CN117363768B - Molecular marker and primer for inhibiting development of broccoli and cauliflower leaves and application of molecular marker and primer - Google Patents

Molecular marker and primer for inhibiting development of broccoli and cauliflower leaves and application of molecular marker and primer Download PDF

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CN117363768B
CN117363768B CN202311122605.3A CN202311122605A CN117363768B CN 117363768 B CN117363768 B CN 117363768B CN 202311122605 A CN202311122605 A CN 202311122605A CN 117363768 B CN117363768 B CN 117363768B
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broccoli
cauliflower
primer
molecular marker
plant
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CN117363768A (en
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程锋
张扬勇
李幸
张磊
韩风庆
张亢
吕红豪
唐琪
王勇
季家磊
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Institute of Vegetables and Flowers Chinese Academy of Agricultural Sciences
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    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
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    • C12Q2600/156—Polymorphic or mutational markers

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Abstract

The invention discloses a molecular marker for inhibiting development of broccoli and cauliflower leaves, a primer and application thereof. The invention develops a molecular marker of a structural variation type based on the structural variation site of the 3' -untranslated region of the genes BoCKX of broccoli, cauliflower and other cabbage vegetables, which is a specific structural variation in the broccoli and the cauliflower, and the molecular marker is used for detecting whether the structural variation exists in the cabbage vegetables, so that whether the detection material is the broccoli and the cauliflower can be rapidly identified, and further the molecular marker can be used for assisting in breeding by a biotechnology.

