Method for improving rice potassium ion efflux antiporter

A technology of antiporter protein and potassium ions, which is applied in the field of improving rice potassium ion efflux and reverse transport, and can solve problems such as yield reduction and photosynthetic efficiency reduction

Inactive Publication Date: 2011-11-23
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Soil salinization is the main reason for the reduction of production of various food crops in the world. Salt stress will reduce the efficiency of photosynthesis, resulting in a reduction in production

Method used

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  • Method for improving rice potassium ion efflux antiporter
  • Method for improving rice potassium ion efflux antiporter
  • Method for improving rice potassium ion efflux antiporter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: Obtaining and morphological observation of kea1 mutant plants

[0029] This mutant was used in our laboratory 60 Japonica rice 9522 seeds were induced by Coγ-rays at a dose of 280Gy. Backcrossing a midrib adaxial-specific whitening mutant in the F2 generation of mutagenesis for three generations obtained a recessive single-gene regulated stably inherited mutant kea1. All plant materials were planted at the experimental base of Shanghai Academy of Agricultural Sciences.

[0030] Morphological observation of kea1 mutant plants. Such as figure 1 , The phenotype comparison between wild type and mutant kea1 shows that the leaf color of wild type 9522 is normal (left) (A, B, C), while the leaves of kea1 mutant are specifically whitened on the adaxial surface of the midrib (right) (A, B, C); the number of chloroplasts on the adaxial surface of the midrib of the wild-type leaves is large, and the arrangement is regular (top) (D); the number of chloroplasts on ...

Embodiment 2

[0031] Example 2: Kea1 mutant and wild-type 9522 ion concentration determination

[0032] (1) Processing of samples. The roots, stems, leaves, leaf sheaths and flowers of the reproductive growth period ( Figure 2A ), and the aerial parts and roots of the 28d seedlings under the conditions of normal growth and salt treatment (treated with NaCl 300mM and KCl 300mM respectively for 3d) ( Figure 2B ), first in 5mM CaCl 2 The solution was quickly washed twice in pure water, once in double distilled water, and three times in ultrapure water. Oven at 90°C until constant weight.

[0033] (2) Determination of potassium and sodium ion concentrations in rice. After the material is baked, weigh it with an electronic balance, then put it into a 50ml centrifuge tube, add 0.1M acetic acid solution, put it in a water bath at 90°C for 3 hours, and mix it upside down for 5-6 times in the water bath. Pour the extract into a 2ml centrifuge tube, centrifuge at 12,000rpm for 5-10min, take th...

Embodiment 3

[0034] Example 3: Location and cloning of the KEA1 gene

[0035] (1) Positioning groups. Kea1 was crossed with the indica rice line Guangluai 4, and the F2 generation was obtained by selfing. Among them, the plants with adaxial-specific whitening of the midrib of the leaves were selected as the positioning population.

[0036](2) Rice DNA extraction. Using the improved CTAB method. The simple steps are as follows: Take 0.1-0.2 grams (about half a piece) of leaves and put them in a small mortar, add an appropriate amount of liquid nitrogen, grind them to powder immediately, put them into a 2ml centrifuge tube, add 700ul of 1.5xCTAB solution preheated at 100℃ in In a centrifuge tube, mix carefully and place in a 56°C water bath. Take out the centrifuge tube after 20 minutes, add an equal volume of chloroform / isoamyl alcohol, mix vigorously, centrifuge (13000rpm) for 10 minutes, take the supernatant into a new tube, add Mix 900ul of absolute ethanol and place at -20°C for more...

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Abstract

The invention belongs to the technical field of biological engineering and discloses a method for improving rice potassium ion efflux antiporter. A function mutant strain of rice potassium ion efflux antiporter protein is obtained by a KEA1 gene for mutation control of rice potassium ion efflux and protein encoded by the gene; meanwhile, gamma rays are utilized to treat 9522 strain of japonica rice, and mutants of which the leaf veins are whitened are selected in F2 generation after the rice is planted so as to obtain the function mutant strain of the rice potassium ion efflux antiporter protein. By utilizing the characteristic that the protein participates in chloroplast potassium ion efflux antiporter and controls rice potassium ion transport by a transgenic technology, a new rice stress-resistant line is generated by inhibiting the expression of the protein, and has important application value in agricultural production.

Description

technical field [0001] The invention relates to an application in the technical field of bioengineering, in particular to a method for improving rice potassium ion efflux and reverse transport by using the KEA1 gene and its encoded protein. Background technique [0002] Rice is one of the most important food crops in my country. More than 60% of the population in China takes rice as the staple food. It is the largest rice producer and consumer in the world. The annual sown area of ​​rice in my country is 30 million hectares, accounting for 20% of the world; the output is 185 million tons, accounting for nearly 1 / 3 of the world, and the output per unit area is 6.35 tons / ha. 65% higher than the global average yield of 3.85 t / ha. Rice has always remained at about 40% of the total grain output in my country, accounting for nearly half of the country. [0003] Photosynthesis is the basis of crop yield. As the site of photosynthesis, chloroplast development regulation mechanism ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N1/21C12N15/82A01H5/00C12R1/01
Inventor 张大兵米华玲时楠梁婉琪袁政
Owner SHANGHAI JIAO TONG UNIV
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