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Application of gene for interfering expression of LCYB and LCYE and simultaneously ectopically expressing TT8 in preparation of brassica plant having red petals

A technology of brassica, flower petals, applied in the field of biology

Inactive Publication Date: 2015-04-22
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the entire Brassica genus including rapeseed, there is no report on the use of molecular breeding for metabolic engineering of rapeseed flower color.

Method used

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  • Application of gene for interfering expression of LCYB and LCYE and simultaneously ectopically expressing TT8 in preparation of brassica plant having red petals
  • Application of gene for interfering expression of LCYB and LCYE and simultaneously ectopically expressing TT8 in preparation of brassica plant having red petals
  • Application of gene for interfering expression of LCYB and LCYE and simultaneously ectopically expressing TT8 in preparation of brassica plant having red petals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1. Microscopic observation of the chromogenic subcellular organelles of Brassica and radish petals

[0042] Take fresh cabbage-type rape yellow petals (BnY); cabbage-type rape milk petals (BnW); mustard-type rape yellow petals (BjY); Chinese cabbage-type rape yellow petals (BrY); kale yellow petals (BoY); kale White petals (BoW); radish red petals (RsR), stored on ice and shipped back to the laboratory, then sliced ​​by hand, observed with low magnification for rapid screening, and selected qualified slices for careful observation and photography. The results are as follows figure 1 shown. The results showed that the yellow substances in the yellow petals of several species of Brassica appeared as many small particles in a cell, distributed unevenly in the cell, and were squeezed by the vacuoles to the adherent position, indicating that the yellow petals were in the form of many small particles. Chromogenic organelles are chromosomes whose pigments are yellow c...

Embodiment 2

[0043] Embodiment 2, detect Brassica and radish petal pigment

[0044] In the morning of full bloom, take the yellow petals of Brassica napus (BnY); Brassica napus milk petals (BnW); Brassica napus yellow petals (BjY); Brassica napus yellow petals (BrY); Brassica napus yellow petals (BrY). BoY); kale white petals (BoW); radish red petals (RsR), immediately cool and fresh, transported back to the laboratory, dried at 50-60 °C, ground into powder, passed through a 60-mesh sieve, and dried in the dark for storage.

[0045] 1. Test results of petroleum ether, hydrochloric acid and ammonia water

[0046] Weigh 0.100g of the preserved petal powder, put them into numbered test tubes with stoppers, add about 10mL each of petroleum ether, 10% hydrochloric acid, and 30% ammonia water, mix gently, filter, and observe the color change, the results are as follows :

[0047] (1) Petroleum ether reaction: Brassica napus, cabbage-type rape, and mustard-type rape all show bright yellow, indi...

Embodiment 3

[0067] Example 3. Molecular identification of Brassica and radish petal pigment biosynthetic pathways

[0068] (1) Expression characteristics of flavonoid pathway genes in Brassica and radish petals

[0069] In order to study the molecular mechanism of the difference between Brassica and radish petal pigments, RT-PCR detection primers for Brassica and radish flavonoid pathway genes were designed, and 5S rRNA was used as an internal reference, as shown in Table 1:

[0070] Table 1. Flavonoid pathway RT-PCR detection primers

[0071]

[0072]

[0073]In the morning of full bloom, take the yellow petals of Brassica napus (BnY); Brassica napus milk petals (BnW); Brassica napus yellow petals (BjY); Brassica napus yellow petals (BrY); Brassica napus yellow petals (BrY). BoY); kale white petals (BoW); radish red petals (RsR), frozen in liquid nitrogen for transportation, and stored in a -80°C refrigerator for later use. Then use RNAprep Pure plant total RNA extraction kit to ...

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Abstract

The invention discloses application of a gene for interfering expression of LCYB and LCYE and simultaneously ectopically expressing TT8 in preparation of a brassica plant having red petals. The invention further discloses an expression vector for preparing the gene for interfering expression of LCYB and LCYE genes and simultaneously ectopically expressing TT8 gene and a preparation method of the vector. Yellow carotenoid is inhibited and red lycopene is accumulated by interfering expression of the LCYB and LCYE genes; accumulation of cyanin can be promoted through ectopic expression of the TT8 gene; therefore, the petals are red; the brassica plant having red petals is successfully prepared; and colours of the brassica plants are enriched.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of interfering LCYB and LCYE expression and ectopic expression of TT8 in preparing red petal Brassica plants. Background technique [0002] Rape is an important oil crop with a long history of cultivation, high economic value, wide application and strong adaptability. It is the largest oil crop in my country and an important oil crop worldwide. Taxonomically, rape is composed of three species of Brassica, namely Brassica napus (Brassica napus, 2n=38, AACC), Brassica rapa ssp.oleifera syn.B.campestris, 2n=20 , AA), Brassica juncea (Brassica juncea, 2n=36, AABB). Brassica napus is a compound species evolved from cabbage (Brassica rapa) and cabbage (Brassica oleracea, 2n=18, CC) through natural interspecies hybridization after double diploidization, although the cultivation history is only a few hundred years. However, due to its strong growth potential and ...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/66C12N15/84A01H5/00A01H4/00
Inventor 柴友荣刘雪董博廖霏霏林呐练剑平王瑞殷家明万华方闻鎏法
Owner SOUTHWEST UNIVERSITY
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