Betaine aldehyde dehydrogenase 2 fragrance gene in paddy rice, primer of molecular marker 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: 2014-01-22
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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  • Betaine aldehyde dehydrogenase 2 fragrance gene in paddy rice, primer of molecular marker and screening method
  • Betaine aldehyde dehydrogenase 2 fragrance gene in paddy rice, primer of molecular marker and screening method
  • Betaine aldehyde dehydrogenase 2 fragrance gene in paddy rice, primer of molecular marker and screening method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Cloning of the upstream sequence of the initiation codon of betaine aldehyde dehydrogenase 2 gene with 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 aroma-type rice, detection of enzyme activity and analysis of aroma substances

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

[0060] The aroma gene was identified by synthesizing two pairs of primers and performing PCR amplification in two sections. 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 the first nucleotide upstream of the start codon of the coding strand of the aroma gen...

Embodiment 2

[0077] Example 2 Identification of molecular marker primers for the aroma gene

[0078] The betaine aldehyde dehydrogenase 2 gene of non-aromatic rice "Nihonbare" was compared with the new betaine aldehyde dehydrogenase 2 gene, and the difference between the two genes was 8 nucleotides at -1314 upstream of the coding sequence. The difference site of the two betaine aldehyde dehydrogenase 2 genes at -1314 was designed as a molecular marker that can distinguish the two betaine aldehyde dehydrogenase 2 genes, and designed as SEQ ID No.3 and SEQ ID No. 4 sequence primers, as primers for identifying two molecular markers of betaine aldehyde dehydrogenase 2 gene.

[0079] According to the upstream sequence of the non-aromatic rice "Nipponbare" betaine aldehyde dehydrogenase 2 gene coding sequence, a pair of primers designed in the present invention: SEQ ID No. 3 and SEQ ID No. The extracted DNA was amplified, and the molecular weight of the PCR product was expected to be 198 bp; th...

Embodiment 3

[0089] Example 3 Cultivation of high-yielding 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 w...

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Abstract

The invention discloses a new betaine aldehyde dehydrogenase 2 gene enabling paddy rice to have the characteristic of light fragrance, and compared with non-fragrant paddy rice nipponbare, the mutation site is not located in coding strand but belongs to the mutation-type betaine aldehyde dehydrogenase 2 gene at the upstream of the coding strand. Three nucleotides are lost at the -81 site at the upstream of the first nucleotide of the initiation codon of the coding strand, and eight nucleotides are inserted into the -1314 site. A new light-fragrance type paddy rice species is cultivated once homozygous progeny is obtained by transferring the gene into non-fragrant paddy rice via hybridization. The invention also discloses a method for screening the paddy rice possessing the light-fragrance type characteristic and a primer of a molecular marker. The new gene is provided for cultivation of the light-fragrance type paddy rice; also the betaine aldehyde dehydrogenase 2 fragrance gene in paddy rice, the primer of molecular marker and the screening method help to perform assistant selection on plants containing light-fragrance gene, are beneficial to improve selection effect, and also help to identify whether the inbred progeny light-fragrance gene formed at the later stage of breeding is in a homozygous state, so that the breeding target can be realized rapidly and effectively.

Description

technical field [0001] The invention relates to the field of molecular genetics and plant breeding, in particular to a new rice aroma type allele, and the use of this new allele, a molecular marker screening method is used to assist conventional breeding to rapidly select and breed new light aroma type rice. Background technique [0002] The price of aromatic rice is generally much higher than that of non-aromatic rice, making it particularly important 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] Fragrant rice produces a special aroma during cooking, which is favored by most consumers. However, the degree of aroma of fragrant rice is not the same for different people. Some people like strong aroma rice, and some people like light aroma rice. The fragrant rice products currently available in the m...

Claims

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Application Information

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Patent Type & Authority Applications(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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