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Extraction of high-quality mitochondrial DNA mtDNA is a prerequisite for mitochondrial genome research. Extracting mtDNA from robust yellowing seedlings can reduce cytoplasmic chloroplast genome contamination and allow more rigorous analysis of the mitochondrial genome. This article uses cabbage as an example to introduce a simple and high-quality culture method for yellow seedlings.
First, a 0.1 to 0.2 cm pore size vermiculite was used as a matrix, washed and then laid flat on a clean seedling tray with a thickness of about 1 cm. Seeds of Chinese cabbage are soaked for 1 to 2 hours after sowing. The amount of seed is about 120g/m2. After sowing, the substrate was covered with 0.5 cm and lightly compacted. Seedling trays were placed in a dark room and cultured at a day/night temperature of 26°C/20°C. Spray water to keep the substrate moist every day, cultured for 1 week to obtain a robust yellowing seedlings.
Due to the lack of mtDNA extraction materials, it is difficult to take, and the workload is large. The use of vermiculite with a pore diameter of 0.1 to 0.2 cm is good for ventilation, and can also reduce the "cap ratio" of seedlings and the quality of the roots of the seedlings when they are taken. This will greatly reduce the workload and improve efficiency. The seeding rate varies according to different crops. The seeding rate is too large and it is easy to cause seedling growth. Thin and thin seedlings are not conducive to the extraction of mtDNA, and too thin seeding affects the yield of mtDNA. The temperature-changing treatment in the culture can appropriately reduce the appetite of the yellowing seedlings, and the moisture control in the growth process is the key to the cultivation. The water spray is performed once a day and the substrate can be kept moist.
Tests have shown that this method can be used for 1 week culture to obtain 750g/m2 of yellowing seedlings. 25μg of mtDNA can be extracted and its purity is high enough for follow-up research of mitochondrial genome.
Cytoplasmic male sterility (CMS) exists widely in higher plants. It not only has important value in the utilization of plant heterosis, but also has important theoretical significance in the study of nuclear and cytoplasmic interactions. The maternally inherited characteristics suggest that it may be related to chloroplasts or mitochondria. Existing studies have shown that the CMS of most plants is mainly related to mitochondria.