Description

Molecular marker and primer for inhibiting development of broccoli and cauliflower leaves and application of molecular marker and primer
Technical Field
The invention relates to the field of molecular breeding, in particular to a molecular marker for inhibiting development of broccoli and cauliflower leaves, a specific primer and application thereof.
Background
Broccoli (Brassica oleracea var. Botrytis) and broccoli (Brassica oleracea var. Itica) are varieties of cabbage, belong to Brassica (Brassica l.) crops of cruciferae, are rich in nutrients, are rich in dietary fibers, multiple vitamins and minerals such as calcium, phosphorus, iron and the like, and have a health care function. The broccoli is called broccoli, the average nutritive value and disease prevention effect of the broccoli are far superior to those of other vegetables, the broccoli contains anticancer substances of glucosinolate, the broccoli can prevent the growth of early cancer cells, and the broccoli has special health care effect and is one of ten health foods recommended by the U.S. J. The flower ball is a nutrient storage organ of the cauliflower and the broccoli and also is an edible part, and the flower ball is planted at the top end of the short stem and is an important economic property of the cauliflower and the broccoli.
Research on the genetic relationship and genetic diversity of cauliflower and broccoli and related species is of great significance to resource evaluation and creation and new variety cultivation. The molecular marker typing technology has the advantages of no environmental influence, short test period, more selectable markers and capability of performing high-throughput sequencing analysis, and is widely used for research and application of genetic diversity and kindred relation analysis, new variety identification, seed purity and variety authenticity detection, molecular marker assisted breeding and the like.
For example, chinese patent application CN114231656a discloses an SSR core primer set for identifying purity of hybrid of cauliflower, and screening method and application thereof, the SSR core primer set is: boGMS0624, OI11G11, boGMS0941; the alternative core primer group is as follows: boE607, boE761, boGMS, 2431, boGMS, 0808, boGMS1530. The method determines the evaluation index of the core primer suitable for the purity identification of the hybrid seeds of the broccoli and the broccoli by DNA fingerprint analysis of 70 hybrid seeds of the broccoli and the broccoli, and determines a set of primers for the purity identification of the hybrid seeds of the broccoli.
At present, molecular markers which are easier to detect, accurate, stable and reliable for cauliflower and broccoli are still lacking.
Disclosure of Invention
In order to solve at least part of the technical problems in the prior art, the invention provides a molecular marker, a primer group and application for identifying broccoli and cauliflower, thereby providing a theoretical basis for molecular assisted breeding of the broccoli and the cauliflower. Specifically, the present invention includes the following.
In a first aspect of the invention, a molecular marker for inhibiting development of broccoli and/or cauliflower leaves is provided, the molecular marker can be used for identifying broccoli and/or cauliflower at the same time, and the molecular marker is a structural variation molecular marker, and is positioned in a 3' untranslated region of 86bp-401bp downstream of BoCKX genes.
In certain embodiments, the molecular marker according to the invention that inhibits development of broccoli and/or broccoli leaves, wherein the molecular marker has the sequence shown in SEQ ID No. 1.
In a second aspect of the invention, there is provided a primer combination for amplifying the molecular marker of the first aspect.
In certain embodiments, a primer combination according to the present invention comprises a forward primer having the sequence set forth in SEQ ID NO.2 and a reverse primer.
In certain embodiments, the primer combination according to the invention, wherein the reverse primer has the sequence shown in SEQ ID No. 3.
In a third aspect of the invention, there is provided a kit for identifying broccoli and/or cauliflower comprising the above-described primer combination.
In a fourth aspect of the invention, there is provided the use of a molecular marker according to the invention, a primer combination according to the invention, or a kit according to the invention for genetic analysis of broccoli and/or cauliflower.
In certain embodiments, the use according to the invention, wherein the genetic analysis comprises genetic diversity analysis, germplasm identification or assisted breeding.
In a fifth aspect of the invention, there is provided a method for identifying whether a plant comprises broccoli and/or cauliflower, comprising the step of detecting in the DNA of the plant to be identified the presence or absence of a molecular marker of structural variation of the 3' untranslated region located 86bp to 401bp downstream of the BoCKX gene.
In certain embodiments, a method for identifying whether a plant comprises broccoli and/or cauliflower according to the invention comprises the steps of:
(1) Extracting sample DNA from a plant sample to be identified;
(2) Amplifying nucleic acid containing the molecular marker from the sample DNA by PCR using a forward primer having a sequence shown as SEQ ID NO.2 and a reverse primer having a sequence shown as SEQ ID NO. 3; and
(3) The plants were identified by detecting the length of the sample PCR products by electrophoresis.
The SV molecular marker is developed based on the structural variation site on the 3' UTR of the broccoli, the cauliflower and other cabbage vegetables BoCKX genes, is specific structural variation in the broccoli and the cauliflower, and can rapidly identify whether the detection material is the broccoli and the cauliflower by detecting whether the SV molecular marker exists in the cabbage vegetables. In addition, the invention also verifies that the SV molecular marker has correlation with inhibition of broccoli and cauliflower leaf development.
Drawings
FIG. 1 is a schematic representation of broccoli and cauliflower and other brassica vegetables.
FIG. 2 shows the distinguishing effect of broccoli and cauliflower and other materials with SV.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in the present invention, it is understood that the upper and lower limits of the ranges and each intermediate value therebetween are specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
