A Molecular Marker Tightly Linked to the Qtl Locus of the Transverse Diameter Trait of Kiwifruit Fruit and Its Application
A technology for kiwi fruit and fruit, which is applied in the fields of molecular biology and genetic breeding, can solve the problems that molecular marker research has not yet been carried out, and achieve the effects of convenient and fast detection methods, saving production costs, and low selection efficiency
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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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