LED不同光对葡萄离体试管苗生理生化特性效应的研究
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摘要
在植物组织培养中,光合光量子通量密度(PPFD:Photosynthetic Photon Flux Density)、光照周期和光谱分布对植物的光合作用和形态建成起重要调控作用。植物组培主要依靠电光源,传统电光源对植物的生物能效低、发热量大,光照用电约占整个电费成本的65%,是植物组织培养中较高的非人力成本之一。因此在植物组织培养中采用新型的LED光源,调控光质和PPFD,不仅能够调控组培植物的生长发育和形态建成,而且能够大大减少能耗,降低成本。同时解决了光质不纯的问题,增加了试验结果的可靠性。
本研究以葡萄砧木品种贝达(Beta)和变叶葡萄(Vitis piasezkii Maxim)试管苗为实验材料,研究其在LED白、红、黄、蓝、绿和蓝红六种光质,24 h·d-1、16 h·d-1、12 h·d-1、8 h·d-1四个光周期试管苗生长发育动态、地上部位(株高、叶面积、生物量、节间长、茎粗)和地下部位(根重、根条数、根长、根粗)增殖以及色素含量、气孔、酶活性的效应。以便得到组织培养最佳的LED光质、光周期。取得的主要研究结果有：
(1)LED红光有利于葡萄试管苗地上地下部位的纵向生长；LED蓝光抑制葡萄试管苗地上地下部位的纵向生长,而促进其横向生长；LED蓝红光对葡萄试管苗的光合速率较普通荧光强。红光、蓝光和蓝红光在16 h·d-1下对生物量的增加有明显的促进作用：在24 h·d-1、蓝光和蓝红光下茎节最粗。
(2)LED黄光、绿光,光周期16 h·d-1、12 h·d-1的光照有利于株高的增加,蓝光和蓝红光在16 h·d-1下有利于试管苗叶面积的增加,绿光和黄光在24 h·d-1下叶面积均较小。节间长在绿光和黄光下较长,而在蓝光下最短；,8 h·d-1、绿光和黄光下茎节最细；试管苗在16 h·d-1,黄光下根粗最细,而光合速率在LED蓝红光下最大,而在LED黄光和绿光下为负值。
(3)在不同的LED光质和光周期下均表现出,气孔频度越大,则气孔长×宽越小,基因型不同所需光质、光周期也不完全相同,变叶葡萄气孔频度在蓝光、16 h·d-1下最高,而贝达试管苗气孔频度在蓝红光和24 h·d-1下最高。红光、蓝光和蓝红光有利于叶绿素的合成,8 h·d-1较其他三种光周期叶绿合成能力差,且光质与光周期效应也因基因型存在差异
SOD活性均在蓝光下最高,红光下最低。贝达试管苗在8 h·d-1下最高,24 h·d-1下最低,变叶葡萄试管苗则在16 h·d-1下最高,12 h·d-1下最低。试管苗的PPO、POD与CAT活性强弱均为：24h.d-1>16 h.d-1>12 h.d-1>8 h.d-1。
Photosynthetic photon flux density (PPFD), photoperiod and spectral distribution play important roles in regulation of plant photosynthesis and morphogenesis during plant tissue culturing. Plant tissue culture mainly depends on electric light source, which has low bioenergy efficiency and high calorific capacity, and its electricity comsuption acount for 65% of the whole power expenditure, that is one of the non-human costs in plant tissue culture. Using light emitting diode as light source in plant tissue culture to regulate light quality and PPFD can not only regulate the plant growth and development and morphogenesis, but also reduce energy comsumption remarkbly, and decrease the cost accordingly. Besides, it solves the weakness of light quality impurity, and increases the reliability of experiment results.
