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A gene and method for rapid screening of Brassica plants with high accumulation of cadmium

A Brassica and plant technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as difficulty in large-scale promotion, achieve good application value, save time and cost , sensitive and specific effects

Active Publication Date: 2020-11-03
HUNAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing reports have shown that some cruciferous plants have a strong ability to accumulate cadmium. For example, it was found that Indian mustard has a strong tolerance and accumulation ability to cadmium, and has a large biomass. However, the growth and distribution of Indian mustard It has strong regional characteristics and is difficult to promote on a large scale

Method used

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  • A gene and method for rapid screening of Brassica plants with high accumulation of cadmium
  • A gene and method for rapid screening of Brassica plants with high accumulation of cadmium
  • A gene and method for rapid screening of Brassica plants with high accumulation of cadmium

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A method for rapid screening of high cadmium-rich Brassica plants, comprising the steps of:

[0031] 1) Using the mustard rapeseed with high cadmium enrichment determined by atomic spectroscopic absorption method as the material, take plump seeds, sterilize the surface with mercuric chloride, blot the water on the surface of the seeds with sterile absorbent paper, and inoculate them on conventional MS solid on culture medium. Set control and treatment, the concentration of cadmium in the treatment group is 30mg·kg -1 , the control group does not contain cadmium, the day-to-night ratio is 16 / 8h, the day-to-night temperature is 22 / 16°C, and the light intensity is 250μmol m -2 the s -1 , grown in the greenhouse for 30 days. The total RNA of roots and leaves of Brassica napus in the treatment and control group was extracted, and the high-throughput sequencing technology platform was used to perform deep sequencing of the transcriptome, and the reads with an average quali...

Embodiment 2

[0039] To determine the primers of cadmium-rich genes whose expression levels are simultaneously increased in roots and leaves of Brassica napus under cadmium induction.

[0040] Select the identified high cadmium-enriched Brassica napus as the material, take the plump seeds, sterilize the surface with mercury chloride, blot the water on the surface of the seeds with sterile absorbent paper, and inoculate them on the conventional MS solid medium . Set control and treatment, the concentration of cadmium in the treatment group is 30mg·kg -1 , the control group does not contain cadmium, the day-to-night ratio is 16 / 8h, the day-to-night temperature is 22 / 16°C, and the light intensity is 250μmol m -2 the s -1 , grown in the greenhouse for 30 days.

[0041] The total RNA of roots and leaves of Brassica napus in the treatment and control group was extracted, and the high-throughput sequencing technology platform was used to perform deep sequencing of the transcriptome, and the rea...

Embodiment 3

[0052] 1) The cadmium enrichment capacity of the mustard rape material with high cadmium enrichment determined by atomic absorption spectrometry is as follows, when the cadmium concentration is 20 mg·kg -1 The average enriched cadmium concentration in the plants reached 217.2 mg·kg when cultured in MS medium for 15 days -1 .

[0053] 2) Take the seeds of the above-mentioned mustard type rapeseed, after surface sterilization with mercuric chloride, blot the moisture on the surface of the seeds with sterile absorbent paper, inoculate them on the conventional MS solid medium medium, set the control group and the treatment group, the cadmium concentration 0, 20mg·kg respectively -1 . When the day-to-night ratio is 16 / 8h, the day-to-night temperature is 22 / 16°C, and the light intensity is 200μmol m –2 the s –1 Cultivate for 15 days under the above conditions, extract the total RNA of the root and leaf of the above-mentioned Brassica napus, perform quantitative PCR experiment af...

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Abstract

The invention discloses a method for rapidly screening brassica plants capable of highly enriching cadmium by utilizing cadmium-enriching gene expression detection. The method comprises the following steps: taking mustard type oilseed rape capable of highly enriching the cadmium as materials and carrying out transcriptome sequencing on roots and leaves of cadmium treatment materials and control materials, so as to obtain genes which correspond to an mRNA (messenger Ribonucleic Acid) sequence and have expression amount, increased by 2.00 times or more, in the roots and the leaves; analyzing correlation between changes of the expression amount of the genes and cadmium enrichment and finding out six genes which have expression amount, increased by 2.00 times or more, in the roots and the leaves of all treatment groups, wherein the expression amount has significant positive correlation with cadmium enrichment amount; determining that the genes are cadmium-enriching genes. According to the method disclosed by the invention, primers of the six genes are used for detecting changes of gene expression in the roots and the leaves of the brassica plants in an MS culture medium with the cadmium concentration of 10mg.kg<-1> to 50mg.kg<-1>, and the brassica plants with the expression amount of the six cadmium-enriching genes in the root and the leaves, which is increased by 2.00 times or more when being compared to a control group, belong to the plants capable of highly the enriching cadmium. The detection method has high sensitivity and strong specificity and can be used for rapidly detecting a cadmium-enriching capability of the brassica plants.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for screening Brassica plants with strong cadmium-enriching ability by using the expression changes of cadmium absorption-related gene mRNA in early plants under the induction of cadmium, and the method can be used for Brassica genus Germplasm identification of plant cadmium uptake capacity. Background technique [0002] With the mining and smelting of metal mines, the extensive use of fertilizers and pesticides has caused many soils to be polluted by cadmium to varying degrees. Phytoremediation of heavy metal-contaminated soil is a relatively feasible method. Hyperaccumulative plants are used to absorb cadmium in moderately or slightly polluted soil, and the plants are harvested and treated to reduce the cadmium content in the soil and achieve the purpose of soil safety utilization. . Existing reports have shown that some cruciferous plants have a strong ability to accumu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2600/13C12Q2545/114C12Q2521/107
Inventor 严明理黄婷张大为向言词冯涛刘丽莉
Owner HUNAN UNIV OF SCI & TECH