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Acridid feeding preference detecting method and kit
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A preference, kit technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as large quantitative errors
Active Publication Date: 2014-07-30
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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These methods have certain limitations when analyzing different types of locusts and eating a variety of plants at the same time, and the quantitative error is large
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Embodiment 1
[0050] 1. Collecting locust crops
[0051] Select a plot with relatively flat terrain and relatively uniform vegetation, and collect locusts at around 15:00 every day. When sampling, detour to the upper air outlet and throw out the bottomless sample frame, and take samples with the fluke machine, and take 10 sample squares for each grassland type. The insect sucker collects locust gnats and adults, quickly extracts their crops, separates the crops from other insects, and stores them in quick-frozen liquid nitrogen tanks on-board after marking.
[0052] 2. Preparation and quantification of DNA in locust crop contents
[0053] Proceed as follows:
[0054] DNA preparation
[0055] 1) Take locust crops and put them into a 1.5ml centrifuge tube, add liquid nitrogen and grind until powdery;
[0056] 2) Add 600 μl 2% CTAB preheated at 65°C, shake well, and place in a 65°C water bath for 1 hour. During this period, turn the centrifuge tube up and down several times, take it out o...
[0115] Mix the locust crops fed by a single plant (Stipa cloverii, Leymus chinensis, and Cryptospermia grassa) obtained by mixing 3 repeated experiments, extract DNA, and detect the copy number of the corresponding plant by fluorescence quantitative PCR with plant-specific primers, and use the above equation to correspond Return the weight of the plant in the locust crop contents. It is compared with the plant weight that the locusts ingested in the body per hour obtained by the traditional method, and the accuracy of the method of measuring the food intake of the locusts by checking the fluorescent quantitative PCR technique (such as Figure 7-1 , 7-2 , 7-3).
[0116] By comparing and analyzing the change trends of the six sets of data and the standard deviations of each set of data, the quality of the locust crop contents at 15 o'clock every day (that is, three hours after the locusts start eating) is selected to repr...
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Abstract
The invention relates to the technical field of biology, and in particular relates to an acridid feeding preference detecting method and kit. The method comprises the step of performing fluorescent quantitative polymerase chain reaction (PCR) by virtue of a stipa krylovii gene specific primer sequence, a Chinese wildrye gene specific primer sequence, a cleistogenes squarrosa gene specific primer sequence, a caragana microphylla gene specific primer sequence, an artemisia frigida willd gene specific primer sequence and a salsola collina pall gene specific primer sequence. The detecting method provided by the invention can be used for quickly and accurately detecting acridid feeding preference, adopts materials which are readily available and easy to buy, is simple to operate, is time-saving and labor-saving, and is suitable for popularization and application.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a method and a detection kit for detecting the feeding preference of locusts. Background technique [0002] Plants provide food and habitat for insects, and herbivorous insects have obvious selectivity for host plants. Locusts are important members of the ecosystem and play an important role in the biological chain. At the same time, it is also one of the consumers, which harms agricultural production and competes with livestock for feed plants. Qualitative and quantitative detection of locust feeding habits and food intake can provide technical guidance for biological control, ecological management, and diversity regulation. [0003] The research methods of locust feeding habits include research on feeding characteristics of locusts in cages, adaptability test of locust mouthparts structure, analysis of morphological characteristics of locust feces, morphological research of locus...
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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 吴惠惠曹广春张泽华
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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