Development On Callus And Regnerable Masses Of Anthurium Andraeanum And Induction Of Tetraploid

Anthuriums are known as an exotic ornamental crop, they consists of about 1000 species, the Anthurium andraeanum is one of the major cultivated species and is grown for cut-flower and potted plant production. In it’s Rapid Propagation of Anthurium andraeanum via callus culture has been established and used commercially, but this method has shortcoming in loss of ability to differentiation of callus after several subcultures and formation of off-types. In order to resolve the aforementioned problems, we investigated the possibility of producing regenrable mass via aerial root in it’s and differentiation of masses. In addition, micro structural difference between different differentiation potential callus and masses were observed. and senescent index changes were also studied. The mass was sensitive to colchicines. therefore colchicines induced tetraploid were also tried in this paper.1. Establishment of Rapid Propagation System of Anthurium andraeanum via aerial root in vitroThe induction of regenrable masses from aerial root were investigated, it was found that 2,4-D had very important effect to induce regenrable masses, and 2,4-D should cooperate with 6-BA. The green tissue was essential to differentiation. The mineral salt concentration of MS played an important role in regenrabole masses induction and differentiation. The 1/2MS medium supplement with 1.0 mg·L-1 6-BA and 0.6 mg·L-12,4-D was suitable to aerial root growth, the optimal induction medium was 1/2MS+6-BA1.0 mg·L-1+2,4-D0.2 mg·L-1 and the best medium for mass differentiation was MS+6-BA1.0 mg·L-1+2,4-D0.2 mg·L-1. The masses did not showed degradation phenomenon after continued reproduction.2. Cytological Study on Callus of leaf and Regenrable Mass of Aerial Root in Anthurium andraeanumIn vitro morphogenesis of three leaf callus and regenrable masses from aerial root in Anthurium andraeanum’Arizona’were compared. It was showed that in the Kelly, yellow and brown callus, the karyoplasm ratio decrease in turn, cell arrangement became looser and looser, the shape of cell did not kept integrity. Some similar apoptotic bodies were appeared in some cells of yellow and brown callus. The morphogenesis of regenrable masses were indirect organogenesis. regenrable masses occurred from the cortex of aerial root ends. There were a lot of small and actively divided cell groups with dense nucleus in regenrable masses and meristic cells were originated from these cell groups indirect. With the proliferation of meristic cells and formation of bud primordium, the outer layer cells apoptosis gradually. Which led to bud emergence on the surface of masses. Organogenesis was the main morphogenesis pathway of masse, although a few embryoid morphogenesis was observed.3. Histochemistry Studies on Callus of Leaf and Regenrable Mass of Aerial Root in Anthurium andraeanumFour kinds of callus from leaf and Regenrable mass of aerial root in Anthurium andraeanum’Arizona’were studied by paraffin section and histochemical coloration. The results showed that the granular saccharide was mainly surrounding the nuclei. In the callus and regenrable mass protein was mainly in or around the nuclei. In kelly callus the levels of saccharide and protein decreased clearly from the outer layer of cells to the inner layer of cells. and in dark green callus the grads was unconspicuous. The contents of saccharidein and the even distribution was observed in the golden callus. The distribution of saccharide was dispersedly in the brown callus. The changes of protein were inconspicuous from the outer layer of cells to the inner layer of cells in the gold callus. However, grads of protein was little in the brown callus. There was also a visible grads of saccharide and protein when regenrable mass occurred, in the expending period of regenrable mass, the saccharide and protein were distribute evenly. In the differentiation stage of regenrable mass, the saccharide was concentrated in the cells which surrounded the dense small cell groups and the primary vascular tissue. The protein mainly concentrated in the nuclei of the cell groups. There were special cells in all types of callus and regenrable mass. The content and the grads of the saccharide and protein and the integration of the nuclei played important roles in the differentiation of callus and regenrable mass.4. Comparative Studies on Senescent Index in Two Kinds of Regenerative System in Anthurium andraeaumThe superoxide and relative enzymes between different types of callus originated from leaves and regenerable mass from aerial roots were compared. It was showed that the orders of the levels of superoxide, the activities of superoxide dismutase (SOD) and catalase (CAT) from high to low were yellow green callus> golden callus>dark green callus> regenrable mass>brown callus. The highest activities of GPX (glutathione peroxidase) were appeared in brown callus, followed by dark green, yellow green callus, golden callus and regenrable mass For malondialdehyde (MDA), the highest contents were observed in golden callus, followed by brown callus, yellow green callus, dark green callus and RMB. It was demonstrated that the activities of SOD and CAT had synergistic effects in eliminating oxidative stress, and the higher activities of SOD and CAT in mass suggested their greater ability of eliminating oxidative stress. The enzyme activities of GPX in RMB were significantly lower than that in all callus, in other hand the activities of GPX were relatively lower in yellow green callus, the lower contents of super-oxides and MDA might be the results of higher antioxidant activities. In short, rapid propagation by regenrable mass had advantages over that by callus because of their higher anti-oxidant activities.5. Induction of Anthurium andraeanum’Arizona’tetraploid by Colchicine in VitroTetraploids plants of Anthurium andraeanum’Arizona’were successfully induced after treating diploid tissue masses with colchicine. Masses originating from diploid aerial roots were treated with colchicine at three different concentrations (i.e.,0.1.0.2,0.3%) for about 3,5 and 7h, and then were transferred into Murashige & Skoog medium containing 3 mg·L-1 6-BA+0.2 mg·L-1 2,4-D. After 60 days, the survival rate and numbers of regenerative shoots were scored. The high concentration and longer duration sharply reduced survival rate. In contrast. the regeneration of plantlets was not noticeably affected by colchicine. Tetraploid plants were obtained in all treatments, but the percentage of induced tetraploids ranged from 0.2% to 7.6%. The best induction was obtained with a 5-h. treatment with 0.3% colchicine. The stomatal size of tetraploid plants was larger than in diploid plants; however, the stomatal density was lower than in diploid plants. Tetraploid plants possessed stronger petioles, thicker and deeper green leaves, and thicker and longer lived spathes in comparison with diploid plants. Abnormal spathes, such as double spathes or those lacking pedicels, were observed in tetraploid plants. Tetraploid plantlets could be regenerated via aerial roots; this technique could be applied to tetraploid plant propagation.