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粉红粘帚霉SWFUYHL 02-03侵染楚雄腮扁叶蜂幼虫的扫描电镜及透射电镜观察

芦俊佳 徐荣 李永和

芦俊佳, 徐荣, 李永和. 粉红粘帚霉SWFUYHL 02-03侵染楚雄腮扁叶蜂幼虫的扫描电镜及透射电镜观察[J]. 北京林业大学学报, 2018, 40(12): 68-75. doi: 10.13332/j.1000-1522.20180230
引用本文: 芦俊佳, 徐荣, 李永和. 粉红粘帚霉SWFUYHL 02-03侵染楚雄腮扁叶蜂幼虫的扫描电镜及透射电镜观察[J]. 北京林业大学学报, 2018, 40(12): 68-75. doi: 10.13332/j.1000-1522.20180230
Lu Junjia, Xu Rong, Li Yonghe. SEM and TEM observations of Clonostachys rosea SWFUYHL 02-03 infecting the body wall of Cephalcia chuxiongica larvae[J]. Journal of Beijing Forestry University, 2018, 40(12): 68-75. doi: 10.13332/j.1000-1522.20180230
Citation: Lu Junjia, Xu Rong, Li Yonghe. SEM and TEM observations of Clonostachys rosea SWFUYHL 02-03 infecting the body wall of Cephalcia chuxiongica larvae[J]. Journal of Beijing Forestry University, 2018, 40(12): 68-75. doi: 10.13332/j.1000-1522.20180230

粉红粘帚霉SWFUYHL 02-03侵染楚雄腮扁叶蜂幼虫的扫描电镜及透射电镜观察

doi: 10.13332/j.1000-1522.20180230
基金项目: 

国家自然科学基金项目 31360156

国家级省级大学省创新创业训练计划项目 516027

详细信息
    作者简介:

    芦俊佳,博士生。主要研究方向:森林病虫害防治。Email:58826911@qq.com  地址:650224云南省昆明市盘龙区白龙寺300号西南林业大学

    责任作者:

    李永和,教授,博士生导师。主要研究方向:森林病虫害防治。Email:swfclyh@126.com  地址:同上

  • 中图分类号: S763.38

SEM and TEM observations of Clonostachys rosea SWFUYHL 02-03 infecting the body wall of Cephalcia chuxiongica larvae

  • 摘要: 目的了解粉红粘帚霉对楚雄腮扁叶蜂幼虫的侵染过程,为粉红粘帚霉对楚雄腮扁叶蜂的致病机理研究提供参考。方法利用扫描电镜和透射电镜技术对粉红粘帚霉SWFUYHL 02-03孢子侵染楚雄腮扁叶蜂幼虫体表过程及其超微结构的变化进行观察研究。结果扫描电镜结果显示,接种6 h后,粉红粘帚霉SWFUYHL 02-03分生孢子成功附着于幼虫体壁的气门、节间膜等部位;接种14 h后,粉红粘帚霉SWFUYHL 02-03孢子萌发出芽管,并侵入体壁;接种26 h后,孢子和菌丝穿透楚雄腮扁叶蜂幼虫体壁,且菌丝迅速生长;接种72 h后,菌丝穿出虫体表面,并长出产孢结构。透射电镜观察发现粉红粘帚霉SWFUYHL 02-03菌丝在穿透幼虫体壁过程中,会伴随酶的参与和组织溶解现象;接种6 h后,粉红粘帚霉SWFUYHL 02-03分生孢子附着于幼虫体壁的两个节间褶间的部位;接种14 h后,分生孢子萌发并进入表皮细胞层分泌物中;接种26 h后,菌丝侵入表皮并扩展蔓延,且在菌丝周围出现电子密度很低的光晕;接种38 h后,皮细胞层细胞排列松弛变成空泡;接种48 h后,菌丝充满虫体,表皮层出现大量菌丝。结论粉红粘帚霉SWFUYHL 02-03孢子及菌丝能寄生楚雄腮扁叶蜂幼虫,并可致其死亡。观察结果准确反映了粉红粘帚霉SWFUYHL 02-03对楚雄腮扁叶蜂幼虫的侵染过程,并证实了侵染过程中酶与代谢产物的存在。

     

  • 图  1  粉红粘帚霉SWFUYHL 02-03侵染楚雄腮扁叶蜂幼虫死亡率变化

    Figure  1.  Mortality change of Ce. chuxiongica larvae infected by Cl. rosea SWFUYHL 02-03

    图  2  粉红粘帚霉SWFUYHL 02-03侵染楚雄腮扁叶蜂幼虫体壁的扫描电镜观察

    a.孢子(co)附着在楚雄腮扁叶蜂幼虫体壁(箭头处) (接种后6 h) Conidias (co) adhered onto the body wall of Ce. chuxiongica larvae (arrow) (6 hours after inoculation); b.附着孢子(co)萌发芽管(gt) (箭头处) (接种14 h后) Adhered conidias germinated germ tubes (gt) (arrow) (14 hours after inoculation); c.孢子(co)侵入幼虫体壁(箭头处) (接种26 h后) Conidias (co) penetrated into the body wall of larvae (arrow) (26 hours after inoculation); d.菌丝(ph)穿透幼虫体壁(箭头处) (接种26 h后) Some hypha (ph) penetrated out from the body wall of the larvae (arrow) (26 hours after inoculation); e.气门处菌丝(ph)(接种38 h) Hypha extended on the spiracles (38 hours after inoculation); f.开始长出新产孢结构(cs)(箭头处)和孢子(箭头处)(接种72 h) Began to generate new conidiogenous structure (cs) (arrow) and conidias (co) (arrow) (72 hours after inoculation)