Unless otherwise indicated, the gene position in the present invention refers to a position relative to cabbage T10.
The term "broccoli and cauliflower" as used herein refers to a variety of Brassica (Brassica l.) of the Cruciferae family. The term "variant" means that a species is initially published on an academic journal without a grade under the species (i.e., variant, subspecies, variant), and later as one gets knowledge of the species, it is found that there are individuals or groups of plants within the species that have a variation from the characteristics of the species that were recognized when the species was initially published, and that the variation is obvious and stable, and it is worth separating them individually to show the difference, the plant scientist would name the type with the variation and publish it as a variant within the species according to circumstances. In contrast, a type that has the original characteristics and does not undergo significant variation is called the original variant of this variant.
Molecular markers
In a first aspect of the invention, a specific molecular marker is provided, which has a characteristic sequence for distinguishing broccoli and/or cauliflower from other cabbage types, and the SV molecular marker has a close correlation with inhibition of broccoli and cauliflower leaf development. The inhibition of leaf development is intended to include inhibition of leaf development processes of broccoli and cauliflower such that its leaves exhibit a suppressed state, such as the case where the number, shape, etc. of leaves are significantly different from those of other brassica oleracea vegetables.
In the invention, the molecular marker is a structural variation with the length of 316bp, and is positioned in a 3' UTR region at the position 86bp-401bp downstream of broccoli and cauliflower BoCKX genes. In a specific embodiment, the sequence has the nucleotide sequence shown in SEQ ID NO. 1.
The molecular markers ("signature sequences") in the present invention refer to, for example, gene fragments which are present only in broccoli and cauliflower vegetables in all subspecies and varieties of brassica oleracea vegetables. In the present invention, other brassica is intended to include any cruciferous species other than broccoli and cauliflower, preferably any brassica species, examples of which include, but are not limited to, common head cabbage, broccoli, cauliflower, cabbage mustard, kohlrabi, edible kale, brussels sprouts, and the like.
Primer combination
In a second aspect of the invention, there is provided a specific primer combination for amplifying a molecular marker of the invention, comprising a forward primer and a reverse primer. In a specific embodiment, the forward primer has the sequence shown in SEQ ID NO.2 and the reverse primer has the sequence shown in SEQ ID NO. 3.
Kit for detecting a substance in a sample
The invention also provides a kit for simultaneously identifying broccoli and cauliflower, comprising the primer combination of the invention and optionally reagents for polymerase chain reaction. Reagents for the polymerase chain reaction include any of those used in conventional PCR, such as a polymerase, a buffer, and the like.
In addition to the components described above, the kits of the present invention may also include precautions related to regulating manufacturing, use, or marketing of the diagnostic kit in a form prescribed by a government agency. In addition, the kits of the invention may also be provided with detailed instructions for use, storage and troubleshooting. The kit may also optionally be provided in a suitable device, preferably for robotic operation in a high throughput setting.
In certain embodiments, the components of the kits of the invention may be provided as dry powders. When the reagents and/or components are provided as dry powders, the powders may be restored by the addition of a suitable solvent. It is contemplated that the solvent may also be disposed in another container. The container will typically include at least one vial, test tube, flask, bottle, syringe, and/or other container means, with the solvent optionally being placed in aliquots. The kit may further comprise means for a second container comprising a sterile, pharmaceutically acceptable buffer and/or other solvent.
In certain embodiments, the components of the kits of the invention may be provided in solution, e.g., in aqueous solution. Where present in aqueous solution, the concentration or amount of these ingredients can be readily determined by one skilled in the art according to various needs. For example, for storage purposes, the concentration of the components may be present in a higher form, and the concentration may be reduced to the working concentration by, for example, diluting the higher concentration solution when in the working state or in use.
Where more than one component is present in a kit, the kit will also typically contain a second, third or other additional container in which additional components may be placed separately. In addition, combinations of various components may be included in the container. Any combination or reagent described herein may be a component in a kit.
Use of the same
The invention further provides the use of molecular markers, substances, primer combinations or kits capable of detecting said molecular markers in genetic analysis of broccoli and/or cauliflower, including but not limited to genetic diversity analysis, germplasm identification, assisted breeding, and the like. Substances capable of detecting the molecular markers include those polynucleotide molecules capable of complementary pairing with the target molecule, and the like.
Method for identifying whether a plant comprises broccoli and/or cauliflower
The present invention provides a method for identifying whether a plant comprises broccoli and/or cauliflower. Preferably, the plant described herein refers to a plant from the Cruciferae family (Cruciferae), more preferably the plant described herein is a plant from the Brassica genus (Brassica), further preferably the plant described herein is a plant from the Brassica class of vegetables. In certain embodiments, a "plant" herein is a type of plant, where the methods of the invention refer to methods for identifying whether the plant is broccoli and/or cauliflower. In certain embodiments, a "plant" herein is a mixture of multiple types of plants or plant components thereof, where the methods of the invention refer to methods for identifying whether broccoli and/or cauliflower are included in a plant.
