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DNA bar code for discriminating aegilops tauschii and aegilops cylindrica and application thereof

A technology of goat grass and goat grass, applied in the field of molecular biology and bioinformatics, can solve the problem of the lack of DNA barcode sequences of goat grass and goat grass, the source of DNA barcode test materials, and no report of goat grass and uploading issues, to achieve the effect of reducing the threat of national food security, good economic and social benefits, and reducing dependence on

Inactive Publication Date: 2017-12-19
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Question 1, the source of DNA barcode test materials
[0011] Question 2: Selection of target genes for plant DNA barcoding
[0012] Question 3: Identification Accuracy of DNA Barcodes
In addition, in the existing GenBank database, there are very few DNA barcode sequences of A. cylindrica and A. cylindrica. There is only one trnH-psbA barcode sequence of A. cylindrica. There is no report or upload of this sequence of A. cylindrical
[0013] Question 4, Data Analysis of DNA Barcode Sequences
The principles of data processing of these three methods are different, so the accuracy and scope of application of the discrimination are different.

Method used

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  • DNA bar code for discriminating aegilops tauschii and aegilops cylindrica and application thereof
  • DNA bar code for discriminating aegilops tauschii and aegilops cylindrica and application thereof
  • DNA bar code for discriminating aegilops tauschii and aegilops cylindrica and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1 Differentiate the DNA barcodes of A. cylindrica and A. cylindrica to obtain

[0052] The core DNA barcode maturation enzyme K coding gene (matK) and ribulose-1,5-bisphosphorylation / oxygenase recommended by the present invention for land plants recommended by CBOL Plant Working Group (CBOL Plant Working Group, 2009) The large subunit encoding gene (rbcL), the auxiliary barcoded nuclear gene (ITS) and the chloroplast intergenic region sequence (trnH-psbA) were screened and verified as follows:

[0053] Experimental Materials:

[0054] Select A. japonica HE-4, HA-8, SN-23, A. cylindrica HE-29, and HE-30 as template DNA. The sample information is shown in Table 1. For the four fragments (matK, rbcL, trnH-psbA and ITS ) primers for PCR amplification and sequencing. The primer information of each fragment is shown in Table 2.

[0055] Table 1 Sample source

[0056]

[0057] Note: HE: Hebei Province; HA: Henan Province; SN: Shaanxi; GB: GenBank

[0058] Ta...

Embodiment 2

[0082] Example 2 Large sample size verification DNA barcode primer trnH-psbA (619) identification accuracy

[0083] Integrate the seeds of A. japonica and A. cylindrica collected by the Weeds Group of the Institute of Plant Protection, Chinese Academy of Agricultural Sciences. The collection time was from 2007 to 2016, and they were stored in the low-temperature seed bank. After a detailed comparison of species characteristics, the species name of each material was determined by weed morphology identification experts. Including 27 parts of Aegilops tauschii and 3 parts of Aegilops cylindrica, see figure 2 : spikelet (A), seedling (B), inflorescence (C) and field hazard map (D). In addition, 2 sequences of the same species (A. japonica) and a related species (A. tachycarpa) were downloaded from GenBank, as shown in Table 4.

[0084] Table 4 Sources of Samples for Verification of Identification Accuracy

[0085]

[0086]

[0087] Note: HE: Hebei Province; HA: Henan Pro...

Embodiment 3

[0102] Example 3 Specificity and Sensitivity Analysis of PCR Primer trnH-psbA (619)

[0103] Sample source:

[0104] The gramineous plants used in this example include wheat and common weeds in wheat fields brome, bluegrass, goosegrass, wild oats, poisonous wheat, chrysanthemum, syringa, Japanese scorpion, and non-wheat field common gramineous Weeds Foxtail, Sansevieria, crabgrass, goosegrass, barnyardgrass (Table 5). Among them, numbers 1 and 2 are controls. The seeds and specimens were stored in the herbarium of the Weeds Group of the Institute of Plant Protection, Chinese Academy of Agricultural Sciences; the DNA used for the test was stored in the ultra-low temperature refrigerator of this group.

[0105] Table 5 Sample sources for PCR-specific detection

[0106]

[0107] DNA extraction:

[0108] The plant seeds (No. 3-16) that will be used for the specific detection test are sown and cultivated in an incubator, and the young leaves when getting 2-3 leaf ages are ex...

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Abstract

The invention provides a DNA bar code for discriminating aegilops tauschii and aegilops cylindrica and an application thereof. A nucleotide sequence of DNA bar code is shown in as SEQ ID NO: 1-2. By designing a pair of novel DNA bar code primers, an optimal identification sequence is obtained, the DNA bar code sequences of aegilops tauschii and aegilops cylindrica in a Genbank database are improved, aegilops tauschii, aegilops cylindrica and aegilops umbellulata can be distinguished by only one pair of the primers, and the unknown aegilops tauschii and aegilops cylindrica can be rapidly identified. Compared with a form characteristic identification method at a growth anaphase, the DNA bar code breaks up the restriction of an identification period, greatly increases the identification efficiency, increases the detection speed and accuracy, is in favor of guidance for timely controlling two wheat field worst weeds for an agriculture production mechanism and peasants, ensures the production safety of wheat and peasant income, reduces the threat to national food security, and has good economic benefit and social benefit.

Description

technical field [0001] The invention relates to the fields of molecular biology and bioinformatics, in particular to a DNA barcode for identifying A. cylindrica and A. cylindrica and application thereof. Background technique [0002] Our country has included the jellygrass and goat grass (goat grass) in the list of quarantine pests that are prohibited from entering the country. Our country's inspection and quarantine department often intercepts Azalea and Goatweed during the quarantine of imported grains. These two quarantine weeds are very similar in plant and seed morphology, and it is difficult to accurately identify and identify them. [0003] Aegilops tauschii Coss. is a vicious weed in the world, originated in Eastern Europe, West Asia, and the Middle East, and is an annual or annual herb. It is a dangerous weed that enters the country. Its distribution was first discovered in the central part of the Yellow River and the banks of the Ili River in Xinjiang. With the in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11G06F19/18G06F19/24
CPCC12Q1/686C12Q1/6895C12Q2600/156G16B20/00G16B40/00C12Q2563/185
Inventor 姜翠兰魏守辉黄红娟张朝贤黄兆峰陈景超王慧敏赵丹丹
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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