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20世纪80年代至今,切梢小蠹Tomicus spp.在中国云南肆意猖獗,对云南松造成了毁灭性的灾难,被称为云南松(Pinus yunnanensis Franch)的“癌症”[1-3]。蠹害不仅是小蠹虫造成的,小蠹虫在其蛀害的过程中还常携带病原真菌即“伴生菌”侵入,蠹害是寄主树木的抗御与小蠹虫及其伴生菌的蛀害侵染之间相互作用的结果[4-6]。研究发现,小蠧虫与其所带的病原真菌关系是相对稳定的,某种小蠹虫总是与某一种或几种病原真菌发生联系[7]。这些病原真菌往往需要借助小蠹虫传带到达寄主树木,完成生命周期;病原真菌对树木的危害削弱了树木的抗性,从而有助于小蠹虫的成功入侵[4, 8]。病原真菌通过小蠹虫的蛀害过程被携带进入寄主树木组织内,在侵染过程中分泌毒素对寄主树木进行危害[9],进一步研究发现,病原真菌分泌的毒素原液中具有蛋白类或氨基酸类物质,它对寄主植物抗性物质代谢有关的过氧化物酶(PO)、多酚氧化酶(PPO)及苯丙氨酸解氨酶(PAL)的活性均有抑制作用,从而削弱了寄主树木的抗性作用,这也是伴生真菌的一个重要致病机制[10-12]。
多年来,对云南发生危害的切梢小蠹种类存在争议[12]。在过去很长时间里,将在云南成灾危害的切梢小蠹被视为纵坑切梢小蠹(Tomicus pinperda),其在欧洲和北美以及我国的北方是一种次期性害虫,主要危害衰弱木,虫口密度大时才侵害健康木。但在云南,切梢小蠹不仅危害衰弱木,也危害健康木,然而,在云南发生的切梢小蠹的生物学和生态学特性明显不同于欧美及我国北方的种类[12-14]。基于分子生物学和形态学的研究,2008年在云南造成严重危害的切梢小蠹被重新认识,并被正式定名为云南切梢小蠹(T. yunnanensis)[15],此后,在云南楚雄危害华山松(P. armandii)的切梢小蠹于2010年被正式定名为华山松切梢小蠹(T. armandii) [16]。2014-2015年松芽小蠹(T. brevipilosus)也被证实在云南多地与云南切梢小蠹和横坑切梢小蠹(T. minor)联合危害松林[17-18]。迄今,全球已知的8种切梢小蠹有4种在中国云南发生危害,云南成为开展切梢小蠹相关研究的最佳区域,也成为其伴生菌研究的最佳区域。
一直以来,为了更好地防控蠹害,学术界对切梢小蠹的生物学及生态学进行了大量研究,但对其伴生菌的研究却很少,尤其是国内少有研究[10-12]。切梢小蠹伴生真菌是导致蠹害的关键因素之一,也是揭示该区域切梢小蠹大发生的研究瓶颈。因此,开展切梢小蠹伴生菌的研究显得极为重要。为了弄清中国云南地区切梢小蠹对伴生菌的携带率的大小(简称带菌率),本研究检测分析了不同切梢小蠹种类携带伴生菌的带菌率差异,比较了切梢小蠹不同虫态以及成虫的头、足、翅的带菌率差异,通过野外随机抽样,分析了切梢小蠹危害程度与切梢小蠹带菌率的关系,旨在揭示切梢小蠹成灾机制,为探索切梢小蠹有效防控新途径提供科学数据支持。
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30天后,对照木段组,均未发现蓝变现象。处理组,将4种切梢小蠹成虫接种到新鲜木段,根据木段接种部位蓝变数量统计得出:云南切梢小蠹带菌率最高,达到80 %,横坑切梢小蠹带菌率次之,为63 %,松芽小蠹带菌率为53 %,华山松切梢小蠹带菌率为56 %(见表 1)。
表 1 切梢小蠹种类带菌率
Table 1. Carrying rate of Tomicus sp. with blue stain fungi
虫种
Tomicus species试验虫量/头
No. of test
insects带菌虫量/头
No. of insect
carrying fungi带菌率
Carrying
rate%云南切梢小蠹
Tomicus yunnanensis30 24 80 横坑切梢小蠹Tomicus minor 30 19 63 松芽小蠹Tomicus brevipilosus 30 16 53 华山松切梢小蠹
Tomicus armandii30 17 56 -
30天后,对照木段组,均未发现蓝变现象。处理组,分别取不同虫态的切梢小蠹,接种到木段中,统计蓝变数量,总供试幼虫的带菌率为90%,其中,云南切梢小蠹幼虫的带菌率最高达到100%,华山松切梢小蠹幼虫的带菌率次之为93%,横坑切梢小蠹和松芽小蠹幼虫的带菌率分别为87%和80%。总供试蛹的带菌率为78%,种间,横坑切梢小蠹蛹的带菌率最高为87%,云南切梢小蠹和松芽小蠹蛹的带菌率均为80%,华山松切梢小蠹蛹的带菌率最低67%。总供试新成虫的带菌率为58%,种间,云南切梢小蠹>松芽小蠹>横坑切梢小蠹=华山松切梢小蠹,新成虫的带菌率分别为67%,60%,53%,53%(表 2)。经数据显著性分析结果表明,4种切梢小蠹不同虫态带菌率种间差异极显著(P < 0.01),但同一虫态间没有差异(P>0.05),均表现为:幼虫的带菌率明显高于蛹,而新成虫的带菌率又明显小于蛹的带菌率。
