内蒙古典型草原优势种蝗虫食物适应性研究
摘要
植物群落结构与蝗虫食物适应性决定蝗虫的时空及营养生态位，人工饲喂和自然取食方法研究了优势种蝗虫取食特性，应用荧光定量PCR技术建立了蝗虫嗉囊内含物植物种类分析和食量测定的方法。研究结果如下：
建立了草原蝗虫嗉囊提取和保存的标准和方法。根据6种优势种植物克氏针茅、羊草、糙隐子草、锦鸡儿、冷蒿、猪毛菜核糖体DNA（ribosome DNA, rDNA）的转录间隔区（Internal TranscribedSpacers，ITS）和5.8S的基因序列，设计了适于用SYBR Green I的特异性引物，建立了定量检测的标准曲线。以6种植物定量饲喂2种蝗虫，定量PCR检测蝗虫嗉囊内含物各植物ITS区的拷贝数，明确了ITS区拷贝数与食量的关系。
植物群落结构影响蝗虫的群落组成，典型相关结果表明蝗虫综合因子与植物综合因子间紧密相关（r=0.955，P<0.05）；优势种植物与优势种蝗虫密度密切相关，羊草为优势种的草地类型中羊草生物量损失率与毛足棒角蝗相对多度显著正相关（r=0.849,p<0.01）；克氏针茅为优势种的草地类型和克氏针茅伴生杂类草的草地类型中，克氏针茅生物量损失率与亚洲小车蝗相对多度显著正相关（r=0.947,0.93,p<0.01）。荧光定量PCR检测证明毛足棒角蝗对于羊草有显著的取食偏好。毛足棒角蝗与亚洲小车蝗营养生态位宽度分别为0.224和0.207，亚洲小车蝗对毛足棒角蝗的生态位重叠数值为0.125，毛足棒角蝗对亚洲小车蝗的生态位重叠数值为0.115，存在食物竞争关系。
依据蝗虫死亡率和食性参数两个指标研究亚洲小车蝗、毛足棒角蝗、鼓翅皱膝蝗密度及样地条件4因素3水平正交试验，结果表明：影响蝗虫死亡率的主要因素是样地条件，次要因素是蝗虫自身的虫口密度；影响蝗虫食性的最主要因素是样地条件，次要因素是与其营养生态位有竞争关系的其他蝗虫的虫口密度。
Temporal, spatial and trophic niches of grasshoppers are dependent on the structure of plantcommunity and food preference of grasshoppers. Articial and natural feeding experiments were used todetermine the feeding characters of grasshoppers, amount of ingested plants in grasshopper’s crop wereanalyzed using RT-PCR. The results are as follows.
1) The standard and method of extracting and storage of grasshopper crop were established. Basedon the ITS of rDNA and5.8s gene sequences of six plants, Stipa krylovii, Leymus chinensis,Cleistogenes squarrosa, Caragana microphylia, Artemisia frigida, and Salsola collina, special primers,fit to SYBR Green I, were designed in those plants. Standard curve for quantitative detection of theamount of plants ingested by grasshopper was also established. The relationship of the copies of ITSand the weight of contents in crops was determined by feeding six kinds of host plants to Oedaleusasiaticus and Dasyhippus barbipes and analyzing plant contents in crops with RT-PCR.
2) Grassland community structure could affect the community of grasshopper. Canonicalcorrelation analysis showed that there was a positive correlation (r=0.955，P <0.05) between thecomprehensive factor of grasshopper and the comprehensive factor of host plant. Density of dominantgrasshopper species depended on dominant host plant speices. There was positive correlations betweenbiomass loss of L. chinensis and relative abundance of D. barbipes in sample plot one, between biomassloss of S. krylovii and relative abundance of O. asiaticus in sample plots two and three. RT-PCR showedthat D. barbipes prefered Leymus chinensis. The width of trophic niches of D. barbipes and O. asiaticuswas0.224and0.207, respectively; the former overlapping the latter by0.115and the latter overlappingthe former by0.125, which suggests that there is food competition in these two grasshopper species.
3) Orthogonal test was employed to analyze the effects of density of O. asiaticus, D. barbipes andAngaracris barabensis and sample plot structure on mortality and food ingestion of the grasshoppers.The results showed that the two most important factors that affected grasshopper mortality weregrassland structure and population density of grasshopper, and the two most important factors that affectfood ingestion of grasshoppers were grassland structure and population density of other grasshopperswith high level of trophic niche overlap.
