Reference gene screening in colocasia esculenta corm development process and application thereof

An internal reference gene and bulb technology, applied in the field of internal reference gene screening, can solve problems such as unstable expression and achieve the effect of reducing differences

Active Publication Date: 2021-12-07
GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, with the deepening of research, the expression of many internal reference genes (suc

Method used

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  • Reference gene screening in colocasia esculenta corm development process and application thereof
  • Reference gene screening in colocasia esculenta corm development process and application thereof
  • Reference gene screening in colocasia esculenta corm development process and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0040] This embodiment is the acquisition of the internal reference gene gene-Taro_030192, which is obtained by the following method:

[0041] One, the test material of present embodiment:

[0042] The taro material used in this example is Guiyu No. 2, a new taro variety bred by the Institute of Biotechnology, Guangxi Academy of Agricultural Sciences. In March 2020, it was planted in Guangxi Academy of Agricultural Sciences to build a scientific research base. Sampling was carried out continuously throughout the corm development period, and regular sampling was performed every 30 days. 30d, 60d, 90d, 120d, 150d, 180d, 210d, and 240d after sowing were collected from the corms, a total of 8 groups of samples were marked as YTS1, YTS2, YTS3, YTS4, YTS5, YTS6, YTS7, and YTS8. Each group of samples had three biological replicates. Peel the collected bulbs, cut them into small pieces of about 2 cm, put them into 50 ml sterile cryovials, freeze them with liquid nitrogen and put th...

Embodiment 2

[0087] According to the gene-Taro_030192 with the best stability screened out in Example 1, further verification was carried out to ensure that true and reliable quantitative data can be obtained. Using gene-Taro_030192 as an internal reference gene, the CeAGPS1 gene, a key enzyme gene of starch synthesis in the corm development process, was selected as the target gene, and the expression of the target gene in the eight stages of taro corm development was analyzed, as follows:

[0088] Different stages were extracted respectively: that is, the entire development period of taro, that is, the bulbs of 30d, 60d, 90d, 120d, 150d, 180d, 210d, and 240d after sowing (marked as YTS1, YTS2, YTS3, YTS4, YTS5, YTS6, YTS7, YTS8) The cDNA of the period was used as the template for quantitative PCR, the gene-Taro_030192 gene was used as the internal reference gene, and the ADP-glucose pyrophosphorylase small subunit gene CeAGPS1 was used as the target gene to design primers. The primers are ...

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Abstract

The invention relates to the technical field of plant genes, in particular to reference gene screening and application in the colocasia esculenta corm development process. RNA-Seq sequencing is conducted on samples in the colocasia esculenta corm development period, genes with stable and medium expression quantity are screened as candidate reference genes, the expression stability of each candidate reference gene in the whole development stage of the colocasia esculenta corm is evaluated and analyzed by utilizing software, and the reference gene gene-Taro-030192 which is more stable than a common housekeeping gene is finally screened out by utilizing target gene verification, so that the difference between samples and the difference inside the samples caused by selection of the reference genes are reduced to the greatest extent, and a solid foundation is laid for exploring key genes related to colocasia esculenta corm development by using q RT-PCR in the later period.

Description

【Technical field】 [0001] The invention relates to the technical field of plant genes, in particular to the screening of internal reference genes in the development process of taro bulbs and its application. 【Background technique】 [0002] Taro (Colocasia esculenta), also known as taro, taro, and hairy taro, is a perennial herb of the genus Colocasia in the family Araceae. Taro was originally produced in Southeast Asia, then spread eastward to Far East Asia and Oceania, and westward to the Mediterranean region. It is now widely planted around the world and is the main food of many African and Asian Pacific island countries. It is related to other tubers and root plants. Compared with taro, taro has higher nutritional value. Taro bulbs contain almost twice as many carbohydrates as potatoes and 11% more protein than yams, cassava, and sweet potatoes. With such a rich nutrient content, it is important to understand the genetic basis of their composition. [0003] However, ther...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/11C12Q1/6869C12Q1/6895
CPCC12Q1/6869C12Q1/6895C12Q2600/166C12Q2600/13C12Q2563/107C12Q2545/101C12Q2561/113C12Q2531/113Y02A50/30
Inventor 何芳练董伟清蒋慧萍黄诗宇唐娟
Owner GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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