Arabidopsis thaliana, also known as Arabidopsis thaliana, belongs to the Cruciferae family and is a biennial herb. It has a small plant shape, with a tile diameter of 5cm and a height of 30cm; the growth area is small and easy to cultivate; the genome is small, only 5 chromosomes, 157 million base pairs, 27000 genes; fast growth, short cycle, about It takes 6 weeks; it can self-pollinate or cross between different germplasm lines. It has many seeds, and each plant can produce thousands of seeds per generation. Therefore, Arabidopsis is one of the model plants in plant science, including genetics and plant development research (Figure 1). In 2000, Arabidopsis became the first plant whose genome was completely sequenced.
Although Arabidopsis thaliana has a short growth period and is easy to cultivate, according to different experimental needs, in the cultivation process, the control of various conditions such as light, moisture and humidity is different. Set different growth environments to meet different experimental needs.
1. Normal cultivation Simple Arabidopsis cultivation can be used for gene expression, organ development, gene mutation and other studies.
1. First put the Arabidopsis seeds on the filter paper, sterilize with 70% alcohol, and then treat with anhydrous alcohol (you can also choose 20% bleach and 0.05% Triton X-100 non-ionic surfactant for sterilization Processing for 20 minutes). Rinse 5 times with sterile water, then spot the seeds on MS medium, pH 5.9, plus 0.9% agar and 2% sucrose.
2. Put the seeds into vernalization under 4 ℃ dark conditions for about 48h-72h, germinate under long-day or short-day conditions, and then transfer them to the PERCIVAL AR series incubator (Figure 2). Use the Intellus controller to edit the simple day and night mode, set the light time 16 hours (dark 8 hours) or light 14 hours (dark 10 hours); optional Q23 + continuous light intensity adjustment device, set the light intensity to 100-150umol / m2.s; set the temperature inside the cabinet to be 20-22 ℃ under light conditions and 18-20 ℃ under dark conditions; the optional humidity control system is used to set the relative humidity in the cabinet to 70%.
3. It takes about 7 to 10 days to emerge, and then it is transferred to the soil for cultivation and watered by bottom irrigation (using the capillary action of the soil). Before the first watering, the soil can be completely dried, and then water every 7 to 10 days.
4. The soil can be configured with a ratio of 10: 1: 1 nutrient soil, vermiculite and perlite, and can also be directly mixed with compost, vermiculite and peat. During the growth process, ordinary flower fertilizer can be used to fertilize once or not.
5. When the plant grows to a certain height, the stem can be cut to produce more inflorescences, which can be used for transformation studies or to obtain more seeds (Figure 3).
2. Special cultivation In order to speed up the growth of Arabidopsis thaliana, after moving into the soil for cultivation, you can set long daylight, use Intellus controller, edit the program to set 24h light, light intensity 100-250umol / m2.s, temperature lasts 22- 23 ℃.
Arabidopsis thaliana root treatment culture can reduce the light time to 8-12h, increase the light intensity to 100-350umol / m2.s, the temperature is 22-23 ° C under light conditions, 18-20 ° C under dark conditions, and the other culture conditions are the same Normal cultivation.
Reference 1, David M. Brown, Leo AHZeef, Joanne Ellis, et al. Identification of Novel Genes in Arabidopsis Involved in Secondary Cell Wall Formation Using Expression Profiling and Reverse Genetics. The Plant Cell 2005, 17, 2281-2295
2.John C. Sedbrook, David W. Ehrhardt, Sarah E. Fisher, et al. The Arabidopsis SKU6 / SPIRAL1 Gene Encodes a Plus End-Localized Microtubule-Interacting Protein Involved in Directional Cell Expansion. The Plant Cell 2004, 16, 1506 -1520
3. Nobuhiro Suzuki, Ludmila Rizhsky, Hongjian Liang, et al. Enhanced Tolerance to Environmental Stress in Transgenic Plants Expressing the Transcriptional Coactivator Multiprotein Bridging Factor 1c. Plant Physiology 2005,139, 1313-1322
4. Sholpan Davletova, Ludmila Rizhsky, Hongjian liang, et al. Cytosolic Ascorbate Peroxidase 1 Is a Central Component of the Reactive Oxygen Gene Network of Arabidopsis. The Plant Cell 2005, 17, 268-281
5. Stephen Tottey, Maryse A. Block, Michael Allen, et al. Arabidopsis Chl27, Located in both envelope and thylakoid membranes, is required for the synthesis of protochlorophyllide. PNAS 2003, 100, 16119-16124
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