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Aspergillus candidus AcGDH protein for improving drought and alkali stress tolerance of rice, gene and application of Aspergillus candidus AcGDH protein

A drought stress and tolerance technology, applied in applications, genetic engineering, plant genetic improvement, etc., can solve problems such as infeasibility and reproductive isolation barriers, and achieve the effects of reducing yield loss, promoting absorption and assimilation, and reducing ammonia toxicity

Active Publication Date: 2021-02-05
HUNAN UNIV
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  • Abstract
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Problems solved by technology

However, due to reproductive isolation barriers between species, it has become simply not feasible to employ conventional breeding methods to substantially increase NUE in rice

Method used

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  • Aspergillus candidus AcGDH protein for improving drought and alkali stress tolerance of rice, gene and application of Aspergillus candidus AcGDH protein
  • Aspergillus candidus AcGDH protein for improving drought and alkali stress tolerance of rice, gene and application of Aspergillus candidus AcGDH protein
  • Aspergillus candidus AcGDH protein for improving drought and alkali stress tolerance of rice, gene and application of Aspergillus candidus AcGDH protein

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Embodiment Construction

[0055] The experimental methods mentioned in the following examples are conventional methods unless otherwise specified.

[0056] 1. Cloning of AcGDH gene

[0057] The conserved region of the GDH gene was found through the sequence of the sequenced fungal GDH gene, and degenerate primers were designed using primer5. The forward primer of the degenerate primer starts from the start codon of the open reading frame, and the sequence is 5'-ATGTCCCACCTGCCTTTCGARCCNGARTT-3'; the reverse primer of the degenerate primer is located behind the 3' end of the open reading frame, and the sequence is 5'-CCCCACCAGTCACCCCTGGTCRTGCATNGC -3'. Then the AcGDH gene was cloned by RT-PCR using the cDNA of the fungus Aspergillus candidus as a template. According to the method of Chen et al. (2006), the AcGDH gene was cloned into the GATEWAY entry vector pGWC and then sequenced (Chen QJ, Zhou HM, Chen J, WangXC.Using a modified TA cloning method to create entry clones[J].Anal Biochem,2006 , 358:120...

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Abstract

The invention belongs to the field of plant genetic engineering, and discloses cloning and application of a fungus Aspergillus candidus NADP (H) dependent glutamate dehydrogenase gene AcGDH capable ofimproving drought and alkali stress tolerance of rice. Studies show that in vitro forward reaction enzyme activity of AcGDH protein is greater than reverse reaction enzyme activity, i.e., AcGDH tendsto convert alpha-ketoglutaric acid into glutamic acid by utilizing NH4+. Meanwhile, it is found that affinity of AcGDH to NH4+ is higher than that of rice endogenous OsGDH4. The AcGDH gene is heterologously overexpressed in the rice through a genetic engineering technology, and tolerance of the transgenic rice under drought and alkali stress conditions is improved. Therefore, the AcGDH gene can be used for cultivating new drought-resistant and alkali-stress-resistant rice varieties.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, in particular, the invention relates to the heterologous expression of the NADP(H)-dependent glutamate dehydrogenase gene AcGDH of the fungus Aspergillus candidus in rice (Oryza sativa L.) , can improve rice drought and alkali stress tolerance, and rice nitrogen use efficiency and yield. Background technique [0002] Rice is the most important food crop in the world. More than half of the world's population uses rice as the staple food. In China, rice is the food crop with the largest planting area and the highest yield. In the process of plant growth, it is inevitable to suffer from various adverse environmental stresses. The most widely affected are abiotic stress factors such as drought, low temperature, and saline-alkali stress (Liu Cong, Dong Laai, Lin Jianzhong, Liu Xuanming. Plants under adversity stress Research progress on active oxygen metabolism and regulation mechanism [J]. L...

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Application Information

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IPC IPC(8): C12N9/06C12N15/53C12N15/11C12N15/82A01H5/00A01H5/10A01H6/46
CPCC12N9/0016C12N15/8273C12N15/8271C12Y104/01002
Inventor 刘选明林建中燕璐唐冬英朱咏华
Owner HUNAN UNIV
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