不同种质番茄材料抗番茄黄化曲叶病毒病特性研究

doi:10.3969/j.issn.1004-1524.2021.11.11

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (11): 2085-2097.DOI: 10.3969/j.issn.1004-1524.2021.11.11

不同种质番茄材料抗番茄黄化曲叶病毒病特性研究

王贤(), 刘放, 魏小红^*(), 朱晓林, 王宝强

甘肃农业大学生命科学技术学院,甘肃省作物遗传改良与种质创新重点实验室,甘肃省干旱生境作物学重点实验室,甘肃兰州 730070

收稿日期:2020-08-22 出版日期:2021-11-25 发布日期:2021-11-26
作者简介:*魏小红,E-mail: weixh@gsau.edu.cn
王贤(1997—),男,甘肃天水人,硕士研究生,研究方向为植物学。E-mail: 1431268954@qq.com
通讯作者: 魏小红
基金资助:
国家自然科学基金(31560663)

Study on resistance of different tomato germplasm materials to yellow leaf curl virus disease

WANG Xian(), LIU Fang, WEI Xiaohong^*(), ZHU Xiaolin, WANG Baoqiang

College of Life Science and Technology, Gansu Agricultural University, Gansu Key Lab of Crop Genetic & Germplasm Enhancement, Gansu Provincial Key Lab of Aridland Crop Science, Lanzhou 730070, China

Received:2020-08-22 Online:2021-11-25 Published:2021-11-26
Contact: WEI Xiaohong

摘要/Abstract

摘要：

以不同种质番茄为实验材料,利用苗期农杆菌接种法接种番茄黄化曲叶病毒(tomato yellow leaf curl virus, TYLCV),采用实时定量PCR和生理生化方法评价对TYLCV的抗性,统计分析不同抗性番茄材料不同发病时期的表型特征、带毒率、抗氧化酶活性、总酚与类黄酮含量及抗性基因表达量的变化情况,并分析各抗性指标之间的关系。结果表明,番茄材料带毒率为100%;853没有任何感病症状,802与803感病最为严重;抗氧化酶活性、总酚与类黄酮含量及抗性基因表达量在各时期均较高的有867、857、853,817与842次之,803最低。SOD活性、CAT活性、APX活性、总酚含量、类黄酮含量、Ty-1基因、Ty-2和Ty-3基因的表达量相互之间均呈极显著正相关(总酚与Ty-1之间为显著正相关,P<0.01)。番茄植株感染TYLCV后,Ty-1的表达相比于其他抗病基因具有延迟性。综合分析筛选出867、857、853三种抗病性较强的材料,植株体内的抗性基因、抗氧化酶、总酚与类黄酮之间呈正相关,共同提高了番茄植株对TYLCV的抗性。

关键词: 番茄, 番茄黄化曲叶病毒病, 抗氧化酶, 次生代谢物, 抗性基因, 抗性

Abstract:

Different tomato materials were inoculated with tomato yellow leaf curl virus (TYLCV) by Agrobacterium tumefaciens infiltration. Real-time quantitative PCR and physiological and biochemical methods were used to evaluate the resistance to TYLCV. The phenotypic characteristics, virus carrying rate, antioxidant enzyme activity, secondary metabolite content and resistance gene expression of tomato materials with different resistance were analyzed. The results showed that the virus infection rates of tomato materials were 100%; the tested variety 853 had no symptoms, while 802 and 803 were the most susceptible; the varieties 867, 857 and 853 had higher antioxidant enzyme activity, contents of total phenols and flavonoids, and higher resistance gene expression, followed by 817 and 842, the lowest was 803. SOD activity, CAT activity, APX activity, total phenol content, flavonoid content, Ty-1 gene, Ty-2 and Ty-3 gene expression levels were significantly positively correlated with each other (total phenol and Ty-1 showed significantly positive correlation, P<0.01). Compared with other disease resistance genes, the expression of Ty-1 was delayed after tomato plants were infected with TYLCV. The resistance genes, antioxidant enzymes, total phenols and flavonoids in plants were significantly positively correlated, which jointly improved the resistance of tomato plants to TYLCV.

Key words: tomato, tomato yellow leaf curl virus disease, antioxidant enzymes, secondary metabolites, resistance gene, resistance

中图分类号:

S641.2

王贤, 刘放, 魏小红, 朱晓林, 王宝强. 不同种质番茄材料抗番茄黄化曲叶病毒病特性研究[J]. 浙江农业学报, 2021, 33(11): 2085-2097.

WANG Xian, LIU Fang, WEI Xiaohong, ZHU Xiaolin, WANG Baoqiang. Study on resistance of different tomato germplasm materials to yellow leaf curl virus disease[J]. Acta Agriculturae Zhejiangensis, 2021, 33(11): 2085-2097.

