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Oryza sativa qVEalpha-t3-1 site molecular marker and application thereof
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A molecular marker and rice technology, which is applied in the fields of rice breeding and molecular biology, can solve the problems of limited research and inability to be directly used by humans, and achieve the effects of speeding up breeding, quick and easy prediction, and improving rice quality
Active Publication Date: 2022-01-04
ZHEJIANG NORMAL UNIVERSITY
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However, tocotrienols only exist in monocots and a few dicots, and are mainly distributed in the seed coat, which cannot be directly used by humans.
[5]
, so cultivating and promoting rice varieties with higher tocotrienol content is of great significance to the improvement of human health
[0004]
Currently, studies on α-tocotrienol in rice grains are limited
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Embodiment 1
[0046] Example 1. Mapping of the main QTL for regulating the content of α-tocotrienol in rice grains
[0047] 1. Acquisition of experimental materials
[0048]Reyan 2 was used as the recipient parent, and the rice variety Huazhan was used as the donor parent for hybridization, and the single-seed method was used (that is, the F1 was treated with bagging a single plant until the phenotype of the offspring lines did not segregate). Finally, 120 stable genetic lines (F12, all lines with stable phenotypes) were obtained to form a recombinant inbred line RIL population, such as figure 1 .
[0049] Select 60 seeds of the parent and each strain (F12), soak the seeds for 2 days after surface disinfection, then wrap them in a moist towel, place them in a 37°C incubator for 48 hours, and select seeds with consistent dew and whiteness for sowing. After 30 days, the parents with similar growth conditions and 24 seedlings of each line were selected for transplantation. All rice materials...
[0058] Set molecular markers Indel Tococ3-1 and molecular markers IndelTococ3-2 at the upstream and downstream of QTL site qVEα-t3-1, and design primers;
[0060] Upstream primer: 5'-GTTGGCTTGGGGGAAATAAT-3' (SEQ ID NO.1);
[0061] Downstream primer: 5'-GGGACAGCCAAGATATGAGC-3' (SEQ ID NO.2);
[0062] The primer pair for molecular marker Indel Tococ3-2 is:
[0063] Upstream primer: 5'-CTCAGCATCTGGCCGCTAT-3' (SEQ ID NO.3);
[0064] Downstream primer: 5'-CTTGGAATCAAATCGCTTGC-3' (SEQ ID NO.4).
[0065] Take the rice leaves of parents Reyan No. 2, Huazhan and its F1 generation and RIL population (obtained in step 1 of Example 1), extract genomic DNA, and use the above-mentioned molecular markers to perform PCR amplification on its genomic DNA;
[0066] PCR reaction system: upstream primer (10 μmol) 1 μL, downstream primer (10 μmol) 1 μL, DNA template (>100 ng...
Embodiment 3
[0075] Example 3, Application of QTLs Related to Rice α-Tocotrienol Content in Rice Breeding
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Abstract
The invention discloses a major QTL for regulating and controlling the content of alpha-tocotrienol in oryza sativa. The major QTL is positioned on a 09 chromosome of the oryza sativa and is named as qVEalpha-t3-1; and the genetic distance is 71.29-98.33 cM, and the physical distance is 16630835-22938953 bp. The invention also discloses a molecular marker of the major QTL for regulating and controlling the content of alpha-tocotrienol in the oryza sativa. According to the major QTL, molecular markers Indel Tococ3-1 and Indel Tococ3-2 are utilized to detect QTL related to the content of alpha-tocotrienol in an oryza sativa variety or strain, so that the oryza sativa with high content of alpha-tocotrienol is bred through the molecular markers.
Description
technical field [0001] The invention belongs to the technical field of rice breeding and molecular biology, and specifically relates to a marker of a main effect QTLs site for regulating the content of α-tocotrienol in rice and its application. Background technique [0002] Rice (Oryza sativa.L) is one of the most important food crops in China and the world, and has made great contributions to solving the problem of food shortage. In order to better solve the problem of food and clothing, the development of rice in my country has gone through stages such as dwarf breeding, utilization of heterosis and super rice breeding programs, and major breakthroughs have been made in high-yield breeding [1] . With the improvement of people's living standards, the breeding goal has shifted from a single yield to a combination of high-quality, multi-resistance and high-yield traits, especially the quality traits of rice have attracted more and more attention. [2] . [0003] Vitamin E (...
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