**7. Shoot culture and micropropagation**

In shoot culture, apical meristem (located at shoot apex and root apex) is cultured, and this culture is also known as meristem culture due to large size of the explant (5–10 mm). Shoot culture is widely applied in horticulture, agriculture and forestry. Murashige, from Morel research team (1960), has significantly established the technique for micropropagation and its further biotechnological application. Due to the small size of explant to be propagated and it occurs *in vitro* as compared to conventional propagation, this technique is also known as micropropagation. Stages in micropropagation are as following:

**Stage I** Selection and preparation of explant – Suitable explant is selected and inoculated into nutrient medium. This step is done is extreme aseptic condition.

**Stage II** Multiplication of cells – Growth of culture takes up to 2 months, followed by repeated subcultures.

**Stage III** Shooting and rooting induction – Culture is transferred to nutrient medium with suitable composition to induce multiple shoots. This step may take about 4 weeks. Subsequently, the culture is transferred on medium suitable for root induction and incubated for about 3 weeks.

**Stage IV** Transfer to soil – After about one month of culture, plantlets are aseptically removed from test tube environment to natural and harsh environment. At this stage, roots should be fully functional in potting soil mix. During this step of transplantation, plantlets have tendency to fail to survive due to desiccation i.e. from 100% humidity of test tubes to low humidity under ambient conditions), unfavorable environment, invasion of soil microorganisms, as well as fails to adapt changes from dependent (artificial medium) to independent environment.
