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A kind of transgenic rice with good eating quality and breeding method

A technology of genetically modified rice and eating quality, applied in botany equipment and methods, plant products, using vectors to introduce foreign genetic material, etc., can solve problems such as functions that have not been reported, and achieve the effect of improving eating quality

Active Publication Date: 2014-10-15
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, SSSIIb is a member of the rice soluble starch synthase subfamily, and its function has not been reported yet.

Method used

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  • A kind of transgenic rice with good eating quality and breeding method
  • A kind of transgenic rice with good eating quality and breeding method
  • A kind of transgenic rice with good eating quality and breeding method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Obtaining of Example 1 DNA Sequence and Construction of RNA Interference Vector

[0027] According to the whole genome information of rice, after obtaining the coding sequence of SSSIIb, the homology of this gene with other isoforms was compared on the Grammne website. The specific primer SSSIIbi (upstream primer 5'-AGGCTGATCATGTTGAG-3', SEQ ID NO.2; downstream primer 5'-G TGTTGATTTTGTGTTT-3', SEQ ID NO.3) was designed for the amplification of RNA interference fragments. The amplified sequence is shown as SEQ ID NO.1.

[0028] The young rice leaves of Nipponbare were used to extract the total RNA of rice by the cold phenol method. After the purity and concentration of the RNA were detected by gel agarose current and spectrophotometer, 1ul of total RNA was reverse-transcribed. The reverse transcriptase was purchased from Fermentas Company (Product No.: 00082399). Use 1ul of the reverse transcription product as a template, and carry out routine PCR amplification with sp...

Embodiment 2

[0029] Embodiment 2 Cultivation and Identification of Transgenic Rice Containing RNA Interference Vector

[0030] Rice tissue culture and Agrobacterium-mediated transformation procedures are all carried out by the method established in the inventor's laboratory (Liu Qiaoquan, etc., the establishment of the rice high-efficiency transformation system mediated by Agrobacterium tumefaciens, Plant Physiology, 1998,24 (3) :259~271). Using Nipponbare mature embryos as explants, callus was induced on N6D2 medium as the transformation recipient, and the constructed RNA interference construct was introduced into rice callus through Agrobacterium-mediated; co-cultured on co-culture medium N6D2C After 3 days, the callus was transferred to N6D2S1 selection medium for screening for about 14 days, and then screened on N6D2S2 selection medium, and the times and days of selection were adjusted according to the actual growth of the callus. A lot of resistant calli were obtained, and after pre-di...

Embodiment 3

[0032] The quality performance of embodiment 3 transgenic rice rice

[0033] Measure rice amylose content according to the document method issued by the Ministry of Agriculture with the number NY147-88. Accurately weigh 50 mg of rice flour on a ten-thousandth balance and put it into a 50 ml test tube, add 0.5 ml of absolute ethanol to disperse the sample, and then Add 4.5ml of 1.0mol / l NaOH solution and mix well, put in boiling water bath for 20min, cool to room temperature, and distilled water to make up volume. Take 5ml of digestive juice, add it to a 100mL volumetric flask filled with half a bottle of distilled water, then add 1.0ml of 1.0mol / l acetic acid solution to acidify the sample; add 1.5ml of 0.02% iodine solution, dilute to volume with distilled water, and shake well After standing still for 15 minutes; add 0.5ml of 95% ethanol to 4.5ml of 1N NaOH to replace the sample, and prepare a blank solution. Use the blank solution to adjust the zero point of the spectropho...

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Abstract

The invention discloses a method for reducing apparent amylose content of rice and improving the starch viscosity so as to improve the eating quality of the rice, and belongs to the technical field of organisms. The method is achieved by disturbing soluble starch synthase SSSIIb gene expression in paddy; a ribonucleic acid (RNA) interference carrier of the SSSIIb gene is built; and the transgenic rice of disturbing the SSSIIb gene is obtained by an agrobacterium mediated method. The polymerase chain reaction (PCR) experiment shows that a target gene is integrated into rice genome; and homozygous transgenic rice is obtained in a breeding manner. The amylose content in rice endosperm of the transgenic rice is obviously reduced in comparison with the unconverted parent Nipponbare. A rapid viscosity tester analysis shows that the starch viscosity of the transgenic rice is reduced; the gelatinization temperature is reduced; and the eating characteristics of the rice are obviously improved.

Description

technical field [0001] The invention relates to a method for reducing the apparent amylose content in rice seeds in the field of plant biotechnology, in particular to a method for reducing the amylose content in rice seeds by using RNA interference technology to improve rice quality. Background technique [0002] Plant transgenic technology transfers the target gene isolated from animals, plants or microorganisms into the genome of plants through various methods to make it stably expressed. This technology is applied in agriculture and plays a huge role in crop insect resistance, stress resistance, disease resistance and quality improvement. [0003] Starch is composed of amylose and amylopectin, which are the most important storage substances in rice endosperm. The composition ratio of the two types of starch is one of the most important factors determining the palatability of rice. Generally speaking, the amylose content (AC) of rice with good eating value is 10-20%. Gen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N15/113A01H5/00
Inventor 刘巧泉李娟张昌泉蒋美艳于恒秀顾铭洪
Owner YANGZHOU UNIV
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