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Method for detecting color characters of rape flowers

A detection method, rapeseed technology, applied in the field of agricultural biology, can solve problems such as the difficulty of parental population construction and the difficulty of locating petal pigment genes

Active Publication Date: 2020-07-17
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the red and orange rapeseed flowers cultivated by our team are distant hybrids of radish and rapeseed, it is very difficult to map the petal pigment gene through the conventional QTL method, because the parents have great difficulties in constructing populations

Method used

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  • Method for detecting color characters of rape flowers
  • Method for detecting color characters of rape flowers
  • Method for detecting color characters of rape flowers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: Analysis of LDOX / ANS gene sequence in different color Brassica napus

[0030] For the sequencing results, the sequencer software was used to analyze the sequences of different copies of the amplification, and the DNAMAN software was used for sequence comparison.

[0031] Sequencing results showed that there were differences in the gene sequences of four different copies of the LDOX / ANS gene in the red, orange and yellow rapeseed lines.

[0032] The four copies of the LDOX / ANS gene are: BnaA01g12530D, BnaA03g45610D, BnaC01g14310D, BnaC07g37670D. The results showed that: the sequence of BnaA01g12530D gene in different flower color materials was consistent with the reference sequence in Brassica napus genome database (SEQ ID NO: 1); BnaA03g45610D gene did not amplify bands in white flower materials, but in the other four The sequences in rapeseed are different, and the alignment results are as follows figure 2 Shown (red material SEQ ID NO: 2, orange mater...

Embodiment 2

[0033] Example 2: Verification of the sequence of the BnaA03g45610D copy of the LDOX / ANS gene in different flower colors

[0034] Take the leaves of red, orange, yellow and white rapeseed lines (planted in October 2018). Among them, the yellow rapeseed was verified by Zheyou 51, Zheyou 50 and B57 varieties.

[0035]Using colored rapeseed as material, DNA was extracted, and PCR amplification was carried out according to the primers provided in Table 1. After the PCR products were sequenced, it was found that there were significant differences in the gene sequences of colored rapeseed BnaA03g45610D with different petal colors ( figure 2 ). These differences can be used to distinguish colored rapeseed of different colors. For example, the yellow flower gene is missing from 32 to 51 bases, and all other colors of rapeseed contain this segment; there are specific SNPs at positions 204, 216, and 280, which can be distinguished from other colored rapeseed. Red rapeseed has specif...

Embodiment 3

[0036] Example 3: Verification of the sequence of the BnaA03g45610D copy of the LDOX / ANS gene in different flower colors

[0037] Red 156, Red 185 and Red 191 (different strains with red petals) planted in October 2019; two kinds of orange rapeseed, numbered Orange 905 and Orange 906 (different strains with orange petals) line); three kinds of yellow rapeseed materials were sampled, namely Zheyou 51, Zheyou 50 and B57 (different varieties with yellow petals). Cloning and sequencing results showed that the BnaA03g45610D gene sequence in red rapeseed was completely consistent with SEQ ID NO: 2; the BnaA03g45610D gene sequence in orange rapeseed was completely consistent with SEQ ID NO: 3; :5 sequence is completely consistent, and the Zheyou 50 sequence is completely consistent with the reference sequence SEQ ID NO:6. It can be seen that the BnaA03g45610D copy of LDOX / ANS gene has the characteristics of distinguishing different flower colors.

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Abstract

The invention discloses a method for detecting color characters of rape flowers. The method comprises the following steps: (1) taking rape leaves, extracting DNA, and amplifying a BnaA03g45610D sequence of an LDOX / ANS gene by using amplification primers; and (2) performing sequencing analysis on an amplification product. According to the result of sequencing analysis, rapes with red flowers, orange flowers, yellowish brown flowers, yellow flowers and white flowers can be completely distinguished, and the method has an important practical value for improving seed selection efficiency and savingbreeding cost.

Description

technical field [0001] The invention relates to a method for detecting flower color traits of rapeseed, which belongs to the field of agricultural biology technology. Background technique [0002] In addition to oil extraction from seeds, rape has many functions, such as vegetable, ornamental, feed, honey, and fertilizer. Under the background of the rural revitalization strategy, as an ornamental or both oil and ornamental colored rapeseed variety, it has broad application prospects in the construction of beautiful countryside. The cultivation of colorful rapeseed can promote the integrated development of primary, secondary and tertiary industries by attracting tourists to watch rapeseed flowers, local characteristic agricultural products and farmhouse entertainment; at the same time, it can also make full and effective use of winter fallow fields and reduce the phenomenon of land abandonment in winter. Therefore, in this case, even if the colored rapeseed does not harvest ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 华水金柳寒林宝刚任韵朱建方张冬青余华胜
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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