The method of the invention comprises the step of detecting in the DNA of the plant to be identified the presence or absence of a characteristic sequence of the BoCKX gene located in broccoli and/or cauliflower. The feature sequences have already been described and will not be described in detail here.
In the present invention, the method of detecting the presence or absence of the signature sequence of the BoCKX gene located in broccoli and/or cauliflower may employ any method known in the art, examples of which include a method of amplifying nucleic acid comprising the signature sequence, a method of directly sequencing a plant genome, and the like.
In certain embodiments, the methods of the invention for identifying whether a plant comprises broccoli and/or cauliflower comprise the step of amplifying a gene sequence comprising a signature sequence from DNA of a plant sample to be identified by PCR using forward and reverse primers.
In an exemplary embodiment, a method for identifying whether a plant comprises broccoli and/or cauliflower comprises the steps of:
(1) Extracting sample DNA from a plant sample to be identified;
(2) Amplifying nucleic acid containing the molecular marker from the sample DNA by PCR using forward primer and reverse primer;
(3) The length of the sample PCR product was checked by electrophoresis to identify whether the plant was broccoli and/or cauliflower.
In step (1), the "plant sample to be identified" refers to the whole plant or a part of the plant, e.g., the plant root, leaf or stem, etc.
In step (2), the ratio of the forward primer to the reverse primer is 1:1.PCR reaction procedure: 85-98deg.C for 1-5min, 85-98deg.C for 10-30s, 40-70deg.C for 10-30s, 50-80deg.C for 10-30s, and 10-40 cycles for 1-10min. Also preferably, the PCR reaction procedure: 90-96 ℃ for 1-3min, 90-96 ℃,12-20s, 50-65 ℃,15-20s, 70-75 ℃ for 10-20s, 10-30 cycles, 65-75 ℃ for 1-5min.
In the step (3), the amplified fragments of broccoli and cauliflower are 600bp in the length of the PCR product of the sample detected by electrophoresis.
Examples
The development of molecular markers of the BoCKX gene 3' untranslated region Structural Variation (SV) associated with inhibition of broccoli and cauliflower leaf development is shown below.
The SV-based patterned flood genome was constructed using high quality genomes of 27 different cabbage varieties, including 2 wild cabbage, 7 common head cabbage, 7 collard, 3 broccoli, 2 kohlrabi, 2 cabbage mustard, 1 brussels sprout, 1 ornamental cabbage, as follows.
Wild cabbage T10 is used as a reference genome, NUCmer (v4.0.0) is utilized for the preparation of the wild cabbageEt al, 2018) to compare other genomes to T10, set parameters: -maxmatch-c100-l50, filtering the comparison results, using delta-filters with the parameters: -m-i90-l100, using show-records for the filtered result conversion format, the parameters being: -T-H-r-d.
SV was identified from the alignment using SyRI (Goel et al., 2019) and pooled for redundancy between different genomes.
The non-redundant SV was combined with the T10 reference genome using VG ToolKit (v1.33.0) (Garrison et al 2018) to construct a patterned broad genome based on SV.
Resequencing 704 parts of a different variety of cabbage material, comprising: 36 parts of wild cabbage, 310 parts of common head cabbage, 153 parts of broccoli, 63 parts of broccoli, 46 parts of kohlrabi, 34 parts of kale, 24 parts of cabbage mustard, 20 parts of brussels sprouts, 18 parts of ornamental cabbage, and the sequencing platform is Illumina HiSeq 2000, and the raw data is filtered by Trimmomatic v 0.39.39 to remove joints, poly-N and low-quality fragments, so that CLEAN DATA is obtained. The SV-based patterned flood genome is indexed in xg and gcsa formats by VG Index, CLEAN DATA is aligned to the patterned flood genome by VG Map, and the low-quality alignment result is filtered by VG Pack, wherein the parameters are as follows: -Q5. The SV genotype of each sample was obtained using VG Call with the parameters: -a-s.
Correlation analysis of SV with cabbage material of different variety types revealed that an SV specific to broccoli and cauliflower was 316bp in length, which was present in 150 parts broccoli and 60 parts cauliflower, which was absent in 3 parts broccoli and 3 parts cauliflower, which was absent in the other 487 parts cabbage material, and which was present in only one broadleaf collard. The SV is located in the 3' UTR region of 86bp-401bp downstream of BoCKX gene.
Extracting genome DNA from seedling of broccoli, cauliflower and common head cabbage, synthesizing SV molecular marker primer, and its sequence is as follows ：CKX3SV-F：TCGTGAAAGATATTACAGAGGAAGTGGAACC(SEQ ID NO.2);CKX3SV-R：GTTGGACATTTTGGTACGAGGTGG(SEQ ID NO.3).
And carrying out PCR reaction by taking the DNA of different types of cabbage materials as templates, and carrying out agarose gel electrophoresis detection on the obtained products, wherein the electrophoresis result is shown in figure 2, the obtained products of broccoli and cauliflower materials are 600bp, and the obtained product of the common head cabbage material is 284bp. Meanwhile, the product is subjected to sanger sequencing, and the sequencing result shows that the SV region sequence is shown as SEQ ID NO.1 and is 316bp.
By using the molecular marker primer, 50 parts of cabbage materials are identified, wherein the molecular marker primer comprises 10 parts of broccoli and 15 parts of broccoli materials, 25 parts of other cabbage materials, and the other cabbage materials comprise 10 parts of common head cabbage, 3 parts of cabbage mustard, 3 parts of kohlrabi, 3 parts of edible collard, 3 parts of ornamental collard and 3 parts of brussels sprouts. Extracting genome DNA of all materials, and performing PCR reaction identification.
The results show that: 25 parts of broccoli and cauliflower materials are 600bp, and the identification accuracy is 100%;25 parts of other cabbage materials are 284bp, and the accuracy is 100%.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the application described herein without departing from the scope or spirit of the application. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present application. The specification and examples of the present application are exemplary only.