表 2 切梢小蠹不同虫态带菌率比较
Table 2. Carrying rate of Tomicus different developing stages with blue stain fungi
虫态和虫种
Stages & Tomicus species试验虫量/头
No. of test
insects带菌虫量/头
No. of insect
carrying fungi带菌率
Carrying
rate%幼虫
Larva云南切梢小蠹
Tomicus yunnanensis15 15 100 横坑切梢小蠹
Tomicus minor15 13 87 松芽小蠹
Tomicus brevipilosus15 12 80 华山松切梢小蠹
Tomicus armandii15 14 93 小计Subtotal 60 54 90 蛹
Pupa云南切梢小蠹
Tomicus yunnanensis15 12 80 横坑切梢小蠹
Tomicus minor15 13 87 松芽小蠹
Tomicus brevipilosus15 12 80 华山松切梢小蠹
Tomicus armandii15 10 67 小计Subtotal 60 47 78 新成虫
New
adults云南切梢小蠹
Tomicus yunnanensis15 10 67 横坑切梢小蠹
Tomicus minor15 8 53 松芽小蠹
Tomicus brevipilosus15 9 60 华山松切梢小蠹
Tomicus armandii15 8 53 小计Subtotal 60 35 58 -
30天后,对照木段组,均未发现蓝变现象。处理组,分别取4种切梢小蠹的头、足和翅,接入木段,统计木段蓝变数量,总供试切梢小蠹成虫的头的带菌率为33 %,种间差异显著(0.01 < P < 0.05),横坑切梢小蠹的带菌率最高,为40%,华山松切梢小蠹带菌率最低27%,云南切梢小蠹和松芽小蠹带菌率均为33%。总供试切梢小蠹成虫的足的带菌率为30%,种间差异显著(0.01 < P < 0.05),横坑切梢小蠹>华山松切梢小蠹>云南切梢小蠹>松芽小蠹,带菌率分别为40%,33%,27%,20%。总供试切梢小蠹成虫的翅的带菌率为32%,种间差异显著(0.01 < P < 0.05),云南切梢小蠹>横坑切梢小蠹>华山松切梢小蠹=松芽小蠹,带菌率分别为40%,33%,27%,27%。(表 3)。经数据显著性分析结果表明,4种切梢小蠹成虫的头、足、翅的带菌率种间存在差异,但总体而言,切梢小蠹成虫的头部带菌率稍高于足和翅的带菌率,但差异并不明显(P>0.05)。由此见,切梢小蠹伴生菌在切梢小蠹虫体上分布较为均匀,未集中于虫体某一部位。
表 3 切梢小蠹成虫头足翅带菌率
Table 3. Carrying rate of Tomicus's head, leg and wing with blue stain fungi
器官和虫种
Organ & Tomicus species试验虫量/头
No. of test
insects带菌虫量/头
No. of insect
carrying fungi带菌率
Carrying
rate%头
Head云南切梢小蠹
Tomicus yunnanensis15 5 33 横坑切梢小蠹
Tomicus minor15 6 40 松芽小蠹
Tomicus brevipilosus15 5 33 华山松切梢小蠹
Tomicus armandii15 4 27 小计Subtotal 60 20 33 足
Leg云南切梢小蠹
Tomicus yunnanensis15 4 27 横坑切梢小蠹
Tomicus minor15 6 40 松芽小蠹
Tomicus brevipilosus15 3 20 华山松切梢小蠹
Tomicus armandii15 5 33 小计Subtotal 60 18 30 翅
Wing云南切梢小蠹
Tomicus yunnanensis15 6 40 横坑切梢小蠹
Tomicus minor15 5 33 松芽小蠹
Tomicus brevipilosus15 4 27 华山松切梢小蠹
Tomicus armandii15 4 27 小计Subtotal 60 19 32 -
