Plant expression vector containing chloroplastic glutamine synthetase genes, construction and use thereof

Abstract

The invention discloses a special plant expression vector pK2-35S-Prbcs-*T-GS2 which contains an arabidopsis chloroplast type glutamine synthetase gene (GS2) and can improve the metabolic ability of plant nitrogen. A RT-PCR method is utilized to clone a GS2 gene from a model plant Arabidopsis, a promoter of a Rubisco small subunit with the light-inducible promoter is used to control the overexpression of the GS2 gene in plant leaves, and the GS2 gene is transformed into wild tobacco through a leaf disk transformation method. The experimental result shows that the GS2 gene can perform normal transcription in transgenic tobacco and the growth condition of a strain with the GS2 gene is better than that of a check plant (un-transformed wild type) under low nitrogen nutrition condition, which indicates that the overexpression of the GS2 gene can improve the efficiency of a GS / GOGAT (glutamine synthetase / glutamate synthetase) way to a great extent, thereby improving the capability of the assimilation of the plant nitrogen. The special vector has the advantages that the special vector can be used for molecular breeding of crops, improves the utilization rate of nitrogen and the tolerance to low nitrogen stress, and obtains higher yield under the condition of small nitrogenous fertilizer application or even no nitrogenous fertilizer application.

Description

technical field [0001] The invention belongs to the field of plant engineering, and in particular relates to a plant expression vector pK2-35S-Prbcs-*T-GS2 of a chloroplast-type glutamine synthetase gene GS2, a construction method thereof and its application in transgenic crops. Background technique [0002] Nitrogen is the mineral nutrient element with the largest demand for plants, and it is one of the nutrient elements necessary for the normal growth and development of plants. At the same time, it is also one of the most restricted nutrient elements for agricultural production and ecosystem growth and development (Jones et al., 2005, Soil Biology and Biochemistry, 37:413–423). In order to obtain as much nitrogen as possible for growth and development, plants have formed several ways to obtain nitrogen: soil nitrogen absorption, atmospheric nitrogen fixation (legumes), dry and wet deposition nitrogen absorption, insect predation, etc. , for general plants (non-legume plan...

AI Technical Summary

Problems solved by technology

However, the nitrogen in the soil is difficult to meet the needs of crop growth and development, so in production, the method of applying nitrogen fertilizer is widely used to meet the needs of crops

However, due to ammonia volatilization, nitrate nitrogen loss, and denitrification, the nitrogen utilization rate of the applied nitrogen fertilizer is very low. The national average nitrogen utilization rate in the current season is less than 30%. Soil and water pollution, causing harm to the environment on which people live

However, because the trait of high nitrogen use efficiency is difficult to evaluate in field conditions, it is difficult to select crop varieties with high nitrogen use efficiency by traditional breeding methods. Ability to become the most natural and environmentally friendly way

At present, there is no plant expression vector containing the chloroplast-type GS2 gene present in the Arabidopsis genome regulated by the tomato 1,5-bisphosphate ribulose carboxylase oxygenase small subunit gene (rbcS) and the 35S double promoter. to report

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