38种储粮昆虫中Wolbachia的共生情况调查及Wolbachia对杂拟谷盗生殖调控的研究
发布时间：2018-08-19 13:36
【摘要】：Wolbachia(沃尔巴克氏体)是共生于昆虫体内的一种革兰氏阴性(Gran-)胞内共生菌,由于其广泛存在,并对宿主昆虫的生殖具有调控作用,包括诱导宿主胞质不融合,孤雌生殖,雄性致死和雄性雌性化等效应,对宿主生物的遗传和进化产生重要影响,Wolbachia可能作为一种潜在的害虫生物防治的策略而倍受研究者重视,在农业昆虫和媒介昆虫中广泛报道,然而在储粮昆虫中的报道却很少。本研究对中国13个省份64个地理种群的38种储粮昆虫采用PCR扩增ftsZ和wsp基因的方法对其体内的Wolbachia进行了检测,检测到杂拟谷盗,烟草甲,药材甲,锯谷盗,绿豆象,黑毛皮蠹,嗜卷书虱,麦蛾,地中海粉螟和麦蛾茧蜂等10种昆虫和一种储粮昆虫的捕食性天敌直腹伪蝎中感染有Wolbachia。但是在调查的数量最多且危害范围更广的赤拟谷盗,谷蠹,米象,玉米象,印度谷螟和嗜虫书虱的所有收集到的品系中,均没有检测到Wolbachia的存在。基于wsp基因序列的进化树分析,这些昆虫中感染的Wolbachia都属于A和B亚群(supergroup),这些调查和研究为将来可能利用Wolbachia防治储粮害虫和研究Wolbachia的生殖调控作用提供一些基本的信息指导。在对中国部分地区储粮害虫中Wolbachia的感染情况调查后,本研究选取了一种常见的储粮害虫杂拟谷盗为研究对象,研究了Wolbachia在杂拟谷盗体内的感染和消除动态及对其生殖行为等的影响。结果表明:(1)杂拟谷盗在收集到的两个地理种群中感染的概率为100%,且雌雄虫全部感染。(2)在饲料中添加0.03%,0.05%,0.15%和0.3%的抗生素(四环素)浓度均显著降低杂拟谷盗体内的Wolbachia感染密度,经过4周以上的处理,可以完全去掉杂拟谷盗体内感染的Wolbachia,去除效率可以达到100%。(3)在杂拟谷盗的不同生长阶段中新羽化的成虫(羽化3~5日达到性成熟后的交配高峰期)时体内的Wolbachia感染密度达到峰值,每头杂拟谷盗体内可达到3.7×107的感染密度,其次为刚羽化的成虫和蛹期,也达到了1×107数量级的感染密度。(4)在杂拟谷盗不同的身体部位和组织中感染的Wolbachia丰度比较为腹部胸部头部,其中腹部Wolbachia的含量可占总体的60%以上,此外,生殖系统内Wolbachia的丰度明显高于消化系统和免疫系统内,且雌性体内Wolbachia的含量高于雄性。(5)本研究设计了4个不同的交配组合检验杂拟谷盗的胞质亲和性,在4个交配组合中,只有♂w+(w+,Wolbachia阳性个体,来自自然种群)×♀w-(w-阴性个体,抗生素饲喂处理消除Wolbachia)不能产生后代,说明感染Wolbachia可以造成杂拟谷盗完全的单向胞质不融合现象。对产卵量和发育历期等参数的统计发现,四个组合的单头雌虫的总产卵量没有显著差异,但是卵的孵化率存在显著差异,表现出母系遗传的特质,雌虫被抗生素消除Wolbachia感染后卵的孵化率显著降低。(6)Wolbachia对雄虫的交配选择行为与繁殖力和生殖力的测定,发现感染Wolbachia与否对杂拟谷盗雄虫的交配选择行为没有显著性差异,消除Wolbachia后雄虫的生殖能力下降。(7)通过同性和异性种间及种内的交配试验也揭示了,不管是在同种的杂拟谷盗之间还是在杂拟谷盗和赤拟谷盗之间,都没有发生Wolbachia的传播。与抗生素饲喂消除的Wolbachia阴性种群相比,在自然种群中,感染Wolbachia的杂拟谷盗具有生殖上的优势,只有雌雄都感染Wolbachia时,对种群的增长才最有利。本研究丰富了Wolbachia在储粮昆虫的感染动态和Wolbachia对储粮昆虫的生殖调控的理论,为是否利用Wolbachia防治储粮害虫提供了基础的理论指导。
[Abstract]:Wolbachia (Wolbachia) is a kind of Gram-negative intracellular symbiotic bacteria in insects. Because of its widespread existence, Wolbachia plays an important role in regulating the reproduction of host insects, including induction of host cytoplasmic incompatibility, parthenogenesis, male lethality and male feminization. Wolbachia plays an important role in the inheritance and evolution of host organisms. Wolbachia may have attracted much attention as a potential biological pest control strategy. It has been widely reported in agricultural insects and vector insects, but rarely in Stored-grain insects. Wolbachia was detected in 10 species of insects and a predatory natural enemy of one kind of stored grain insects, including Corydalis heterophyllus, Tobacco beetle, Medicinal Material A, Sawmillet Steller, Mung bean weevil, Black fur beetle, Bookworm addict, wheat moth, Mediterranean meal borer and wheat moth cocoon wasp. Wolbachia was not detected in any of the collected strains of Rhododendron oryzae, Rhododendron oryzae, Oryzae oryzae, Corn weevil, Indian millet borer and insectivorous bookworm. Evolutionary tree analysis based on the WSP gene sequence showed that Wolbachia