基于蠹害轻、中、重3个林区的调查发现,由表 4、表 5分析可知,在云南陆良县,横坑切梢小蠹在云南松上的危害发生数量高于云南切梢小蠹。横坑切梢小蠹的带菌率存在一定的随机性,轻度林区为50%,中度林区为39.29%,重度林区为66.67%(表 4)。云南切梢小蠹的带菌率与林分的蠹害程度同样存在一定的随机性,轻度林区为68.18%,中度林区为86.36,重度林区为23.53%(表 5)。但分析蛀害的成功率(有效坑道率)发现,不管是云南切梢小蠹还是横坑切梢小蠹,检测出带伴生菌(Ophiostoma canum)的切梢小蠹的坑道,其蛀害成功率(有效坑道率)均显著的高于未检测出伴生菌的切梢小蠹的蛀害成功率,而且云南切梢小蠹和横坑切梢小蠹带菌组在轻、中、重3种蠹害林区有效坑道率均大于无菌组,由此表明,伴生菌的存在显著提高了切梢小蠹的蛀干繁殖成功率,进而加速了松树的死亡进程(表 4,表 5)。
表 4 野外横坑切梢小蠹与伴生菌带菌调查情况
Table 4. Carrying rate of accociated fungi by Tomicus minor in Yunnan pine trunks in forests
危害类型
Damage types处理
Treatments带菌种类
Fungi species统计坑道数量
No. of survey
galleries入侵孔数
No. of entry
holes带菌率
Carrying rate
of fungi/%有效侵入孔数
No. of successful
entry holes有效坑道率
percent of successful
entry holes/%轻度A
Mild level A带菌carrying fungi O.canum 28 14 50 10 71.43 不带菌No fungi 14 50 8 57.14 中度B
Middle level B带菌Carrying fungi O.canum 28 11 39.29 7 63.64 不带菌No fungi 17 60.71 7 41.18 重度C
Severe level C带菌Carrying fungi O.canum 33 22 66.67 17 77.27 不带菌No fungi 11 33.33 3 27.27 (云南陆良; Luliang, Yunnan) 表 5 野外云南切梢小蠹与伴生菌带菌调查情况
Table 5. Carrying rate of accociated fungi by Tomicus yunnanensis in Yunnan pine trunks in forests
危害类型
Damage types处理
Treatments带菌种类
Fungi species统计坑道数量
No. of survey
galleries入侵孔数
No. of entry
holes带菌率
Carrying rate
of fungi/%有效侵入孔数
No. of successful
entry holes有效坑道率
percent of successful
entry holes/%轻度A
Mild level A带菌Carrying fungi O.canum 22 15 68.18 10 66.67 不带菌No fungi 7 31.82 3 42.86 中度B
Middle level B带菌Carrying fungi O.canum 22 19 86.36 10 52.63 不带菌No fungi 3 13.64 1 33.33 重度C
Severe level C带菌Carrying fungi O.canum 17 4 23.53 3 75.00 不带菌No fungi 13 76.47 3 23.08 (云南陆良; Luliang, Yunnan)
4种切梢小蠹携带伴生菌的带菌率差异
Carrying Rates and Differences of Fungi Associated with Four Tomicus Species in Yunnan
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摘要:
目的 为了弄清中国云南4种切梢小蠹伴生菌的携带率差异,了解伴生菌在蠹害过程中的互作关系。 方法 采用人工接种法和野外调查法,检测松树韧质部是否蓝变,判断切梢小蠹带菌情况,统计分析小蠹试验材料的带菌率。 结果 云南切梢小蠹成虫带菌率(80%)明显高于横坑切梢小蠹(63%)、松芽小蠹(53%)、华山松切梢小蠹(56%)。切梢小蠹幼虫的带菌率最高(90%),其次是蛹的带菌率(78%),新成虫的带菌率最低(58%)。切梢小蠹头(33%)、足(30%)、翅(32%)3个重要器官带菌率差异不明显。林间调查表明,切梢小蠹伴生菌提高了小蠹蛀干繁殖的成功率。 结论 首次报道了中国云南4种切梢小蠹伴生菌的带菌特性及其差异,研究结果为揭示云南省切梢小蠹成灾机制及探索切梢小蠹有效防控新途径提供了科学数据支持。 