稻飞虱肠道细菌多样性分析
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摘要
本研究主要采用传统培养与16S rRNA基因序列分析研究的方法对白背飞虱[Sogatella furcifera (Horvath)]和褐飞虱[Nilaparvata lugens (Stal)]肠道中的细菌菌落进行了初步分离与鉴定研究,对其生理生化特性进行了分析。
①采用16S rRNA基因序列分析的方法从白背飞虱和褐飞虱的肠道中鉴定出属于11个属的细菌,它们是Wolbachia、短波单胞菌属(Brevundimonas sp.)、定氮螺旋菌属(Azospirillum sp.)、苍白杆菌属( Ochrobaclrum sp.)、草螺菌属(Herbaspirillumsp.)、丛毛单胞菌属(Comamonas sp.)、不动杆菌属(Acinetobacler sp.)、窄食单胞菌属(Stenotrophomonas sp.)、白色杆菌(Leucobater sp.)、Cardium菌、以及暂定菌群TM7菌。
②根据已鉴定到的菌种的基因序列设计引物,检测各细菌在室内及田间白背飞虱与褐飞虱体内的感染率。实验结果表明：白背飞虱和褐飞虱的室内种群与田间种群个体之间各种细菌的感染率差别不大；白背飞虱与褐飞虱各细菌的感染率差别比较大。在白背飞虱的室内和田间种群中Wolbachia和Cardinium的感染率最高,是白背飞虱的优势共生菌种；在褐飞虱的室内种群和田间种群中,不动杆菌和TM7的感染率却是最高的,是褐飞虱的优势共生菌种。
③采用传统的培养方法,对白背飞虱与褐飞虱肠道细菌进行了培养分离,对分离到的纯化菌株进行16S rRNA基因序列分析,序列分析表明通过培养纯化共分离得到属于10个属的细菌和一种不可培养的微生物,它们是芽孢杆菌属(Bacillus sp.)金黄杆菌属(Chryseobacterium sp.)、葡萄球菌属(Staphylococcus sp.)、微杆菌属(Microbacterium sp.)、放线菌属(Actinobacterium sp.)、短波单胞菌属(Brevundimonas sp.)、红球菌属(Rhodococcus sp.)、不动杆菌属(Acinetobactersp.)、肠杆菌属(Enterobacter sp.)、欧文氏菌属(Erwinia sp.)。从白背飞虱肠道中得到的菌株主要为金黄杆菌属、芽孢杆菌属、葡萄球菌属、放线菌属、短波单胞菌属、微杆菌属、不动杆菌属以及一株不可培养的微生物,而从褐飞虱肠道中得到的菌株主要是不动杆菌属、欧文氏菌属、芽孢杆菌属、肠杆菌属、微杆菌属以及金黄杆菌属。
④对可培养细菌的培养性状、染色反应、生理生化反应等进行了系统研究。对各种可培养菌的特性有初步的了解。
⑤从室内的白背飞虱吡虫啉筛选种群、噻嗪酮筛选种群以及褐飞虱的氟虫腈筛选种群中各分离筛选到一个菌株。这三个菌株能分别以吡虫啉、噻嗪酮、氟虫腈为唯一碳源和能源生长。
This study is mainly about intestinal bacterium of the white-backed planthopper[Sogatella furcifera (Horvath)] and brown planthopper [Nilaparvala lugens (Stal)]. We initially studied the intestinal microorganism of white-backed planthooper and brown planthopper by traditional and molecular gene anlysis methods. and studied the physiology and biochemistry characteristic of bacterium we got from planthopper.
①We got 11 genuses bacterium from white-backed planthopper and brown planthopper by 16S rRNA molecular gene anlysis method, they were Wolbachia t.Brevundimonas sp. Azospirillum sp, Ochrobactrum sp. Herbaspirillum sp, Comamonas sp. Acinelobacter sp. Stenotrophomonas sp. Leucobaler sp,. Cardium. TM7 bacterium.
②Designing primers according to gene sequence that we have identified, and to detect the infection rate of every bacterium in laboratory colonies and farm colonies. The results show that there is a little difference intersubject laboratory and farm colonies of white-backed planthopper and brown planthopper. and there is a great difference between laboratory and farm colonies. Wolbachia and Cardium had high infection rate in the white-backed planthopper from laboreatory and farm colonies:Acinelobacter and TM7 bacterium had high infection rate in brown planthopper from laboratory and farm colonies.
③We isolated 10 genuses bacterium ang one uncultured bacterium from white-backed planthopper and brown planthopper by traditional cultural method, they were Bacillus sp.Chryseobacrerium sp, Staphylococcus sp, Microbacterium sp, Actinobacterium sp, Brevundimonas sp. Rhodococcus sp. Acinetobacter sp. Enterobacter sp, Erwinia sp. We got Bacillus sp., Chryseobacterium sp.. Staphylococcus sp., Microbacterium sp., Actinobacterium sp., Brevundimonas sp., Rhodococcus sp.,Acinetobacter sp and one uncultured bacterium from white-backed planthopper. and we got Bacillus sp..Chryseobacterium sp.. Actinobacterium sp., Microbacterium sp..Enterobacter sp.,Erwinia sp from brown planthopper.
④We have studied the cultivatied character, the staining reaction and physiology and biochemistry reaction of cultivatied bacterium. We have initially knowed these bacterium.
