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A plant growth negative regulatory cis-element se1 and its application in regulating plant growth

A cis-element, negative regulation technology, applied in the field of genetic engineering

Active Publication Date: 2019-02-01
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has not been found Eui1 Genes have negative regulatory cis-elements (silencers) and methods for improving rice plant height using silencers

Method used

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  • A plant growth negative regulatory cis-element se1 and its application in regulating plant growth
  • A plant growth negative regulatory cis-element se1 and its application in regulating plant growth
  • A plant growth negative regulatory cis-element se1 and its application in regulating plant growth

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Rice dwarf mutant dEui1 Identification and cloning of related genes

[0029] In the process of tissue culture, the inventor obtained a dwarf mutant with gain-of-function of the long-necked panicle gene, which was named as dEui1 ( figure 1 A). dEui1 at T 0 The generation showed a typical dwarf phenotype, while in T 1 In generations, only 1 / 4 of the plants exhibited normal plant height, which indicates that dEui1 is a dominant dwarf mutant. It was found by analysis dEui1 is a T-DNA insertion mutant. The adjacent sequence was obtained by TAIL-PCR, and the T-DNA locus was inserted into a long-necked spike gene encoding cytochrome P450 monooxygenase (CYP450) located on chromosome 5 Eui1 (LOC_Os05g40384). Eui1 Consisting of two exons and one intron, through further sequence analysis, T-DNA replaced Eui1 The intron is close to the 135 base sequence of the first exon ( figure 1 B). for further confirmation dEui1 It is caused by T-DNA insertion,...

Embodiment 2

[0031] Example 2 Build and carry Eui1 Genetic Transformation of ΔNS1 or ΔNS2 Functional Complementary Vectors Deleting Small Intron Fragments eui1 Mutants to obtain transgenic plants of different plant heights

[0032] Genomic DNA of japonica rice Zhonghua 11 was used as a template, and a segmented amplification strategy was used to construct a construct containing Eui1 Functional complementation transformation vectors ΔNS1 and ΔNS2 with small intron deletions ( figure 2 A), and then transform these plasmids into Agrobacterium, and introduce them into eui1 in mutants. T 0 Generation of transgenic detection ( figure 2 B, 2C). The inventors found that transforming with pCAMBIA1300-ΔNS1 expression vector eui1 The plant height of the mutant transformants returned to the level of wild-type plant height ( figure 2 D). Transformed with pCAMBIA1300-ΔNS2 expression vector eui1 The plant height of the mutant transformants showed a dwarf plant height phenotype compared...

Embodiment 3

[0033] Example 3 Transformation of rice without introns Eui1 dwarf transgenic plants

[0034] Using pCAMBIA1300 and the primers in Table 2, the intron-removed Eui1 The coding region cDNA was transformed into wild-type rice Zhonghua 11, and dwarf transgenic positive plants were also obtained ( image 3 A). Transcript expression analysis showed that, Eui1 -cDNA in transgenic plants Eui1 expression level and dEui1 Similar to the wild type in roots, stems and leaves were significantly increased ( image 3 B and 3C).

[0035]

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Abstract

The invention belongs to the technical field of genetic engineering, and specifically discloses a negative-regulated cis element SE1 (Silencer of Eui1) for plant growth and an application of the cis element SE1 in regulation of plant height. According to the negative-regulated cis element SE1 for the plant growth, a dwarfing plant into which one rice T-DNA is inserted is utilized, and the T-DNA is inserted in the intron of an Eui1 gene through TAIL-PCR (Thermal Asymmetric Interlaced Polymerase Chain Reaction) treatment of amplification flanking sequences. Eui1 is one of four genetic materials in heterosis-utilized rice breeding. The negative-regulated cis element SE1 for the plant growth and the application of the cis element SE1 in the regulation of the plant height have the advantages that a silencer SE1 of the Eui1 gene in the intron section of the Eui1 gene is determined, and a novel dwarfing plant related to the Eui1 is obtained through a functional complementation assay of utilizing the Eui1 including the intron with small deletions to genetically transform an eui1 mutant, so that the SE1 element is proved to have better application potentialities in control of plant height of rice, construction of ideal plant types for hybrid rice breeding and genetic improvement for improving rice yield.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, more specifically, to a plant growth negative regulation cis-element SE1 And its application in regulating plant strain high school. Background technique [0002] Plant height is one of the important agricultural traits that can affect crop yield. The first "green revolution" doubled the yield of rice on the basis of semi-dwarf breeding. The heterosis utilization breeding technology centered on male sterility traits set off the wave of the second "green revolution", and achieved remarkable results in crops such as corn, rice, and rapeseed. Male sterile lines of rice generally have the phenomenon of "neck wrapping" where the tassels cannot be pulled out, which brings certain difficulties to hybrid rice breeding. At the same time, in order to further increase rice yield, breeders have proposed the concept of ideal strain breeding. For example, the plant height should be controlled at...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82A01H5/00A01H6/46
CPCC12N15/8261C12N2830/00
Inventor 陈乐天谢勇尧张雅玲黄建乐李元元田庆玮陈远玲刘耀光
Owner SOUTH CHINA AGRI UNIV
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