【摘要】：蝗總科昆蟲是直翅目的重要組成部分,也在食物鏈中扮演著多種角色,對維持生態(tài)系統(tǒng)穩(wěn)定有不可替代的作用�；瓤偪评ハx對農業(yè)、牧業(yè)和林業(yè)的發(fā)展有巨大影響,與人類生活息息相關,所以對蝗總科的研究意義重大。在蝗總科的研究中,系統(tǒng)學一直是研究的熱點。傳統(tǒng)分類學主要以形態(tài)為依據,已經不能滿足現代研究的要求。本研究采用酚-氯抽提、PCR擴增和DNA測序等手段,輔助以Bio Lign、MEGA5.05、PAUP4.0、Mr Bayes3.1.2等軟件,對劍角蝗科、斑翅蝗科、槌角蝗科和斑腿蝗科16種蝗蟲的16Sr DNA和COII基因,進行序列組成分析、堿基替換分析、密碼子使用分析、氨基酸組成分析、遺傳距離分析和堿基替換飽和度分析,并最后以短額負蝗為外群,采用ML、MP、NJ、BI法,分別對16Sr DNA基因、COII基因和兩個基因聯合構建系統(tǒng)發(fā)育樹,進行分子系統(tǒng)學研究。通過研究分析,得到以下主要結論:(1)蝗總科線粒體基因具有明顯A+T偏向性,其中16Sr DNA基因A+T含量為71.2%,COII基因A+T含量為71.8%。(2)16Sr DNA基因保守性為66.5%,COII基因保守性為53.2%。16Sr DNA基因比COII基因更保守。同時,基因中的第三位點的變異容易被保留下來,所以第三位點變異率更高。(3)COII基因中出現頻率較高的密碼子為AUU(21.3)、UUA(17)和ACA(12.1),而CUG、UCG和GCG沒有出現,密碼子出現頻率的差異可能與線粒體內相應種類t RNA含量有關。含量較高的氨基酸為異亮氨酸(11.57)、亮氨酸(11.49)和絲氨酸(9.72)。(4)16種蝗蟲的16Sr DNA基因的非矯正遺傳距離在0.006-0.151之間。16種蝗蟲的COII基因的非矯正遺傳距離范圍在0.007-0.236之間。(5)通過建立遺傳距離與R值、TS、TV的關系,證明該兩個基因的堿基替換沒有達到飽和。(6)在系統(tǒng)發(fā)育關系上,劍角蝗科、斑翅蝗科、槌角蝗科和斑腿蝗科互為單系群。紅翅皺膝蝗與鼓翅皺膝蝗親緣關系較近,但不是一個物種(支持率95)。黃脛小車蝗并沒有和亞洲小車蝗形成姊妹群,而是與輪紋異痂蝗親緣關系更近。本論文通過測定16種蝗蟲的16Sr DNA和COII基因序列,對其進行系統(tǒng)發(fā)育分析,為蝗蟲的系統(tǒng)分類提供了分子依據。后續(xù)工作中將增加蝗蟲種類,加大擴增片段長度,以獲得更可靠的系統(tǒng)發(fā)育信號。
[Abstract]:Acridae is an important part of Orthoptera and plays a variety of roles in the food chain, which plays an irreplaceable role in maintaining ecosystem stability. Acridae insects have great influence on the development of agriculture, animal husbandry and forestry, and are closely related to human life, so the research on Acridae is of great significance. Systematics has always been a hot topic in the studies of Acridoidea. Traditional taxonomy is mainly based on morphology and can not meet the requirements of modern research. In this study, the 16Sr DNA and COII genes of 16 species of grasshopper of the family Acridoidea, Acridae, Acridae and Acridae were sequenced and analyzed by means of phenol-chlorine extraction PCR amplification and DNA sequencing, and Bio Lign,MEGA5.05,PAUP4.0,Mr Bayes3.1.2 software was used to analyze the 16Sr DNA and COII genes of 16 species of grasshopidae. Base substitution analysis, codon usage analysis, amino acid composition analysis, genetic distance analysis and base substitution saturation analysis were used. Finally, ML,MP,NJ,BI method was used. The phylogenetic tree was constructed by 16Sr DNA gene and two genes, and the molecular phylogeny was studied. The main conclusions are as follows: (1) Mitochondrial genes of Acridoidea have obvious A-T bias. The A T content of 16Sr DNA gene was 71.2 and that of COII gene was 71.8. (2) the conserved nature of 16Sr DNA gene was 66.5%. 53.2%.16Sr DNA gene was more conserved than COII gene. At the same time, the variation of the third locus in the gene is easy to be preserved, so the mutation rate of the third locus is higher. (3) the codon with higher frequency in COII gene is AUU (21.3) UUA (17) and ACA (12.1), but CUG,UCG and GCG do not appear. The difference of codon frequency may be related to the content of t RNA in mitochondria. The higher amino acids were isoleucine (11.57), leucine (11.49) and serine (9.72). (4). The uncorrected genetic distance of 16Sr DNA gene of 16 locust species was 0.006-0.151. The uncorrected genetic distance of COII gene of 16 species locust was 0.007-0.236. The relationship between genetic distance and R value of TSTV. It is proved that the base substitution of the two genes is not saturated. (6) the phylogenetic relationships of the two genes are monophyletic. The phylogenetic relationship between Gymnochus rotundii and A. typhus was close, but it was not a species (support rate 95). It was not a sister group with the Asiatic locust, but more closely related to the whorl heteroglyphagus locust. In this paper, the 16Sr DNA and COII genes of 16 species of locusts were sequenced and phylogenetic analysis was carried out, which provided molecular basis for the systematic classification of locusts. In the following work, the species of locusts and the length of amplified fragments will be increased to obtain more reliable phylogenetic signals.
【學位授予單位】：東北師范大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S433.2

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