**3.4 Perform regeneration of the rescued** *Bothriocline auriculata* **species specimens into many seedlings and propagation of the species seedlings for in-situ conservation**

The extracted BAT plant parts and seeds were regenerated into mass seedlings using tissue culture and in-vitro seed germination techniques respectively [14]. But because the species is endemic and critically endangered, a few plants were got whose seedling and multiplication rates by the natural process of germination, − to raise the required number of seedlings for in-situ conservation, is not 100% guaranteed.

Thus, artificial micropropagation of the collected seedlings or plant tissues by tissue culture [15], were included as a backup process just in case the seeds generated by germination are not enough to raise the required number of seedlings over successive generations. But because the tissue culture method despite multiplying seedlings in mass numbers at a faster rate and producing disease-free plantlets, the method is challenged by the production of genetically identical seedlings [15]. To increase genetic diversity, artificial crossings between the tissue cultured plantlets with those produced from the natural germination process were made. Afterwards, the progeny/ F1 generation of the crossed seedlings was back-crossed with germinated parents [21], to increase genetic diversity and encourage additional crossovers during recovery and regeneration in the natural environment.

The *Bothriocline auriculata* species specimens were collected and rescued from their fragile ecosystems along the slopes of Mount Elgon for multiplication into many seedlings for propagation and conservation. The collected specimens were put into specialized aerated specimen sampling bags. The bags containing the species specimens were in a cool box and taken to the Makerere University Tissue Culture laboratory for processing. In the lab, the specimens were cleaned with tap water, disinfected with ethanol and cut into replicate 25 smaller vegetative portions of length 2–3 cm. The specimens were regenerated and multiplied into many seedlings using plant tissue culture and micropropagation protocols. The vegetative species specimens were successfully regenerated and multiplied into at least 150 juvenile seedlings under aseptic conditions, artificial lighting and growth hormones. The seedlings produced are identical to the parent donor *Bothriocline auriculata* plants. **Figure 4** shows some of the pictures for species tissue cultured *Bothriocline auriculata* seedlings.

**Figure 4.** *Photos of the* Bothriocline auriculata *seedlings tissue cultured in situ.*

*Integrated Conservation Approaches for Rescuing, Regeneration and Adaptive Management… DOI: http://dx.doi.org/10.5772/intechopen.106893*

Over three tissue cultures, seedling propagation cycles were performed during the generation of BAT specimens. A total of 150 seedlings were propagated and regenerated during tissue culture propagation cycles.
