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A Molecular Marker Method for Rapid Identification of Tea Germplasm with Early Bud and Leaf Growth

A germplasm and tea tree technology, which is applied in biochemical equipment and methods, microbiological measurement/inspection, DNA/RNA fragments, etc., can solve the problems of long identification period, complicated identification procedures, large manpower, material and financial resources, etc., to achieve Rapid identification, high identification accuracy, and the effect of reducing investment

Active Publication Date: 2022-03-22
TEA RES INST OF FUJIAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The direct identification method has the disadvantages of long identification period, complex identification procedures, and a large amount of manpower, material and financial resources.

Method used

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  • A Molecular Marker Method for Rapid Identification of Tea Germplasm with Early Bud and Leaf Growth
  • A Molecular Marker Method for Rapid Identification of Tea Germplasm with Early Bud and Leaf Growth
  • A Molecular Marker Method for Rapid Identification of Tea Germplasm with Early Bud and Leaf Growth

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1, the judgment of new tea variety Fuyun No. 6

[0038] 1. Genomic DNA extraction of tea tree germplasm leaves

[0039] 1) Add 3mL of 1.5%CTAB (1.5%CTAB, 0.1M Tris, 20 mmol EDTA, 1.4M NaCl, pH8.0) solution into the mortar, take 2.0g sample into the mortar and grind evenly, draw 1.0mL Transfer the grinding solution into a 2.0mL centrifuge tube, place in a 65°C water bath for 1 hour, and mix gently once every 15 minutes.

[0040] 2) After cooling, add 600uL of chloroform:isoamyl alcohol (24:1), mix well, centrifuge at 12000r / min for 15min.

[0041] 3) Aspirate the supernatant, transfer to a new 1.5mL centrifuge tube, repeat step 2) once.

[0042] 4) Aspirate the supernatant, transfer to a new centrifuge tube, add 1 / 3 volume of NaAc (3mol / L) and 1mL -20°C pre-cooled anhydrous ethanol, mix well and place at -20°C for 30min.

[0043] 5) 12000 r / min, 10min, discard the supernatant.

[0044] 6) Add 75% ethanol to wash twice, place it on absorbent paper and dry i...

Embodiment 2

[0059] Embodiment 2, the judgment of new tea tree variety Chuntaoxiang

[0060] 1. Genomic DNA extraction of tea tree germplasm leaves

[0061] 1) Add 3mL of 1.5%CTAB (1.5%CTAB, 0.1M Tris, 20 mmolEDTA, 1.4M NaCl, pH8.0) solution into the mortar, take 2.0g of the sample and grind it evenly in the mortar, draw 1.0mL for grinding The solution was transferred into a 2.0mL centrifuge tube, placed in a 65°C water bath for 1 hour, and mixed gently once every 15 minutes.

[0062] 2) After cooling, add 600uL of chloroform:isoamyl alcohol (24:1), mix well, centrifuge at 12000r / min for 15min.

[0063] 3) Aspirate the supernatant, transfer to a new 1.5mL centrifuge tube, repeat step 2) once.

[0064] 4) Aspirate the supernatant, transfer to a new centrifuge tube, add 1 / 3 volume of NaAc (3mol / L) and 1mL -20°C pre-cooled anhydrous ethanol, mix well and place at -20°C for 30min.

[0065] 5) 12000 r / min, 10min, discard the supernatant.

[0066] 6) Add 75% ethanol to wash twice, place it o...

Embodiment 3

[0081] Embodiment 3, the determination of tea tree new variety Purple Guanyin

[0082] 1. Genomic DNA extraction of tea tree germplasm leaves

[0083] 1) Add 3mL of 1.5%CTAB (1.5%CTAB, 0.1M Tris, 20 mmolEDTA, 1.4M NaCl, pH8.0) solution into the mortar, take 2.0g of the sample and grind it evenly in the mortar, draw 1.0mL for grinding The solution was transferred into a 2.0mL centrifuge tube, placed in a 65°C water bath for 1 hour, and mixed gently once every 15 minutes.

[0084] 2) After cooling, add 600uL of chloroform:isoamyl alcohol (24:1), mix well, centrifuge at 12000r / min for 15min.

[0085] 3) Aspirate the supernatant, transfer to a new 1.5mL centrifuge tube, repeat step 2) once.

[0086] 4) Aspirate the supernatant, transfer to a new centrifuge tube, add 1 / 3 volume of NaAc (3mol / L) and 1mL -20°C pre-cooled anhydrous ethanol, mix well and place at -20°C for 30min.

[0087] 5) 12000 r / min, 10min, discard the supernatant.

[0088] 6) Add 75% ethanol to wash twice, pla...

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Abstract

The invention belongs to the field of identification and utilization of tea tree germplasm resources and new variety selection and breeding, and specifically relates to a molecular marker method for quickly identifying tea tree germplasm with early bud and leaf growth. A pair of dCAPS marker primers are designed by using the DNA sequence near the SNP marker. Different tea tree germplasm DNA can be screened out by using primers for PCR amplification, electrophoresis separation and genotype analysis, which realizes the molecular marker-assisted breeding of tea tree varieties with early bud and leaf growth.

Description

technical field [0001] The invention belongs to the field of identification and utilization of tea tree germplasm resources and new variety selection; in particular, it relates to the rapid identification of tea tree germplasm with early bud and leaf growth by using molecular markers. Background technique [0002] Tea tree is a foliage crop, and the germination period (or mining period) of its buds and leaves is one of the important economic traits of tea tree. The temperature in early spring is low, pests and diseases basically do not occur, no chemical pesticides are used in tea gardens, and the overwintering buds of tea trees accumulate a large amount of organic compounds. Therefore, "first spring tea" is of the highest quality and often has a higher price, and the sooner it is released, the better the price . The germination period of tea tree buds and leaves is related to the growth environment, cultivation management measures and varieties, among which the variety fac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2531/113C12Q2521/313C12Q2565/125
Inventor 王让剑高香凤杨军孔祥瑞
Owner TEA RES INST OF FUJIAN ACADEMY OF AGRI SCI
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