Calabash gourd anti-plague gene segment or gene marker and application

A gene fragment and gene marker technology, applied in the field of plant biotechnology science, can solve problems affecting the growth and development process of gourd plants, economic losses, etc., and achieve the effects of improving breeding efficiency, shortening breeding years, and shortening excavation cycle.

Inactive Publication Date: 2012-10-03
常熟市支塘镇新盛技术咨询服务有限公司
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Problems solved by technology

The disease has seriously affected the normal growth and development

Method used

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  • Calabash gourd anti-plague gene segment or gene marker and application
  • Calabash gourd anti-plague gene segment or gene marker and application
  • Calabash gourd anti-plague gene segment or gene marker and application

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Embodiment Construction

[0017] The identification of disease-resistant genes plays an important role in the theoretical research of crop disease-resistant genetics and the selection of disease-resistant varieties. The method can quickly identify the powdery mildew gene of japonicus japonicus. The specific implementation process is as follows:

[0018] 1. DNA extraction: take an appropriate amount of young gourd leaves, dry them with silica gel, grind them into powder, and extract the total genomic DNA by CTAB method. The extracted DNA was detected by an ultraviolet spectrophotometer, and samples with an OD260 / OD280 value of 1.8-2.0 were selected and stored at -20°C for future use.

[0019] 2. Amplification and cloning of the homologous fragment of the disease resistance gene: according to the amino acid sequence GG(L / I / V / M / S)GKTT and G(L / N) of the conserved region of the known NBS-LRR type disease resistance gene ) PL(A / T / S)(L / V), designed a pair of degenerate primers: NBS1 5'-GKTT GGIGGIWSIGGIAARA...

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Abstract

The invention mainly relates to a calabash gourd anti-plague gene segment or gene marker and application. Three calabash gourd anti-plague gene segments or gene markers are rapidly separated by adopting an anti-disease gene homologous sequence method, and by sequencing, the nucleotide lengths of the gene segments or gene markers are 510bp, 498bp and 501bp. According to the calabash gourd anti-plague gene segment or gene marker and application provided by the invention, the excavation period of the calabash gourd anti-plague gene is effectively shortened, and the rapid identification of plague gene is facilitated; the identified plague gene can be used for developing corresponding coseparation functional markers (SNP (single nucleotide polymorphism), SCAR (sequence-characterized amplified region) and the like), and can also be rapidly used for assisted selection of a molecule marker of the anti-plague gene, and the accuracy is high; development of multi-resistance breeding materials can be carried out by combining with other anti-disease gene molecule markers, the breeding time is shortened and the breeding efficiency is improved; and foundation is established for expounding the molecular mechanism of the anti-plague gene.

Description

technical field [0001] The present invention clones gourd anti-epidemic disease gene fragments or transforms them into molecular markers by means of the homologous sequence method of disease-resistant genes. , the cloning of disease-resistant genes and the analysis of the regulatory mechanism of disease-resistant genes belong to the field of plant biotechnology science. Background technique [0002] Blight is one of the most important diseases in gourd production. Mainly damage the stems, leaves and fruits of gourds. Infected stems: Dark green water-soaked spots often appear at the base of the stem or tender stem nodes, and then become soft and constricted, causing the leaves above the diseased part to wilt or die. Leaf infection: large round to irregular water-soaked spots are produced, the leaves are soft and sagging, and they are bluish white to light brown thin paper in dry conditions, easy to break. Fruit infection: water-soaked dark green spots are produced, gradual...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 钱孝英
Owner 常熟市支塘镇新盛技术咨询服务有限公司
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