| Open tissue culture in an open environment with bacteria,the culture medium is bacteriostatic by adding bacteriostatic agent to the medium instead of high-temperature and high-pressure sterilizer and ultra clean worktable,which simplifies the tissue culture link,reduces the production cost,and provides a new model for factory seedling raising.In this study,Xuxiang kiwifruit was used as material to screen the types and concentrations of antimicrobial agents.By adjusting the addition amount of agar powder,sucrose,mancozeb concentration and hormone concentration to optimize the culture link.Including explant disinfection,induction of axillary bud germination,proliferation culture and rooting culture.An open culture system of Xuxiang kiwifruit was established to provide a reference for its industrialized seedling raising.The main results are as follows:1.The optimum explant disinfection combination of traditional tissue culture of Xuxiang kiwifruit was 75% alcohol at 30 s + 0.1% mercuric chloride at 9 min,and the survival rate was 94.67%.The optimum medium for inducing axillary bud germination was MS + 6 g/L agar powder + 30 g/L sucrose + 0.3 mg/L NAA + 2.0 mg/L 6-BA,and the germination rate was 90%.The optimum proliferation medium was MS + 6 g/L agar powder + 30 g/L sucrose + 0.1 mg/L NAA + 1.5 mg/L 6-BA,the survival rate was 90%,the proliferation coefficient was 4.72,and the average plant height was 1.58 cm.The optimum rooting medium was 1/2 MS + 6 g/L agar powder + 30 g/L sucrose + 0.7 mg/L IBA,the rooting rate was 100%,the average number of roots was 11.58,and the average root length was 6.55 cm.The optimum transplanting medium is humus: vermiculite = 4:1,and the survival rate is 100%.These provide basis and control measures for the open cultivation of Xuxiang kiwifruit.2.The addition amount of screening agar powder is 3 g/L.Through the screening of the type and concentration of antimicrobial agents,changing the amount of sucrose,screening the concentration of antimicrobial agents in MS and 1/2 MS medium,it is concluded that the concentration of antimicrobial agents increases with the increase of the amount of sucrose.The optimum concentration of mancozeb in the proliferation and rooting medium of Xuxiang kiwifruit was tested.The results showed that the optimum concentration of mancozeb in proliferation culture was determined as follows: 30 g/L sucrose + 0.15 g/L mancozeb concentration,the survival rate was 86.67%,and the proliferation coefficient was 2.0;the survival rate of 20 g/L sucrose + 0.1 g/L mancozeb concentration was 83%,and the proliferation coefficient was 2.79;the survival rate of 10g/L sucrose + 0.08 g/L mancozeb concentration was 66.66%,and the proliferation coefficient was 2.77.The optimum concentration of mancozeb in rooting culture was determined as follows: 30 g/L sucrose + 0.09 g/L mancozeb concentration,the survival rate was 100%,and the rooting rate was 6.67%;the amount of 20 g/L sucrose + 0.08 g/L mancozeb concentration was 73.33%,and the rooting rate was 13.33%;the survival rate of10 g/L sucrose + 0.04 g/L mancozeb concentration was 93.33%,and the rooting rate was13.33%.3.The optimum combination of open explant disinfection was 75% alcohol at 30 s +0.1% mercuric chloride at 10 min,and the survival rate was 73.33%.The optimum medium for open induction of axillary bud germination was MS + 3 g/L agar powder + 30g/L sucrose + 0.15 g/L mancozeb concentration + 0.2 mg/L NAA + 2.0 mg/L 6-BA,and the germination rate was 70%.The optimum medium for open proliferation culture was MS + 3 g/L agar + 20 g/L sucrose + 0.1 g/L mancozeb concentration + 0.2 mg/L NAA +1.5 mg/L 6-BA.The survival rate was 93.33% and the proliferation coefficient was 6.539.The optimum medium for open rooting culture was 1/2 MS + 3 g/L agar powder + 10 g/L sucrose + 0.04 g/L mancozeb concentration + 0.3 mg/L IBA.The rooting rate was 93.33%,the average number of roots was 4 and the average root length was 2cm.4.Compare the links and costs between traditional tissue culture and open tissue culture.Both open tissue culture and traditional tissue culture take the production of 1million Xuxiang kiwifruit seedlings as an example for preliminary calculation.The total cost of open tissue culture is 631406.56 yuan,and the total cost of traditional tissue culture is 1033060.12 yuan.The traditional tissue culture is about 1.64 times that of open tissue culture.The unit price of each Xuxiang kiwifruit seedling in open tissue culture is about0.63 yuan.The unit price of each Xuxiang Kiwifruit in traditional tissue culture is about1.03 yuan.The cost per plant was saved by 0.4 yuan. |
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