抗毒素Camalexin对大白菜抗根肿病的作用研究

doi:10.16420/j.issn.0513-353x.2021-0591

摘要/Abstract

摘要：

对易感根肿病大白菜自交系‘BJN3-1’在接种根肿菌（Plasmodiophora brassicae）的同时施用了5个不同剂量的抗毒素camalexin进行处理,研究camalexin在大白菜中抗根肿菌的作用。结果表明,接种根肿菌的对照材料感病症状明显,施用不同剂量camalexin的5个处理在前21 d可明显抑制根肿菌的增殖,随着camalexin施用量的增加对根肿菌生长的抑制作用增强。茉莉酸、水杨酸、乙烯合成途径和病程相关蛋白基因在camalexin处理14 ~ 21 d后表达显著高于对照处理。这表明外源施用camalexin能够诱导寄主响应根肿菌胁迫,从而提高抗病性。施用camalexin后28 d各处理均严重感病,camalexin对根肿菌增殖的抑制作用减弱,这可能与根肿菌的解毒机制有关。

关键词: 大白菜, 根肿病, 根肿菌, camalexin

Abstract:

To investigate the resistant effects of camalexin to Plasmodiophora brassicae in Chinese cabbage,treatments with five amount of camalexin were carried out by P. brassicae inoculation on a Chinese cabbage susceptible inbred line‘BJN3-1’. Results indicated that the control showed the visually susceptible symptoms. But the treatments with different amount of camalexin after 21 days treatment can significantly inhibit the invasion of P. brassicae to reduce the disease index of clubroot. The inhibition of P. brassicae infection was enhanced with the increase of camalexin application. Furthermore,the genes involved in synthesis pathways of jasmonic acid,salicylic acid,ethylene and pathogenesis-related proteins were significantly up-regulated after 14-21 days under camalexin treatment. These indicated that the application of camalexin can induce the host responses to P. brassicae invasion,and improve resistance. However,the effects of all camalexin treatments on clubroot development were reduced after 28 days treatment which may be related to the microbial detoxification activities of P. brassicae.

Key words: Chinese cabbage, clubroot disease, Plasmodiophora brassicae, camalexin

中图分类号:

S634.1

王钰, 张雪, 张学颖, 张思雨, 闻婷婷, 王迎君, 甘彩霞, 庞文星. 抗毒素Camalexin对大白菜抗根肿病的作用研究[J]. 园艺学报, 2022, 49(8): 1689-1698.

WANG Yu, ZHANG Xue, ZHANG Xueying, ZHANG Siyu, WEN Tingting, WANG Yingjun, GAN Caixia, PANG Wenxing. The Effect of Camalexin on Chinese Cabbage Resistance to Clubroot[J]. Acta Horticulturae Sinica, 2022, 49(8): 1689-1698.

图/表 7

表1 茉莉酸、水杨酸、乙烯信号途径中相关基因和病程相关蛋白基因扩增引物

Table 1 Primers of genes involved in jasmonic acid,salicylic acid,ethylene synthesis pathway and pathogenesis-related proteins

用途 Usage	基因 Gene	引物序列（5′-3′） Primer sequence
内参基因 Internal genes	GAPDH	F：CAGGTTTGGAATTGTCGAGG	R：GAGCTGTGGAAGCACCTTTC
茉莉酸途径	PDF1.2	F：ACCATGTCCCACTTGGCTTCT	R：ATCACCCTTATCTTCGCTGCTC
Jasmonic acid synthesis	AOS	F：CCTAGTCGCAAGCGGTTGAT	R：AGGGATTGAGCAGATGGAGC
pathway	COI1	F：CCTCTTTGTGAGACTAAGCCTTCT	R：AACTATGTTGTCTGGGCCTTTC
	JAZ	F：GGCCACCTTCATAATGTCCTC	R：TGCTAACCCTTCCGATACCTC
水杨酸途径	NPR1	F：TTGGACCTCACTGATCCTGTTTT	R：ACGGTATTTCCCACGCTGCTC
Salicylic acid synthesis pathway	ESD5	F：CTTGGGAAGGTTACGGTTCG	R：CATCATCTACTGTGCTACCTCCAT
	WRKY70	F：ACCACAACCATAAATAGCCTCA	R：AACACTAAAGAAGTATCACCCAAGA
	TGA	F：TTGCGATGATTGCTGGAGGT	R：TATCGGGCATGTGGCGAACT
乙烯途径	CTR1	F：ACAGGAGATGTATTGGGAGGGA	R：GGACAGTGAAGGTTTGCGATTT
Ethylene synthesis pathway	ETR	F：CAACAGGATGCTGCTGACGAA	R：GCTGGACAGGGAGATGGGATT
	EIN2	F：CCACTGCTAGTGTTGTCCCTTA	R：TGTCTGCGATTCTTGATGAGTT
病程蛋白相关基因 Pathogenesis related proteins	PR1	F：CCCTCGAGAGCTCAAAACAG	R：TCTCGTTGACCCAAAGGTTC
	PR2	F：CTCGGAGTTCCACCCTTCACA	R：CAAACCTCTTCGACGCCAATC
	PR5	F：AATGAACCAGGATGGTCAGG	R：CGGTGACGGTGCATTTTAG
	PR9	F：AGCTGCTGGATGTAACCTGAAT	R：CCATGATATGGGCAATGACTGA

表2 大白菜幼苗接种根肿菌并外源施用camalexin后的病情指数

Table 2 Disease index of Chinese cabbage after camalexin treatments with Plasmodiophora brassicae inoculation

Camalexin/（ng · mL^-1）	病情指数 Disease index
Camalexin/（ng · mL^-1）	7 d	14 d	21 d	28 d
0	0	0	74.33 ± 4.00 a	97.00 ± 1.00 a
250	0	0	74.67 ± 2.50 a	95.67 ± 1.35 a
500	0	0	65.68 ± 2.90 b	95.33 ± 1.68 a
1 000	0	0	63.22 ± 2.55 b	96.58 ± 2.58 a
2 000	0	0	44.50 ± 1.76 bc	93.05 ± 2.31 a
5 000	0	0	38.12 ± 2.21 bc	81.25 ± 3.50 b

图1 大白菜幼苗不同处理时期植株形态和显微观察红色箭头指示为被根肿菌侵染的细胞,圆形显微观察图为※号区域放大。

Fig. 1 Clubroot symptoms and microscopic observation of Chinese cabbage at different stage Red arrow indicates the cells infected by Plasmodiophora brassicae,the circular microscopic observation image is enlarged for the area marked with ※.

图2 接种根肿菌并外源施用不同剂量camalexin后不同时期茉莉酸合成途径基因表达

Fig. 2 Genes expression of jasmonic acid synthesis pathway during clubroot development at different amount of camalexin treatments

图3 接种根肿菌并外源施用不同剂量camalexin后不同时期水杨酸合成途径基因表达

Fig. 3 Genes expression of salicylic acid synthesis pathway during clubroot development at different amount of camalexin treatments

图4 接种根肿菌并外源施用不同剂量camalexin后不同时期乙烯途径关键基因表达

Fig. 4 Genes expression of ethylene synthesis pathway during clubroot development at different amount of camalexin treatments

图5 接种根肿菌并外源施用不同剂量camalexin后不同时期病程相关蛋白基因表达

Fig. 5 Genes expression of pathogenesis related proteins during clubroot development at different amount of camalexin treatments

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