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Application and application method of OsGPT1 gene

A genome and gene overexpression technology, applied in application, genetic engineering, plant gene improvement and other directions, can solve the problem of OsGPT2 non-expression, and achieve the effect of improving rice varieties and increasing 1000-grain weight.

Active Publication Date: 2019-09-20
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, through our fluorescent quantitative PCR analysis, it was found that only OsGPT1 was highly expressed in rice seeds, while the expression level of OsGPT2 in mature rice seeds was much lower than that of OsGPT1, and OsGPT2 was not expressed in endosperm ( figure 1 )

Method used

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  • Application and application method of OsGPT1 gene
  • Application and application method of OsGPT1 gene
  • Application and application method of OsGPT1 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Localization of OsGPT1

[0033] Although GPTs have been isolated, purified or cloned in rice, Arabidopsis, maize, sorghum, potato and poplar, the subcellular localization of GPTs in living cells is rarely reported. We transformed the P OsGPT1 : OsGPT1-eYFP and fluorescent probe plastid marker (pt-rk CD3-999) (Nelson et al., 2007) were co-transformed into rice protoplasts, and the photos of subcellular localization are shown in Figure 7 . The specific experimental method is as follows:

[0034] (1) Preparation and transformation of rice protoplasts.

[0035] The preparation and transformation of rice protoplasts were to isolate rice protoplasts from the stems and leaf sheaths of yellowed rice seedlings by enzymatic hydrolysis, and transform them into plasmids mediated by PEG (Zhang et al., 2011). The specific method is as follows:

[0036]1. Sterilize the rice seeds and sow them in the solid medium prepared by aseptic Phytagel and MES, and sow about 2-3 ...

Embodiment 2

[0084] Example 2: Obtain OsGPT1 overexpression material, the seeds become larger, and the thousand-grain weight increases

[0085] In planting P OsGPT1 :OsGPT1-eYFP Nip#5 material, it was found that the seeds of this material increased, and the thousand-grain weight increased by about 12.7%. detected POsGPT1 : The expression of OsGPT1 in OsGPT1-eYFP Nip#5 is up-regulated, which is about 19 times the expression of OsGPT1 in Nip ( Figure 6 ). Furthermore, we will P OsGPT1 The :OsGPT1-eYFP Nip#5 material was used as the female parent, pollinated with Nip pollen, and it was found that the hybrid seed of this seed had a large increase compared with the hybrid seed with Nip as the female parent and Nip as the male parent, and has broken through The size of the original chaff ( Figure 6 ).

[0086] The specific experimental method is as follows:

[0087] (1) Extraction method of RNA in starch-rich rice seeds

[0088] TransGen TransZol Plant (catalogue number: ET101-01 ) reag...

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Abstract

The invention belongs to the technical field of biology and relates to application and application method of OsGPT1 gene. In one aspect, the invention provides OsGPT1 gene. In other aspect, the invention also provides application and application of the OsGPT1 gene. The application and the application method have the advantages that it is discovered through studies on the OsGTP1 gene that overexpression of the OsGPT1 gene in rice allows rice seeds to be enlarged and thousand seed weight to be increased; on such basis, the overexpression of the OsGPT1 gene in rice helps regulate the size of rice and thousand seed weight so that improvement of rice varieties is achieved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application and application method of OsGPT1 gene. Background technique [0002] Rice is the most consumed food crop on earth. It is estimated that by 2025, the output of rice must reach 800 million tons to solve the problem of food and clothing for the population on the earth. In 2003, the annual output of rice was 585 million tons (Virk PS, 2004), so from 2003 to 2025, more than two dozen The annual world average rice yield per hectare needs to increase by 5-8.5 tons. As the area of ​​arable land decreases year by year, the area of ​​rice planting is also decreasing year by year. Therefore, the increase in output per hectare by 2025 should be much higher than the expected output in order to solve the problem of food and clothing for the world's population. [0003] The organelle that stores starch in the endosperm of higher plants is the highly specialized plastid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/29A01H5/10A01H6/46
CPCC07K14/415C12N15/8261
Inventor 曲爱丽徐娟
Owner ZHEJIANG UNIV