Claims (2)

1. The application of a primer combination in preparing a kit for identifying whether a plant is broccoli or cauliflower is characterized in that the primer combination is used for amplifying a structural variation molecular marker, is positioned in a 3' untranslated region downstream of BoCKX genes and has a sequence shown as SEQ ID NO.1, and comprises a forward primer and a reverse primer, wherein the sequence of the forward primer is shown as SEQ ID NO.2, and the sequence of the reverse primer is shown as SEQ ID NO. 3.
2. A method for identifying whether a plant is broccoli or cauliflower, characterized in that the presence of a structurally mutated molecular marker located in the 3' untranslated region downstream of the BoCKX gene, the sequence of which is shown in SEQ ID No.1, is detected in the DNA of the plant to be identified, the method comprising the steps of:
(1) Extracting sample DNA from a plant sample to be identified;
(2) Amplifying the sample DNA by PCR by using a forward primer and a reverse primer, wherein the sequence of the forward primer is shown as SEQ ID NO.2, and the sequence of the reverse primer is shown as SEQ ID NO. 3; and
(3) The plants were identified by detecting the length of the sample PCR products by electrophoresis, and when the length of the obtained PCR products was 316bp, the plants were broccoli or cauliflower.
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113528698A (en) * 2021-07-14 2021-10-22 中国农业科学院油料作物研究所 InDel molecular marker for identifying and/or distinguishing cabbage vegetables and application thereof
KR20220056806A (en) * 2020-10-28 2022-05-06 서울대학교산학협력단 DNA marker for discriminating genotype of Brassica rapa, Brassica oleracea and their interspecific hybrid and uses thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20220056806A (en) * 2020-10-28 2022-05-06 서울대학교산학협력단 DNA marker for discriminating genotype of Brassica rapa, Brassica oleracea and their interspecific hybrid and uses thereof
CN113528698A (en) * 2021-07-14 2021-10-22 中国农业科学院油料作物研究所 InDel molecular marker for identifying and/or distinguishing cabbage vegetables and application thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
HG994373.1.《GenBank》.2021,第1页. *
LR031875.1.《GenBank》.2018,第1-2页. *

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