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林业科学 ›› 2020, Vol. 56 ›› Issue (7): 63-69.doi: 10.11707/j.1001-7488.20200707

所属专题: 森林有害生物

• 论文与研究报告 • 上一篇    下一篇

无细菌松材线虫对马尾松的致病性

朱丽华,章欣月,夏馨蕊,万羽,代善俊,叶建仁*   

  1. 南方现代林业协同创新中心 江苏省有害生物入侵预防与控制重点实验室 南京林业大学林学院 南京 210037
  • 收稿日期:2019-07-01 出版日期:2020-07-25 发布日期:2020-08-11
  • 通讯作者: 叶建仁
  • 基金资助:
    国家重点研发计划项目(2017YFD0600104);国家自然科学基金项目(31971659);江苏高校优势学科建设工程资助项目(PAPD)

Pathogenicity of Aseptic Bursaphelenchus xylophilus on Pinus massoniana

Lihua Zhu,Xinyue Zhang,Xinrui Xia,Yu Wan,Shanjun Dai,Jianren Ye*   

  1. Co-Innovation Center for Sustainable Foresty in Southern Chain Jiangsu Key Laboratory of Pest Invasion Prevention and Control College of Forset, Nanjing Foresty University Nanjing 210037
  • Received:2019-07-01 Online:2020-07-25 Published:2020-08-11
  • Contact: Jianren Ye

摘要:

目的: 探讨松材线虫及其伴生细菌在松材线虫病中的作用,为其致病机制研究及病害防治提供理论依据。方法: 以5年生温室健康马尾松盆栽苗为材料,人工接种无菌松材线虫和带菌松材线虫(8 000条·株-1),观察松苗萎蔫情况,并于接种5、10和30天后取茎段横切面观察松脂分泌情况,对接种20天后的茎段组织解剖结构进行显微观察,接种后30天对植株取样进行线虫再分离,统计线虫在树体内种群增殖情况。结果: 无菌松材线虫和带菌松材线虫均能引起5年生马尾松盆栽苗发病,且症状一致,接种35天后致萎蔫率均达到90%,对照植株保持健康状态;茎段横切面观察结果显示,无菌松材线虫接种5天后松脂停止分泌,10天后茎段开始出现少量空洞,30天时空洞化严重;带菌线虫接种5天后尚有少量松脂分泌,10天松脂分泌停止,30天后茎段产生空洞;对照松树松脂分泌正常,茎段横切面完整平滑。石蜡切片结果显示,2组线虫接种致萎蔫松树茎段的组织解剖结构均发生显著变化,皮层及木质部细胞、髓心细胞大多破碎变形,树脂道上皮细胞破碎解体。无菌松材线虫在松苗体内种群增殖速率高于带菌松材线虫,萎蔫松苗体内线虫数量分别为(8 105±5 661)条·g-1和(4 317±1 896)条·g-1,但二者差异不显著(P=0.361 9)。结论: 无菌松材线虫和带菌松材线虫接种均能使5年生马尾松苗发病,无菌化处理未使松材线虫失去对马尾松苗的致病性,松材线虫的致病性与其伴生细菌无关。

关键词: 松材线虫, 无细菌, 致病性, 马尾松, 组织病理学

Abstract:

Objective: The aim of this study is to explore the roles of Bursaphelenchus xylophilus (pine wood nematode,PWN) and the bacteria associated with PWN in the development of pine wilt disease (PWD),to provide theory gist for research on PWN pathogenic mechanism and PWD prevention. Method: Five-year-old healthy Pinus massoniana seedlings grown in green house were artificially inoculated with aseptic PWNs and non-aseptic PWNs,including 8000 PWNs per seedling,and then the status of seedling wilting was observed. The cross section was taken to detect the resin secretion after 5,10 and 30 days of inoculation. The anatomical structure of stem section was observed under a microscope after 20 days of inoculation. The PWNs were re-isolated from wilting seedlings after 30 days of inoculation. Result: Both aseptic PWNs and non-aseptic PWNs wilted 5-year-old seedlings of P. massoniana, expressing similar symptom.The wilting rate reached 90% after 35 days of inoculation,while the control seedlings remained healthy. The observation of cross sections showed that resin stopped secreting after 5 days of aseptic PWNs inoculation,a small amount of cavities began to appear after 10 days,and serious cavitation appeared after 30 days. The observation of cross sections showed that resin decreased secretion but not stopped yet after 5 days of non-aseptic PWNs inoculation,the secretion stopped after 10 days and cavities formed after 30 days. While the control pines secreted normally resin and there is no cavity in their stems. The results of paraffin section showed that the histological and anatomical structure of the stem segments of wilted seedlings changed remarkably in the both treated groups. Most of the cells,in cortex,xylem and pith,were deformed and the epithelial cells near the resin canal were broken and disintegrated. The multiplication rate of aseptic PWNs in wilted seedlings was higher than that of non-aseptic PWNs. The average number of nematodes re-isolated from wilted seedlings was (8 105±5 661)·g-1 for aseptic PWNs and (4 317±1 896)·g-1 for non-aseptic PWNs,with no significant difference between the two groups (P = 0.3619). Conclusion: Both of the aseptic and non-aseptic PWNs are able to infect P. massoniana. Asepsis does not make PWNs lose their pathogenicity on P. massoniana. PWNs are necessary factor in the PWD development while,the pathogenicity has nothing to do with the associated bacteria.

Key words: Bursaphelenchus xylophilus, asepsis, pathogenicity, Pinus massoniana, histophology

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