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Specific primer pair, allele and molecular marker for screening thea section with low caffeine trait and screening method

A specific primer pair and allele technology, applied in the biological field, can solve the problems of long time consumption and low efficiency, and achieve the effect of accelerating the pace of breeding

Active Publication Date: 2016-04-20
TEA RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method requires a certain amount of tea tree leaves, and the identified plants need to grow up to 3-4 years before they can be identified, which takes too long and is inefficient

Method used

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  • Specific primer pair, allele and molecular marker for screening thea section with low caffeine trait and screening method
  • Specific primer pair, allele and molecular marker for screening thea section with low caffeine trait and screening method
  • Specific primer pair, allele and molecular marker for screening thea section with low caffeine trait and screening method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Discovery of TCS1 allele differential sequence, primer pair and development of functional marker InDel-TCS1:

[0039] 1. Test materials

[0040] Select 5 parts of tea (C.sinensisvar.sinensis), 2 parts of Assam tea (C.sinensisvar.assamica), 3 parts of white hair tea (C.sinensisvar.pubilimba), 2 parts of thick shaft tea (C.crassicolumna), 1 15 tea group plant resources with large genetic background differences, including one Dali tea (C.taliensis), one Dachang tea (C.tachangensis) and one unclassified wild tea tree (Camellia sp.), were studied. The specific names are shown in the table 1.

[0041] Table 1 The 15 tea group plant resources studied in this experiment

[0042]

[0043]

[0044] 2. Genomic DNA extraction:

[0045] Take 1g of fresh young shoots, add liquid nitrogen and grind to powder. Put 0.2g of the powder into a 1.5mL centrifuge tube, add 700 μL CTAB extract, mix well, put in a water bath at 65°C for 1 hour, and stir once every 20 minutes. Add an ...

Embodiment 2

[0056]Genotype Analysis of Tea Plant Resources by Functional Marker InDel-TCS1

[0057] 1. Test materials

[0058] The materials studied in this experiment are listed in Table 2, including 10 decaffeinated (caffeine content less than 10 mg / g) and 17 conventional caffeine resources (caffeine content greater than 10 mg / g).

[0059] Table 2 The 10 low caffeine and 17 conventional caffeine resources studied in this experiment

[0060]

[0061]

[0062] 2. Identification of caffeine content in 27 materials

[0063] Caffeine identification method high-performance liquid chromatography is the same as described in Example 1. The results showed that 10 were decaffeinated resources and 17 were regular resources.

[0064] 3. Genotype detection of different tea group plant resources by functional markers

[0065] DNA extraction, PCR amplification system and procedures are the same as in Example 1. Amplified products were separated by 2% agarose gel electrophoresis. Such as f...

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Abstract

The present invention belongs to the technical field of biology, and relates to a specific primer pair, an allele and a molecular marker for screening thea section with low caffeine trait and a screening method. The present invention discloses allelic variation TCS1a, TCS1b, TCS1c, TCS1d, TCS1e and TCS1f sequence fragments of thea section TCS1 gene, and provides a functional marker for identification of low caffeine thea section, and the functional marker is used for molecular marker-assisted selection for quickly screening a low caffeine thea section material so as to speed up the pace of breeding of low caffeine tea tree varieties. The molecular marker-assisted selection of the low caffeine tea tree varieties has important theoretical significance and economic value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an allele, a molecular marker and a screening method for screening tea group plants with low caffeine properties. Background technique [0002] Tea tree originated in Yunnan Province of my country, and belongs to Theaceae (Theaceae), Camellia (Camellia L.), and Tea Group (Sect. Thea (L.) Dyer) in plant classification. Because of its rich content, tea is popular all over the world and is one of the three major non-alcoholic beverages in the world. At present, there are more than 50 countries in the world to cultivate tea trees, mainly concentrated in China, India, Sri Lanka, Japan and Kenya in Asia and other countries in Africa. [0003] Caffeine is an important flavor substance and physiologically active characteristic component of tea, and its content is generally 2% to 5%. Purine alkaloids such as caffeine use xanthosine as a substrate and S-adenosylmethionine (SAM) as a methyl don...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12Q1/68C12N15/11
Inventor 金基强陈亮姚明哲马春雷马建强
Owner TEA RES INST CHINESE ACAD OF AGRI SCI
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