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Molecular marker in close linkage with transverse-diameter QTL (quantitative trait loci) of kiwifruit fruits and application of molecular marker

A technology for kiwifruit and fruit, applied in the fields of molecular biology and genetics and breeding, can solve the problem that molecular marker research has not yet been carried out, and achieve the effects of convenient and fast detection method, clear location, and improved selection efficiency.

Active Publication Date: 2017-08-11
WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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AI Technical Summary

Problems solved by technology

[0004] At present, the QTL mapping of the important quality traits of kiwifruit fruit diameter and the discovery of molecular markers related to fruit diameter have not yet been carried out.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Acquisition of the main effect QTL and molecular markers linked to the development of kiwi fruit transverse diameter:

[0023] ① In this example, the cross-breeding of Actinidia 'MT570001' with a small transverse diameter of the fruit and Actinidia sinensis 'Guihai No. 4' with a large transverse diameter was used to construct F 1 Separate groups. In the 6th year of fruiting of the population, select 150 F 1 The individual plants of the offspring of the population were the research objects.

[0024] ②A total of 150 individual plant leaves were collected from the population, and the total DNA was extracted by the CTAB method. According to the RADseq method, the libraries of Chinese kiwifruit 'Guihai 4', Shanli kiwifruit 'MT570001' and 150 offspring were constructed and sequenced.

[0025] ③ Filter the sequencing data to remove sequences with low-quality base content greater than 50%, and remove sequences with sequencing adapter contamination and a large number of repet...

Embodiment 2

[0035] The application of a molecular marker closely linked to the QTL locus of the transverse diameter trait of kiwifruit in kiwifruit breeding, the steps are:

[0036] (1) F 1 Generation, select F 1 60 individual plants in the generation population were the research objects.

[0037] (2) At the ripening stage of the fruit, collect F 1 The fruit of 60 single plants of the generation group was collected, and 40 fruits were collected from each plant, and the transverse diameter of the fruit was measured with a vernier caliper.

[0038] (3) Extract F 1 Generation of total DNA of 60 individual plants, Premix Ex Taq TM II (Tli RNaseH Plus), ROXplus kit instruction, design 20uL reaction system: 5ng·uL -1 Kiwifruit genomic DNA template 2ul, 2×SYBR Premix 10ul, 50×ROX Dye II 0.4ul, 10uM TD-F and TD-R each 0.8ul, double distilled water to make up the rest. The amplification program was: pre-denaturation at 95°C for 5 s, annealing at 60°C for 1 min; denaturation at 95°C for 5 s...

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PUM

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Abstract

The invention discloses a molecular marker in close linkage with transverse-diameter QTL (quantitative trait loci) of kiwifruit fruits and application of the molecular marker. Primers designed according to the molecular marker are TD-F:5'-GATTGCCCCTTTGCTCTATGA-3' and TD-R:5'-CATGGTCTTGTGCCACCAAT-3. The molecular marker and the application have the advantages that the major effective QTL for controlling the transverse diameters of the fruits are positioned in kiwifruits for the first time, 14.61% of phenotypic variance can be explained, the molecular marker related to transverse-diameter traits of the fruits is detected, and accordingly the transverse diameters of the kiwifruit fruits can be predicted and further can be accurately and quickly screened.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and genetic breeding, and more specifically relates to a molecular marker closely linked with a QTL site for the transverse diameter trait of kiwi fruit and its application. Background technique [0002] Kiwi fruit is rich in vitamin C, dietary fiber and various mineral nutrients, and is a healthy fruit favored by consumers. China is the original origin center of the genus Actinidia. Actinidia is extremely rich in resources, with high morphological and genetic diversity. China has about 96% of the total resources of Actinidia species in the world. Relying on abundant natural resources, breeding new varieties with excellent traits plays a key role in the healthy and stable development of the kiwifruit industry. Fruit size is one of the important factors affecting fruit quality. The transverse diameter is an important index to measure fruit size, which directly affects fruit size, and the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 张琼黄宏文钟彩虹韩飞刘小莉
Owner WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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