CN-120866347-B - ZmARF6 protein for regulating and controlling nitrogen absorption of plants, biological material, application and method for improving plant characteristics

CN120866347BCN 120866347 BCN120866347 BCN 120866347BCN-120866347-B

Abstract

The invention relates to the technical field of agricultural biology, in particular to ZmARF6 protein and biological material for regulating and controlling nitrogen absorption of plants, and application and a method for improving plant properties. The amino acid sequence of the ZmARF6 protein is selected from any one of 1) an amino acid sequence shown as SEQ ID NO. 1, 2) an amino acid sequence which has at least 91% of identity with the amino acid sequence shown as SEQ ID NO. 1, 3) a fusion polypeptide obtained by connecting a tag at the N end or/and the C end of the amino acid sequence shown as SEQ ID NO. 1, and 4) an amino acid sequence derived from one or more amino acid residues deleted, substituted, inserted and/or added in the amino acid sequence shown as SEQ ID NO. 1. According to the invention, nitrogen absorption phenotype identification is carried out on the over-expression material and the gene editing knockout material of the ZmARF6 gene, and the over-expression material is found to have the advantages of increased lateral root elongation, enhanced low nitrogen tolerance, higher nitrogen content and grain weight per spike.

Inventors

ZHANG MINGCAI
Wan Jiachi
ZHANG YUSHI
YANG JIA
ZHANG QINGYI
LI ZHAOHU
LIU JIFENG

Assignees

中国农业大学

Dates

Publication Date: 20260512
Application Date: 20250805

Claims (6)

The application of ZmARF6 related biological material in regulating plant lateral root growth, low nitrogen tolerance and/or grain weight per ear, which is characterized in that the biological material is any one of the following: b1 The amino acid sequence of the ZmARF6 protein is shown as SEQ ID NO. 1, or fusion polypeptide obtained by connecting a tag at the N end or/and the C end of the sequence shown as SEQ ID NO. 1; b2 A nucleic acid molecule encoding a ZmARF6 protein, wherein the nucleic acid molecule has a base sequence as shown in SEQ ID NO.2 or a sequence which hybridizes with the sequence shown in SEQ ID NO.2 under stringent conditions and encodes the amino acid sequence shown in SEQ ID NO. 1; B3 An expression cassette comprising B2) said nucleic acid molecule; b4 A recombinant vector comprising the nucleic acid molecule of B2) or a recombinant vector comprising the expression cassette of B3); B5 A recombinant microorganism comprising B2) said nucleic acid molecule, or a recombinant microorganism comprising B3) said expression cassette, or a recombinant microorganism comprising B4) said recombinant vector; b6 A transgenic plant cell line comprising the nucleic acid molecule of B2), or a transgenic plant cell line comprising the expression cassette of B3), or a transgenic plant cell line comprising the recombinant vector of B4); b7 A transgenic plant tissue comprising B2) said nucleic acid molecule, or a transgenic plant tissue comprising B3) said expression cassette, or a transgenic plant tissue comprising B4) said recombinant vector; B8 A transgenic plant organ containing the nucleic acid molecule of B2), or a transgenic plant organ containing the expression cassette of B3), or a transgenic plant organ containing the recombinant vector of B4); B9 A transgenic plant containing the nucleic acid molecule of B2), or a transgenic plant containing the expression cassette of B3), or a transgenic plant containing the recombinant vector of B4); b10 A tissue culture produced by regenerable cells comprising the transgenic plant of B9); B11 Containing B10) protoplasts produced by said tissue culture; the application is any one of the following: D1 Use in transgenic plants with elongation of lateral roots, increased low nitrogen tolerance and/or increased grain weight per ear; D2 Use in products with elongation of lateral roots, increased low nitrogen tolerance and/or increased grain weight per ear; The plant is corn.
2. The zmahf 6 protein of claim 1, wherein said tag comprises a Flag tag, a His tag, an MBP tag, an HA tag, a myc tag, a GST tag, and/or a SUMO tag.
3. The biomaterial of claim 1, wherein the recombinant vector comprises a nucleic acid molecule encoding a zmahf 6 protein and a backbone plasmid; The construction method of the recombinant vector comprises the steps of amplifying nucleic acid molecules for encoding ZmARF6 protein, recovering PCR products, and connecting the recovered PCR products with a backbone plasmid to obtain the recombinant vector.
4. The biomaterial of claim 1, wherein the recombinant microorganism is obtained by transforming a recombinant vector into the microorganism.
5. The biomaterial of claim 4, wherein the microorganism is yeast or agrobacterium.
6. A method for improving plant traits, which is characterized in that the expression level of a nucleic acid molecule encoding ZmARF6 protein, the activity of the ZmARF6 protein and/or the content of the ZmARF6 protein in plants is increased to obtain improved plants, wherein the improved traits refer to promotion of lateral root elongation and/or enhancement of low nitrogen tolerance; the amino acid sequence of the ZmARF6 protein is shown as SEQ ID NO. 1.