The test-tube plantlets of grape variety Beta and Vitis piasezkii Maxim were used as the material in this experiment, and their dynamic growth, propagation of the aboveground parts (plant height, leaf size, biomass, internode length and stem diameter) and underground parts (root weight, root numbers, root length and root diameter), pigment content, stoma and enzyme activity were studied with LEDs irradiation (white, red, yellow, blue and green) on four photoperiods (24 h·d-1、16 h·d-1、12 h·d-1、8 h·d-1), so as to get the optimum LED light quality and photoperiod to the plantlets growth.The results are as follows:
1. Red LED was favorable to the longitudinal growth of the plantlets aboveground and underground parts, however, blue LED had opposite effect, but promoted their cross growth; blue-red LED was better than ordinary fluorescence to increase photosynthetic rate of the plantlets. Red, blue and blue-red LEDs enhanced the biomass obviously with 16 h·d-1 irradiation; the stem nodes were the thickest under blue and blue-red LEDs with 24 h·d-1 irradiation.
2. Yellow, green LEDs with 16 h·d-1、12 h·d-1 irradiation were favorable to increase plant height; blue and blue-red LEDs with 16 h·d-1 were benefit to leaf size increase, but the leaf size was smaller under green and yellow LEDs with 24 h·d-1 irradiation. The internodes were longer under green and yellow LEDs, but the shortest under blue LED; the stem nodes were the thinnest under green and yellow LEDs with 8 h·d-1 irradiation; the root was the thinnest under yellow LED with 16 h·d-1 irradiation; however, the photosynthetic rate was the maximum under blue-red LED, but showed negative value under yellow and green LEDs.
3. With each LED and photoperiod irradiation, it was showed that the bigger the stomatal frequency is, the smaller the length X width of the stomata is. The different genotypes needed different LED and photoperiod:the stomatal frequencies of Vitis piasezkii Maxim and Beta were the biggest with blue,16 h·d-1 irradiation and blue-red 24 h·d-1 irradiation, respectively. Red, blue, blue-red LEDs were suitable to chlorophyll synthesis, but 8 h·d-1 was inferior to the other three photoperiods to synthesize chlorophyll; light quality and photoperiod effect varied due to different genotypes.
SOD activities were the highest under blue LED, but the lowest under red LED, and the activity of Beta plantlet was the highest with 8 h·d-1 irradiation, the lowest with 24 h·d-1 irradiation; but that of Vitis piasezkii Maxim was the highest with 16 h·d-1 irradiation, the lowest with 12 h·d-1 irradiation. The order of PPO,POD and CAT activities in test tube plantlets were all 24h.d-1>16 h.d-1 >12h.d-1>8h.d-1。
本研究以葡萄砧木品种贝达(Beta)和变叶葡萄(Vitis piasezkii Maxim)试管苗为实验材料,研究其在LED白、红、黄、蓝、绿和蓝红六种光质,24 h·d-1、16 h·d-1、12 h·d-1、8 h·d-1四个光周期试管苗生长发育动态、地上部位(株高、叶面积、生物量、节间长、茎粗)和地下部位(根重、根条数、根长、根粗)增殖以及色素含量、气孔、酶活性的效应。以便得到组织培养最佳的LED光质、光周期。取得的主要研究结果有：
(1)LED红光有利于葡萄试管苗地上地下部位的纵向生长；LED蓝光抑制葡萄试管苗地上地下部位的纵向生长,而促进其横向生长；LED蓝红光对葡萄试管苗的光合速率较普通荧光强。红光、蓝光和蓝红光在16 h·d-1下对生物量的增加有明显的促进作用：在24 h·d-1、蓝光和蓝红光下茎节最粗。
(2)LED黄光、绿光,光周期16 h·d-1、12 h·d-1的光照有利于株高的增加,蓝光和蓝红光在16 h·d-1下有利于试管苗叶面积的增加,绿光和黄光在24 h·d-1下叶面积均较小。节间长在绿光和黄光下较长,而在蓝光下最短；,8 h·d-1、绿光和黄光下茎节最细；试管苗在16 h·d-1,黄光下根粗最细,而光合速率在LED蓝红光下最大,而在LED黄光和绿光下为负值。
(3)在不同的LED光质和光周期下均表现出,气孔频度越大,则气孔长×宽越小,基因型不同所需光质、光周期也不完全相同,变叶葡萄气孔频度在蓝光、16 h·d-1下最高,而贝达试管苗气孔频度在蓝红光和24 h·d-1下最高。红光、蓝光和蓝红光有利于叶绿素的合成,8 h·d-1较其他三种光周期叶绿合成能力差,且光质与光周期效应也因基因型存在差异
SOD活性均在蓝光下最高,红光下最低。贝达试管苗在8 h·d-1下最高,24 h·d-1下最低,变叶葡萄试管苗则在16 h·d-1下最高,12 h·d-1下最低。试管苗的PPO、POD与CAT活性强弱均为：24h.d-1>16 h.d-1>12 h.d-1>8 h.d-1。
