Application of loc663462 gene in pest control
A technology for preventing and controlling pests and genes, which is applied in the field of genetic engineering, can solve problems such as inedibility, discoloration and odor, and moldy odor of victims, so as to reduce the body length and width, the number of eggs laid and the hatching rate Effect
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Embodiment 1
[0039] Example 1 The expression of the LOC663462 gene in different periods of the red beetle
[0040] The expression of the LOC663462 gene was detected in the whole worm of Trichobolus chinensis by quantitative PCR, and the quantitative primers were:
[0041] F': GCAGTTCAAAGCGCAAAGACA (SEQ ID NO: 4),
[0042] R': TTTCACCGGTTGAGAGACGG (SEQ ID NO: 5);
[0043] The internal reference gene is RP3, and the primer sequences are:
[0044] F': TCAAATTGATCGGAGGTTTG (SEQ ID NO: 6),
[0045] R': GTCCCACGGCAACATAATCT (SEQ ID NO: 7);
[0046] The detection period includes the 1st day of the egg stage (EggD1), the 4th day of the egg stage (EggD4), the 18th-20th day of the larval stage (L-D18~20), the 1st-5th day of the pupal stage (P1~5), 0.5-1 hour after adult eclosion (Ad 0.5-1h), 1-6 days after adult eclosion (Ad1-6).
[0047] Such asfigure 1 Shown is the expression graph of the LOC663462 gene in different periods of Tribetium chinensis. It can be seen that the expression level of t...
Embodiment 2
[0048] Example 2 Synthesis of LOC663462 gene dsRNA and its interference efficiency
[0049] The primers for synthesizing dsRNA are:
[0050] F': TAATACGACTCACTATAGGGATGGATACGATTGTGCAAGG (SEQ ID NO: 2),
[0051] R': TAATACGACTCACTATAGGGCCGTTTCCTGGTCAGTTAT (SEQ ID NO: 3);
[0052] Such as figure 2 Shown is the electropherogram of the synthesized dsRNA; from left to right, the first lane is the marker, the second lane is the DNA template used for the synthesis of dsRNA, and the third lane is the synthetic dsRNA, where the dsRNA is 500bp.
[0053] Using GFP dsRNA as a control, the primers for synthesizing GFP dsRNA are:
[0054] F': TAATACGACTCACTATAGGGTGGTCCCAATTCTCGTGGAAC (SEQ ID NO: 8),
[0055] R': TAATACGACTCACTATAGGGCTTGAAGTTGACCTTGATGCC (SEQ ID NO: 9);
[0056] According to the expression of the LOC663462 gene in different stages of Tribulus chinensis, the dsRNA injection was performed on the second day of the pupal stage. 200ng of dsRNA was injected into each of the...
Embodiment 3
[0058] Example 3 Effect of dsRNA of LOC663462 gene on the phenotype of Tribulus chinensis
[0059] Pupae emerged into adults 96 hours after dsRNA injection.
[0060] Observe the adults 1h after eclosion, such as Figure 4 As shown, no significant difference in appearance was found from the control group.
[0061] Observe the adults 1 day after eclosion, such as Figure 5 As shown, whether it is female or male, the appearance of the worms injected with LOC663462dsRNA is not significantly different from that of the control group, but the body size is slightly smaller than that of the control group.
[0062] The body length and body width of adults 1 day after eclosion are counted, and the results are as follows: Figure 6 As shown, it can be seen that among the males, the body size of wild-type (WT) and GFP dsRNA (dsGFP) injected is relatively similar, and the body length and body width of adults injected with LOC663462 dsRNA (dsLOC663462) are smaller than those injected with...
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