图/表 13

表1 不同番茄材料的基因型

Table 1 Genotypes of different tomato materials

编号 Number	基因型 Genotype	编号 Number	基因型 Genotype
801	ty-1/ty-1+ty-2/ty-2+ty-3/ty-3	822	ty-1/ty-1+ty-3/ty-3
802	ty-1/ty-1+ty-2/ty-2+ty-3/ty-3	841	ty-1/ty-1+ty-2/ty-2+ty-3/ty-3
803	ty-1/ty-1+ty-2/ty-2+ty-3/ty-3	842	ty-1/ty-1+ty-2/ty-2+ty-3/ty-3
805	ty-1/ty-1+ty-2/ty-2+ty-3/ty-3	853	Ty-1/Ty-1+ty-2/ty-2+Ty-3/Ty-3
806	ty-1/ty-1+ty-2/ty-2+ty-3/ty-3	857	Ty-1/Ty-1+ty-2/ty-2+Ty-3/Ty-3
817	ty-1/ty-1+ty-2/ty-2+ty-3/ty-3	867	Ty-1/Ty-1+ty-2/ty-2+Ty-3/Ty-3
818	Ty-1/Ty-1+ty-2/ty-2+ty-3/ty-3	齐达利Qidali	Ty-1/Ty-1+ty-2/ty-2+Ty-3a/ty-3a
819	ty-1/ty-1+ty-2/ty-2+ty-3/ty-3	MM	无
820	Ty-1/Ty-1+ty-2/ty-2+ty-3/ty-3

表2 抗病基因及引物序列

Table 2 Disease resistance genes and primer sequences

基因 Gene	引物序列 Primer sequence(5'-3')
Ty-1	F:GGCAAAATATGCAGCCAGGCTTTCC R:TCAGTATGTATACGAGGTTCGCCGT
Ty-2	F:TGCGGCTGTTTCCTATGGTT R:TGGTTCTGGAAAGGGATCGAA
Ty-3	F:TGAGCGGAGAAATGTGACAG R:CGGGGCTTTGATGAAACGAA
actin	F:GAAATAGCATAAGATGGCAGACG R:ATACCCACCATCACACCAGTAT

图1 部分品种感病后的表型 A,对照;B,感病后表型。

Fig.1 The phenotypes of some varieties after virus infection A, The control; B, The phenotype after infection.

图2 不同番茄材料接种20 d后的PCR扩增结果

Fig.2 PCR amplification results of different tomato materials after virus inoculation for 20 days

图3 不同番茄材料接种病毒后超氧化物歧化酶(SOD)活性的变化情况柱上没有相同小写字母表示差异显著(P<0.05)。下同。

Fig.3 Changes of superoxide dismutase (SOD) activity in different tomato materials after virus inoculation The bars without the same lowercase letters showed significant difference(P<0.05). The same as below.

图4 不同番茄材料接种病毒后过氧化氢酶(CAT)活性的变化情况

Fig.4 Changes of catalase (CAT) activity in different tomato materials after virus inoculation

图5 不同番茄材料接种病毒后抗坏血酸过氧化物酶(APX)活性的变化情况

Fig.5 Changes of ascorbate peroxidase (APX) activity in different tomato materials after virus inoculation

图6 不同番茄材料接种病毒后总酚含量的变化情况

Fig.6 Changes of total phenol content in different tomato materials after virus inoculation

图7 不同番茄材料接种病毒后类黄酮含量的变化情况

Fig.7 Changes in the content of flavonoids in different tomato materials after virus inoculation

图8 不同番茄材料接种病毒后Ty-1基因的表达情况

Fig.8 Expression of Ty-1 gene in different tomato materials after virus inoculation

图9 不同番茄材料接种病毒后Ty-2基因的表达情况

Fig.9 Expression of Ty-2 gene in different tomato materials after virus inoculation

图10 不同番茄材料接种病毒后Ty-3基因的表达情况

Fig.10 Expression of Ty-3 gene in different tomato materials after virus inoculation

表3 各抗性指标的相关性分析

Table 3 Correlation analysis of resistance indexes

	SOD	CAT	APX	总酚 Total phenols	类黄酮 Flavonoids	Ty-1	Ty-2	Ty-3
SOD	1
CAT	0.757^**	1
APX	0.867^**	0.690^**	1
总酚Total phenols	0.802^**	0.699^**	0.739^**	1
类黄酮Flavonoids	0.702^**	0.797^**	0.682^**	0.798^**	1
Ty-1	0.635^**	0.658^**	0.568^**	0.356^*	0.527^**	1
Ty-2	0.734^**	0.753^**	0.702^**	0.631^**	0.705^**	0.698^**	1
Ty-3	0.754^**	0.762^**	0.703^**	0.631^**	0.786^**	0.732^**	0.822^**	1

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不同种质番茄材料抗番茄黄化曲叶病毒病特性研究

Study on resistance of different tomato germplasm materials to yellow leaf curl virus disease

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