A molecular marker closely linked to kiwi fruit soluble solids trait qtl locus and its application
A kiwifruit, soluble technology, applied in the field of molecular biology and genetic breeding, can solve the problems of molecular marker research that has not yet been carried out, and achieve the effects of convenient and fast detection methods, accurate and fast screening, and saving production costs
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Embodiment 1
[0022] Acquisition of molecular markers linked to main effect QTL and development of kiwifruit soluble solids:
[0023] ① In this example, the kiwifruit 'MT570001' with a small soluble solid content and the Chinese kiwifruit 'Guihai No. 4' with a large soluble solid content were used to construct F 1 Separate groups. In the 6th year of the group's results, 125 F 1 The individual plants of the offspring of the population were the research objects.
[0024] ②A total of 125 individual plant leaves were collected from the population, and the total DNA was extracted by CTAB method. According to the RADseq method, the libraries of Chinese kiwifruit 'Guihai 4', Shanli kiwifruit 'MT570001' and 125 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 repetitions. Use the software SOAP2 to compare the sequenci...
Embodiment 2
[0035] The application of a molecular marker closely linked to the kiwifruit soluble solids trait QTL site in kiwifruit breeding, the steps are:
[0036] (1) F 1 Generation, choose 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 individual plants in the generation group, 20 fruits were randomly collected for each sample, and the soluble solids content was measured using an ATAGO Palette PR-32α digital refractometer.
[0038] (3) Extract F 1 Generation of total DNA of 60 individual plants, according to Premix Ex Taq TM II (Tli RNaseHPlus), ROX plus 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 SSC-F and SSC-R 0.8ul each, 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...
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