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
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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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