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Tobacco microRNA detection method under phytophthora parasitica stress

A technology of blackleg bacteria and detection method, applied in the field of tobacco microRNA detection under the stress of blackleg bacteria

Inactive Publication Date: 2021-04-09
KUNMING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many scholars have used high-throughput sequencing technology to study the miRNA expression characteristics of tobacco before and after topping, drought stress and chromium stress, and to identify the miRNA transcription profile of the characteristic variety Hongda. Stress response miRNAs have been systematically studied

Method used

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Experimental program
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Effect test

Embodiment

[0033] The present invention adopts the tobacco seed provided by Yunnan Academy of Tobacco Agricultural Sciences to implement, and the pathogenic bacteria is No. 0 physiological race of flue-cured tobacco highly pathogenic black shank bacteria.

[0034] First prepare the spore suspension:

[0035] Prepare oat culture medium, the oat culture medium is calculated according to weight percentage, take 30g of oatmeal, 15g of agar powder, 1000ml of deionized water, mix well and set aside. The isolated and purified tobacco black shank physiological race 0 was cultured on oat medium for 21 days with 0.1% KNO 3 Soak the mycelium with 1% glucose mixture, and put it in a 26°C incubator for 72h, then place it in a refrigerator at 8°C for 20min and take it out. A large number of zoospores will be released in 10-30min at room temperature, filtered, and examined under a microscope. Adjust the concentration of zoospores to 1 x 10 3 ~1×10 4 pieces / ml.

[0036] Tobacco Stem Sample Processin...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a tobacco microRNA detection method under phytophthora parasitica stress. The method is as follows: preparing a spore suspension of phytophthora parasitica physiological race, manually injecting the spore suspension of phytophthora parasitica physiological race into the basal part of a tobacco stem to obtain a detection sample, detecting phytophthora parasitica stress response miRNA, constructing a miRNA library in response to tobacco phytophthora parasitica stress through the research of the detection technology, adopting an Illumina / Solex deep sequencing technology to mine and analyze miRNA information related to tobacco phytophthora parasitica resistance, so that the understanding of the tobacco miRNA function is further enriched.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for detecting tobacco microRNA under the stress of blackleg pathogen. Background technique [0002] In plants, gene expression is regulated by multiple mechanisms at the transcriptional and post-transcriptional levels to ensure normal plant growth and development and to respond to changing environmental conditions. Post-transcriptional gene regulation mediated by 21-24nt long small RNA (sRNA) is one of the most important regulatory mechanisms in plants. Small RNAs in plant cells are very complex and can be classified into different regulatory types according to their sources and biological functions, which also implies that they have a wide range of regulatory functions in plants. Small interfering RNAs (siRNAs) and miRNAs are two of the most widely studied regulatory small RNAs. siRNAs originate from double-stranded RNA (dsRNA) formed by different mechanisms. Another typ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895
CPCC12Q1/6895C12Q2600/13C12Q2600/178
Inventor 黄飞燕叶贤文余磊刘佳妮陈泽斌柯艳果魏环宇方宇王启宇赵娅红薛至勤
Owner KUNMING UNIV
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