| Acacia melanoxylon is native to Australia, introduced into our country lately, it has a high economic, ecological and social benefits. In this experiment, five superior clones of Acacia melanoxylon tissue culture seedlings were used as materials, the type and concentration of plant growth regulator and the adjustment control of external environment condition as well as using new LED light source were studied, to further explore the influencing factors on proliferation of Acacia melanoxylon; the effect of different types and concentrations of auxins on test-tube rooting of Acacia melanoxylon was studied, and to study a better way for the root of Acacia melanoxylon,the way of ex vita) rooting of Acacia melanoxylon was studied, the tissue culture technique of Acacia melanoxylon was optimized, to provide the basis and technical support for cost savings of industrialized seedling production. The main researches were summarized as follows:1. Effects of different types and concentrations of hormones on propagation coefficient of five clones.This experiment considered the effects of the different concentrations of 6-BA, NAA, IBA and GA3 on propagation coefficient of five clones with orthogonal design. The average propagation coefficients of five clones were as follows:FM2(7.148)> FM4(7.088)>FM1(6.637)> FM10(6.440)>FM5(6.337). There were differences in the proliferation ability between five clones, and the demands for the concentration of the same hormone were also different,6-BA had the largest influence on proliferation of five clones. Different factors were considered synthetically, the best formula for multiplication culture of five clones was as follows:FM1 is MS+6-BA0.3mg·L-1+NAA0.05mg·L-1+IBA0.3mg·L-1+GA30.1mg·L-1;FM2 is MS+6-BAO.1mg·L-1+BA0.2mg·L-1+NAA0.05mg·L-1+GA30.1mg·L-1;FM4 is MS+6-BA0.1mg·L-1+BA0.3mg·L-1+NAA0.05mg·L-1+GA30.2mg·L-1;FM5 is MS+6-BA0.3mg·L-1+GA30.2mg·L-1+IBA0.3mg·L-1+NAA0.1mg·L-1;FM10 is MS+6-BA0.1mg·L-1+GA30.1mg·L-1+NAA0.1mg·L-1+IBA0.3mg·L-1.2. Effect of different seal materials on propagation coefficient of five clones. The experiment considered the effects of different seal materials on propagation coefficient of five clones, the five clones showed a consistent result:the propagation coefficient with breathable paper was slightly higher than the plastic cover.3. Effects of different light environment on propagation coefficient of five clones. The results showed that the light intensity had reached the extremely significant influence on proliferation of FM2, FM4 and FM5, with the significant effect on FM1, while it hadn’t significant influence on FM10.Five clones all had maximum average propagation coefficient in 45μmol·m-2·s-1, tissue culture seedlings grew good as the light intensity increased, but it was not conducive to the proliferation and differentiation. To study the effects of different light quality on propagation coefficient of five clones, three kinds of combination of red and blue LED lights was designed, 20% red LED+80% blue LED,80% red LED+20% blue LED, and 50% red LED+50% blue LED. The results showed that the effects of all the combinations of LED on proliferation of five clones were not significant difference, and differentiation was worse than the fluorescent lamp processing. Two photoperiods were designed:16 h light and 8 h dark,12 h light and 12 h dark. The results showed that the effect of the photoperiod on proliferation of five clones was different, five clones reduced with varying degrees in average multiplication coefficient when illumination time increased to 16 h, the difference of two photoperiods about propagation coefficient in FM2 and FM4 reached extremely significant level.lt reached significant level in FM5. The long daily photoperiod was not conducive to the proliferation of tissue culture seedlings of Acacia melanoxylon.4. Effects of different types and concentrations of hormones on rooting of five clones. This experiment considered the effects of the different concentrations of NAA, DBA and 2,4-D on rooting of five clones with orthogonal design. The results showed that the effects of NAA and 2,4-D on rooting of five clones were larger in the three kinds of auxin. With the range analysis of the rooting rate, the best formula for rooting of five clones was as follows:FM1 is 1/2MS+NAA0.2mg·L-1+2,4-D0.5mg·L-1+IBA0.5mg·L-1;FM2 is 1/2MS+NAA0.3mg·L-1+2,4-D0.5mg·L-1+IBA1.5mg·L-1;FM4 is 1/2MS+2,4-D0.5 mg·L-1+IBA1.5 mg·L-1+NAA0.3 mg·L-1;FM5 is 1/2MS +2,4-D0.5 mg·L-1+NAAO.1 mg·L-1+TBA1.5 mg·L-1;FM10 is 1/2MS+2,4-D0.5 mg·L-1+NAA0.2 mg·L-1+IBA1.5 mg·L-1.5. Effects of different types, concentrations and treatment time of root-inducing regulators on ex vitro rooting of five clones. This experiment considered the effects of the different concentrations and treatment time of NAA, IB A and ABT on ex vitro rooting of five clones. It was found that the difference of the effects of different types, concentrations and treatment time of root-inducing regulators on ex vitro rooting of five clones were significant. The suitable type, concentration and treatment time of root-inducing regulator on ex vitro rooting of five clones were showed as follows:FM1 was processed in ABT 100mg·L-1 for 5 minutes, and the rooting rate reached 100%, the average root number was 8.83, the average root length was 1.42cm; FM2 was processed in ABT 50mg·L-1 for 5 minutes, and the rooting rate reached 100%, the average root number was 10.66, the average root length was 1.33cm; FM4 was processed in ABT 100mg·L-1 for 5 minutes, and the rooting rate reached 100%, the average root number was 13.66, the average root length was 1.77cm; FM5 was processed in ABT SOmg·L-1 for 15 minutes, and the rooting rate reached 100%, the average root number was 9.50, the average root length was 1.89cm; FM10 was processed in ABT100mg·L-1 for 25 minutes, and the rooting rate reached 100%, the average root number was 18.86, the average root length was 1.79cm.6. Effects of different transplanting mediums on ex vitro rooting of five clones.In this experiment,it considered three kinds of transplanting mediums:(1) peat soil; (2) peat soil+ vermiculite (1:1); (3) peat soil+vermiculite+perlite (5:4:1). The test results showed that the difference of different transplanting mediums on ex vitro rooting of five clones was significant All things considered,the suitable transplanting medium for ex vitro rooting of Acacia melanoxylon plantlets was peat soil+vermiculite+perlite (5:4:1).7. Effects of different moisture content of transplanting medium on ex vitro rooting of five clones. The test results showed that the best moisture content of transplanting medium was 40%.8. Effects of different light intensity on ex vitro rooting of five clones.The experiment considered three kinds of light intensity in this test:45μmol·m-2·s-1,90μmol·m-2·s-1,135μmol·m-2·s-1. With the multiple comparison analysis, the results of 5 clones were consistent:rooting rate and average root number increased as the light intensity increased, but the average root length decreased.9. Effects of different photoperiod on ex vitro rooting of five clones. It considered two kinds of photoperiod in this test:12h/12h (light/dark) and 16h/8h (light/dark). The test results showed that the difference of the different photoperiod on rooting rate and rooting length of five clones was not significant, while it had significant effects on average root number.The average root length decreased as the illumination time extended, but the average root number increased. |
PDF Full Text Request