infected by these insects belonged to subgroups A and B. These investigations and studies suggest that Wolbachia may be used to control the disease in the future. After investigating the infection of Wolbachia in Stored-grain Pests in some areas of China, this study selected a common stored-grain pest, Cortex mimetica, as the research object, and studied the infection and elimination dynamics of Wolbachia in Cortex mimetica. The results showed that: (1) The probability of infection in the two geographic populations was 100%, and both sexes were infected. (2) The concentrations of 0.03%, 0.05%, 0.15% and 0.3% antibiotics (tetracycline) in the diet significantly decreased the Wolbachia infection density in the two geographic populations, after more than four weeks. (3) Wolbachia infection density reached a peak at different growth stages of newly emerged adults (the peak of mating time after 3-5 days of emergence) and reached 3.7 *107 infection density in each of the adults. (4) The abundance of Wolbachia in different body parts and tissues was higher than that in abdominal chest and head, in which the content of Wolbachia in abdomen could account for more than 60% of the total. In addition, the abundance of Wolbachia in reproductive system was significantly higher than that in elimination. The content of Wolbachia in female was higher than that in male. (5) Four different mating combinations were designed to test the cytoplasmic compatibility of C. variegata. Of the four mating combinations, only w + (w +, Wolbachia positive individuals, from the natural population) - (w - negative individuals, antibiotic feeding eliminates Wolbachia) did not. The results showed that Wolbachia infection could cause complete unidirectional cytoplasmic non-fusion. There was no significant difference in egg production and development duration among the four combinations, but there was significant difference in egg hatching rate, showing maternal inheritance and female resistance. The hatching rate of eggs decreased significantly after Wolbachia was eliminated. (6) The mating selection behavior, fecundity and fecundity of the males were determined. It was found that there was no significant difference between the males infected with Wolbachia and those infected with Wolbachia. The male reproductive ability decreased after Wolbachia was eliminated. (7) The male reproductive ability decreased through the same and heterosexual species. In addition, intraspecific mating tests also revealed that no Wolbachia transmission was observed among the same species or between the same species and between the two species. This study enriched Wolbachia's infection dynamics in stored-grain insects and Wolbachia's theory of reproductive regulation on stored-grain insects, and provided basic theoretical guidance for the use of Wolbachia to control stored-grain insects.
