Method for screening MicroRNAs of nilaparvata lugens with effect on oryza sativa resistance adaptability

A brown planthopper, adaptive technology, applied in the field of bioengineering

Active Publication Date: 2018-09-07
INST OF FOOD CROPS HUBEI ACAD OF AGRI SCI
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  • Method for screening MicroRNAs of nilaparvata lugens with effect on oryza sativa resistance adaptability
  • Method for screening MicroRNAs of nilaparvata lugens with effect on oryza sativa resistance adaptability
  • Method for screening MicroRNAs of nilaparvata lugens with effect on oryza sativa resistance adaptability

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[0069] 1) Synthesis of miRNA mimic, miRNA inhibitor, and control strands Through differential analysis of miRNA expression, we found 26 miRNAs with large differential expression. Among them, the miRNA expression level of the BPH population (biotype I population) on the susceptible rice TN1 was higher, and the miRNA expression level of the NPH population (biotype Y population) on the insect-resistant rice YHY15 was lower. The differentially expressed miRNAmimic, inhibitor, negative control for mimic (NCm) and negative control for inhibitor (NCi) synthesized in this experiment were designed and synthesized by Guangzhou Saizhe Biotechnology Co., Ltd.

[0070] 2) Microinject and artificially feed miRNA mimic and miRNA inhibitor to BPH to verify the function of miRNAs.

[0071] The purified miRNA mimic and miRNA inhibitor were slowly injected into the intersegmental membrane of the prothorax and mesothorax ventral surface of the newly eclosioned brown planthopper using the instrume...

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Abstract

The invention discloses a method for screening MicroRNAs of nilaparvata lugens with effect on oryza sativa resistance adaptability, and relates to the bioengineering technology. The method comprises the following steps: (1) sample collection; (2) sequencing of small RNA; (3) pretreatment of sequencing raw data; (4) sequence alignment; (5) new miRNA prediction; (6) analysis of miRNA differential expression; (7) prediction of differential miRNA target genes; (8) synthesis of miRNA mimic, miRNA inhibitor and control chain; (9) microinjection and artificial feeding of miRNA mimic and miRNA inhibitor to nilaparvata lugens and verification of the function of miRNAs; (10) preliminary study on oryza sativa resistance adaptability change of the nilaparvata lugens after microinjection and artificialfeeding. Small RNAs of two nilaparvata lugens groups are deeply sequenced with a high-throughput sequencing technology, and are analyzed, identified and predicted according to subsequent bioinformatics, differences between the two nilaparvata lugens groups on insect-susceptible oryza sativa TN1 and insect-resistant oryza sativa YHY 15 are compared, the miRNA associated with host resistance adaptability is analyzed, screened and discovered, and the new method is provided for oryza sativa insect-resistant breeding.

Description

technical field [0001] The invention relates to bioengineering technology, in particular to a method for screening the adaptability of MicroRNAs in brown planthopper's resistance to rice. Background technique [0002] Rice (Oryza sativa) is one of the world's major food crops, and more than half of the world's population uses rice as a staple food. Experts estimate that by 2025, the world's rice production will have to increase by 60% to meet the demand. my country is a big country in rice production. The annual rice planting area is about 30 million hectares, accounting for 1 / 4 of the world's total rice planting area. Therefore, the safety of rice planting is inseparable from my country's economic development and people's lives. [0003] Brown planthopper (Nilaparvata lugens BPH) is an insect belonging to the family Homoptera (Homoptera: Delphacidae), also known as brown rice lice. The brown planthopper mainly absorbs the sap of the rice phloem, and the first to be dam...

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6869G06F19/22G06F19/18
Inventor 闸雯俊游艾青徐华山李培德李三和周雷刘凯陈志军杨国才刘林
Owner INST OF FOOD CROPS HUBEI ACAD OF AGRI SCI
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