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Br-or-Indel molecular maker and application thereof

A DNA molecule, a pair of technology, applied in the field of Br-or-Indel molecular markers, can solve the problems of not being able to distinguish heterozygous and homozygous, poor repeatability, and inability to distinguish heterozygous and homozygous

Inactive Publication Date: 2014-01-29
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Liu Xiucun et al. (2003) found a RAPD molecular marker linked to the orange-red ball color gene, with a genetic distance of 3.8 cM. However, as a dominant marker, the RAPD marker cannot distinguish heterozygous and homozygous types, and is susceptible to Poor repeatability of environmental influences
Wang Qi et al. (2006) obtained a SCOR marker with a genetic distance of 3.7 cM from the orange leaf bulb gene through the screening of molecular markers linked to orange leaf bulb traits in Chinese cabbage. However, this SCOR marker is also a dominant marker and is not very good. The distinction between heterozygous and homozygous
Zhang Fenglan et al. (2008) and Feng Hui et al. (2010) obtained SCAR markers and co-dominant markers through segregation population screening and located the orange-red spherical color gene or between 8cM and 13cM and between 1.3cM ​​and 3.3cM, but the genetic distance still relatively far
Zhang Junxiang et al. (2013) used two Indel molecular markers to locate the or gene within the range of 0.1cM and 0.2cM, but there is still a certain recombination rate

Method used

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  • Br-or-Indel molecular maker and application thereof
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  • Br-or-Indel molecular maker and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1, the acquisition of Br-or-Indel molecular marker

[0057] 1. Artificially synthesize the following primers:

[0058] Br-or-HF: 5'-AATCCCGTAACGTGTGCTGAT-3'; (SEQ ID No.1)

[0059] Br-or-HR: 5'-CCATAGTGCCTGTCACATAAAACC-3'. (SEQ ID No.2)

[0060] 2. Genomic DNA was extracted from the paternal parent Chinese cabbage 91-112 and the female parent orange-red Chinese cabbage T12-9 leaves.

[0061] 3. Using two kinds of Chinese cabbage genomic DNAs as templates and using Br-or-HF and Br-or-HR as primers to carry out PCR amplification.

[0062] 4. Sequence the PCR amplification products, compare the sequence differences of the two PCR products with DNAMAN software, and design the following primers according to the two ends of the difference sequences:

[0063] Br-or-IndelF: 5'-TTGCTCTCGTAAAGACTCTCCCT-3'; (SEQ ID No.3)

[0064] Br-or-IndelR: 5'-ATCTCACGACCAACTGCCTTCA-3'. (SEQ ID No.4)

[0065] 5. Using the genomic DNAs of the two cabbage obtained in step 2 as t...

Embodiment 2

[0071] Embodiment 2, BC 1 Br-or-Indel analysis of segregating populations

[0072] 1. BC 1 Population construction and phenotyping

[0073] F 1 The generation (white heart) is used as the male parent, and it is backcrossed with orange red heart Chinese cabbage T12-9 to produce a backcross generation, which is BC 1 Separate groups. to BC 1 The phenotypes of orange-red hearts and white hearts were identified and verified by Chi-square analysis. The results showed that the segregation of phenotypes (Chinese cabbage heart color) conformed to Mendel's 1:1 segregation rule.

[0074] 2. Adopt the CTAB method to extract the parents (i.e. Chinese cabbage with white heart 91-112, Chinese cabbage with orange red heart T12-9 and Chinese cabbage with white heart 91-112 as the male parent, Chinese cabbage with orange red heart T12-9 as the female parent) f 1 Chinese cabbage) and BC 1 The genomic DNA of the leaves of mature plants was isolated and detected by agarose gel electrophore...

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Abstract

The invention discloses a Br-or-Indel molecular maker and application thereof. A pair of primers disclosed by the invention consists of two primers shown by (1) Br-or-IndelF and (2) Br-or-IndelR, wherein the sequence of Br-or-IndelF is shown in SEQ ID No. 3, and the sequence of Br-or-IndelR is shown in SEQ ID No. 4. The Br-or-Indel molecular maker disclosed by the invention can be applied to the increase of the seed selection efficiency of orange-heart Chinese cabbage.

Description

technical field [0001] The invention relates to a Br-or-Indel molecular marker and its application. Background technique [0002] Lycopene is a natural pigment contained in plants, a type of carotenoid, and one of the strongest antioxidants found in plants in nature. Lycopene is far more effective at scavenging free radicals than other carotenoids and vitamin E. It can not only effectively delay aging, but also has anti-cancer and tumor-suppressing effects, and is also of great significance for strengthening the human immune system. [0003] Orange red heart Chinese cabbage is a characteristic variety of Chinese cabbage, and the inner leaves of the bulb are orange-red. Studies have shown that the orange-red formation of the center of Chinese cabbage is closely related to the accumulation of lycopene. Li Juan (2007) showed that the accumulation of lycopene in orange Chinese cabbage is characterized by a decrease first and then an increase, and no lycopene was detected in o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 王姣于拴仓张凤兰余阳俊张德双赵岫云卢桂香苏同兵汪维红
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES