Method for detecting parasitism rate and ovipositing quantity of aphelinus mali in eriosoma lanigerum

An apple aphid and sunbee technology, applied in the field of agricultural biotechnology detection, can solve problems such as failure to hatch or die before developing to the pupal stage, inaccurate egg production of sunbees, time-consuming and labor-intensive, etc., and achieve clear control. Harmful effect, overcoming the effect of low parasitic rate

Active Publication Date: 2017-09-01
QINGDAO AGRI UNIV
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Problems solved by technology

[0004] The sun bee achieves parasitic effect on the apple aphid by laying eggs in the apple cotton aphid. The eggs develop into larvae in the apple cotton aphid and absorb their nutrients. Before and after the sun bee pupates, the apple aphid turns black and dies. In the past, the statistics of the parasitism rate of the sun bee were based on the blackening and death of the apple aphid, and the number of apple aphids parasitized by the sun bee was counted. After the sun bee parasitized the apple cotton aphid in the field, some of the apple aphids were parasitized by the sun bee during the development process of the sun bee. The apple cotton aphid parasitized by the sun bee will be lost due to the predation of other natural enemies and the erosion of rainwater. More importantly, in the field, not all eggs of the sun bee can develop to the larval or pupal stage, and some apple

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  • Method for detecting parasitism rate and ovipositing quantity of aphelinus mali in eriosoma lanigerum
  • Method for detecting parasitism rate and ovipositing quantity of aphelinus mali in eriosoma lanigerum
  • Method for detecting parasitism rate and ovipositing quantity of aphelinus mali in eriosoma lanigerum

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: the design of primer

[0038]The applicant's previous research found that, based on the 700bp gene in the second half of the mitochondrial COI, my country's sun bee is divided into three haplotypes, Hap1, Hap2, and Hap3, which can be divided into two sun bee genetic branches, the Liaoning branch and the Shandong branch. There are differences in the biological characteristics and pest control ability of these two genetic branches, and the research found that YL (Yili), ZT (Zhaotong), HZ (Heze), TA (Tai'an), LC (Liaocheng) and QD (Qingdao) It is a pure population of Shandong branch, DL (Dalian), HLD (Huludao), QHD (Qinhuangdao) and SJZ (Shijiazhuang) is a pure population of Liaoning branch, YT (Yantai), WF (Weifang), BD ( Baoding), JZ (Jinzhong), CC (Changzhi) and YC (Yuncheng) are mixed populations of two clades. Therefore, when studying the parasitism rate of sunbee on the apple aphid and the detection method of egg production, we should fully consider the t...

Embodiment 2

[0057] Example 2 Detection of sunbee, apple aphid and apple aphid parasitized by sunbee by PCR amplification technology

[0058] (1) In the apple orchard of Chengyang District, collect 5 sunbee adults, apple aphids and apple cotton aphids parasitized by sunbees;

[0059] (2) Preparation of lysate

[0060] Prepare a reagent bottle with a capacity of more than 200ml, wash it and use it for the experiment

[0061] First need to equip 0.05mol / L Tris-HCl (pH 8.4)

[0062] 1) 0.1mol / L Tris: 1.211g Tris for 100ml distilled water

[0063] 2) 0.1mol / L HCl: 0.862mL (the concentration of common industrial hydrochloric acid is 12M) dissolved in 100mL distilled water

[0064] 3) Mix 50mL of 0.1mol / L Tris alkali solution with 17.2mL of 0.1mol / L HCl, add water to make up to 100mL, and obtain 0.05mol / L Tris-HCl (pH 8.4) (sterilize after preparation)

[0065] Add the following reagents to 0.05mol / L Tris-HCl (pH 8.4)

[0066]

[0067] After adding everything, put it in the refrigerator ...

Embodiment 3

[0085] Embodiment 3 detects the parasitism rate of the apple cotton aphid parasitized by the sun bee in the field

[0086] (1) On May 20, 2016, an apple branch with apple aphids (some of which had been parasitized by sun bees) was collected in the field, and brought back to the room with tweezers to gently remove all the apple aphids, a total of 13;

[0087] (2) Put one head of the collected cotton aphid into a 0.2 mL centrifuge tube, add 10 μL of lysate, grind and homogenize the sunbee with a 10 μL sealing tip, and then add 40 μL of lyse. Place in a water bath at 65°C for 15 minutes, then at 95°C for 10 minutes, and store below -20°C;

[0088] (3) using the genomic DNA obtained in step (2) as a template, performing PCR amplification on the target gene in the genomic DNA to obtain a PCR amplification product;

[0089] The PCR amplification system is 13 μl:

[0090] Genomic DNA solution 2μl, 20μM primer 0.26μl, 5U / μl Taq enzyme 0.2μl, 10×Taq Buffer (buffer) 1.3μl, 10mM dNTP 1...

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Abstract

The invention provides a method for detecting parasitism rate and ovipositing quantity of aphelinus mali in eriosoma lanigerum. An invented primer pair is used for conducting PCR amplification, the aphelinus mali and target genes of the aphelinus mali in genomic DNA of the eriosoma lanigerum can be detected, and accordingly, whether the eriosoma lanigerum is parasitized by the aphelinus mali or not is distinguished. By adoption of the method provided by the invention, the ovipositing quantity of the aphelinus mali can be quite clearly calculated, and calculation is more accurate and easier than an anatomical method; the parasitism rate of the parasites in the eriosoma lanigerum can be accurately calculated, the pest control effect of the aphelinus mali on the eriosoma lanigerum is confirmed, and the phenomenon that certain aphelinus mali eggs die in the development process of the aphelinus mali or part of the eriosoma lanigerums parasitized by the aphelinus mali fall preys or are lost, and consequentially the parasitism rate is low is avoided.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology detection, and in particular relates to a method for detecting the parasitism rate and egg laying rate of the apple cotton aphid by sun bees. Background technique [0002] The apple aphid Eriosoma lanigerum (Hausmann) is an important pest of the genus Apple. It has strong reproductive ability. Because it has a layer of cotton wool on its body, it will cause little damage when spraying liquid medicine. Ordinary chemicals cannot kill it at all. it. In recent years, the harm in our country has become increasingly serious, and there is a tendency to further expand and spread. The apple aphid mainly parasitizes the adults and nymphs on apple trees throughout the year, and often forms tumor-like galls on the branches and roots of apples, directly affecting the growth and development of apple trees, the differentiation of flower buds, and the deterioration of fruit quality; Gradually widening,...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2565/125
Inventor 周洪旭鲁莎谭秀梅苏敏
Owner QINGDAO AGRI UNIV
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