| In order to achieve the large-scale cultivation of Taxus chinensis var.mairei callus cells to produce taxol,the research group had invented the curtain-like anchoring culture bioreactor and culture method in the early stage of the research,the quality of the culture bed net-nap fiber in the bioreactor is the key to obtain the nutrition and realize the transition from the physical anchoring growth to the biological anchoring growth for the callus in the system.In this paper,16 kinds fibers from phloem,wood,bamboo wood and straw of 14 kinds plants,which were representative,were prepared by self-made,and the factors influence the mass transfer process that plant fibers transferred MS nutrient solution by infiltration were studied by optical microscopy observing,HPLC,atomic absorption spectrometry and"Mass transfer simulation device for fiber infiltration",which was made simulating the mass transfer process of curtain-like anchored culture bioreactor by self-made,it establishedthe the basis for the determination of the main factors that should be investigated for screening the net-nap fiber of the culture bed in the system.The results and conclusions are as follows:Four methods were used to prepare respectively the fibers from 16 kinds materials.It was found that the fiber yield was the highest with lime method,73.06%on average,but the time-consuming was the longest,more than 14 times of the other three methods,and the non fiber material and non dispersed bundle fiber remained higher.that of Populus alba wood was the highest,was 95.02%.The sulphate method was 52.87%,was more suitable for preparing the fiber of Boehmeria nivea phloem,Bambusa emeiensis,Bambusa textilis,Phyllostachys heterocycla bamboo wood and bagasse;AS-AQ method and enzyme method was 50%respectively,AS-AQ method is more suitable for preparing the fiber of Broussonetia kazinoki phloem and wood,B.nivea,P.alba and Pinus massoniana wood,Triarrhena sacchariflora,Triticum aestivum,Oryza sativa and Zea mays straw;the enzyme method is more suitable for preparing the fiber of Arundo donax,Miscanthus floridulus straw and P.massoniana wood.The results of fiber microscopic observation showed that the fiber cell length was phloem>bamboo wood>straw>wood according to the material type,by species（mm）:B.nivea phloem（48.4）>B.kazinoki phloem（13.2）>A.donax straw（3.4）>B.emeiensis wood（3.3）>P.heterocycla wood（3.2）>B.textilis wood（2.8）>M.floridulus straw（2.7）>T.sacchariflora straw（2.3）>bagasse（2.0）>Z.mays straw（1.8）)>T.aestivum straw（1.6）>O.sativa straw,P.alba,P.massoniana and B.kazinoki wood（0.8）>B.nivea wood（0.7）.The fiber cell width was phloem>bamboo wood>wood>straw according to the material type,by species（μm）:B.nivea phloem（38.7）>bagasse（28.6）>P.alba（28.3）>P.heterocycla bamboo wood（25.6）>B.kazinoki phloem（21.9）>B.textilis bamboo wood（21.8）>A.donax and M.floridulus straw（20.3）>Z.mays straw（19.7）>B.kazinoki wood（19.2）>B.emeiensis bamboo wood（17.1）>O.sativa straw（13.0）)>P.massoniana wood（12.2）>T.sacchariflora straw（11.3）>T.aestivum straw（11.1）>B.nivea wood（10.4）.The wall width was bamboo wood>phloem>wood>straw according to the material type,by species（μm）:B.nivea phloem and P.alba wood（8.5）>P.heterocycla bamboo wood（5.1）>B.emeiensis bamboo wood（4.8）>bagasse and B.kazinoki wood（4.3）>B.kazinoki phloem（3.1）>Z.mays straw（3.0）>B.textilis bamboo wood（2.6）>O.sativa straw（2.6）>T.sacchariflora straw（2.3）>M.floridulus straw（1.7）>P.massoniana wood（1.6）>B.nivea wood（1.3）>A.donax straw（1.2）>T.aestivum straw（1.1）.The lumen diameter of fiber cell was phloem>bamboo wood and straw>wood according to the material type,according to the species（μm）:B.nivea phloem（21.7）>bagasse（20.0）>A.donax straw（17.9）>M.floridulus straw（16.9）>B.textilis bamboo wood（16.6）>B.kazinoki phloem（15.7）>P.heterocycla bamboo wood（15.4）>Z.mays straw（13.7）>P.alba wood（11.3）>B.kazinoki wood（10.6）>P.massoniana wood And T.aestivum straw（9.0）>O.sativa straw（8.0）>B.nivea wood（7.8）>B.emeiensis bamboo wood（7.5）>T.sacchariflora straw（6.7）.From high to low,the rough and complexity level of the fibers cell surface structure was phloem:B.nivea phloem>B.kazinoki phloem;bamboo wood:B.textilis>P.heterocycla>B.emeiensis;wood:B.nivea>B.kazinoki>P.alba>P.massoniana;straw:Z.mays>T.aestivum>O.sativa>bagasse>T.sacchariflora>A.donax and M.floridulus.The rate of MS medium mass transfer in the fibers by infiltration in fiber wetting stage was as follows:B.kazinoki phloem and O.sativa straw>bagasse>A.donax straw>Z.mays straw>B.nivea wood>B.nivea phloem>P.heterocycla bamboo wood>B.textilis bamboo wood>T.sacchariflora straw>M.floridulus straw>T.aestivum straw>B.kazinoki wood>P.alba wood>B.emeiensis bamboo wood>P.massoniana wood,the highest was nearly 8 times of the lowest,the difference was very significant.There was no significant difference between the orderly and disordered groups in fiber arrangement direction,between 0.02-1.06 cm·min-1.After saturated water absorption,the continuous water column system formed in the micro space of fiber bundle partially counteracted the effect of fiber surface difference,resulting in a significant decrease