Photosynthetic photon flux density (PPFD), photoperiod and spectral distribution play important roles in regulation of plant photosynthesis and morphogenesis during plant tissue culturing. Plant tissue culture mainly depends on electric light source, which has low bioenergy efficiency and high calorific capacity, and its electricity comsuption acount for 65% of the whole power expenditure, that is one of the non-human costs in plant tissue culture. Using light emitting diode as light source in plant tissue culture to regulate light quality and PPFD can not only regulate the plant growth and development and morphogenesis, but also reduce energy comsumption remarkbly, and decrease the cost accordingly. Besides, it solves the weakness of light quality impurity, and increases the reliability of experiment results.
The test-tube plantlets of grape variety Beta and Vitis piasezkii Maxim were used as the material in this experiment, and their dynamic growth, propagation of the aboveground parts (plant height, leaf size, biomass, internode length and stem diameter) and underground parts (root weight, root numbers, root length and root diameter), pigment content, stoma and enzyme activity were studied with LEDs irradiation (white, red, yellow, blue and green) on four photoperiods (24 h·d-1、16 h·d-1、12 h·d-1、8 h·d-1), so as to get the optimum LED light quality and photoperiod to the plantlets growth.The results are as follows:
1. Red LED was favorable to the longitudinal growth of the plantlets aboveground and underground parts, however, blue LED had opposite effect, but promoted their cross growth; blue-red LED was better than ordinary fluorescence to increase photosynthetic rate of the plantlets. Red, blue and blue-red LEDs enhanced the biomass obviously with 16 h·d-1 irradiation; the stem nodes were the thickest under blue and blue-red LEDs with 24 h·d-1 irradiation.
2. Yellow, green LEDs with 16 h·d-1、12 h·d-1 irradiation were favorable to increase plant height; blue and blue-red LEDs with 16 h·d-1 were benefit to leaf size increase, but the leaf size was smaller under green and yellow LEDs with 24 h·d-1 irradiation. The internodes were longer under green and yellow LEDs, but the shortest under blue LED; the stem nodes were the thinnest under green and yellow LEDs with 8 h·d-1 irradiation; the root was the thinnest under yellow LED with 16 h·d-1 irradiation; however, the photosynthetic rate was the maximum under blue-red LED, but showed negative value under yellow and green LEDs.
3. With each LED and photoperiod irradiation, it was showed that the bigger the stomatal frequency is, the smaller the length X width of the stomata is. The different genotypes needed different LED and photoperiod:the stomatal frequencies of Vitis piasezkii Maxim and Beta were the biggest with blue,16 h·d-1 irradiation and blue-red 24 h·d-1 irradiation, respectively. Red, blue, blue-red LEDs were suitable to chlorophyll synthesis, but 8 h·d-1 was inferior to the other three photoperiods to synthesize chlorophyll; light quality and photoperiod effect varied due to different genotypes.
SOD activities were the highest under blue LED, but the lowest under red LED, and the activity of Beta plantlet was the highest with 8 h·d-1 irradiation, the lowest with 24 h·d-1 irradiation; but that of Vitis piasezkii Maxim was the highest with 16 h·d-1 irradiation, the lowest with 12 h·d-1 irradiation. The order of PPO,POD and CAT activities in test tube plantlets were all 24h.d-1>16 h.d-1 >12h.d-1>8h.d-1。
引文
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