Anther culture is a plant tissue culture technique. The anthers developed to a certain stage are inoculated on artificial medium by aseptic technique to change the developmental process of pollen grains in the anther, induce differentiation, and continuously undergo mitosis. The cell mass, which in turn forms a mass of undifferentiated parenchyma, callus, or differentiates into embryoid bodies, and then differentiates the callus into intact plants. Factors affecting anther culture include the following five aspects. NAA http:// 2,4-D http:// MS Culture Media http://
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Main factors affecting anther culture
1. Basic medium <br> The choice of basic medium varies depending on the plant species and variety. MS medium and H medium are suitable for dicotyledonous anther culture; B5 medium is suitable for anther culture of legumes and cruciferous plants; Nitsch medium is suitable for anther culture of Brassica and Mandala; and anthers of cereals Culture media such as N6, C17 and W14 are often used for culture.
2. Inorganic Salts <br> High concentrations of ammonium ions in the medium significantly inhibited the formation of pollen callus. Iron salts are important for the development of pollen embryoid bodies. For example, on low-iron or iron-free medium with a Fe-EDTA concentration of less than 40 μmol/L, tobacco pollen embryos only form multicellular protoplasts (globular embryos) and stop developing.
3. Growth regulators <br> The types and concentrations of growth regulators play a key role in the initiation, division and differentiation of pollen. Cytokinin also promotes pollen differentiation into embryoid bodies, and auxins, especially 2,4-D, promote callus formation, but 2,4-D inhibits callus differentiation into embryoid bodies. High concentrations of auxin can even cause the transformation of Solanaceae pollen embryos into callus. Therefore, callus induction into seedlings should be transferred to differentiation medium without 2,4-D or low concentrations of IAA, NAA and higher concentrations of cytokinin. Anthers of a few plants such as tobacco and rice can form callus or pollen embryos on a medium containing no growth regulator.
4, sucrose <br> sucrose as a carbon source and adjust the osmotic potential of the medium, its concentration has a certain impact on pollen callus induction rate, such as the induction of rapeseed, tobacco pollen callus or embryoid body suitable sucrose concentration It is 2%-3%, rice is 4%-8%, sugar cane is 20%, but for most plants it is 2%-4%. In many plant anthers and pollen cultures, it is preferred to use a higher concentration of sucrose in the callus formation of pollen, and a lower concentration of sucrose should be used in the callus differentiation.
5, add-on <br> Add natural organic substances such as hydrolyzed milk protein, hydrolyzed casein, coconut juice, yeast extract, etc., is a Supplement to the basic medium components and growth regulators, can improve pollen callus and embryoid The body induction rate has a good effect on promoting its growth. In addition, activated carbon can also promote the development of embryoid bodies and increase the yield of pollen plants, and has been confirmed in plants such as tobacco, rapeseed and potato.