Studies On Tissue Culture And Induction Of Moringa Oleifera Lam.

Plant tissue culture and colchicines induction technology were applied to the induction of autotetraploid of optimal improved variety with Moringa oleifera PKM-1, and systematically researched on rapid identification, multiplication, the rooting and transplanting of test tube seedling.The result indicated that:Aseptic germination was fundmental to the experiments. On condition that disinfecting 25s with 50% alcohol, and 9minute dealing with 0.1% HgCl₂ was economical,efficient and high qualitative way.There are many factors influence the rate of regeneration: The explant startup culture with 2.0mg/L6-BA and 0.2mg/LNAA is the appropriate medium for inducing buds,induction rate reached 86.7% and mean of axilla buds was 3.17.The best media was MS+0.6mg/L6-BA for differentiation and MS+0.6mg/L6-BA+0.2mg/L NAA for callus induction in darkness. The multiplication of adventitious was best on MS medium with the addition of BA 0.6 mg/L the multiplication coefficient reached 3.1.The result that appended different density of PP₃₃₃ showed that, adding 1.5mg/L PP₃₃₃ to medium strengthening can be finished with multiplication of seeding in vitro,and was propitious to take root.The basal medium MS adding 7% sucrose is optimal for rooting, the induction rate was 100%, the states of plantlets and roots were well growth.The ploidy of plantlets in vitro were identified and the numbers of chromosome were 28(2n=2x=28). Under the condition of tissue culture, the inducing polyploid of Moringa oleifera was carried out by colchicine. The tetraploid plantlets were obtained by immersing buds in various concentrations of colchicine solution,the result showed that,It was the most effectively induced by immersing buds in 0.1% colchicine solution for 24 hours, which had the most tetraploid plantlets, the inductive frequency was 19%. Tetraploid plants and a few chimeras were obtained by colchicine inducing. Polyploid Moringa oleifera had a good multiplication and rooting effect, which cultivated on the best mediums for diploid plant. Tetraploid plantlets of Moringa oleifera were transplanted and survived transitorily.Morphology, physiology and biochemical characteristics,cytology were used and compard for identifying ploidy level of tetraploids arising from tube cultured seedlings,the result expressed that, tetraploids leaves were thicker, but shorter and with lower leaf index. The thickness and breadth of leaves of tetraploids were 325% and 165% or the length was 94% of those of the diploids respectively, on the small side of diploid. The tetraploids with fewer stoma densities which were 75.8% of the diploids respectively.The guard cell obviously augment.The length and breadth of tetraploids were 121.8% and 138.7% of those of the diploids respectively. The numbers of their chloroplast in the guard cells increased evidently and there were 182.3% of the diploids respectively. By the analysis, between tetraploid and diploid, leaf thickness, stoma density, guard cell size, chloroplast number were mighty significantly different, leaf length was significantly different. So, leaf thickness, leaf length, stoma density, guard cell size, chloroplast number can be used to identify tetraploid and diploid of Moringa oleifera.The tetraploid plantlets and diploid plantlets of Moringa oleifera were carried out the low-temperature treatment, then their exosmosis rates of electrolyte and catalase activities were compared. Under the same temperature, the electrolyte exosmosis rate of tetraploid plants were lower than diploid plants. Especially, when in-5℃and-10℃condition, electrolyte exosmosis rates of diploid were 26.79% and 39.04% respectively, but their tetraploid's electrolyte exosmosis rates were 20.78% and 35.23% respectively. Comparison of drought and heat resistance on different ploidies Moringa oleifera for the first time,the result indicated that,the drought and heat resistance of tetraploid all stronger than diploid,it has fully manifested the superiority of polyploidy.The rearch on photosynthesis characteristic of different ploidies Moringa oleifera has found that, the six Photosynthesis indexes except Tleaf are all significantly different,and all positive relation to ploidy levels. The chlorophyll content was closely related to the photosynthesis,the mensurate result find that, it was increasing along with the ploidy.This result can provide important data for planting and breeds selectively.