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Wuchang Roseau nematodes and Ovomermis mitochondrial genome polymorphism analysis

Author: HeFang
Tutor: WangGuoXiu
School: Central China Normal University
Course: Zoology
Keywords: Nematodes Wuchang Roseau nematodes Ovomermis Mitochondrial genome Rolling circle replication Long PCR
CLC: S476.15
Type: Master's thesis
Year: 2009
Downloads: 62
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Abstract


Entomopathogenic Mermithidae widely parasitic on agriculture, forestry and health pests in vivo, and its parasitic insect mortality rate is equal to, is a kind of insect predators valuable resources, protect the environment and achieve farmland biodiversity is of great significance. Mitochondria are found in most eukaryotic cells, the exercise of oxidative phosphorylation function of important organelles. Mitochondrial DNA (mtDNA) is a double-stranded genetic material outside the core, rich in animals, with a small molecular weight, molecular evolution speed, conserved structural genes and other characteristics, are widely used in molecular genetic evolution, classification and identification, developmental biology and variation with some diseases. At present, foreign countries have been on cutworms six cable nematode (Hexamermis agrotis), Dosso nematode Pending kind BH-2006 (Agamermis sp.BH-2006), mosquito Roseau nematodes (Romanomermis culicivorax), 尼尔森里斯罗 cable nematode (R. nielsenni), Erichsen Roseau nematodes (R.iyengari) and a total of six kinds Mermithidae Thaumamermiscosgrovei mitochondrial genome was sequenced and research in this area has just started domestic. In order to improve Mermithidae mitochondrial genome sequence database, a more systematic study of mitochondrial genome characteristics and system evolution, and thus to improve the potential biocontrol nematodes lay the foundation of the laboratory for the first time in the country to carry out Mermithidae mitochondrial genome research . The experiment will be rolling circle replication and long PCR amplification technology combine successfully completed Wuchang Roseau nematode (Romanomermis wuchangensis) and Ovomermis (Ovomermis sinensis) mitochondrial genome sequencing, and the latter exists intraspecific genetic polymorphism of the reasons for the inquiry. In addition, with the six already in Genbank Mermithidae mitochondrial genome sequences were compared, summarized Mermithidae basic characteristics of the mitochondrial genome and analyzed. These findings will now be reported as follows. 1, Wuchang Roseau nematode mitochondrial genome by rolling circle replication, restriction enzyme digestion and agarose gel electrophoresis, and its spectrum shows the mitochondrial genome of each monomer digested bands consistent with its size is basically the same, total length is about 15kb. After the two-step on a long nematode PCR amplification and sequencing stitching get their mitochondrial genome sequence of a total length of 14832bp, consistent with the restriction map. Its mitochondrial genome, including 12 (12 kinds) protein-coding genes, two rRNA genes, 23 tRNA genes and a section of the AT-rich D-loop region. 2, Ovomermis mitochondrial genome by rolling circle replication, restriction enzyme digestion and agarose gel electrophoresis, and its spectrum shows each monomer of the mitochondrial genome fragments have shown specificity, length changes in the larger, 16.5 to 24.5 kb, showed Ovomermis mitochondrial genome exists intraspecific genetic polymorphism. In a Ovomermis mitochondrial genome sequencing based on the success of this experiment, using multi-step PCR (four steps) for another one representative nematode sequences were amplified and sequenced, stitching get their mitochondrial genome sequence a total length of 16777 bp, consistent with the restriction map. Its mitochondrial genome, including 13 (12 kinds) protein-coding genes, two rRNA genes, 20 tRNA genes and a section of the AT-rich D-loop region. 3, through two Ovomermis mitochondrial genome sequence comparison of polymorphism and its causes were discussed. Its mainly by mitochondrial genome polymorphism in the variable part caused by, in this section there is a different gene order and gene duplication. We cited the doubling - random deletion model and mechanism of mitochondrial gene rearrangements be a reasonable explanation. Sequence multiplication and intramolecular, intermolecular recombination is Ovomermis mitochondrial genome polymorphism generated the most likely cause. 4, with the six already in Genbank Mermithidae mitochondrial genome sequence comparison generalize Mermithidae mitochondrial genome has the following characteristics: ① mitochondrial gene order varies; ② some nematode mitochondrial genes duplication and repetition Views differ; ③ mitochondrial length there is a big difference. In summary, the present study, rolling circle replication and long PCR amplification method of combining the successful completion of a nematode and a Wuchang Roseau Ovomermis mitochondrial genome sequencing, and Ovomermis mitochondrial genome species Within polymorphism discussed. With the six already in Genbank Mermithidae mitochondrial genome sequences were compared, summarized Mermithidae basic characteristics of the mitochondrial genome, and its analyzes. Accumulated research Mermithidae mitochondrial genome valuable methodological and theoretical experience to better carry out a comprehensive mitochondrial genome of Mermithidae study provides some reference information.

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CLC: > Agricultural Sciences > Plant Protection > A variety of control methods > Biological control > The use of microbial pathogens > Nematodes
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