Description

ZmARF6 protein for regulating and controlling nitrogen absorption of plants, biological material, application and method for improving plant characteristics Technical Field The invention relates to the technical field of agricultural biology, in particular to ZmARF6 protein and biological material for regulating and controlling nitrogen absorption of plants, and application and a method for improving plant properties. Background Nitrogen is a key limiting factor for plant growth and development and is critical to crop productivity. Over the past few decades, a large number of chemical fertilizers have been applied to the field to achieve higher crop yields. However, in the agroecosystem, nitrate, the major form of available inorganic nitrogen, is readily dissolved and lost by leaching, resulting in temporal and spatial variations in nitrate concentration, further resulting in increased nitrosation, reduced nitrogen fertilizer availability, and environmental pollution. For example, corn is one of the crops that responds most to nitrogen supplementation, and can only effectively obtain 25% to 50% of the nitrogen during the season of use during which nitrogen fertilizer is utilized. Plant root architecture has a significant impact on N perception and absorption. Plants have evolved the ability to monitor and respond to external nutrient availability by adjusting root system configuration in an attempt to maximize the uptake of nitrogen in the soil in response to highly varying NO 3 concentrations in soil, time and space. Thus, a framework of mechanisms that reveal the response root development in response to nitrate concentration is a key goal to improve agricultural sustainability. Currently, combined phenotypic verification by histology screening is an effective technical means for corn gene research. Auxins act at each stage of root development through localization patterns and signal transduction. Nitrate supply has been shown to affect auxin accumulation and distribution to regulate corn root development, and remodeling of this root system configuration can promote better plant growth while enhancing NUE. In addition, extensive studies have found that the Auxin Response Factor (ARF) is a key mediator of the auxin signaling pathway and can regulate the expression of downstream auxin signaling genes to regulate growth and development. It has been reported that moderate levels of nitrate can promote auxin signaling and lateral root initiation by regulatory module AtARF, while nitrogen deficiency also activates AtARF6 to promote root hair development to gain more nitrogen. Furthermore, osARF6 has been identified as a key regulator of nitrogen absorption. However, researches on ZmARF6 in aspects of plant regulation and root system development are freshly reported at present, and the function of ZmARF6 in corn nitrogen absorption is unknown. Therefore, the role of ZmARF6 in lateral root elongation and nitrogen absorption is revealed to provide excellent regulation targets and theoretical guidance for corn single-fly efficient cultivation. Disclosure of Invention In order to solve the problems, the invention provides a ZmARF6 protein for regulating and controlling nitrogen absorption of plants, a biological material, application and a method for improving plant properties. The invention is realized by the following technical scheme: a zmahf 6 protein that modulates nitrogen absorption in a plant, the amino acid sequence of the zmahf 6 protein selected from any one of the following: 1) The amino acid sequence is shown as SEQ ID NO. 1. 2) An amino acid sequence having at least 91% identity to the amino acid sequence shown in SEQ ID NO. 1. 3) And (3) connecting a tag to the N end or/and the C end of the amino acid sequence shown in SEQ ID NO. 1 to obtain the fusion polypeptide. 4) An amino acid sequence derived from one or more amino acid residues deleted, substituted, inserted and/or added in the amino acid sequence shown in SEQ ID NO. 1. Preferably, the amino acid sequence has 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity to the amino acid sequence shown in SEQ ID NO. 1. SEQ ID NO:1: MKLSPSASGGLQDQPASPEEAEEHKCLNSELWHACAGPLVSLPSVGSRVVYFPQGHGEQVAASTNKEMEAQIPNYPSLPPQLICQLHNVTMHADAETDEVYAQMTLQPLSPQELKDPFLPAELGTASNQPTNYFCKTLTASDTSTHGGFSVPRRAAEKVFPPLDFNQQPPAQELIAKDLHGNDWKFRHIFRGQPKRHLLTTGWSVFVSAKRLVAGDSVLFIWNDNNQLLLGIRRANRPQTVMPSSVLSSDSMHIGLLAAAAHAASTNSRFTIFYNPRASPCEFVIPMAKYVKAVYHTRISVGMRFRMLFETEESSVRRYMGTITGISDLDPVRWPNSHWRSVKVGWDESTAGEKQPRVSLWEIEPLTTFPMYPSPFALGLKRPWPAGLPSLYGGRGDGLTSSLMWLRDRANPGFQSLNFSGLGMSPWMQPRLDNSLLGLQSDMYQTIAAAAALQSTTKQVPPSAMQFQQPQNIADRSALLSSQILQQVQPRFQQIYPQNLNENKIQGHTQPEYLQVQQQLQRCQSFNEQKPPMHPQQQQQEPQQQQCVQTPQDQQMQEQKHLHNFHSLPDALSAFSQLSPATHSPPSALQTVPAFSHQQNFPDTNISSLSPSTGPSMHGMLGRLPSEAASSLPCVAMNAPVSVSDPWSSKRVAVESVNPCRPHVSPHIEHLDMATCNMPQSSALAPLPGRECLVDEDGCSDPQNHLLFGVHIDSHSLLMQGGIPALQNDNSSGTIPYSTSNFLSPSENDFPLNQPLRSAGCLDESDYLPCAENAEQANQQFATFVKVYKSGTVGRLLDITRFSSYDELRSEVGRLFGLEGQLEDPLRSGWQLVFVDRE