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Molecular marker linked to green retardation gene of pepper and application thereof

A technology of molecular markers and green-sustaining genes, which is applied in the fields of capsicum breeding and molecular biology, can solve the problems of unsatisfactory green-keeping effects and high costs, and achieve the goal of reducing planting scale, reducing workload, and improving efficiency and accuracy. Effect

Active Publication Date: 2019-01-15
HUNAN VEGETABLE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In actual production, although there are many ways to keep vegetables green, some methods cost too much, and some methods are restricted by the Food Sanitation Law due to the residue of heavy metals in the processing process, and the green keeping effect is not very ideal.

Method used

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  • Molecular marker linked to green retardation gene of pepper and application thereof
  • Molecular marker linked to green retardation gene of pepper and application thereof
  • Molecular marker linked to green retardation gene of pepper and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Acquisition of the Molecular Marker SGR Molecular Marker Linked to the Capsicum Chlorophyllin Gene

[0045] BSR positioning method

[0046] 1. Group construction

[0047] The material 'CT2' (such as figure 2 shown on the left), and the material 'CS17' whose fruit color is brown is the parent (such as figure 2 Shown on the right side of the middle), all are resequenced pepper germplasm resources, the brown parent and the red parent were crossed to obtain the F1 generation, and the F1 generation was self-crossed to obtain the F2 population.

[0048] 2. Fruit color identification

[0049] After the fruit is ripe, the brown / red color of the fruit is identified.

[0050] 3. Fruit color main effect QTL analysis

[0051] F in CT2 x CS17 2 In the group, 30 plants with red fruit color and 30 mature fruits with brown fruit color were selected, and the fruits of each plant were mixed in equal amounts to construct a red fruit / brown fruit RNA mixed pool. The total...

Embodiment 2

[0056] Example 2 Verification of mature capsicum fruit color marking SGR

[0057] Using the molecular marker primers obtained for the color of the fruit, F 2 90 individual plants of the population carry out fruit color identification, and its steps are as follows:

[0058] (1) Carry out touchdown PCR reaction with pepper genomic DNA as a template; add 3 primers labeled with SGR to the reaction system, the sequence of the universal reverse primer labeled with SGR is SGR-1 (SEQ ID NO.1), and the two primers labeled with SGR A forward primer sequence is SGR-2 (SEQ ID NO.5) connected with a fluorescent linker sequence, and SGR-3 (SEQ ID NO.6) connected with a fluorescent linker sequence. Touchdown PCR reaction system: the total volume is 3 μl, and the specific components are as follows: ddH 2 O 1.4983μl, 2×KASPMasterMix 1.4792μl, two forward primers SGR-2 (SEQ ID NO.5) and SGR-3 (SEQ ID NO.6) connected with the fluorescent adapter sequence each 0.005μl (100uM), reverse To prime...

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Abstract

The present invention discloses a molecular marker linked to green retardation gene of hot pepper and its application. Two highly homozygous materials are used to construct the F2 population. The chromosomal regions closely linked to the brown (green-stagnant) gene of pepper fruits are obtained by BSR population mapping, and the candidate genes are identified by molecular markers in the candidateregion. A KASP molecular marker is designed according to the single base mutation of candidate gene. The genotype identification of 90 individual plants in F2 population by using this marker showed that the coincidence rate is 100%. The results not only contribute to the color identification and assisted breeding of pepper fruits, but also provide a basis for map-based cloning of green retardationgene and molecular mechanism of green retardation, which is worth popularizing widely.

Description

technical field [0001] The invention belongs to the fields of capsicum breeding and molecular biology, and relates to a molecular marker linked with a capsicum (Capsicum annuum L) green-sustaining gene and the application of the molecular marker in capsicum fruit color identification and breeding. Background technique [0002] Capsicum, Latin name: Capsicum annuum L., Solanaceae, Capsicum genus annual or limited perennial herb. Capsicum mainly includes 5 cultivars: C.annuum L, C.frutescens L, C.baccacum L, C.pubescensKeep, C.chinese Jacquin. Chili peppers are native to the tropical regions of Central Latin America, and the country of origin is Mexico. In China, it is mainly distributed in Sichuan, Guizhou, Hunan, Yunnan, Shaanxi, Henan (Xichuan County), Jize County of Hebei Province and Tuoketuo County of Inner Mongolia. According to the statistics of the bulk vegetable system of the Ministry of Agriculture, the cultivation area of ​​pepper in my country is increasing cont...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156C12Q2600/158
Inventor 欧立军刘宇华杨博智陈文超张竹青周书栋杨莎李雪峰马艳青戴雄泽邹学校
Owner HUNAN VEGETABLE RES INST
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