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InDel molecular marker for identifying pepper maturity and application thereof

A technology of molecular markers and hot peppers, which is applied in recombinant DNA technology, microbial detection/testing, DNA/RNA fragments, etc., can solve the problems of low accuracy of phenotype selection, large limitations in use, and long genetic distance of target genes, etc. problems, to achieve the effect of speeding up the breeding process, high accuracy, and stable amplification

Active Publication Date: 2018-06-15
VEGETABLE & FLOWER INST JIANGXI ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In natural variation, the flowering node position of pepper is controlled by a few major genes and some minor genes, and is also affected by the environment such as temperature and light. It is a quantitative trait, and the accuracy of traditional phenotypic selection is not high.
Most of the currently available molecular markers for flowering initiation in pepper are far away from the target gene, or have relatively limited use

Method used

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  • InDel molecular marker for identifying pepper maturity and application thereof
  • InDel molecular marker for identifying pepper maturity and application thereof
  • InDel molecular marker for identifying pepper maturity and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Development of Molecular Marker InDel at the Initiation Node of Capsicum

[0031] 1. Population material construction and genetic analysis

[0032] Extra Early Pepper B 9431 (depending on the planting environment, the first flowering node is 1-4 nodes) as the female parent, late-maturing pepper (depending on the planting environment, the first flowering node is 13-18 nodes)A 145 Crossed for the male parent to get F 1 Generation, F 1 Plants selfed to get F 2 group. Plant this F in spring 2017 2There are 297 plants in the population, and the node position phenotype of flower initiation is measured by the number of true leaves (node ​​positions) between the cotyledon and the first flower on the main stem. The statistical results show that the node position of flower initiation in this population is bimodal, indicating that There are 215 late-maturing plants with high flower-start nodes (≥5) and 82 ultra-early-maturing plants with low flower-start nodes (≤4)...

Embodiment 2

[0037] Example 2 Application of Molecular Marker InDel at the Initiation Node of Capsicum 1

[0038] Extract parent B by CTAB method 9431 , parent A 145 , F 1 Plant (B 9431 ×A 145 ) and 297 F 2 For plant DNA, the DNA concentration was detected with a Nanodrop spectrophotometer, and ddH was added 2 O Adjust all DNA concentrations to 100 ng / μL. Perform PCR according to the following conditions: the total reaction volume is 10 μL, including 5 μL of Kangwei Century Biotechnology Co., Ltd. 2×Es Taq MasterMix (Dye) (CW0690), forward primer FFN1-5UF5 (10 μM) and reverse primer FFN1-e1R1 ( 10 μM) 0.08 μL each, reverse primer FFN1-PA-R (10 μM) 0.4 μL, 100 ng / μL DNA template 0.6 μL, ddH 2 O 3.84 μL. PCR reaction program: pre-denaturation at 95°C for 2min; denaturation at 95°C for 30s, annealing at 56°C for 30s, extension at 72°C for 40s, 38 cycles; extension at 72°C for 10min. Take 3 μL of the PCR reaction product for electrophoresis detection, see image 3 . It was found tha...

Embodiment 3

[0039] Example 3 Application of Molecular Marker InDel at the Initiation Node of Capsicum Pepper 2

[0040] Extra Early Pepper B 9431 (first flower nodes 1-4 nodes) and late-maturing pepper B 025 F 1 Generation, F 1 Plants selfed to get F 2 group. Plant this F in spring 2017 2 Population plants, randomly select wherein 61 strains extract DNA, press the same condition in embodiment 2 to parent, F 1 and F 2 The plants were subjected to PCR, and 3 μL of PCR reaction products were taken for electrophoresis detection, and the electrophoresis results of some amplified products were shown in Figure 4 . The results showed that the genotype identified by this marker was consistent with the plant phenotype, that is, the band type was the same as that of the extra precocious parent B 9431 Consistent with the ultra-early-maturing peppers at the first flower node ≤ 4, the belt type is the same as that of the late-maturing parent A 025 or F 1 The relatively late-maturing peppers...

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Abstract

The invention provides an InDel molecular marker for identifying pepper maturity and application thereof. The InDel molecular marker is characterized in that a F2 group is built based on a very early-maturing hot pepper self-line B9431 and a late-maturing self-line A145; a chromosome site at the node of a first flower is rapidly identified and controlled by using a BSR-Seq (Bulked Segregant RNA-Seq) technology; early-maturing gene hot pepper at the nodes 1 to 4 of the first flower and late-maturing gene hot pepper at the nodes great than 4 of the first flower can be accurately and rapidly screened specific to a specific InDel developing molecular marker (SEQ ID NO:4) on the site, auxiliary selection of the hot pepper maturing seedling molecular marker is realized, and the breeding selection efficiency is improved; the InDel molecular marker has an important application value in the hot pepper maturing breeding.

Description

technical field [0001] The invention relates to a molecular marker detection technology, in particular to an InDel molecular marker for identifying the ripeness of pepper and its application. Background technique [0002] Pepper (Capsicum spp.) is an important economic crop of Solanaceae, which is widely planted all over the world. China is an important producer of pepper, and pepper is one of the main vegetable crops in my country. The pepper plant is a symaxial branch structure, the main stem grows to a certain point and stops growing, and its top meristem is transformed into an inflorescence meristem. The lower part develops as a solitary flower, and the flowering marks the termination of the growth of the main stem. The lateral meristem grows and develops into a new branch, and soon the new branch stops growing, and the lateral meristem at the top of the new branch continues to replace the apical meristem. Repeat to form symaxial branches. [0003] The insertion node o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895
Inventor 袁欣捷陈学军方荣周坤华
Owner VEGETABLE & FLOWER INST JIANGXI ACADEMY OF AGRI SCI
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