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CN-121992001-A - Gene OsARF1 for regulating and controlling ability of rice root system to penetrate hard soil and application thereof

CN121992001ACN 121992001 ACN121992001 ACN 121992001ACN-121992001-A

Abstract

The invention discloses a gene OsARF1 for regulating and controlling the capacity of a rice root system to penetrate hard soil and application thereof. The amino acid sequence of the gene OsARF1 is shown as SEQ ID No. 2. The nucleotide sequence of the gene OsARF1 is shown as SEQ ID No. 1. According to the invention, through over-expressing the OsARF1 gene in rice, the expression level of the OsARF1 gene in rice varieties is improved, and further, the rice strain with enhanced root penetration capacity in hard soil is obtained. The OsARF1 gene is up-regulated under the condition of hard soil, the expression of a downstream OsCESA gene is inhibited, the cellulose synthesis and the cell wall thickening of the cortical cell wall are reduced, the thickening of root systems is promoted, and the capability of penetrating through the hard soil of root systems is improved. The application of the invention can cultivate new rice varieties suitable for the growth of hard soil, and has important significance for rice planting under the condition of improving bad soil such as saline-alkali soil and the like.

Inventors

  • LIANG WANQI
  • ZHANG JIAO
  • ZHANG DABING

Assignees

  • 上海交通大学

Dates

Publication Date
20260508
Application Date
20241105

Claims (10)

  1. 1. The application of the gene OsARF1 in regulating and controlling the capacity of the root system of the rice to penetrate hard soil is characterized in that the amino acid sequence coded by the gene OsARF1 is shown as SEQ ID No. 2.
  2. 2. The use according to claim 1, wherein the nucleotide sequence of the gene OsARF1 is shown in SEQ ID No. 1.
  3. 3. The use according to claim 1, wherein the use is to overexpress the OsARF1 gene in rice, thereby obtaining a rice line with enhanced root penetration in hard soil.
  4. 4. The use according to claim 1, wherein the rice variety is japonica rice variety No. 7.
  5. 5. The recombinant expression vector comprises a gene OsARF1, and is characterized in that the amino acid sequence coded by the gene OsARF1 is shown as SEQ ID No. 2; And/or the nucleotide sequence of the gene OsARF1 is shown as SEQ ID No. 1.
  6. 6. The construction method of the recombinant expression vector is characterized by comprising the following steps: S1, amplifying an OsARF1 gene sequence, namely performing PCR amplification by taking rice root cDNA as a template to obtain a PCR product of the OsARF1 gene shown as SEQ ID NO. 2; s2, enzyme digestion treatment, namely enzyme digestion treatment is carried out on the PCR product of the OsARF1 gene in the step S1 to generate a proper sticky end, and enzyme digestion treatment is carried out on pTCK carriers to enable the carrier to have the sticky end matched with the PCR product; S3, connecting the fragment of the PCR product of the OsARF1 gene after the cleavage with the fragment of the pTCK vector after the cleavage by using In-fusion enzyme, and obtaining the pTCK-OsARF 1 plasmid after sequencing verification.
  7. 7. The method of constructing a recombinant expression vector according to claim 6, wherein in step S1, the primer sequences for PCR amplification are as set forth in SEQ ID No.3 and SEQ ID No. 4.
  8. 8. The method of claim 6, wherein in S2, bamHI is used to cleave the PCR product of OsARF1 gene and pTCK vector, respectively.
  9. 9. A method for regulating and controlling the root morphology of rice is characterized in that the expression of OsCESA gene is regulated down by up-regulating the expression of OsARF1 gene to promote thickening of root system, the amino acid sequence of the gene OsARF1 is shown as SEQ ID No.2, and/or the nucleotide sequence of the gene OsARF1 is shown as SEQ ID No. 1.
  10. 10. The method of regulating rice root morphology of claim 9, wherein the regulation is positive regulation.

