Primer and method for detecting molecular marker of rice aroma allele

An allele and rice technology, which is applied in the fields of genetics and plant breeding, can solve problems such as heavy workload, and achieve the effect of simplifying preliminary operation steps and achieving breeding goals.

Inactive Publication Date: 2014-12-10
SHANGHAI NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

So far, 13 allelic types of aroma genes have been found to be caused by the mutation of the betaine aldehyde dehydrogenase 2 gene. Therefore, it is necessary to explore what kind of mutation the betaine aldehyde dehydrogenase 2 gene selected as the parent of the fragrant rice belongs to. The type of workload is still very large

Method used

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  • Primer and method for detecting molecular marker of rice aroma allele
  • Primer and method for detecting molecular marker of rice aroma allele
  • Primer and method for detecting molecular marker of rice aroma allele

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 The establishment and detection method of molecular markers capable of identifying various types of aroma genes

[0036] The fragrant rice "Yinxiang 18" with the mutation type of the second exon and the fragrant rice "Wxiang 99075" with the mutation type of the seventh exon were used as representatives of the fragrant rice varieties with the mutation type of the coding chain, and the rice "Nanhai 138" was used as the coded Representative of aromatic rice varieties with upstream mutation types, extracted 3 kinds of rice genome DNA, designed 8 pairs of primers, and cloned the betaine aldehyde dehydrogenase 2 gene (badh2) directly related to the aroma of these 3 kinds of rice The 7666bp sequence was sequenced and analyzed, and the betaine aldehyde dehydrogenase 2 gene sequence (AP004463.2) of the non-aromatic rice "Nipponbare" published in Genbank was also checked.

[0037] Comparing the betaine aldehyde dehydrogenase 2 gene sequences of 4 kinds of rice, the arom...

Embodiment 222

[0042] Example 222 Identification of molecular markers of aromatic rice CAPS (Alu Ⅰ)

[0043] In order to illustrate that the CAPS (Alu Ⅰ) molecular marker designed by the present invention can be applied to the identification of aroma genes in more aromatic rice, we have collected 19 varieties from China, the United States, and Thailand, and the mutation sites of aroma genes are the second respectively. Fragrant rice with exon and exon 7: "Suhu Xiangjaponica", "Dahua Xiangjaponica", "Tongyun Japonica", "Rosin Zaojaponica", "Gehu Xiangnuo", "99983 Fragrant Rice ", "Xiangjing 05-7", "Wuxiangjing 14", "49② Fragrant Rice", "Yinxiang 18", "Wuyun 2645", "Zixiangnuo 861", "07-08 Fragrant Rice", "Xiangdao No. 1", "Zhongxiang No. 1", American fragrant rice "Della" naked rice, "Thailand fragrant rice", "Cxiang 517", "Dalixiang", and the 3 used in Example 1 Fragrant rice: "Wxiang 99075", Japanese fragrant rice "Nanhai 138" and "B 1 There are 22 kinds of fragrant rice in total, and two...

Embodiment 3

[0045] Example 3 Identification of Non-perfume Rice CAPS (Alu Ⅰ) Molecular Marker

[0046] Since the yield of most aromatic rices is low, in the molecular marker-assisted breeding of new aromatic rices, non-aromatic rices with higher yields are generally selected for hybridization with aromatic rices, and then the hybrid F1 plants are combined with High-yielding non-perfume rice is backcrossed for several generations. Before each backcross, molecular markers for identifying aroma genes should be used to detect the offspring of the backcross, and heterozygous plants containing aroma genes should be selected to continue backcrossing with non-perfume rice. Therefore, for the molecular marker loci to identify aroma genes, it should be possible to distinguish the aroma genes and non-aroma genes of the aroma rice and non-aroma rice as parents.

[0047] For the identification of whether the +898 site of the first nucleotide downstream of the initiation codon of the non-perfume rice b...

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Abstract

The invention discloses a primer and a detection method which can detect a screened molecular marker aiming at aromatic rice with multiple types of aroma genes and amplify a molecular marker of multiple types of betaine aldehyde dehydrogenase genes 2 (Badh2 / badh2) which are directly related to rice aroma formation. The detection method comprises the following steps: (1) extracting total DNAs of the rice as a template, and adding the primer for performing polymerase chain reaction (PCR) amplification; (2) performing restriction endonuclease Alu I digestion on a PCR product; (3) performing gel electrophoresis detection on an enzyme-digested product. The situation that only a mutation type of the aroma allele can be identified in the process of detecting a molecular marker of aroma genes in the past is broken, plants containing different types of the aroma genes can be selected in an assisted mode, a mutation type of the aroma allele of a parent aromatic rice variety is not required to determined before the operation of performing novel aromatic rice variety breeding on the molecular marker in an assisted mode, and pre-operation steps of molecular marker-assisted breeding are simplified, so that a breeding objective for breeding a novel aromatic rice variety is rapidly and effectively realized.

Description

technical field [0001] The invention relates to the fields of genetics and plant breeding, specifically screening molecular markers of aromatic rice, primers of the molecular markers and a detection method, which are used to assist conventional breeding in rapid, convenient and accurate selection of aromatic rice. Background technique [0002] Basmati rice is favored by consumers because of its pleasant fragrance during cooking. The price of fragrant rice is generally higher than that of non-perfume rice varieties. With the improvement of people's living standards, the current market demand for fragrant rice is increasing. These have greatly promoted people's genetic research on the aroma characteristics of rice, and also accelerated the process of breeding new varieties of aromatic rice. [0003] Although there were differences in the research results on the genetics of rice aroma in the past, most scholars believe that rice aroma is controlled by a recessive major gene (...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 李建粤时亚琼许言福张建中王英存
Owner SHANGHAI NORMAL UNIVERSITY
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