３） The sterilized plating |
| You have to deal seeds and/or seed pod in sterile in the clean Bench or the clean box basically. You may make it by yourself. But you have to sterilized inside of the box enough by medicine and you had better use a gas burner or a spirit lamp before your using. In the case of closed system of the clean box, you don't need a gas burner because you can work through rubber gloves without opening door. In this case, after you put the materials you are going to use including seeds and/or young plants, you sterilize inside of the box completely by medicine like chloroform or chlorine and wait about 30 min until the medicine still down.In case of a clean bench, you had better use a gas burner and you had better work beside the burner. Don't trust the clean bench too much. |
（Ａ）The plating for un-burst pod |
ａ；How to sterilize the seed pod |
|
|
| （１）Take off the abract covered the seed pod, （２）Cut the stem leaving about 3 cm from the pod. （３）Burn out the fine fiber on the pod by fire. This procedure will be very important to excluding the fiber and wax on the pod. You will be much easier to sterilize it latter. |
|
|
| （４）Immerse the pod in 75% of Ethanol for about
1 min. This procedure will make it easier to sterilize latter and not is sterilizing procedure itself. Further more, Alcohol is not useful for sterilizing for fungi and some kinds of bacteria. （５）Immerse the pod in a few % of chlorine solution for 5 min. There is the relationship between the time and the concentration of chlorine solution. But for the beginners, you had better use the higher concentration of chlorine solution in this case. You may use the original dense solution. Be careful, you have to keep it in sterile after this procedure. |
| （６）Open the autoclaved filter paper covered by aluminum] foil. You may open the foil by your dirty fingers but don't touch the inside of the foil and filter papers. Take the autoclaved tweezers by dirty fingers , take the sterilized pod to the center of filter papers. You take off the chlorine solution from the pod by the filter paper. |
ｂ；Cutting the seed pod |
 |
（７）Open the filter paper and cut the center of pod by the autoclaved knife like in the figure. Be careful not to cut the filter paper down the pot. |
ｃ；Continue to plating |
