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Soybean bi-1 gene and its application

A technology of BI-1 and gene, which is applied in the field of cell apoptosis inhibitor BI-1 gene and its application, can solve the problems of unreported functional research, few separation and identification, etc., to reduce chemical fertilizers, strengthen nitrogen fixation ability, and increase root nodules The effect of the number of

Active Publication Date: 2021-03-12
XINYANG NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the research on the function of the apoptosis inhibitor BI-1 gene is mainly concentrated in the model plant Arabidopsis thaliana, and there are few studies on the isolation and identification of the BI-1 gene in soybean, and its role in the process of symbiotic nodulation Functional studies have not been reported yet

Method used

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  • Soybean bi-1 gene and its application
  • Soybean bi-1 gene and its application
  • Soybean bi-1 gene and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] A method for preparing the apoptosis inhibitor GmBI-1 gene involved in symbiotic nodulation of leguminous plants, the steps of which are:

[0033] Cloning of the candidate gene GmBI-1: the fusion of the intramembrane kinase domain GmNFR1α-pk of the nodulation factor receptor protein GmNFR1α of the leguminous plant soybean and the yeast protein expression vector pGBKT7 was used as the bait plasmid, and the soybean was screened by the method of yeast two-hybrid AD-cDNA library, after the colonies grown on the SD / -Leu-Trp-Ade-His (auxotrophic yeast medium) plate were verified by rotation, the plasmid was extracted from the positive transformant and sequenced to obtain GmBI -1 gene, its sequence is the nucleotide sequence shown in SEQ NO:1. The implementation steps are preparation of yeast competent cells by LiAc method, yeast transformation, and library screening, as follows:

[0034] (1) Streak the frozen yeast Y187 (purchased from clontech company) on the YPDA plate fro...

Embodiment 2

[0086] A kind of application of the apoptosis suppressing factor GmBI-1 gene participating in the symbiotic nodulation of legumes in regulating the number of root nodules of legumes (Lotus japonicus, milk vetch, alfalfa, soybean and peanut), the steps are:

[0087] A. Detection of tissue-specific expression of GmBI-1 in soybean:

[0088] Using cultivated soybean W82 as material, harvest soybean root tissues at different times before and after inoculation, and root nodules at 12, 30, 42, 64 and 84 days after inoculation, and extract total RNA from different tissues at different times (Trizol reagent, purchased from Invitrogen Corporation ), and then use fluorescent quantitative PCR technology to detect its expression (attached figure 2 shown). The specific implementation steps are: reverse transcribe the extracted total RNA into cDNA, use the kit PrimeScriptTM RT reagent Kit (Perfect RealTime) for the synthesis of the first strand of cDNA, reverse transcribe 500ng Total RNA i...

Embodiment 3

[0119] Embodiment 3 is basically the same as Embodiment 2, the difference is that

[0120] B. Construction and genetic transformation of GmBI-1 gene overexpression vector:

[0121] In concrete implementation step (1), plant material cultivation is to germinate alfalfa 5 days in advance respectively; In step (7), rhizobium inoculation is alfalfa Rhizobium Sinorhizobium meliloti 1021 (this bacterial strain is the bacterial strain that has announced in the world, Juan etc., Comparison of Developmental and Stress-Induced Nodule Senescence in Medicagot runcatula. Plant Physiol, 152:1574-1584, 2010).

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Abstract

The invention discloses a gene BI-1 of cell withering inhibition factors for regulating and controlling symbiotic nodulation of leguminous plants, and provides application of the gene in regulating and controlling number of root nodules of leguminous plant. A nucleotide sequence of the gene is shown in SEQ ID NO:1 in a sequence list. The gene has the advantages that a yeast two-hybrid technique isutilized to screen in a soybean root nodule AD-cDNA library, a kind of new protein which interacts with receptor protein of nodulation factor is found, and the gene is proofed to be a cell apoptosisinhibition factor gene through methods of sequence comparison, function prediction and the like; an over-expression technique is utilized to study the function of the gene in the symbiotic nodulationprocess of crowtoe of the leguminous plant; proofed by results, the protein encoded by the gene participates in the symbiotic nodulation process, the number of the root nodules is obviously increasedafter over-expression, and the application prospect on the symbiotic nitrogen immobilizing of the leguminous plant is realized.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to an apoptosis inhibitory factor BI-1 gene involved in the regulation of symbiotic nodulation in leguminous plants and its application. Background technique [0002] BI-1 gene is a cell apoptosis inhibitor cloned and identified in 1998, and its protein product can inhibit cell apoptosis caused by Bax. The BI-1 protein contains 6-7 transmembrane domains, and the hydrophilic region at the C-terminus may be a key segment of protein interaction. Its protein family members are mainly located in the inner membrane of mammalian and plant cells, and in higher plants and Animals are highly conserved. Studies have shown that the BI-1 gene can enhance plant tolerance to abiotic stresses such as low temperature, drought, high salinity, and oxidative stress, as well as biotic stresses such as pathogen infection. The BI-1 gene is involved in a variety of stress signal respons...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N1/21C07K14/415A01H5/06A01H6/54C12R1/01
CPCC07K14/415C12N15/8261
Inventor 柯丹霞舒勇袁松丽彭昆鹏
Owner XINYANG NORMAL UNIVERSITY
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