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Herba myc3 gene and its application

A technology of sagegrass and genes, applied in the field of genetic engineering, can solve problems such as the difficulty of sagegrass management

Active Publication Date: 2020-07-28
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Long-term continuous and unscientific application of herbicides can easily lead to the emergence of weed resistance. In some areas of my country, wheat and rapeseed have developed serious resistance to herbicides, and with the emergence of non-target resistance, the weed Governance is more difficult

Method used

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  • Herba myc3 gene and its application
  • Herba myc3 gene and its application
  • Herba myc3 gene and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Acquisition of MYC3 gene

[0075] The test selects sensitive population WC10-4 and resistant population WC10-10 collected from Jiangsu Province.

[0076] 1. Transcriptome sequencing

[0077] Select 3 samples of sensitive and drug-resistant Smilax 24 hours after treatment with methylsulfuron-methyl and 3 samples without treatment, and send them to the company for transcriptome sequencing.

[0078] 2. Acquisition of MYC3 gene

[0079] According to the transcript sequencing results, differentially expressed genes were screened, and the differentially expressed genes involved in the jasmonic acid signaling pathway could be determined through significant enrichment of pathways, and gene information and gene sequences were obtained from the sequencing results.

[0080] The nucleotide sequence of the MYC3 gene of Lemongrass is shown in SEQ ID NO:1.

Embodiment 2

[0081] The extraction of embodiment 2 total RNA

[0082] The test was collected from sensitive population WC10-4 and drug-resistant population WC10-10 in Jiangsu Province.

[0083] 1. Extraction of total RNA

[0084] Select 3 samples of Sensitive and drug-resistant Smilax 24 hours after treatment with Mesosulfuron-methyl and 3 samples without treatment, and extract total RNA respectively according to the instructions of the kit to obtain the corresponding total RNA solution.

[0085] 2. RNA agarose gel electrophoresis

[0086] Take 3uL total RNA solution, use 1% agarose gel for electrophoresis detection, the voltage is 160V, the electrophoresis time is 20min, and use gel imaging to scan. like figure 1 As shown, channel 1 from the left is Maker, channels 2 and 3 are both WC10-4; channels 4 and 5 are both WC10-10, and two clearly distinguishable main bands (28S and 18S) can be seen in ultraviolet detection, Indicates that the RNA is intact without degradation.

Embodiment 3

[0087] Example 3 Screening of primers for detecting the expression level of Smilax MYC3 gene

[0088] 1. Reverse transcription (synthesis of cDNA first strand)

[0089] (1) Prepare the reverse transcription system shown in Table 1:

[0090] Table 1

[0091]

[0092] (2) Carry out the reverse transcription reaction on the PCR instrument according to the following conditions:

[0093] 15min at 42°C, 5s at 85°C, and keep warm at 12°C.

[0094] (3) Dilute the product of (2) 3 times and set aside.

[0095] 2. Primer Design

[0096] (1) Obtain the MYC3 gene sequence of Smilax chinensis by transcriptome sequencing.

[0097] (2) Design fluorescent quantitative PCR primers.

[0098] In step (2), use Oligo 7.0 software and Primer5.0 software to design primers. The designed primers are screened for specific primers at NCBI, and 3 pairs of primers are obtained after screening. The primer sequences, amplified fragment lengths and annealing temperatures are shown in Table 2:

[00...

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Abstract

The present invention provides a beckmannia syzigachne MYC3 gene and an application thereof. A nucleotide sequence of the beckmannia syzigachne MYC3 gene is shown in SEQ ID NO:1. The present inventionalso provides fluorescent quantitative PCR primers (SEQ ID NO:2-3) used for detecting an expression level of the beckmannia syzigachne MYC3 gene and an established method for detecting the expressionlevel of the beckmannia syzigachne MYC3 gene based on the primers, thereby judging resistance of beckmannia syzigachne to herbicides of sulfonylurea (especially mesosulfuron). The method has advantages of simplicity, high sensitivity and strong specificity, and has important application prospects in detection and research of mesosulfuron non-target resistance of the beckmannia syzigachne. The present invention discloses for the first time a relationship between the beckmannia syzigachne MYC3 gene and the resistance to sulfonylurea herbicides. Detection of the MYC3 gene expression level can beused to identify whether the beckmannia syzigachne produces metabolic resistance and has a guiding significance for timely detection of the resistance, guidance of scientific drug use of farmland andfurther spread of alleviation of the drug resistance.

Description

technical field [0001] The present invention relates to the technical field of genetic engineering, in particular, relates to the MYC3 gene and application thereof. Background technique [0002] Long-term continuous and unscientific application of herbicides can easily lead to the emergence of weed resistance. In some areas of my country, wheat and rapeseed have developed serious resistance to herbicides, and with the emergence of non-target resistance, the weed Governance is more difficult. Currently, studies have shown that transcription factors act as regulators in non-target resistance (Christophe Délye. Unravelling the genetic bases of non-target-site-based resistance (NTSR) toherbicides: a major challenge for weed science in the forthcoming decade.PestManag Sci 2013;69:176–187), studies have shown that the transcription factor MYC2 has a positive regulatory role in plant antioxidant capacity (Dombrecht B, Xue G P, Sprague S J, et al.Myc2 differentially modulates divers...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/11C12Q1/6895
Inventor 崔海兰王京京李香菊陈景超黄兆峰
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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