密闭式人工光组培室的环境控制与洁净技术的研究
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摘要
光独立组织培养能够促进植株的生长发育、减少污染，是植物种苗工厂化生产中的一个应用方向。针对我国光独立组织培养迅速发展与组培环境综合调控技术相对落后的问题，本文采用环境控制与空气净化相结合的方法来综合调控组培室内的物理环境，并对试制的环境控制系统进行了试验研究，期待为工厂化组培苗生产提供综合环境调控的方法与硬件设备。
1．通过对环境控制和空气净化的系统研究，试制了一套适用于密闭式人工光组培室内的温度、相对湿度、CO_2浓度和光照环境等综合调控的环境控制系统，同时能控制室内空气洁净度。
2．试验结果表明，室内控制点的温度、相对湿度和CO_2浓度的控制精度分别在±0.4℃，±7.9％，±34μmolmol~(-1)以内。当荧光灯开启总数(120盏，4.4kW)的1／4和1／2时，室内温度和相对湿度的最大差值分别为2.3℃，4.0％；荧光灯全开时，室内温差达3.9℃，控制点的相对湿度控制在65.04±17.5％，与控制目标65.0±5.0％相差较大。该组培室正式运转前和运转一年后，室内的悬浮粒子和沉降菌指标均达到洁净度国家标准7级。
3．根据对加湿过程的理论计算与试验分析，指出本系统中降温与加湿共存阶段加湿效果不明显，同时由于加湿器的加湿容量不足，顶部送风和下部两侧回风的通风方式造成了室内的相对湿度无法在荧光灯全开时达到控制要求。
4．利用该环境控制系统，在三角瓶容器中采用光独立组织培养的生姜组培苗长势粗壮、品质高，其新增叶片数、干鲜重和株高的增加分别为常规组织培养的1.4，1.5和1.7倍。
5．综合分析该环境控制系统的试验结果，提出了该环境系统与洁净技术的不足及改进点。
As a new technique for tissue culture, photoautotrophic micropropagation has many advantages such as promoting growth and development of leafy explants, and minimizing microbial contamination. It will be widely used for the large-scale plantlets production in the near future. The rapid development of the photoautotrophic micropropagation in China requires integrated environment control and air purification simultaneously. In this study, an environmental control system combined with clean technology was developed to control air condition and lighting environment in a closed tissue culture system under artificial lighting. The experiment results are shown as following.
1. The environmental control system could achieve to control temperature, relative humidity, CO2 concentration, light intensity, and cleanliness, simultaneously.
2. The control point showed an uniformity in themperature, relative humidity, CO2 concentration within ±0.4℃,±7.9%, ±34 mol mol-1, respectively. The gradient of temperature and relative humidity were 2.3 ℃ and 4.0% in the closed tissue culture system when 1/4 and 1/2 of all fluorescent lamps were turn on. However, when all of the fluorescent lamps were turn on, the gradient of temperature was 3.9℃ and relative humidity was 65.0±17.5% even the setpoint was 65.0±3.0%. Before the system running and after one year, the cleanliness measured by airborne particles and settling microbe were both 7 grade according to GB50073-2001 and GB/T 16294-1996.
3. Based on theoretical analysis and experimental results, the high relative humidity was diffcult to maintain in the closed tissue culture system when all of the fluorescent lamps were turn on, because of deficiency of the humidifier's capacity and air ventilation by top air nozzle outlets and return air in the bottom.
4. The ginger (Zingiber officinale) plantlets cultured in triangle vessels under photoautotrophic micropropagation were vigorous and 1.4, 1.5, 1.7 times in leaf number, dry weight, and stem elongation compared to those culutred under conventional micropropagation, respectively.
5. Some improved suggestions were shown based on the disadvantages of the environmental control system in the closed tissue culture system.
