Callus Induction And Differentiation From Leaf And Rooting Differentiate From Stem Tip In Reaumuria Trigyna

Reaumuria trigyna (Reaumuria Linn genus, family Tamaricaceae), an endangered and endemic small shrubs, distributing in western Inner Mongolia desert, has a vital ecological function due to its remarkable capability of salt and drought tolerance. To screening of resistance genes and investigate gene function, the genetic transformation system including callus induction, differentiation and rooting cultivation of stem cuttage in R. trigyna should be performed. The main results are as follows:1. Induction rate of leaf callus induced by MS culture medium containing different hormones were counted in the trial of callus induction. The results showed that:the highest induction rate of leaf callus was 96.99%, which is induced by MS+0.2 mg·L-1TDZ+2 mg·L-1 NAA. Induction rate of the leaf callus (93.77%) were induced by MS+0.5 mg·L-16-BA+1mg·L-1IBA. Induction rate of the leaf callus (82.60%) were induced by MS+0.1 mg·L-16-BA+1mg·L-12,4-D. Induction rate of leaf callus (79.53%) were induced by MS+0.5 mg·L-16-BA+2 mg·L-1 NAA. Lowest induction rate of leaf callus (77.31%) were induced by MS+0.2 mg·L-1CPPU+2 mg·L-1NAA. Overall, MS culture medium containing TDZ(0.2 mg·L-1)+NAA (2 mg·L-1) might be appropriate for induce leaf callus.2. MS culture medium containing five different hormones were prepared in the callus differentiation experiments:①0.2,0.5,1,2,3 mg·L-1 CPPU;②With 0.02 mg·L-1NAA as the fixed factor, adding CPPU 0.2,0.5,1,2,3 mg·L-1 respectively; ③0.05,0.1 mg·L-1 TDZ and 0.02,0.05,0.1,0.2,0.5 mg·L-1 GA3 respectively; ④0.02, 0.05,0.1,0.5,1 mg·L-1 TDZ and 0.05,0.2 mg-L"1 IBA respectively; ￤ith 0.02 mg·L-1 IAA as the fixed factor, adding 0.1 mg·L-1 CPPU+0.1 mg·L-16-BA,0.25 mg·L-1 CPPU+0.25 mg·L-1 6-BA、0.5 mg·L-1 CPPU+0.5 mg·L-1 6-BA、1 mg·L-1 CPPU+1 mg·L-16-BA,1.5 mg·L-1 CPPU+1.5 mg·L-1 6-BA respectively. In conclusion, the bud could not differentiate by callus from leaf. Bud can be differentiated by the callus from stem segment with axillary, but it might be existed that the axillary differentiate bud directly.3. The roots could be induced by the certain concentration of salt and IBA in stem tip cuttage of R. trigyna. The suitable rooting medium were MS+0.05 mg·L-1 IBA+80 mmol·L-1 NaCl, its induction rate was 53.33%.