【学位授予单位】：河南工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S379.5
[Abstract]:Wolbachia (Wolbachia) is a kind of Gram-negative intracellular symbiotic bacteria in insects. Because of its widespread existence, Wolbachia plays an important role in regulating the reproduction of host insects, including induction of host cytoplasmic incompatibility, parthenogenesis, male lethality and male feminization. Wolbachia plays an important role in the inheritance and evolution of host organisms. Wolbachia may have attracted much attention as a potential biological pest control strategy. It has been widely reported in agricultural insects and vector insects, but rarely in Stored-grain insects. Wolbachia was detected in 10 species of insects and a predatory natural enemy of one kind of stored grain insects, including Corydalis heterophyllus, Tobacco beetle, Medicinal Material A, Sawmillet Steller, Mung bean weevil, Black fur beetle, Bookworm addict, wheat moth, Mediterranean meal borer and wheat moth cocoon wasp. Wolbachia was not detected in any of the collected strains of Rhododendron oryzae, Rhododendron oryzae, Oryzae oryzae, Corn weevil, Indian millet borer and insectivorous bookworm. Evolutionary tree analysis based on the WSP gene sequence showed that Wolbachia infected by these insects belonged to subgroups A and B. These investigations and studies suggest that Wolbachia may be used to control the disease in the future. After investigating the infection of Wolbachia in Stored-grain Pests in some areas of China, this study selected a common stored-grain pest, Cortex mimetica, as the research object, and studied the infection and elimination dynamics of Wolbachia in Cortex mimetica. The results showed that: (1) The probability of infection in the two geographic populations was 100%, and both sexes were infected. (2) The concentrations of 0.03%, 0.05%, 0.15% and 0.3% antibiotics (tetracycline) in the diet significantly decreased the Wolbachia infection density in the two geographic populations, after more than four weeks. (3) Wolbachia infection density reached a peak at different growth stages of newly emerged adults (the peak of mating time after 3-5 days of emergence) and reached 3.7 *107 infection density in each of the adults. (4) The abundance of Wolbachia in different body parts and tissues was higher than that in abdominal chest and head, in which the content of Wolbachia in abdomen could account for more than 60% of the total. In addition, the abundance of Wolbachia in reproductive system was significantly higher than that in elimination. The content of Wolbachia in female was higher than that in male. (5) Four different mating combinations were designed to test the cytoplasmic compatibility of C. variegata. Of the four mating combinations, only w + (w +, Wolbachia positive individuals, from the natural population) - (w - negative individuals, antibiotic feeding eliminates Wolbachia) did not. The results showed that Wolbachia infection could cause complete unidirectional cytoplasmic non-fusion. There was no significant difference in egg production and development duration among the four combinations, but there was significant difference in egg hatching rate, showing maternal inheritance and female resistance. The hatching rate of eggs decreased significantly after Wolbachia was eliminated. (6) The mating selection behavior, fecundity and fecundity of the males were determined. It was found that there was no significant difference between the males infected with Wolbachia and those infected with Wolbachia. The male reproductive ability decreased after Wolbachia was eliminated. (7) The male reproductive ability decreased through the same and heterosexual species. In addition, intraspecific mating tests also revealed that no Wolbachia transmission was observed among the same species or between the same species and between the two species. This study enriched Wolbachia's infection dynamics in stored-grain insects and Wolbachia's theory of reproductive regulation on stored-grain insects, and provided basic theoretical guidance for the use of Wolbachia to control stored-grain insects.
【学位授予单位】：河南工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S379.5
【参考文献】
相关期刊论文 前10条
1 张艳凯;张开军;谢蓉蓉;赵冬晓;洪晓月;;共生菌Wolbachia引起宿主细胞质不亲和的研究进展[J];昆虫学报;2015年12期
2 程超;沈加飞;明庆磊;;赤拟谷盗和杂拟谷盗体内Wolbachia的感染状况和种间传播[J];江苏农业学报;2015年02期
3 王阿e，
本文编号：2191809
本文链接：https://www.wllwen.com/shoufeilunwen/zaizhiyanjiusheng/2191809.html
