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A rice betaine aldehyde dehydrogenase 2 fragrance gene, primers for molecular markers and screening method

A technology of betaine aldehyde dehydrogenase and aroma gene, applied in the fields of molecular genetics and plant breeding

Inactive Publication Date: 2016-07-06
SHANGHAI NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no research report on the mutation of the upstream sequence of the coding region of the betaine aldehyde dehydrogenase 2 gene

Method used

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  • A rice betaine aldehyde dehydrogenase 2 fragrance gene, primers for molecular markers and screening method
  • A rice betaine aldehyde dehydrogenase 2 fragrance gene, primers for molecular markers and screening method
  • A rice betaine aldehyde dehydrogenase 2 fragrance gene, primers for molecular markers and screening method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Cloning of the upstream sequence of the start codon of the betaine aldehyde dehydrogenase 2 gene with a mutation in the upstream of the coding chain and comparative analysis with the corresponding sequence of non-aromatic rice; Gene expression of betaine aldehyde dehydrogenase 2 in aromatic rice, detection of enzyme activity and analysis of aroma substances

[0059] Firstly, using the conventional methods of molecular biology, taking "Shangshixiang 2" rice as an example, the upstream sequence of the coding chain of betaine aldehyde dehydrogenase 2 gene was cloned and sequenced.

[0060] The aroma gene was identified by synthesizing two pairs of primers and amplifying in two stages of PCR. The sequences of the first pair of upstream and downstream primers are: 5'-GATGTATAATATCGGTCG-3' and 5'-GGGAGTAGTATAGCCAAC-3' (SEQ ID No.5 and SEQ ID No.6). These two primers are homologously paired with -1442 and -511 of the first nucleotide upstream of the initiation codon...

Embodiment 2

[0077] Embodiment 2 identifies the molecular marker primer of this fragrance type gene

[0078] Compared with the betaine aldehyde dehydrogenase 2 gene of the non-fragrant rice "Nipponbare" and the new betaine aldehyde dehydrogenase 2 gene, there is a difference of 8 nucleotides at -1314 upstream of the coding sequence. The difference site of the two betaine aldehyde dehydrogenase 2 genes at -1314 is designed as a molecular marker that can distinguish the two betaine aldehyde dehydrogenase 2 genes, and primers such as SEQIDNo.3 and SEQIDNo.4 are designed , as primers for the identification of two molecular markers of the betaine aldehyde dehydrogenase 2 gene.

[0079] According to the sequence deduction of the upstream sequence of the betaine aldehyde dehydrogenase 2 gene coding sequence of non-fragrant rice "Nipponbare", a pair of primers designed by the present invention: SEQIDNo. Amplified, the molecular weight of the PCR product is expected to be 198bp; the novel betaine ...

Embodiment 3

[0089] Example 3 Cultivation of high-yield light-scented rice resistant to stripe leaf blight

[0090] "Shangshixiang No. 2" rice is bred from the hybridization of "Nanhai 138" fragrant rice and "Jiahua No. 1" non-perfume rice. Although the yield of "Shangshixiang 2" rice is much higher than that of "Nanhai 138", its yield is the same as that of the conventional rice "Xiushui 128" mainly planted in Shanghai a few years ago. However, the yield of "Xiushui 128" rice was significantly lower than that of "Xiushui 134" conventional rice. Both "Nanhai 138" fragrant rice and "Jiahua 1" do not contain the rice stripe blight resistance gene, so the "Shangshixiang 2" rice produced by hybridization is also not resistant to rice stripe blight. "Xiushui 134" rice has stripe leaf blight resistance gene qSTV11 KAS , resistant to rice stripe disease. To this end, we used the "Shangshixiang 2" rice as the light-flavored fragrant rice parent, and crossed and multi-generation backcrossed with...

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Abstract

The present invention discloses a new betaine aldehyde dehydrogenase 2 gene that can make rice show a light-flavored characteristic. Compared with the non-perfume rice "Nipponbare", the mutation site is not located on the coding chain, but belongs to the upstream mutation of the coding chain. Types of betaine aldehyde dehydrogenase 2 genes. There was a deletion of 3 nucleotides at position -81 upstream of the first nucleotide of the initiation codon of the coding strand, and an insertion of 8 nucleotides at -1314. By crossing this gene into non-perfume rice and obtaining homozygous offspring, a new light-scented rice variety can be bred. The invention also discloses a method for screening rice with the light-scented characteristic gene and molecular marker primers. The invention provides new genes for cultivating light-scented rice, and can also assist in selecting plants containing light-scented genes, which is conducive to improving the selection effect, and can also determine whether the light-scented genes of self-bred offspring formed in the later stage of the breeding process are in a homozygous state Identification to achieve breeding goals quickly and efficiently.

Description

technical field [0001] The invention relates to the fields of molecular genetics and plant breeding, specifically a new rice aroma allele, and using the new allele, a molecular marker screening method is used to assist conventional breeding to rapidly select new light aroma rice. Background technique [0002] The price of fragrant rice is generally much higher than that of non-fragrant rice, which makes it occupy a particularly important position in the international rice trade. With the improvement of people's living standards, the current market demand for fragrant rice is increasing, which also promotes the genetic and breeding research of fragrant rice. [0003] Basmati rice produces a special aroma during cooking, so it is favored by most consumers. However, for the degree of fragrance that fragrant rice has, different people's acceptance is not the same. Some people like strong-flavored rice, while others like light-flavored rice. At present, the fragrant rice produ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/53C12N15/11C12Q1/68
Inventor 李建粤时亚琼王俊峰窦兰兰黄菊
Owner SHANGHAI NORMAL UNIVERSITY
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