Studies On Vitro Micropropagation Technique Of Origanum Marjorana

In this study, Labiatae aromatic Origanum marjorana for the material, through the sterilization time, exogenous hormones and the screening of environmental hardening transplant, to observe the performance of Origanum marjorana in primary culture, subculture, rooting and transplanting stages. Researched the endogenous hormones (IAA, ZRs, iPAs) and the changes of enzyme activity (POD, PAL, protease)during the subculture of Origanum marjorana, then set up a fast, efficient and stable propagation techniques of Origanum marjorana, the results are as follows:(1) The screening of sterilization timeThe best sterilization time was 6min with 0.10% mercuric chloride while the pollution was only 5.76% and the browning rate was 4.17%.(2)The screening of initial mediumIn the screening of initial medium, the number of adventitious buds and the growth status of adventitious buds splitting was observed. IBA with a concentration of 0.05mg/L to screening the type and concentration of cytokinin(6-BA,TDZ,6-BA +KT). The results show that:the best of cytokinin is 6-BA with the concentration of 3.00 mg/L when the number of adventitious buds split up to 5.00, and leaves the normal grayish green with a short pitch. Use 6-BA with concentration of 3.0mg/L to screening the type and concentration of auxin. Results showed that:the auxin of IBA to induce adventitious buds got the best result, when the concentration is 0.1 Omg/L the number of adventitious buds split up to 5.55, leaves showed greyish green with length pitch.(3) The screening of subculture mediumSubculture medium chosen on the basis of initial medium bud number and growth conditions, as 6-BA was cell division number in the concentration of 3.00mg/L, selected IBA as growth hormone, the results show that the:when the IBA concentration was 0.10 mg/L, the number of adventitious buds split were more than other treatments which up to 5.55, with the normal grayish green leaves. (4) The screening of root mediaResearch shows that rooting on Marjoram:1/2MS as the basic medium, and NAA concentration of 0.10 mg/L lead a 97.62% of rooting rate, a muscular and longer root.(5)The screening of transplanting stage of hardened matrix and the shading of different degreeOn different substrates and the degree of shading show that:in the shade of 95% and perlite:vermiculite ratio of 1:2 up to 92.86% survival rate.(6) The process of Origanum marjorana in vitro micropropagation techniquesThe best sterilization time was 6min with 0.10% mercuric chloride,the optimum initial medium and subculture medium with 6-BA3.00mg/L+IBAO.10mg/L of MS medium, rooting for the add 0.10mg/L NAA's 1/2MS medium, transplanting hardening stage under 95% shading, perlite:vermiculite 1:2 ratio have the highest survival rate.(7) The study of endogenous hormones in subcultureThrough the stduy of IAA, iPAs and ZRs during subculture, the results show that: IBA concentration was 0.05,0.10,0.15 mg/L, three endogenous hormones trend was consistent that was first increased and then decline, IAA, and iPAs peak at the time 7d, ZRs peak in the 14d, while the IBA was 0.00mg/L treatment trends inconsistent with the other three treatment.(8) The activity of enzyme in subcultureThrough the stduy of peroxidase (POD), phenylalanine lyase (PAL) and the protease in subculture, the results showed that:with the IBA concentration of 0.05 mg/L and 0.10mg/L, POD and PAL increased firstly and then increased less tendency to peak in the 14d, the other two deal with inconsistent trends. Changes in protease activity, IBA concentration of 0.00mg/L and 0.10mg/L increased firstly and then decreased after the two treatment tended to increase, the other two deal with inconsistent trends.