10 are from stem-tip culture seedlings, and samples 11‒20 are from bud-tip culture seedlings. E‒F: Sample 1‒20 were from bud tip culture seedlings. M: DNA molecular marker. N: Sterile water negative control. P: Positive control with SCSMV virus cDNA

Fig. 5 Symptoms of mosaic disease and agronomic traits of virus-infected and virus-free chewing cane

Table 2 Comparison of agronomic traits between virus-infected and virus-free chewing cane

材料

Material

株高

Plant height (cm)

茎径

Stem diameter (cm)

节间长度

Internode length (cm)

有效茎

Effective stems (Stems/pot)

ingle stem weight (kg)

单茎重 S

锤度

Brix (%)

蔗糖分

Sucrose content (%)

非脱毒果蔗

Virus-infected chewing cane

脱毒果蔗

Virus-free chewing cane

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