Abstract:Objective The purpose of the study is to clarify the carrying rates and the differences of fungi associated with four Tomicus species in Yunnan, China and understand the relationships between the beetles and these fungi during Tomicus attacking process. Method Fungal carrying rate was analyzed by detecting stained phloem based on artificial inoculation and field survey. Result The fungal carrying rate of Tomicus yunnanensis (80%) was distinctly higher than that of the other three Tomicus species:T. minor (63%), T. armandii (56%), and T. brevipilosus (53%); the fungal carrying rate of Tomicus-larva (90%) was higher than its pupae (78%) and new adult (58%); No significant difference in fungal carrying rate was observed among heads (33%), wings (32%) and legs (30%) of Tomicus-adults. According to field survey, the associated fungi significantly improved the breeding success rate of Tomicus on the trunk. Conclusion This is the first report on the carrying rates and the differences of fungi associated with four Tomicus species in Yunnan, China. The results provide new data for reveal outbreak mechanism of Tomicus species in Yunnan, China. -
Key words:
- Tomicus
- / fungi associated with Tomicus
- / fungal carrying rate
- / damage level
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表 1 切梢小蠹种类带菌率
Table 1. Carrying rate of Tomicus sp. with blue stain fungi
虫种
Tomicus species试验虫量/头
No. of test
insects带菌虫量/头
No. of insect
carrying fungi带菌率
Carrying
rate%云南切梢小蠹
Tomicus yunnanensis30 24 80 横坑切梢小蠹Tomicus minor 30 19 63 松芽小蠹Tomicus brevipilosus 30 16 53 华山松切梢小蠹
Tomicus armandii30 17 56 表 2 切梢小蠹不同虫态带菌率比较
Table 2. Carrying rate of Tomicus different developing stages with blue stain fungi
虫态和虫种
Stages & Tomicus species试验虫量/头
No. of test
insects带菌虫量/头
No. of insect
carrying fungi带菌率
Carrying
rate%幼虫
Larva云南切梢小蠹
Tomicus yunnanensis15 15 100 横坑切梢小蠹
Tomicus minor15 13 87 松芽小蠹
Tomicus brevipilosus15 12 80 华山松切梢小蠹
Tomicus armandii15 14 93 小计Subtotal 60 54 90 蛹
Pupa云南切梢小蠹
Tomicus yunnanensis15 12 80 横坑切梢小蠹
Tomicus minor15 13 87 松芽小蠹
Tomicus brevipilosus15 12 80 华山松切梢小蠹
Tomicus armandii15 10 67 小计Subtotal 60 47 78 新成虫
New
adults云南切梢小蠹
Tomicus yunnanensis15 10 67 横坑切梢小蠹
