Porphyra yezoensis ueda TPS (trehalose-6-phosphate synthase) gene and application thereof in enhancing salt tolerance of rice

A technology of seaweed and salt tolerance, applied in the directions of application, genetic engineering, plant genetic improvement, etc., can solve the problems of poor salt tolerance and no simultaneous problems, and achieve the effect of effective transformation system and improved salt tolerance.

Inactive Publication Date: 2012-12-19
QINGDAO AGRI UNIV
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  • Claims
  • Application Information

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Problems solved by technology

[0005] In view of the situation in the prior art that rice has poor salt tolerance and does not have natural gene transformation plants with both TPS and TPP domains, the present invention provides Porphyra zebra TPS gene and its application in improving rice salt tolerance, the TPS The gene has both a TPS domain and a TPP domain. It is a natural dual-domain TPSP gene. Transferring the TPS gene into rice can significantly improve the salt tolerance of rice.

Method used

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  • Porphyra yezoensis ueda TPS (trehalose-6-phosphate synthase) gene and application thereof in enhancing salt tolerance of rice
  • Porphyra yezoensis ueda TPS (trehalose-6-phosphate synthase) gene and application thereof in enhancing salt tolerance of rice
  • Porphyra yezoensis ueda TPS (trehalose-6-phosphate synthase) gene and application thereof in enhancing salt tolerance of rice

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Embodiment 1

[0043] 1. Experimental materials

[0044] 1. Genes, strains and rice varieties

[0045] The present invention has cloned Porphyra versicolor 6-phosphate trehalose synthase (TPS) gene (PyTPS), Escherichia coli DH5α and Agrobacterium tumefaciens strain AGL1 are preserved by the Laboratory of Genetics Laboratory of Qingdao Agricultural University, and the recipient material of the transgene is rice (Oryza sativa L.) variety "Taipei 309" (TP309) (provided by Hunan Hybrid Rice Research Center).

[0046] 2. Plant medium

[0047] There are six media used in rice transformation:

[0048] 1) Callus induction medium: MS+2,4-D 2mg / L+CH 0.3g / L+proline 2.8g / L+sucrose 45g / L+phytage 3.0g / L, pH5.8;

[0049] 2) Callus subculture medium: MS+2,4-D 2mg / L+CH 0.3g / L+proline 0.5g / L+sucrose 30g / L+phytage 3.0g / L, pH 5.8;

[0050] 3) Co-cultivation medium: CC+AS 20mg / L+CH 1g / L+proline 0.5g / L+KT0.8mg / L+NAA 0.2mg / L+maltose 25mg / L+phytage 3.0g / L, pH 5.2 ;

[0051] 4) Screening medium: CC+2,4-D 2mg / L...

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Abstract

The invention discloses a PyTPS (porphyra yezoensis ueda trehalose-6-phosphate synthase) gene which is cloned to construct a plant expression vector of the gene, and the plant expression vector is transferred into the agrobactrium tumefaciens strain, so that an engineering strain is obtained. A rice variety is transformed by utilizing agrobacterium-mediated transformation, so that genetically modified rice is obtained; after growing and acclimatization, a T0 individual plant is obtained, and a T1 seed is collected; a seedling germinated from the T1-generation genetically modified seed receives PCR (polymerase chain reaction) detection and a salt tolerance assay, so that a T1-generation genetically modified salt-resistant strain is obtained, a T2 generation receives a kanamycin resistance assay, PCR detection and Southern hybridization, and results indicate that the PyTPS gene is already integrated into the genome of the genetically modified strain. The PyTPS gene in the invention has a TPS domain as well as a TPP (thiamine pyrophosphate) domain; the PyTPS gene source is safe; the development of the plant morphology of the PyTPS genetically modified rice is normal; the PyTPS gene can be stably inherited in genetically modified plants; and moreover, the transformation system is highly effective.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the Porphyra zebra TPS gene and its application in improving the salt tolerance of rice. Background technique [0002] Rice is the second largest food crop in the world. Rice is the staple food of about half of the world's population. In my country, rice is the largest food crop. The salt tolerance characteristics of rice have been basically explained ([1] Hu Shikai, Tao Hongjian, Qian Qian, et al. Research progress in genetics and molecular breeding of rice salt tolerance. Molecular Plant Breeding, 2010, 8(4) : 629-640; [2] Zhang Suhong, Liu Lixin, Liu Zhongzhuo. Advances in Rice Salt Tolerance Research and Breeding. Northern Rice, 2009, 39(3): 118-121; [3] Qi Dongling, Han Longzhi, Zhang Sanyuan. Rice Identification and evaluation method of salt / alkaline tolerance. Journal of Plant Genetic Resources, 2005, 6(2): 226-231.). The salt-tolerant characteristics of rice are: 1) rice is...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/52C12N15/84A01H5/00C12Q1/68C12N15/11
Inventor 翁曼丽郭宝太王斌王力乔利仙冯艳宾王玮金德敏王晶珊隋炯明
Owner QINGDAO AGRI UNIV
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