    Figure  2.  Observation of infection process of Cl. rosea SWFUYHL 02-03 on Ce. chuxiongica larvae by SEM

    图  3  粉红粘帚霉SWFUYHL 02-03侵染楚雄腮扁叶蜂幼虫的透射电镜观察

    分生孢子(co)(箭头处)附着于楚雄腮扁叶蜂幼虫节间褶之间的分泌物上(接种6 h) Conidia (co)(arrow) adhered onto the secretion between Ce. chuxiongica's intersegmental fold(6 hours after inoculation); b.菌丝(ph)(箭头处)侵入到楚雄腮扁叶蜂幼虫外表皮(exo)细胞层分泌物中(接种14 h) Hypha (ph)(arrow) penetrated into the secretion of Ce. chuxiongica's exocuticle (exo) (14 hours after inoculation); c.大量菌丝(ph)(箭头处)侵入到外表皮细胞层(exo)中(接种26 h) A lot of hypha (ph)(arrow) penetrated into the exocuticle(exo)(26 hours after inoculation); d.菌丝周围出现电子密度很低的光晕(箭头处) (接种26 h)Halo with low electron density was observed around penetrated hypha (arrow)(26 hours after inoculation); e.皮细胞层(epid)细胞排列松弛变成空泡(vac)(箭头处) (接种38 h) Epidermis (epid) cell arranged loose and became vacuole (vac) (arrow) (38 hours after inoculation); f.表皮层中的大量菌丝(接种48 h) Massive hypha (ph) in cuticle (48 hours after inoculation)

    Figure  3.  Observation of infection process of Cl. rosea SWFUYHL 02-03 on Ce. chuxiongica larvae by TEM

    表  1  扫描电镜观察结果数量统计表

    Table  1.   Number statistics in SEM observation results

    接种后时间Time after inoculation/h 孢子附着体壁Conidias adhering onto body wall 孢子萌发芽管Conidias germ 菌丝穿透体壁Hypha penetrating the body wall 体表菌丝增多Hypha on body wall increased 出现产孢结构Generating conidiogenous structure
    幼虫Larvae 视野Vision 幼虫Larvae 视野Vision 幼虫Larvae 视野Vision 幼虫Larvae 视野Vision 幼虫Larvae 视野Vision
    数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/%
    6 5 100.00 25 100.00 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00
    14 3 60.00 10 21.74 3 60.00 36 78.26 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00
    26 4 80.00 8 50.00 3 60.00 9 18.00 3 60.00 33 66.00 0 0.00 0 0.00 0 0.00 0 0.00
    38 4 80.00 7 14.58 2 40.00 6 12.50 5 100.00 12 25.00 4 80.00 23 47.92 0 0.00 0 0.00
    48 0 0.00 0 0.00 0 0.00 0 0.00 5 100.00 4 20.00 5 100.00 16 80.00 0 0.00 0 0.00
    72 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 5 100.00 20 100.00 5 100.00 20 100.00
    下载: 导出CSV

    表  2  透射电镜观察视野数量统计表

    Table  2.   Statistics in view number observed by TEM

    接种后时间Time after inoculation/h 孢子附着体壁Conidias adhering onto body wall 菌丝进入外表皮层Hypha penetrating into the exocuticle 菌丝周围出现光晕Halo appearing around hypha 皮细胞层细胞排列疏松并形成空泡Epidermis cell arranging loose and becoming vacuole 菌丝段进入表皮层Hypha penetrating into cuticle
    幼虫Larvae 视野Vision 幼虫Larvae 视野Vision 幼虫Larvae 视野Vision 幼虫Larvae 视野Vision 幼虫Larvae 视野Vision
    数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/% 数量Quantity 占比Proportion/%
    6 3 60.00 10 33.33 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00
    14 3 60.00 4 25.00 3 60.00 12 75.00 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00
    26 3 60.00 4 25.00 3 60.00 12 75.00 5 100.00 16 100.00 0 0.00 0 0.00 0 0.00 0 0.00
    38 0 0.00 0 0.00 0 0.00 0 0.00 5 100.00 12 100.00 4 80.00 10 83.33 0 0.00 0 0.00
    48 0 0.00 0 0.00 0 0.00 0 0.00 5 100.00 34 100.00 4 80.00 10 29.41 4 80.00 24 70.59
    72 0 0.00 0 0.00 0 0.00 0 0.00 5 100.00 36 100.00 3 60.00 12 33.33 5 100.00 24 66.67
    下载: 导出CSV
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  • 收稿日期:  2018-07-19
  • 修回日期:  2018-10-01
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