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Cultivation method for flood-resistant and yield-increasing transgenic wheat and related biological materials thereof

A technology of biomaterials and transgenic plants, which is applied in the cultivation of transgenic wheat with waterlogging tolerance and increased yield and related biomaterials, and can solve the problems of slow progress in the research of waterlogging-tolerant wheat varieties

Inactive Publication Date: 2018-05-25
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Slow progress in conventional breeding methods for breeding waterlogging-tolerant wheat varieties

Method used

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  • Cultivation method for flood-resistant and yield-increasing transgenic wheat and related biological materials thereof
  • Cultivation method for flood-resistant and yield-increasing transgenic wheat and related biological materials thereof
  • Cultivation method for flood-resistant and yield-increasing transgenic wheat and related biological materials thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Example 1. Discovery of wheat waterlogging tolerance and yield-increasing protein TaERFVII.1 and its coding gene TaERFVII.1

[0082] The inventors of the present invention used transcriptomics and transcript expression analysis profiles, wheat genome data mining and gene cloning to analyze the differential expression data of genes in response to waterlogging stress of the flood-tolerant wheat Nonglin 46 and the waterlogging-sensitive wheat Yangmai 16, Combined with virus-induced gene silencing (VIGS) analysis, the important gene TaERFVII.1, which is important for waterlogging stress tolerance and yield increase in wheat, was isolated from Nonglin 46. The specific cloning method is as follows:

[0083] The leaves of wheat Nonglin 46 seedlings treated with waterlogging stress for 6 days were treated with liquid nitrogen, and the total RNA of the leaves was extracted according to the instructions of the InvitrogenTRIZOL Reagent total RNA extraction reagent. According to t...

Embodiment 2

[0084] Embodiment 2, the induced expression analysis of TaERFVII.1 gene

[0085] 1. Response expression analysis after waterlogging stress

[0086] Wheat Nonglin 46 seedlings were treated with waterlogging stress, and the wheat leaf tissues were collected 0h, 1, 2, 3, 6, and 9 days after the treatment, and were quickly frozen in liquid nitrogen and stored in a -80°C ultra-low temperature refrigerator to extract RNA. The leaves of Nonglin 46 before treatment were used as the control (0h).

[0087] According to the procedure of Invitrogen's First Strand cDNA Synthesis Kit, it was reverse transcribed into cDNA. Using the constitutively expressed actin gene of wheat as an internal reference, the cDNA concentrations of the samples were normalized. Then carry out real-time quantitative RT-qPCR analysis with the specific primer of TaERFVII.1 gene, use 2 -△△CT Method (Livak KJ, Schmittgen TD.2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2 ...

Embodiment 3

[0095] Example 3, the acquisition of transgenic wheat and identification of waterlogging tolerance and yield

[0096] 1. Construction of recombinant expression vector

[0097] 1. Harvest 46 leaves of wheat agriculture and forestry after 48 hours of waterlogging stress, extract RNA, reverse transcribe into cDNA; use cDNA as a template, and perform PCR amplification with a primer pair composed of TaERFVII.1-transfF and TaERFVII.1-transfR, The PCR amplification product (TaERFVII.1 gene carrying SpeI and SacI sites) was obtained.

[0098] TaERFVII.1-transfF:5-ATC ACTAGT ATGTGCGGCGGCGCCA-3 (SpeI enzyme recognition site is underlined); TaERFVII.1-transfR:5-ATC GAGCTCTCAGAGCCCCAGAGGC-3 (the SacI enzyme recognition site is underlined) PCR reaction program: first 94°C for 3 minutes; then 94°C for 30s, 56°C for 30s, 72°C for 1min, 15 cycles; 94°C for 30s, 58°C for 30s, 72°C 1min, 20 cycles; the last 10min at 72°C.

[0099] 2. The PCR amplified product was recovered, and connected ...

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Abstract

The invention discloses a cultivation method for flood-resistant and yield-increasing transgenic wheat and related biological materials thereof. The method includes a step of introducing a coding geneTaERFVII.1 of protein TaERFVII.1 into a recipient plant to obtain a transgenic plant having higher plant yield and / or flooding tolerance than that of the recipient plant, and the amino acid sequenceof the protein is shown in SEQ ID No.2. A transgenic experiment for introducing TaERFVII.1 into wheat proves that the flooding tolerance and yield of TaERFVII.1 overexpressed transgenic wheat are significantly improved compared with the recipient wheat and shows that TaERFVII.1 is protein related with plant flooding tolerance and yield, and TaERFVII.1 and coding genes thereof can be used for improving the flooding tolerance and yield of plants.

Description

technical field [0001] The invention relates to a cultivation method of waterlogging-resistant and yield-increasing transgenic wheat and related biological materials in the fields of molecular biology and genetic engineering. Background technique [0002] Wheat (Triticum aestivum) is one of the four major crops that humans rely on for survival. More than one-third of the world's population uses wheat as a staple food. The yield and quality of wheat directly affect human survival and quality of life. In recent years, with global warming, frequent rainfall has caused serious waterlogging disasters. At the same time, the backwardness of farmland drainage system has also aggravated the hazards of waterlogging. Globally, up to 10% of agricultural land is flooded, resulting in yield losses of 15-80%. Wheat production in the US, Europe, Australia and Asia has been severely limited by waterlogging. In the wheat areas of the middle and lower reaches of the Yangtze River and Sichua...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N5/10A01H5/00A01H6/46
CPCC07K14/415C12N15/8261C12N15/8271
Inventor 魏学宁张增艳荣玮徐惠君
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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