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Rice seedling stage biomass regulation gene SBM1 and application thereof

A technology for regulating genes and biomass, applied in the field of plant genetic engineering

Active Publication Date: 2021-08-24
CHINA NAT RICE RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it can be noticed that nitrogen use efficiency is not only a characteristic of a certain crop, but a characteristic of the entire crop system. The available nitrogen content in the soil, field management of crops, different crop rotation methods, and environmental conditions will all affect the crop's response to crops. Nitrogen uptake and utilization

Method used

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  • Rice seedling stage biomass regulation gene SBM1 and application thereof
  • Rice seedling stage biomass regulation gene SBM1 and application thereof
  • Rice seedling stage biomass regulation gene SBM1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: Mapping of Rice Seedling Biomass QTL SBM1

[0056] 1. Rice materials and targeting groups

[0057] The present invention utilizes BC comprising 132 strains derived from the combination of 93-11 and PA64s 1 f 13 Recombinant inbred line populations were used for initial QTL mapping analysis of seedling biomass. In order to quickly obtain a near-isogenic line containing the target QTL SBM1, carry out verification and fine mapping, use molecular markers closely linked to SBM1 in the existing chromosomal substitution line population derived from Nipponbare (NPB) and Kasalath matching M1 and M4, the line SBM1 containing the Kasalath introgression fragment in the SBM1 interval under the NPB genetic background was screened Kasalath , and backcrossed the introgression line with the recurrent parent NPB for 4 consecutive generations, the genetic background was highly consistent with that of NPB, and the SBM1 interval was the BC of the heterozygous fragment of NPB an...

Embodiment 2

[0067] Embodiment 2: the method for gene knockout SBM1 gene improvement rice biomass and yield

[0068] Using the cDNA of NPB as a template, the primers SBM1-cDNA-F and SBM1-cDNA-R were used to amplify, and the fragments were sequenced for sequence comparison analysis.

[0069] SBM1-cDNA-F: 5'-ATGGAATCCAGCGAACAATGCGCCCTC-3',

[0070] SBM1-cDNA-R: 5'-TTACACCACCGCGTACATCACGGTGGT-3'.

[0071] The SBM1 gene contains a 4819bp nucleotide sequence (ie, the SBM1 gDNA sequence), as shown in SEQ ID No:1. The 885bp full-length cDNA sequence (SEQ ID No: 2) of the SBM1 gene was obtained by PCR amplification. The protein encoded by the gene has a sequence of 294 amino acids shown in SEQ IS No: 3, and the stop codon (TAA) is omitted at the end. To construct the CRISPR / Cas9 vector, insert the sgRNA target sequence of the target gene into the CRISPR / Cas9 vector to obtain the vector 65110-cas9 and 65120-cas9 vector, where the primer sequences are:

[0072] 65110-U3F:GGCAACCTGATCACGTACCTGAC,...

Embodiment 3

[0083] Example 3: SBM1 participates in the regulation of nitrogen use efficiency and affects rice yield

[0084] In order to investigate whether SBM1 affects nitrogen use efficiency and rice yield, the present invention utilizes NPB and NIL-SBM1 Kasalath , a comparative test of two nitrogen application rates was carried out under field conditions. The nitrogen fertilizer treatment experiment in the field was designed to plant three plots for each material, the plant-to-row spacing was 15×25 cm, and the plot size was 4.0×4.0 m. Different nitrogen treatments were repeated 3 times. Urea was used as the only nitrogen source, and the application rates of urea in the two treatments were 180kg / ha (High nitrogen, HN) and 90kg / ha (Lownitrogen, LN), respectively, and were applied three times at the seedling stage, tillering stage and jointing stage, each accounting for the total 30%, 40%, 30% of the amount of urea. Superphosphate as phosphate fertilizer (90kg / ha) and potassium chlorid...

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Abstract

The invention relates to a QTL SBM1 related to biomass accumulation and having significant difference between indica and japonica subspecies, functions of the gene is confirmed by using a rice seedling stage biomass regulation gene SBM1 cloned by using forward genetics and map-based cloning technology and a Crispr Cas9 knockout technology and a transgenic complementary test. Meanwhile, the invention also relates to a cooperative regulation relation of the gene on nitrogen utilization efficiency and yield. The invention discloses a rice seedling stage biomass regulation gene SBM1, the nucleotide sequence of which is as shown in SEQ ID No: 1, and the CDS nucleotide sequence of which is as shown in SEQ ID No: 2. The gene knockout SBM1 increases the biomass and yield of rice.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and in particular relates to rice biomass regulating gene SBM1 and its application. Background technique [0002] Rice is one of the most important food crops in the world. Fully exploiting the high-yield potential of rice is the key to solving the food security problems brought about by the continuous increase of world population. The breeding and application of semi-dwarf rice varieties, the promotion of hybrid rice and the use of chemical fertilizers have promoted two major leaps in rice yield (Khush 2001), and the method of increasing rice yield by increasing nitrogen fertilizer has been widely used in production (Peng et al. al. 2002). The establishment of ideal plant type and nutrient uptake and utilization complement each other, and they are synergistically regulated in the process of yield formation. However, the nitrogen use efficiency of the main cultivars currently used in pr...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8261
Inventor 张光恒钱前徐静王佳佳曾大力陈敏敏王梓安胡江朱丽高振宇董国军任德勇沈兰张强李清
Owner CHINA NAT RICE RES INST