Description

Gene OsARF1 for regulating and controlling ability of rice root system to penetrate hard soil and application thereof Technical Field The invention belongs to the technical field of agricultural biology, and particularly relates to a gene OsARF1 for regulating and controlling the capability of a rice root system to penetrate hard soil and application thereof. Background Rice is a main grain crop in China, and the yield and the quality of the rice are directly related to national grain safety. With the reduction of cultivated land area and the aggravation of soil degradation, how to improve the stress resistance of rice and improve rice varieties to adapt to the growth under poor soil conditions becomes one of the important directions of current rice breeding research. In the growth process of rice, the root system is used as a main organ for absorbing moisture and nutrients, and the growth condition of the root system directly influences the growth and development of plants. Under the condition of hard soil, the penetration capability of the rice root system is particularly important, so that plants can be promoted to absorb deep nutrients in the soil, the aim of biological soil loosening can be achieved, and the soil structure and environment are further improved. However, the current molecular mechanism for regulating the capacity of the rice root system to penetrate hard soil is limited, the reports on the related gene functions for regulating the root penetration capacity are few, and effective gene resources are lacking to cultivate new rice varieties suitable for the growth of the hard soil. Therefore, the identification and utilization of the key genes capable of improving the capacity of the rice root system to penetrate through the hard soil has important theoretical significance and application value for rice planting under the conditions of cultivating new varieties of hard soil-resistant rice, improving saline-alkali soil and other bad soil. Disclosure of Invention In view of the above, the invention provides a gene OsARF1 for regulating and controlling the capability of the root system of rice to penetrate hard soil and application thereof, so as to solve the problem that an effective way for improving the capability of the root system of rice to penetrate hard soil is lacking in the prior art. The invention aims at realizing the following technical scheme: < first aspect > The application of the gene OsARF1 in regulating and controlling the capacity of the root system of the rice to penetrate hard soil is provided, and the amino acid sequence of the gene OsARF1 is shown as SEQ ID No. 2. The nucleotide sequence of the gene OsARF1 is shown as SEQ ID No. 1. The application is that the OsARF1 gene is over-expressed in rice varieties to obtain rice varieties with enhanced root penetration capacity in hard soil. The rice variety is japonica rice variety Wuyujing No. 7. < Second aspect > The invention also provides a recombinant expression vector comprising a gene OsARF1, wherein the amino acid sequence of the gene OsARF1 is shown as SEQ ID No. 2; And/or the nucleotide sequence of the gene OsARF1 is shown as SEQ ID No. 1. < Third aspect > A construction method of a recombinant expression vector comprises the following steps: S1, amplifying an OsARF1 gene sequence, namely performing PCR amplification by taking rice root cDNA as a template to obtain a PCR product of the OsARF1 gene shown as SEQ ID NO.2, wherein the accession number of the rice root cDNA is EU847008, LOC_Os01g13520; s2, enzyme digestion treatment, namely enzyme digestion treatment is carried out on the PCR product of the OsARF1 gene in the step S1 to generate a proper sticky end, and enzyme digestion treatment is carried out on pTCK carriers to enable the carrier to have the sticky end matched with the PCR product; S3, connecting the fragment of the PCR product of the OsARF1 gene after the cleavage with the fragment of the pTCK vector after the cleavage by using In-fusion enzyme, and obtaining the pTCK-OsARF 1 plasmid after sequencing verification. In step S1, the primer sequences for PCR amplification are as described in OsARF1-F (SEQ ID NO. 3) and OsARF1-R (SEQ ID NO. 4). In S2, the PCR product of OsARF1 gene and pTCK vector were digested singly with BamHI. < Fourth aspect > A method for regulating the morphology of rice root system features that the expression of OsCESA gene is down regulated by up regulating the expression of OsARF1 gene, the cellulose synthesis of the cell wall of cortex of rice root system is reduced, the thickening of cell wall is inhibited, the thickening of root system is promoted and penetrability is enhanced. The regulation is positive regulation. The amino acid sequence of the gene OsARF1 is shown as SEQ ID No. 2; And/or the nucleotide sequence of the gene OsARF1 is shown as SEQ ID No. 1. < Fifth aspect > The invention also provides a method for screening the rice hard soil resistant varieties, which compris