1．通过对环境控制和空气净化的系统研究，试制了一套适用于密闭式人工光组培室内的温度、相对湿度、CO_2浓度和光照环境等综合调控的环境控制系统，同时能控制室内空气洁净度。
2．试验结果表明，室内控制点的温度、相对湿度和CO_2浓度的控制精度分别在±0.4℃，±7.9％，±34μmolmol~(-1)以内。当荧光灯开启总数(120盏，4.4kW)的1／4和1／2时，室内温度和相对湿度的最大差值分别为2.3℃，4.0％；荧光灯全开时，室内温差达3.9℃，控制点的相对湿度控制在65.04±17.5％，与控制目标65.0±5.0％相差较大。该组培室正式运转前和运转一年后，室内的悬浮粒子和沉降菌指标均达到洁净度国家标准7级。
3．根据对加湿过程的理论计算与试验分析，指出本系统中降温与加湿共存阶段加湿效果不明显，同时由于加湿器的加湿容量不足，顶部送风和下部两侧回风的通风方式造成了室内的相对湿度无法在荧光灯全开时达到控制要求。
4．利用该环境控制系统，在三角瓶容器中采用光独立组织培养的生姜组培苗长势粗壮、品质高，其新增叶片数、干鲜重和株高的增加分别为常规组织培养的1.4，1.5和1.7倍。
5．综合分析该环境控制系统的试验结果，提出了该环境系统与洁净技术的不足及改进点。
As a new technique for tissue culture, photoautotrophic micropropagation has many advantages such as promoting growth and development of leafy explants, and minimizing microbial contamination. It will be widely used for the large-scale plantlets production in the near future. The rapid development of the photoautotrophic micropropagation in China requires integrated environment control and air purification simultaneously. In this study, an environmental control system combined with clean technology was developed to control air condition and lighting environment in a closed tissue culture system under artificial lighting. The experiment results are shown as following.
1. The environmental control system could achieve to control temperature, relative humidity, CO2 concentration, light intensity, and cleanliness, simultaneously.
2. The control point showed an uniformity in themperature, relative humidity, CO2 concentration within ±0.4℃,±7.9%, ±34 mol mol-1, respectively. The gradient of temperature and relative humidity were 2.3 ℃ and 4.0% in the closed tissue culture system when 1/4 and 1/2 of all fluorescent lamps were turn on. However, when all of the fluorescent lamps were turn on, the gradient of temperature was 3.9℃ and relative humidity was 65.0±17.5% even the setpoint was 65.0±3.0%. Before the system running and after one year, the cleanliness measured by airborne particles and settling microbe were both 7 grade according to GB50073-2001 and GB/T 16294-1996.
3. Based on theoretical analysis and experimental results, the high relative humidity was diffcult to maintain in the closed tissue culture system when all of the fluorescent lamps were turn on, because of deficiency of the humidifier's capacity and air ventilation by top air nozzle outlets and return air in the bottom.
4. The ginger (Zingiber officinale) plantlets cultured in triangle vessels under photoautotrophic micropropagation were vigorous and 1.4, 1.5, 1.7 times in leaf number, dry weight, and stem elongation compared to those culutred under conventional micropropagation, respectively.
5. Some improved suggestions were shown based on the disadvantages of the environmental control system in the closed tissue culture system.
