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CN-116254273-B - Gene for regulating and controlling plant flowering phase, recombinant vector and application

CN116254273BCN 116254273 BCN116254273 BCN 116254273BCN-116254273-B

Abstract

The application belongs to the field of plant growth, relates to a gene for regulating and controlling plant flowering phase and disease resistance, and in particular relates to a gene for regulating and controlling plant flowering phase, a recombinant vector and application thereof. In the research process of disease-resistant breeding, a NBS-LRR gene NbLOV of Nicotiana benthamiana is cloned. The result of the belief analysis shows that the gene is closely related to the response of the plant to fungal pathogen, and the experiment of knocking out the gene verification gene function through gene editing discovers that the knock-out vector aiming at NbLOV1 constructed by the method and the plant created by the agrobacterium-mediated transformation method have obvious differences compared with the control, namely, after NbLOV is knocked out, the bud and flowering time of the Nicotiana benthamiana are obviously earlier than those of the control. The mutant plant with NbLOV gene knocked out shows stronger resistance to fusarium, only leaves are slightly yellow and slightly wilted, and the tobacco of the control shows damage to vascular bundles, serious wilting and necrotic spots on leaf surfaces.

Inventors

  • PING WENLI
  • LI XUEJUN
  • SUN JIPING
  • SUN HUAN
  • LI XUHUI
  • ZU HUANXIN
  • HOU YONG
  • ZHANG XUEKE
  • Geng Shengna

Assignees

  • 河南省农业科学院烟草研究所

Dates

Publication Date
20260508
Application Date
20221110

Claims (7)

  1. 1. A protein for regulating the flowering phase of plants, characterized in that the protein belongs to a protein comprising a nucleotide binding site and a leucine-rich repeat; the amino acid sequence of the protein is shown as SEQ ID No. 2.
  2. 2. A gene encoding the protein of claim 1, wherein the nucleotide coding sequence is shown in SEQ ID No. 1.
  3. 3. A recombinant vector comprising the gene of claim 2.
  4. 4. The recombinant vector according to claim 3, wherein the recombinant vector is a CRISPR gene-based recombinant vector.
  5. 5. The method of shortening the flowering phase of a plant comprising the recombinant vector of claim 4, wherein the plant is Nicotiana benthamiana.
  6. 6. The use of the recombinant vector according to claim 4 for enhancing fusarium disease resistance of plants, wherein the plants are Nicotiana benthamiana.
  7. 7. The use according to claim 5 or 6, characterized in that the application step is to transfer the recombinant vector into a sample plant by means of agrobacterium-mediated genetic transformation, creating a reduced flowering period and/or fusarium disease resistant line.

Description

Gene for regulating and controlling plant flowering phase, recombinant vector and application Technical Field The invention belongs to the field of plant growth, relates to a gene for regulating and controlling the flowering phase of plants, in particular to a gene, a recombinant vector and application for regulating and controlling the flowering phase of plants. Background Plant flowering is affected by a variety of intrinsic factors including photoperiod, endogenous hormones, etc., and the external environment. The flowering phase is one of key factors influencing the growth period of tobacco and determining the effective leaf number of tobacco, is an important link in the life cycle of flowering plants, and marks the transformation of plants from vegetative growth to reproductive growth. Flowers, fruits, seeds are the main products of many crops, and thus, the time to market and the economic value of the crops are determined by the flowers. The plants undergo long-term selective evolution to form a complex and fine regulation network to respond to various internal and external signals (such as photoperiod, temperature, age, gibberellin and the like), so that flowering is regulated, and the plants can bloom in the optimal time. The flowering phase regulation is to artificially change the external environment of the plant or change the genes or internal components of the plant by biotechnology according to the flowering habit and growth and development rule of the plant so as to realize the early or late flowering. And the flowering phase is regulated, so that the economic value of the plant is improved. The application of the flowering phase regulating technology can prevent early-flowering fruit trees from being damaged by or less by frost in early spring and late spring, and can supply vegetables, fruits, flowers and crops in reverse seasons, such as cucumbers, tomatoes, strawberries and the like in winter. In addition, for the cross breeding work, the flowering phase control technology can lead the hybrid parents which are not encountered in the original flowering phase to bloom in the same period, solve the contradiction between cross pollination and time, be beneficial to the development of the breeding work and have important agricultural and economic values in the regulation of the plant flowering phase. Patent 202110374691.1 discloses a transcription factor LbNAP for delaying lily florescence, which is characterized in that the expression quantity of LbNAP transcription factor is reduced to prolong plant florescence, a virus induction gene silencing vector containing LbNAP transcription factor gene segments based on tobacco brittle virus TRV is constructed, patent 201911317235.2 discloses a gene NtDUF599 related to low-temperature early flowering of tobacco, the gene is used for realizing early flowering period under low-temperature stress, the gene for shortening plant flowering period is always a research hot for breeding work, the growth period of germplasm materials is shortened by means of leading the florescence to advance, and further more than one year is realized, and the breeding efficiency is improved. Disclosure of Invention In order to solve the technical problems, the invention provides a gene, a recombinant vector and application for regulating and controlling the flowering phase of plants. The technical scheme of the invention is realized as follows: In the research process of disease-resistant breeding, the NBS-LRR gene NbLOV1 in Nicotiana benthamiana is cloned, and the sequence is shown as SEQ ID No. 1. The result of the biological analysis shows that the gene is closely related to the response of plants to fungal pathogens, and the experiment of verifying the gene function by gene editing and knocking out the gene shows that the bud and flowering time of the Nicotiana benthamiana are obviously earlier than that of a control after NbLOV a is knocked out. In one aspect, the application claims a gene for regulating flowering phase in plants, the gene being a protein comprising a nucleotide binding site and a leucine-rich repeat. Further, the amino acid sequence of the gene is shown as SEQ ID No. 2. The nucleotide sequence of the gene is shown as SEQ ID No. 1. Recombinant vector with the above gene as target gene. Preferably, the recombinant vector is a CRISPR-based gene editing recombinant vector. Further, the recombinant vector is a pORE-Cas9 editing vector or a pDC45 editing vector. The application of the recombinant vector in shortening plant florescence. The recombinant vector is applied to cultivation of new varieties for more than one year. The recombinant vector is applied to enhancing the fusarium disease resistance of plants. The application steps are that the recombinant vector is transferred into a sample plant through agrobacterium-mediated genetic transformation technology to prepare a variety with shortened flowering period and/or fusarium disease resistance. Preferab