SNP functional molecular marker of lodging-resistant gene of japonica rice and application thereof

A functional molecule and anti-lodging technology, applied in the field of plant molecular biology, can solve the problems of long time-consuming identification of lodging resistance phenotypes, inaccurate manual identification results, and unsafe experimental operations, so as to achieve accurate and reliable detection results and shorten the breeding cycle , the effect of a wide range of application prospects

Inactive Publication Date: 2017-10-03
YANGTZE NORMAL UNIVERSITY
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  • Abstract
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Problems solved by technology

[0005] In view of the above-mentioned deficiencies in the prior art, the purpose of the present invention is to provide a SNP functional molecular marker of the lodging resistance gene of japonica rice, so as to solve the problems of long time-consuming and high manpower input in identification of the lodging resistance phenotype of japonica rice
[0006] The invention also provides a primer set and KASP method for detecting SNP functional molecular markers, which solves the problems of unsafe experimental operation, complicated operation and inaccurate manual identification results, etc.

Method used

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  • SNP functional molecular marker of lodging-resistant gene of japonica rice and application thereof
  • SNP functional molecular marker of lodging-resistant gene of japonica rice and application thereof
  • SNP functional molecular marker of lodging-resistant gene of japonica rice and application thereof

Examples

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Effect test

Embodiment 1

[0033] Example 1: Phenotypic verification of natural genetic material of SNP marker K_06g_SCM2-1

[0034] 23 natural genetic materials were selected for verification, including 11 lodging-resistant materials and 12 lodging-sensitive materials.

[0035] 1. Genomic DNA was extracted from japonica rice leaves by simplified CTAB method.

[0036] (1) Freeze-dry or dry the japonica rice leaves, put an appropriate amount of leaves into a 2.0mL centrifuge tube, add two steel balls, and grind them on a tissue grinder;

[0037] (2) Add 750 μL of CTAB solution, oscillate to homogenate, and incubate at 65°C for 0.5-1 h;

[0038] (3) Cool to room temperature, add 750 μL of chloroform:isoamyl alcohol (24:1) in the fume hood and mix by inverting 3-4 times;

[0039] (4) Centrifuge at 12000rmp for 10min, transfer 500μL supernatant to a new 1.5mL centrifuge tube;

[0040] (5) Add an equal volume of isopropanol solution and shake gently to mix, precipitate at -20°C for more than 1 hour, centr...

Embodiment 2

[0055] Example 2 Phenotype verification of F2 segregation population of SNP marker K_06g_SCM2-1

[0056] Detect the individual plant genotype of the F2 segregation population crossed by the donor parent Habataki and the recipient parent Sasanishiki;

[0057] 1. The genotyping method is the same as in Example 1. For the genotyping results of 80 F2 isolated populations, see figure 2 .

[0058] 2. Select two homozygous genotypes for phenotypic identification. During the period from the booting stage to the heading of rice, remove the top leaves of the plants, measure the mechanical strength and diameter of the plant stems, and comprehensively evaluate the lodging resistance of the plants.

[0059] The genotype and phenotype data of homozygous individual plants are shown in Table 3.

[0060] Table 3 The phenotype identification results of the F2 segregation population of marker K_06g_SCM2-1

[0061]

[0062]

[0063] Note: "R" indicates that the plant phenotype is resist...

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Abstract

The invention discloses an SNP functional molecular marker of a lodging-resistant gene of japonica rice and application of the marker. The lodging-resistant gene is SCM2; the SNP functional molecular marker is K_06g_SCM2-1 and is positioned at the 27481645th basic group of the No. 6 chromosome of rice; the polymorphism of the SNP molecular marker is T/G; the sequence of the marker is shown in SEQ ID No.1; the base group at the 55bp site is T or G and is positioned in a promoter region of the gene SCM2. The SNP functional molecular marker disclosed by the invention can be used for predicting the lodging resistance of a japonica subspecies material of rice in an early stage (seedling stage or embryonic stage), and can be used for performing accurate screening, thereby promoting genetic improvement and breeding of lodging resistance traits of the japonica rice. According to the invention, a KASP method is adopted to detect an SNP site, and the detection method is accurate and reliable, easy and convenient to operate, and suitable for application in high-throughput commercialized molecular breeding.

Description

technical field [0001] 本发明涉及植物分子生物学技术领域,具体涉及一种粳稻抗倒伏基因的SNP功能分子标记及应用。 Background technique [0002] 传统系谱法选育是水稻育种家们普遍采用的方法,通过育种家们的多年努力,培育和创制了大量高产、高抗和优质的水稻新品种。然而,结合表型鉴定的传统育种模式存在表型把控不准、遗传群体需求量大、人力成本高、育种周期长等缺陷,很难实现大规模商业化集成育种或材料遗传改良。伴随着分子生物学和生物信息学的发展,分子标记辅助选择育种(Molecular Assisted Selectoin,MAS)显现了巨大的技术优势,实现了遗传基础与目标性状的结合,选择含有目标基因的材料(单株)进行组配,实现了目标性状的精准改良,能有效回避传统育种方法的技术壁垒。 [0003] 单个核苷酸变异(Single Nucleotide Polymorphism,SNP)在物种的基因组上广泛存在,包括:单个碱基的转换或颠换、单个碱基(片段)插入或缺失等形式,针对这些多态性位点逐步兴起了新一代的SNP分子标记技术。 针对控制目标性状的基因开发功能SNP标记,将性状与基因型相关联进而实现高效、精准育种。 现阶段,适用于SNP检测的手段主要有凝胶电泳和荧光定量PCR,但检测过程需要酶切、电泳及测序等,操作复杂,而且PCR产物的气溶胶和EB对人体的危害,对环境的 pollute. 竞争性等位基因PCR(Kompetitive AlleleSpecific PCR,KASP)已陆续应用于分子辅助育种、目标性状基因定位、种子纯度及真实性鉴定等工作,具有成本低、通量高、实验操作安全和荧光信号采集数据准确等优势。 [0004] 种植水稻的时候,水稻倒伏是一个极常见的问题,水稻倒伏严重影响水稻生产品质和制约产量的提高:一是植株早衰,产量降低。水稻倒伏后,植株相互重叠,受光面积减少;同时由于透风透光差,加之田间湿度较大,下部叶片枯黄腐烂,功能叶面积下降。此外,由于机械损伤,植株营养物质交换受阻,进一步加强了根系和叶片的早衰,影响光合作用,光合物质减少,导致灌浆不饱满,千粒重下降,秕谷增加,结实率降低。二是米质下降。由于灌浆不足,稻米心腹白比例较大,碎米率增加。稻穗长期处于高湿环境中,部分稻粒发生霉变、发芽,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 李文博曾秀红姚启伦李昌满谭永忠刘义华
Owner YANGTZE NORMAL UNIVERSITY
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