不同抗性松树家系中松材线虫致病力和繁殖力比较

王俊伟; 胡龙娇; 吴小芹

doi:10.12302/j.issn.1000-2006.202302017

王俊伟, 胡龙娇, 吴小芹

南京林业大学学报（自然科学版） ›› 2025, Vol. 49 ›› Issue (1) : 21-27.

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南京林业大学学报（自然科学版） ›› 2025, Vol. 49 ›› Issue (1) : 21-27. DOI: 10.12302/j.issn.1000-2006.202302017

专题报道：松材线虫病绿色防控研究（执行主编叶建仁骆有庆）

不同抗性松树家系中松材线虫致病力和繁殖力比较

王俊伟 ¹^,² ,
胡龙娇 ³ ,
吴小芹 ¹^,^*

作者信息 +

Comparison of pathogenicity and reproduction of Bursaphelenchus xylophilus in pine families with different disease resistance

WANG Junwei ¹^,² ,
HU Longjiao ³ ,
WU Xiaoqin ¹^,^*

Author information +

文章历史 +

摘要

【目的】探究在自然条件下松材线虫(Bursaphelenchus xylophilus)与不同抗病力松树家系互作后,是否对松材线虫的致病力和繁殖力产生影响,并分析致病力差异明显的松材线虫虫株与松树互作后其细胞色素 P450(Cytochrome P450)家族相关基因表达是否发生变化。【方法】对前期引自日本的抗松材线虫病赤松(Pinus densiflora)和黑松(P. thunbergii)基因库松树进行田间发病状况调查,发现不同松树家系的抗病能力存在显著差异。对松树基因库中各家系枯死木取样,经松材线虫自动化分子检测后,将确定为松材线虫致死的5个赤松家系和5个黑松家系样木进行线虫分离,采用单异活体培养共获得10个松材线虫虫株,检测这些虫株的致病力和繁殖力,并收集其中4个致病力差异明显的松材线虫虫株,采用RT-qPCR技术检测其与黑松互作后的2个细胞色素P450相关基因相对表达量差异。【结果】同一地块同一树种中抗病能力越强的家系,获得的松材线虫虫株在单异活体培养下繁殖量越低,相反抗病能力越弱的家系,获得的松材线虫繁殖量越高;将各虫株接种2年生黑松发现,5个黑松家系病死木中获得的线虫虫株致病力测定结果与松树家系的抗性强弱呈正相关,其中抗病能力最强的家系34病死木中获得的松材线虫虫株JYK-34的致病力最强。而赤松家系的抗性强弱与松材线虫的致病力并不呈正相关,其中来自抗病能力较强的家系18病死木分离的松材线虫虫株JYK-18的致病力却较弱;与对照线虫相比,致病力差异明显的4个虫株与2年生黑松苗互作30 d后,它们的2个细胞色素P450相关基因BxCYP33C4和BxCYP33C9的相对表达量增长倍数与虫株的致病力强弱呈正相关,各虫株依次为JYK-34>JYK-17>JYK-18>JYK-31。【结论】松材线虫繁殖力的分化在赤松和黑松两个树种不同抗病家系中规律相同;抗性松树基因库中同一地块不同抗病力松树分离的松材线虫的致病力并非单一类型,存在致病力差异的虫株;松材线虫细胞色素P450相关基因BxCYP33C4和BxCYP33C9不仅参与松材线虫与松树的互作过程,还是松材线虫重要的致病相关基因。

Abstract

【Objective】 To explore whether the interaction of pine families with varying levels of disease resistance in the same plot with pine wood nematode (Bursaphelenchus xylophilus) under natural conditions affects the pathogenicity and reproduction of B. xylophilus, and to determine if the expression level of cytochrome P450 i.e. CYP family-related genes of B. xylophilus strains with noticeable differences in pathogenicity different after interaction with pine trees, the following study was conducted.【Method】This study monitored the gene pools of several families of Pinus densiflora and P. thunbergii, which were introduced from Japan previously in the lab. Field monitoring was conducted over several years, revealing significant differences in disease resistance among different pine families. Dead wood samples from each family in the pine tree gene pool were collected, and automated molecular detection was used to identify B. xylophilus. Nematode samples from five P. densiflora families and five P. thunbergii families, confirmed to have died from B. xylophilus, were isolated. A total of 10 B. xylophilus strains were obtained through monoheterotic in vivo culture, and their pathogenicity and reproduction were tested. The relative expression of two cytochrome P450 genes in four B. xylophilus strains with significantly different pathogenicity was detected by RT-qPCR. 【Result】The results indicated that in the same tree species within the same plot, the stronger the disease resistance of the pine families, the lower the reproduction rate of B. xylophilus strains obtained under monoheterotic living culture. Conversely, weaker disease resistance in pine families corresponded with higher reproduction rates of B. xylophilus strains. Inoculating each strain with 2-year-old P. thunbergii revealed that the pathogenicity test results of nematode strains obtained from diseased and dead trees of five P. thunbergii families were positively correlated with the resistance of pine families. The B. xylophilus strain JYK-34, isolated from the most resistant family (34), showed the highest pathogenicity. However, for P. densiflora families, there was no positive correlation between resistance and the pathogenicity of B. xylophilus. The B. xylophilus JYK-18, isolated from a highly resistant family (18), exhibited weak pathogenicity. Compared with the control B. xylophilus, the increased expression of two cytochrome P450 genes, BxCYP33C4 and BxCYP33C9, was positively correlated with the pathogenicity of the four B. xylophilus strains, namely JYK-34>JYK-17>JYK-18>JYK-31, after interacting with the P. thunbergii seedlings for 30 days. 【Conclusion】The differentiation in the reproduction of B. xylophilus was consistent among different disease-resistant families of P. densiflora and P. thunbergii. The pathogenicity of B. xylophilus isolated from pine trees with different resistance levels within the same plot in the resistant pine tree gene pool was not uniform, with different strains exhibiting varying pathogenicity. The cytochrome P450 genes BxCYP33C4 and BxCYP33C9 in B. xylophilus are involved in the interaction between B. xylophilus and pines and are also important genes related to the pathogenicity of B. xylophilus.

导出引用

王俊伟, 胡龙娇, 吴小芹. 不同抗性松树家系中松材线虫致病力和繁殖力比较[J]. 南京林业大学学报（自然科学版）. 2025, 49(1): 21-27 https://doi.org/10.12302/j.issn.1000-2006.202302017

WANG Junwei, HU Longjiao, WU Xiaoqin. Comparison of pathogenicity and reproduction of Bursaphelenchus xylophilus in pine families with different disease resistance[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2025, 49(1): 21-27 https://doi.org/10.12302/j.issn.1000-2006.202302017

中图分类号： S763

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