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Application of kale albino gene wkl1 in plant albinism

A technology of kale and cabbage, applied in the field of plant engineering, can solve the problems of unanalyzed, uncloned, and unstudied genetic laws

Active Publication Date: 2020-06-05
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The applicant found that the white color trait of white kale is an unreported white dominant trait, its genetic law has not been analyzed, and the gene controlling the white color of white kale has not been cloned, and the function of this gene has not been studied

Method used

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  • Application of kale albino gene wkl1 in plant albinism

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Experimental program
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Effect test

Embodiment 1

[0020] The kale albino gene WKL1( W hit K ale L The acquisition of ocus 1):

[0021] 1. Genetic analysis of dominant mutant WKL1 phenotype

[0022] In this study, a genetic analysis of the mutant WKL1 phenotype was performed. F 1 , continue to backcross with the green headed cabbage haplotype to obtain BC 1 , after genetic analysis, in its BC 1 Medium white leaves: green leaves = 304: 299 ≈ 1: 1 (P>0.05), while in BC 1 f 2 White leaves:light white leaves:green leaves=27:52:20≈1:2:1(P>0.05). In summary, the white trait is incompletely dominant inheritance controlled by a single gene, named WKL1( W hit K ale L ocus 1). As an extremely rare dominant gene controlling white leaves, it is of great theoretical significance to understand how it participates in inhibiting the formation of chloroplasts.

[0023] 2. Map-based cloning of WKL1 gene

[0024] The method of BSA combined with RNA-seq, that is, BSR-seq (Liu et al., 2012), was used to initially map the WKL1 gene. ...

Embodiment 2

[0026] Application of kale albino gene WKL1 in plant albinism:

[0027] The WKL1 gene CDS sequence (shown in SEQ ID NO.1) was homologously recombined into the Hellsgate2 vector, and transformed into Agrobacterium, overexpressed with the 35S promoter, transformed into tobacco, and obtained 10 overexpressed T 1 Generation of positive seedlings, 4 of which produced white phenotype ( figure 1 Middle a), and the phenotype is consistent with the genotype. Homologous recombination of the WKL1 gene's own promoter and the full-length sequence of the WKL1 gene (shown in SEQ ID NO.1 and SEQ ID NO.3) into the Hellsgate8 vector, using the white kale WKL1 gene's own promoter and the full-length WKL1 gene functional complementary transfection into green cabbage, three positive plants were obtained, and the phenotype was whitening or severe yellowing of leaves ( figure 1 Middle b). The experimental results proved that the WKL1 gene, that is, CYP708A4, has the function of turning leaves whi...

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Abstract

The present application belongs to the technical field of plant genetic engineering, and specifically relates to the cloning, functional identification and an application of a key dominant gene WhiteKale Locus 1 (WKL1) which controls the low-temperature albino of white kale. A nucleotide sequence of the gene is shown in SEQ ID NO: 1. The invention also relates to a promoter of the WKL1 gene and the cloning of its recessive allele promoter. Through the agrobacterium-mediated transformation of Solanaceae tobacco and green cabbage, the functional verification shows that the gene has the significant function to control the albino of white kale at low temperature. The discovery of the KWL1 gene provides new genetic resources for the production of new white ornamental varieties in cruciferous vegetables, and provides new genetic resources for ornamental horticulture. The utilization of the resource is conducive to the cultivation of new ornamental plant varieties.

Description

technical field [0001] This patent belongs to the technical field of plant engineering. It specifically relates to the cloning, functional identification and application of a key gene WKL1 unique to Cruciferae that controls the albinism of white kale, and the gene can be used to prepare albino ornamental plants. Background technique [0002] White kale is a lovely autumn-winter ornamental that functions somewhat as a flower. White kale has many varieties, such as "White Pigeon", "White Gull", "White Crane" and so on. Previous studies have shown that only at low temperature (≤15°C) can white kale be induced in the new leaves of white kale. Under the low temperature condition, no chloroplast structure was seen in the white area of ​​white kale, and then the white phenotype was produced. In 2014, the genome of green cabbage was released, which facilitated the genetic mapping and cloning of the target traits of cabbage vegetables (white kale and green cabbage are different va...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N15/11
CPCC12N15/8222C12N15/8261
Inventor 陈炯炯严承欢匡汉晖安光辉张维奕朱婷
Owner HUAZHONG AGRI UNIV
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