Method for identifying silkworm silk color in larval stage

Abstract

The invention belongs to the field of biotechnology, and discloses a method for identifying a silkworm silk color in a larval stage. The method provided by the invention comprises the following steps of: extracting the ribonucleic acid (RNA) of a silkworm silk gland of a 5th instar stage to be detected by utilizing a genetic engineering technology; performing reverse transcription to synthesize complementary deoxyribonucleic acid (cDNA); taking the obtained cDNA as a template, performing reverse transcription-polymerase chain reaction (RT-PCR) amplification reaction by using a specific primer; judging whether a silkworm spins yellow silk or green silk or white silk by detecting the type and the length of a PCR amplification product by agarose electrophoresis; and further judging whether the silkworm which spins the yellow silk is a homozygous yellow silk spinning silkworm or a heterozygous yellow silk spinning silkworm, and other silkworms spin green silk or white silk by analyzing the content of the RT-PCR amplification product relative to an electrophoresis gel product. By the method provided by the invention, the pedigree with consistent internal cocoon color and external cocoon color and high color fastness also can be identified and separated from a mixed group with different cocoon layer colors and chromaticities. The identification method provided by the invention is simple, convenient and rapid to operate; and the accuracy for identifying the color of the silkworms is high.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for identifying the silk color of silkworm silkworm, specifically using the specific sequence of mRNA transcribed from the Cbp gene and the Cbp-like gene of silkworm, to specifically identify the silk silk color of silkworm, and at the same time distinguish the silkworm silk The silkworm with yellow silk and the silkworm with part of yellow silk, the silkworm with homozygous yellow silk and the silkworm with heterozygous yellow silk, and the difference in chromaticity and color fastness of yellow silk. Background technique [0002] Silkworm (Bombyx mori), also known as silkworm, is an insect that is industrially used to produce silk protein fibers in large quantities. Silk is a continuous long fiber that is solidified by secreting silk liquid when cooked silkworms make cocoons, also known as cocoon silk. The modern cocoon and silk industry mainly uses the white ...

AI Technical Summary

Problems solved by technology

[0008] At present, domestic and foreign silkworm colored cocoon breeding methods are based on cocoon color evaluation and selection of families and individuals. However, due to the aforementioned yellow and red cocoon colors, homozygous and heterozygous families cannot be judged. Consistency and high color fastness The selection of families and individuals can only be achieved through cocoon color measurement and cocoon silk color fastness testing, resulting in a lack of accuracy in the selection of silkworm varieties with yellow and red cocoons, and it is difficult to breed strains Stable cocoon color, long breeding cycle, high cost and poor effect

Although theoretically by examining the mRNA structure of the Cbp gene and Cbp-like gene transcription, or detecting the CBP protein product, the carotenoid-binding protein genotype of the individual can be confirmed, but the Cbp gene and Cbp-like gene of the silkworm are mainly found in the 5th instar larvae. Silk gland and digestive tract transcription and expression, identification of Cbp gene and Cbp-like gene transcription and expression product types need to kill larvae to take out silk gland or digestive tract tissue, it is impossible to achieve both identification of gene type and seed retention, and it is also impossible to extract from silkworm cocoons The identification and separation of the mixed populations of yellow-red cocoon systems with different cocoon layer colors and different chromaticity can not meet the needs of natural color cocoon variety breeding.

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