⑤We isolated each bacteria from laboratory imidacloprid screening population, buprofezin screening population of white kacked planthopper and fiproni screening population of brown planthopper. Each of the three strains can grow by imidacloprid. buprofezin and fiproni as sole carbon source and energy source.
①采用16S rRNA基因序列分析的方法从白背飞虱和褐飞虱的肠道中鉴定出属于11个属的细菌,它们是Wolbachia、短波单胞菌属(Brevundimonas sp.)、定氮螺旋菌属(Azospirillum sp.)、苍白杆菌属( Ochrobaclrum sp.)、草螺菌属(Herbaspirillumsp.)、丛毛单胞菌属(Comamonas sp.)、不动杆菌属(Acinetobacler sp.)、窄食单胞菌属(Stenotrophomonas sp.)、白色杆菌(Leucobater sp.)、Cardium菌、以及暂定菌群TM7菌。
②根据已鉴定到的菌种的基因序列设计引物,检测各细菌在室内及田间白背飞虱与褐飞虱体内的感染率。实验结果表明：白背飞虱和褐飞虱的室内种群与田间种群个体之间各种细菌的感染率差别不大；白背飞虱与褐飞虱各细菌的感染率差别比较大。在白背飞虱的室内和田间种群中Wolbachia和Cardinium的感染率最高,是白背飞虱的优势共生菌种；在褐飞虱的室内种群和田间种群中,不动杆菌和TM7的感染率却是最高的,是褐飞虱的优势共生菌种。
③采用传统的培养方法,对白背飞虱与褐飞虱肠道细菌进行了培养分离,对分离到的纯化菌株进行16S rRNA基因序列分析,序列分析表明通过培养纯化共分离得到属于10个属的细菌和一种不可培养的微生物,它们是芽孢杆菌属(Bacillus sp.)金黄杆菌属(Chryseobacterium sp.)、葡萄球菌属(Staphylococcus sp.)、微杆菌属(Microbacterium sp.)、放线菌属(Actinobacterium sp.)、短波单胞菌属(Brevundimonas sp.)、红球菌属(Rhodococcus sp.)、不动杆菌属(Acinetobactersp.)、肠杆菌属(Enterobacter sp.)、欧文氏菌属(Erwinia sp.)。从白背飞虱肠道中得到的菌株主要为金黄杆菌属、芽孢杆菌属、葡萄球菌属、放线菌属、短波单胞菌属、微杆菌属、不动杆菌属以及一株不可培养的微生物,而从褐飞虱肠道中得到的菌株主要是不动杆菌属、欧文氏菌属、芽孢杆菌属、肠杆菌属、微杆菌属以及金黄杆菌属。
④对可培养细菌的培养性状、染色反应、生理生化反应等进行了系统研究。对各种可培养菌的特性有初步的了解。
⑤从室内的白背飞虱吡虫啉筛选种群、噻嗪酮筛选种群以及褐飞虱的氟虫腈筛选种群中各分离筛选到一个菌株。这三个菌株能分别以吡虫啉、噻嗪酮、氟虫腈为唯一碳源和能源生长。
This study is mainly about intestinal bacterium of the white-backed planthopper[Sogatella furcifera (Horvath)] and brown planthopper [Nilaparvala lugens (Stal)]. We initially studied the intestinal microorganism of white-backed planthooper and brown planthopper by traditional and molecular gene anlysis methods. and studied the physiology and biochemistry characteristic of bacterium we got from planthopper.
①We got 11 genuses bacterium from white-backed planthopper and brown planthopper by 16S rRNA molecular gene anlysis method, they were Wolbachia t.Brevundimonas sp. Azospirillum sp, Ochrobactrum sp. Herbaspirillum sp, Comamonas sp. Acinelobacter sp. Stenotrophomonas sp. Leucobaler sp,. Cardium. TM7 bacterium.
②Designing primers according to gene sequence that we have identified, and to detect the infection rate of every bacterium in laboratory colonies and farm colonies. The results show that there is a little difference intersubject laboratory and farm colonies of white-backed planthopper and brown planthopper. and there is a great difference between laboratory and farm colonies. Wolbachia and Cardium had high infection rate in the white-backed planthopper from laboreatory and farm colonies:Acinelobacter and TM7 bacterium had high infection rate in brown planthopper from laboratory and farm colonies.
③We isolated 10 genuses bacterium ang one uncultured bacterium from white-backed planthopper and brown planthopper by traditional cultural method, they were Bacillus sp.Chryseobacrerium sp, Staphylococcus sp, Microbacterium sp, Actinobacterium sp, Brevundimonas sp. Rhodococcus sp. Acinetobacter sp. Enterobacter sp, Erwinia sp. We got Bacillus sp., Chryseobacterium sp.. Staphylococcus sp., Microbacterium sp., Actinobacterium sp., Brevundimonas sp., Rhodococcus sp.,Acinetobacter sp and one uncultured bacterium from white-backed planthopper. and we got Bacillus sp..Chryseobacterium sp.. Actinobacterium sp., Microbacterium sp..Enterobacter sp.,Erwinia sp from brown planthopper.
④We have studied the cultivatied character, the staining reaction and physiology and biochemistry reaction of cultivatied bacterium. We have initially knowed these bacterium.
⑤We isolated each bacteria from laboratory imidacloprid screening population, buprofezin screening population of white kacked planthopper and fiproni screening population of brown planthopper. Each of the three strains can grow by imidacloprid. buprofezin and fiproni as sole carbon source and energy source.
引文
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