引文
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[2] 钱迎倩，孙敬三．植物组织培养．北京：人民教育出版社，1987，124-167
[3] 江苏植物组培研究协会编．经济植物组织培养技术．南京：江苏科技出版社，1988，21-76
[4] 王文静，袁道强，高松洁．植物组织培养的应用现状[J]．河南师范大学学报(自然科学版)，2000(3)：137-139
[5] 阙国宁，张健如．组培苗商品化：现状与对策[J]．林业科技开发，1996，(3)：3-5
[6] Kozai, T. Autotrophic (sugar free) tissue culture for promoting the growth of planlets in vitro and for reducing biological contaminacation. Proc. Bangkok. Thailand: International Symposium on Application of Biotechnology for Small Industries, 1990.39-51
[7] 曲英华，胡秀婵，吴毅明．植物组织培养新技术：光独立培养法[J]．农业工程学报，2001，17(6)：90-92
[8] Kozai, T., C. Kubota, B.R. Jeong. Environmental control for the large-scale production of plants through in vitro techniques. Plant Cell. Tissue and Organ Culture, 1997.51:49-56
[9] 史跃林．组培苗的生育环境与调节[J]．植物生理学通讯，1990，30(3)：65-67
[10] Desjardins Y. Photosynthesis in vitro-on the factors regulating CO_2 assimilation in micropropagation systems[J]. Acta Horticulturae, 1995.393:45-61
[11] Honjo, T. and T. Takarura. Measurement of humidity profiles in micropropagation vessels. ASAE Paper St. Joseph. MI:ASAE, 1991.91-151
[12] Ghasshghaie, J., F. Brenckmann and B. Saugier. Water relations and growth of rose plants cultured in vitro under various relative humidities. Plant Cell, Tissue and Organ Culture, 1992.30:51-57
[13] Preece, J.E. and E.G. Sutter. Acclimatization of micropropagated plants to the greenhouse and field. In Micropropagation: technology and application. Ed. Debergh, P.C. and R.H. Zimmerman. Kluwer Academic Publishers, 1990.71-93
[14] Ziv, M. Vitrification: morphological and physiological disorders of in virto plants. In Micropropagation: technology and application. Ed. Debergh, P.C. and R.H. Zimmerman. Kluwer Academic Publishers, 1990.45-69
[15] Fujiwara K, T. Kozai, I. Watanabe. Messurements of carbondioxide gas concentration in closed vessels containing tissue cultured plantlets and estimates of net photosynthetic rates of the plantlets. J. Agric. Meteorol. 1987.43:21-30
[16] Infante R., E. Magnanini, B. Righetti. The role of light and CO_2 in optimizing the conditions for shoot proliferation of Actinidia deliciosa in vitro. Physiol. Plant, 1989.77:191-195
[17] 吴毅明．国外设施园艺环境控制新技术的发展[J]．农业工程学报，1988，(3)：80-91
[18] Kozai, T., Chieri, Byoung R.J. Environmental control for the large-scale production of plants through in vitro techniques. Plant Cell, Tissue and Organ Culture, 1997.51:49-56
[19] 古在丰树．植物组织培养的新阶段[M]．日本农文协出版，1998(3)：50-54
[20] Kozai, T., C. Kubota and I. Watanabe. The growth of carnation plantlets in vitro cultured photoautotrophically and photomixotrophically on different media. Environ. Control Biol, 1990.28(1):21-27
[21] Kozai, T., K. Iwabuchi, K. Watanabe, et al. Photoautrophic and photomixotrophic growth of strawberry plantlets in vitro and changes in nutrient composition of the medium. Plant Cell, Tissue and Organ Cult, 1991.25:107-115
[22] Nakayama, M., T, Kozai, K. Watanabe. Effect of the presence/absence of sugar in the medium and natural/forced ventilation on the net photosynthetic rates of potato explants in vitro. Plant Tissue Cult. Lett. 1991.8(2):105-109
[23] Heo, J., T. Kozai. Development of a forced ventilation micropropagation system for enhacement of the photoautotrophic growth of sweetpotato plug plants cultured in vitro. Jap. J. Trop. Agr. 42. Extra issue, 1998. 1:21-22
[24] 肖玉兰，张立力，张光怡，等．非洲菊无糖组织培养技术的应用研究[J]．园艺学报，1998，25(4)：408-410
[25] Nguyen, Q.T., T. Kozai, K.L. Nguyen, et al. Growth promotion of in vitro woody plants under photoautotmphic cnditons. Abstr. The 6th Ann. Conf. of IPPS, Japan region. Tsukuba, Japan, 1999. 121-132
[26] Niu G.H., T. Kozai. Simulation of CO_2 concentration in the culture vessel and growth of plantlets in micropropagation. Acta. Hort. 1998, 456:37-43
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