建立了草原蝗虫嗉囊提取和保存的标准和方法。根据6种优势种植物克氏针茅、羊草、糙隐子草、锦鸡儿、冷蒿、猪毛菜核糖体DNA（ribosome DNA, rDNA）的转录间隔区（Internal TranscribedSpacers，ITS）和5.8S的基因序列，设计了适于用SYBR Green I的特异性引物，建立了定量检测的标准曲线。以6种植物定量饲喂2种蝗虫，定量PCR检测蝗虫嗉囊内含物各植物ITS区的拷贝数，明确了ITS区拷贝数与食量的关系。
植物群落结构影响蝗虫的群落组成，典型相关结果表明蝗虫综合因子与植物综合因子间紧密相关（r=0.955，P<0.05）；优势种植物与优势种蝗虫密度密切相关，羊草为优势种的草地类型中羊草生物量损失率与毛足棒角蝗相对多度显著正相关（r=0.849,p<0.01）；克氏针茅为优势种的草地类型和克氏针茅伴生杂类草的草地类型中，克氏针茅生物量损失率与亚洲小车蝗相对多度显著正相关（r=0.947,0.93,p<0.01）。荧光定量PCR检测证明毛足棒角蝗对于羊草有显著的取食偏好。毛足棒角蝗与亚洲小车蝗营养生态位宽度分别为0.224和0.207，亚洲小车蝗对毛足棒角蝗的生态位重叠数值为0.125，毛足棒角蝗对亚洲小车蝗的生态位重叠数值为0.115，存在食物竞争关系。
依据蝗虫死亡率和食性参数两个指标研究亚洲小车蝗、毛足棒角蝗、鼓翅皱膝蝗密度及样地条件4因素3水平正交试验，结果表明：影响蝗虫死亡率的主要因素是样地条件，次要因素是蝗虫自身的虫口密度；影响蝗虫食性的最主要因素是样地条件，次要因素是与其营养生态位有竞争关系的其他蝗虫的虫口密度。
Temporal, spatial and trophic niches of grasshoppers are dependent on the structure of plantcommunity and food preference of grasshoppers. Articial and natural feeding experiments were used todetermine the feeding characters of grasshoppers, amount of ingested plants in grasshopper’s crop wereanalyzed using RT-PCR. The results are as follows.
1) The standard and method of extracting and storage of grasshopper crop were established. Basedon the ITS of rDNA and5.8s gene sequences of six plants, Stipa krylovii, Leymus chinensis,Cleistogenes squarrosa, Caragana microphylia, Artemisia frigida, and Salsola collina, special primers,fit to SYBR Green I, were designed in those plants. Standard curve for quantitative detection of theamount of plants ingested by grasshopper was also established. The relationship of the copies of ITSand the weight of contents in crops was determined by feeding six kinds of host plants to Oedaleusasiaticus and Dasyhippus barbipes and analyzing plant contents in crops with RT-PCR.
2) Grassland community structure could affect the community of grasshopper. Canonicalcorrelation analysis showed that there was a positive correlation (r=0.955，P <0.05) between thecomprehensive factor of grasshopper and the comprehensive factor of host plant. Density of dominantgrasshopper species depended on dominant host plant speices. There was positive correlations betweenbiomass loss of L. chinensis and relative abundance of D. barbipes in sample plot one, between biomassloss of S. krylovii and relative abundance of O. asiaticus in sample plots two and three. RT-PCR showedthat D. barbipes prefered Leymus chinensis. The width of trophic niches of D. barbipes and O. asiaticuswas0.224and0.207, respectively; the former overlapping the latter by0.115and the latter overlappingthe former by0.125, which suggests that there is food competition in these two grasshopper species.
3) Orthogonal test was employed to analyze the effects of density of O. asiaticus, D. barbipes andAngaracris barabensis and sample plot structure on mortality and food ingestion of the grasshoppers.The results showed that the two most important factors that affected grasshopper mortality weregrassland structure and population density of grasshopper, and the two most important factors that affectfood ingestion of grasshoppers were grassland structure and population density of other grasshopperswith high level of trophic niche overlap.
引文
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