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Application of non-coding gene capable of controlling corn ear thickness heterosis and precursor sequence of non-coding gene

A heterosis, non-coding technology, applied in the application of non-coding genes and their precursor sequences, can solve the problems of low probability, poor predictability, and large blindness of screening excellent combinations, and achieve the improvement of ear diameter and ear size. yield effect

Active Publication Date: 2021-05-14
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Since the molecular mechanism of heterosis genetic formation is still unclear, in the process of breeding new varieties of crops using heterosis such as corn and rice, effective prediction of excellent hybrid combinations cannot be achieved, and breeders still need to combine and identify a large number of hybrids in the field. The hybrid combination is highly blind and poor in predictability, which not only wastes a lot of manpower and material resources, but also has a very low probability of screening out an excellent combination, thus restricting the process of breeding excellent new varieties

Method used

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  • Application of non-coding gene capable of controlling corn ear thickness heterosis and precursor sequence of non-coding gene
  • Application of non-coding gene capable of controlling corn ear thickness heterosis and precursor sequence of non-coding gene
  • Application of non-coding gene capable of controlling corn ear thickness heterosis and precursor sequence of non-coding gene

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Discovery of maize ear coarse heterosis non-coding gene zma-miR390 involved in the determination of maize ear heterosis:

[0026] A set of single-segment substitution line populations was constructed with the excellent maize inbred line Chang 7-2 as the donor parent and lx9801 as the recipient parent; the corresponding testcross population was constructed by crossing with the testcross parent Zheng 58 (Zheng 58× SSSL); By comparing the phenotypic differences between the test cross and the control Zheng 58×lx9801 (Ludan 9002), a single-segment substitution line containing the heterosis locus of panicle diameter was identified and named lx9801 hlEW2b ;For Zheng 58×lx9801, Zheng 58×lx9801 hlEW2b Ears at different developmental stages were subjected to transcriptome sequencing, degradome sequencing and miRNA-seq;

[0027] The results of miRNA-seq showed that there were significant differences in the expression of zma-miR390 between the two materials; the results of maize e...

Embodiment 2

[0031] Confirmation of maize ear coarse heterosis non-coding gene zma-miR390 regulating maize ear heterosis:

[0032] The pCAMBIA3301 vector was digested with Bgl II and BstE II enzymes, and then the artificially synthesized Zma-miR390a fragment was connected to the pCAMBIA3301 vector using T4 DNA ligase to construct an overexpression vector of zma-miR390a (pCAMBIA3301::35S::Zma-miR390a , figure 2), the RNA sequence of the Zma-miR390a gene is shown in SEQ ID NO.2, and the primers used to amplify Zma-miR390a include CUB-mir390a-F primers and CUB-mir390a-R primers, and the CUB-mir390a-F primers The DNA sequence of the CUB-mir390a-R primer is shown in SEQ ID NO.3, the DNA sequence of the CUB-mir390a-R primer is shown in SEQ ID NO.4, and the DNA sequence of the overexpression vector pCAMBIA3301::35S::Zma-miR390a is shown in SEQ ID NO.4 As shown in ID NO.5;

[0033] With the sequence of zma-miR390 as the target, construct the STTM390 vector (such as image 3 ), the DNA sequence...

Embodiment 3

[0036] Evolutionary conservation of the non-coding gene zma-miR390 in maize ear coarse heterosis:

[0037] MiR390 is an ancient and highly conserved miRNAs. At present, most of the research on plant miR390 genes is focused on model plants such as Arabidopsis and rice, and high-throughput sequencing technology combined with bioinformatics is often used to identify target genes and action sites. Point and function forecasting and more. Download the precursor sequences of gene family members of maize, Arabidopsis, rice, and soybean miR390 from miRBase (http: / / www.mirbase.org / ), and use the ClustalW method of MEGAX software to compare the precursor sequences of miR390 Yes, choose the Neighbor-Joining method to construct the phylogenetic tree (such as Figure 6 ).

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Abstract

The invention relates to the technical field of gene engineering, and particularly discloses application of a non-coding gene capable of controlling female ear thickness heterosis of corn and a precursor sequence of the non-coding gene. The non-coding gene is a zma-miR390 gene, wherein the RNA sequence of the non-coding gene is as shown in SEQ ID NO. 1, and the precursor RNA sequence of the non-coding gene is as shown in SEQ ID NO. 2. The zma-miR390 gene provided by the invention can be used for improving the female ear thickness and yield of corn, and provides help for breeding practice aiming to increase the yield of corn.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the application of a non-coding gene and its precursor sequence for controlling coarse heterosis of maize ears. Background technique [0002] As one of the most important food, feed and energy crops in the world, corn plays an irreplaceable role in ensuring world food security, promoting economic development and alleviating energy crisis. Maize is a monoecious plant with different flowers. The ear development needs to go through axillary bud meristem (AM), inflorescence meristem (IM), spikelet paired meristem (SPM), spikelet meristem (SM), A series of differentiation processes such as floret meristem (FM). The number of corn ear rows (ear diameter) is mainly determined by the size of the IM. The increase of IM can provide enough space for new SPM differentiation, resulting in an increase in the number of SPMs and finally the number of ear rows. The number of grains ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/6895
CPCC12N15/113C12Q1/6895C12Q2600/178C12Q2600/13Y02A40/146
Inventor 汤继华丁冬万炯谢惠玲付志远张稳王洪秋
Owner HENAN AGRICULTURAL UNIVERSITY
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