Gene and method for rapidly judging brassica plants with low cadmium accumulation

A low-accumulation, Brassica technology, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of differences and differences in the enrichment capacity of heavy metal cadmium, and save time and cost , good application prospects, sensitive and specific effects

Active Publication Date: 2017-09-05
HUNAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing studies have shown that there are interspecific differences in the ability of plants to accumulate heavy metal cadmium, and there are also significant differences among different strains within the same species.

Method used

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  • Gene and method for rapidly judging brassica plants with low cadmium accumulation
  • Gene and method for rapidly judging brassica plants with low cadmium accumulation
  • Gene and method for rapidly judging brassica plants with low cadmium accumulation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] To determine the primers of cadmium-suppressing genes whose expression levels are simultaneously increased in roots and leaves of Brassica napus induced by cadmium.

[0030] Select rapeseed mustard with low accumulation of cadmium 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 15mg·kg -1 , the control group does not contain cadmium, the day and night length ratio is 16 / 8h, the day and night temperature is 22 / 16 ℃, and the light intensity is 250μmol m -2 the s -1 , grown in the greenhouse for 30 days.

[0031] The total RNA of roots and leaves of Brassica napus in treatment and control group was extracted, and the high-throughput sequencing technology platform was used for deep sequencing of the transcriptome, and the reads w...

Embodiment 2

[0041] Take the seeds of mustard rape material with low accumulation of cadmium determined by atomic absorption spectrometry, surface sterilize with mercuric chloride, dry the water on the seed surface with sterile absorbent paper, inoculate on the conventional MS solid medium medium, A control group and a treatment group were set up with cadmium concentrations of 0 and 20 mg·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 200 μmol 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 after reverse transcription, and take the Actin gene as the reference gene, repeat 3 times, and take the average number. The reaction conditions and result analysis of quantitative PCR amplification were carried out according to conventional methods.

[0042] Analysis of gene expression ...

Embodiment 3

[0047] The seeds of Brassica napus with low accumulation of cadmium as determined by atomic absorption spectrometry were used as material. After the surface of the seeds was sterilized, blot the water on the surface of the seeds with sterile absorbent paper, inoculate them on the conventional MS solid medium medium, set up the control group and the treatment group, and the cadmium concentrations were 0 and 25 mg 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 200 μmol m –2 the s –1 Cultivate for 15 days under certain conditions, extract the total RNA from the roots and leaves of Brassica napus, perform quantitative PCR experiments after reverse transcription, and use the Actin gene as a reference gene, repeat 3 times, and take the average. The reaction conditions and result analysis of quantitative PCR amplification were carried out according to conventional methods. The primers provided in Ex...

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Abstract

The invention discloses a method for rapidly judging brassica plants with low cadmium accumulation by detecting the up-regulated expression of cadmium inhibition genes. Mustard type rape with low cadmium accumulation is taken as the material, the transcriptomes of roots and leaves of the cadmium processed material and a control material are subjected to sequencing analysis, the mRNA sequences, whose expression amount in leaves and roots is increased by 2.00 times or more, are selected, the functions of the genes corresponding to the mRNA sequences are analyzed, and the genes that can inhibit the absorption and transportation of heavy metals such as cadmium are screened out, namely cadmium inhibition genes. Quantitative PCR is used to analyze the relationship between the gene expression change and the cadmium processing concentrations, 5 cadmium inhibition genes are determined, and the expression amounts of the five genes in the leaves and roots of all processed groups are all increased by 2.00 times or more. The primer of the five genes is utilized to detect the change of gene expression of a brassica plant in a MS culture medium under the induction of cadmium with a concentration of 5-25 mg/kg. If the expression amounts of the five cadmium inhibition genes in roots and leaves are all increased by 2.00 times or more compared with the control group, the brassica plant is a plant with low cadmium accumulation.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a method of screening brassicas with strong cadmium-inhibiting ability by using mRNA expression changes of genes that inhibit the absorption and transport of heavy metals such as cadmium in Brassica plants (abbreviated as "cadmium-inhibiting genes") under cadmium induction. A method for plants of the genus Brassica, which can be used for screening and identification of germplasm resources of plants of the genus Brassica with low accumulation of cadmium. Background technique [0002] With the mining and smelting of mines, the extensive use of fertilizers and pesticides has caused many soils to be polluted by cadmium to varying degrees. Large-scale fallow in lightly polluted soil is not conducive to the sustainable development of agricultural production. In order to ensure the safe production and utilization of lightly polluted soil with cadmium, it is an extremely important wa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6851C12Q1/6895C12Q2600/13C12Q2531/113C12Q2521/107
Inventor 严明理周静张大为向言词冯涛刘丽莉
Owner HUNAN UNIV OF SCI & TECH
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