Tomicus minor15 8 53 松芽小蠹
Tomicus brevipilosus15 9 60 华山松切梢小蠹
Tomicus armandii15 8 53 小计Subtotal 60 35 58 表 3 切梢小蠹成虫头足翅带菌率
Table 3. Carrying rate of Tomicus's head, leg and wing with blue stain fungi
器官和虫种
Organ & Tomicus species试验虫量/头
No. of test
insects带菌虫量/头
No. of insect
carrying fungi带菌率
Carrying
rate%头
Head云南切梢小蠹
Tomicus yunnanensis15 5 33 横坑切梢小蠹
Tomicus minor15 6 40 松芽小蠹
Tomicus brevipilosus15 5 33 华山松切梢小蠹
Tomicus armandii15 4 27 小计Subtotal 60 20 33 足
Leg云南切梢小蠹
Tomicus yunnanensis15 4 27 横坑切梢小蠹
Tomicus minor15 6 40 松芽小蠹
Tomicus brevipilosus15 3 20 华山松切梢小蠹
Tomicus armandii15 5 33 小计Subtotal 60 18 30 翅
Wing云南切梢小蠹
Tomicus yunnanensis15 6 40 横坑切梢小蠹
Tomicus minor15 5 33 松芽小蠹
Tomicus brevipilosus15 4 27 华山松切梢小蠹
Tomicus armandii15 4 27 小计Subtotal 60 19 32 表 4 野外横坑切梢小蠹与伴生菌带菌调查情况
Table 4. Carrying rate of accociated fungi by Tomicus minor in Yunnan pine trunks in forests
危害类型
Damage types处理
Treatments带菌种类
Fungi species统计坑道数量
No. of survey
galleries入侵孔数
No. of entry
holes带菌率
Carrying rate
of fungi/%有效侵入孔数
No. of successful
entry holes有效坑道率
percent of successful
entry holes/%轻度A
Mild level A带菌carrying fungi O.canum 28 14 50 10 71.43 不带菌No fungi 14 50 8 57.14 中度B
Middle level B带菌Carrying fungi O.canum 28 11 39.29 7 63.64 不带菌No fungi 17 60.71 7 41.18 重度C
Severe level C带菌Carrying fungi O.canum 33 22 66.67 17 77.27 不带菌No fungi 11 33.33 3 27.27 (云南陆良; Luliang, Yunnan) 表 5 野外云南切梢小蠹与伴生菌带菌调查情况
Table 5. Carrying rate of accociated fungi by Tomicus yunnanensis in Yunnan pine trunks in forests
危害类型
Damage types处理
Treatments带菌种类
Fungi species统计坑道数量
No. of survey
galleries入侵孔数
No. of entry
holes带菌率
Carrying rate
of fungi/%有效侵入孔数
No. of successful
entry holes有效坑道率
percent of successful
entry holes/%轻度A
Mild level A带菌Carrying fungi O.canum 22 15 68.18 10 66.67 不带菌No fungi 7 31.82 3 42.86 中度B
Middle level B带菌Carrying fungi O.canum 22 19 86.36 10 52.63 不带菌No fungi 3 13.64 1 33.33 重度C
Severe level C带菌Carrying fungi O.canum 17 4 23.53 3 75.00 不带菌No fungi 13 76.47 3 23.08 (云南陆良; Luliang, Yunnan) -
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