Fluorescent quantitative reference genes of different tissues of siberian wildrye and primers and application thereof

An internal reference gene, fluorescence quantitative technology, applied in the field of plant molecular biology, can solve the problem of less research on gene specific expression

Inactive Publication Date: 2019-09-27
LANZHOU UNIVERSITY
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Problems solved by technology

[0004] At present, there are few studies on the specific expression of genes in different tissues of old awn wheat

Method used

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  • Fluorescent quantitative reference genes of different tissues of siberian wildrye and primers and application thereof
  • Fluorescent quantitative reference genes of different tissues of siberian wildrye and primers and application thereof
  • Fluorescent quantitative reference genes of different tissues of siberian wildrye and primers and application thereof

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Embodiment 1

[0035] 1. Test materials

[0036] The test material was ZHN03 wild sage wheat germplasm collected from Zhuoni County, Gansu Province (see Table 1 for specific information). Place the seeds of the ZHN03 germplasm in a petri dish to germinate for 2-3 weeks, and then transplant the seedlings into a flowerpot (10cm in diameter). The flowerpot is filled with turfgrass soil, vermiculite and sand (volume ratio: 2: 2: 1). Plant materials are cultivated in the artificial intelligence greenhouse on the Yuzhong campus of Lanzhou University, where the day and night temperature is 28 / 14°C, the relative air humidity is 30%, and the conventional management methods are adopted to cultivate until the flowering period. Select healthy plants with similar growth, and sample the roots, stems, leaves, flowers and tiller buds of the plants. After sampling, the samples are quick-frozen with liquid nitrogen, and then transferred to the ultra-low temperature refrigerator (-80℃) for storage for RNA extrac...

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Abstract

The invention provides fluorescent quantitative reference genes of different tissues of siberian wildrye: TBP2 and PP2A, the nucleotide sequences of which are as shown in sequence tables SEQ ID NO.1 and SEQ ID NO.2. The two reference gene sequences are originated from siberian wildrye transcriptome sequences and the reference genes are higher in specificity and stability compared with those of universal reference genes on other species. Meanwhile, the two reference genes screened are the most stable reference genes of siberian wildrye in different tissues. Data is true and reliable, thereby laying a foundation for deeply digging the functional genes of siberian wildrye in later period. The fluorescent quantitative reference genes not only solve the current situation that no reference genes are available in current siberian wildrye gene expression analysis, but also can improve the reliability of detection efficiency and detection result. The two reference genes are newly developed genes, so that the quantity of reference genes of plants is enriched, and reference can be also provided for associated researches on other wheat families and lyme grass plants.

Description

Technical field [0001] The invention belongs to the technical field of plant molecular biology, and specifically relates to fluorescence quantitative internal reference genes of different tissues of the old sage wheat and their primers and applications. Background technique [0002] Elymus sibiricus L. (Elymus sibiricus L.) is a perennial, self-pollinated gramineous forage. As a model species of Elymus, it is mostly distributed in temperate regions of the northern hemisphere. The germplasm resources of old awn wheat in our country are also very rich, which are distributed in many provinces in China. Because of its strong adaptability, high grass quality, and good palatability, it is often used in artificial grassland establishment and ecological restoration projects. [0003] Real-time fluorescent quantitative PCR (qRT-PCR) has become an effective tool for detecting gene expression levels because of its fast, efficient, and sensitive advantages. However, this method is also susce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/6895C12Q2600/166C12Q2563/107C12Q2531/113
Inventor 张俊超谢文刚张宗瑜郑玉莹王彦荣
Owner LANZHOU UNIVERSITY
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