in the range of difference,but it was still significant,the highest is more than twice the lowest,and the order became:O.sativa straw>T.aestivum straw>A.donax straw>M.floridulus straw>B.nivea wood>B.kazinoki wood>B.nivea phloem>P.heterocycla bamboo wood>P.massoniana Wood>T.sacchariflora straw>bagasse>B.kazinoki phloem>Z.mays straw>P.alba wood>B.emeiensis bamboo wood>B.textilis bamboo wood.The difference between the orderly and disordered groups in the fiber arrangement direction increased significantly,which was11.56～68.90 ml·h-1·g-1.Compared with the results of microscopic observation,there was no corresponding relationship between the ordering rules of fiber species differences,which proved that there was no correlation between the differences of the fiber infiltration mass transfer rate and the differences of the fibers microscopic structure,and the main factors affecting the mass infiltration transfer process of plant fiber were the differences between the chemical composition of fiber types and the order of fiber arrangement direction in fiber bundles.According to the concentration analysis of each component in MS mass transfer liquid by HPLC and AAS,it was found that there were different amplitude short-time fluctuations in the concentration of each component in about initial 100 min.All of 16kind fibers absorded stronger 6-BA,and the initial detection concentration was mostly zero,while B.kazinoki phloem,T.sacchariflora,O.sativa,T.aestivum straw and bagasse fibers were slightly higher,about 1/2 of the original concentration.The concentration of NAA increased significantly in initial 40～60 min,the fiber of B.nivea phloem and P.heterocycla bamboo wood was more than 10 times,which concentration of NAA in MS mass transfer liquid was of the MS stock solution,the fiber of B.emeiensis and B.textilis bamboo wood,and M.floridulus straw was more than 8 times,the fiber of B.kazinoki phloem,O.sativa and T.sacchariflora straw,bagasse was more than 7 times,the fiber of A.donax and Z.mays straw was more than 4 times,it should be that the fiber absorbed and enriched NAA in the stage of saturated water absorption,and the concentration increased due to NAA eluteed after saturated water absorption.The detection concentration of 2,4-D fluctuated up and down,in which the fluctuation range of B.nivea phloem was more than 2 times of the original concentration,and the lowest detection concentration of four kinds of fibers,namely,B.kazinoki phloem,P.massoniana wood,A.donax and T.aestivum straw,was zero,all of which were instantaneous zero point.The concentration of nicotinic acid also fluctuated up and down for a short time.The fluctuation of nicotinic acid concentration was more obvious,which of B.emeiensis and B.textilis bamboo wood fiber,between 0.02 and 2.58 mg·L-1,the detection concentration increased duo to B.nivea phloem fiber was the most significant,which was 3.17 mg·L-1,more than 5 times of the original concentration,followed by the fiber of B.kazinoki phloem,B.textilis bamboo wood and A.donax straw,which was more than 3 times.The concentration of vitamin B6 in the fiber mass transfer solution of B.textilis and P.heterocycla bamboo wood also detected the instantaneous zero point,and that of B.nivea phloem was detected the high point,which was more than two times of the original concentration,and that of other kinds fiber fluctuated little.The results of AAS showed that the concentration of Na,Mg,Ca,Zn,Mn,Mo element also fluctuated up and down in a short time,and the lowest was zero（Zn,in the initial mass transfer solution of B.kazinoki phloem,B.textilis bamboo wood,B.nivea wood,T.sacchariflora and Z.mays straw fiber.Mo,B.kazinoki phloem and M.floridulus straw）,and the highest was more than 8 times of the original concentration（Mo,B.nivea phloem and wood,P.massoniana wood fiber,the former is close to 10times）.The Na and Ca elements in the initial mass transfer solution of various fibers are higher than the original concentration,in which Na is generally 2～6 times higher,and the variation of Fe and K concentration is lesser.That is to say,there was also a very significant difference among species in the short-term fluctuation of the concentration of each component in the mass transfer solution of plant fiber.The fiber types that have the weakest effect on the concentration of each component of the MS culture medium during the mass transfer process are T.aestivum straw,bagasse,and A.donax straw fibers.By comparing the differences of mass transfer rate and the concentration of each component in the mass transfer solution and the differences of fiber microscopic observation results,it is proved that the main factors affecting the process of plant fiber infiltration MS culture medium mass transfer werw the chemical composition of plant fiber and the order of fiber arrangement direction in the fiber bundle,and The correlation with the micro size structure of fiber was weak.Therefore,the chemical composition of plant fiber should be the main factor for the selection reference on the culture bed net-nap fiber. |
PDF Full Text Request