【作者】 杨金华；

【导师】 杜克久；

【作者基本信息】 河北农业大学 ， 林木遗传育种， 2011， 硕士

【摘要】 毛白杨(Populus tomentosa Carr.)属杨柳科(Salicaceae)杨属(Populus L.)白杨派(Sect.Populus)树种,因其生长快、适应性强、树干通直挺拔、树型优美而在城乡环境绿化美化和工农业用材中发挥了重要作用,通过组织培养快速繁殖良种已是目前广泛应用的技术,但试管苗长期在高湿、弱光、恒温、无风的条件下生长,其气孔的形态和生理与自然条件下生长的植株明显不同,并直接影响试管苗的移栽成活率。本文利用指甲油涂抹撕取法对转基因毛白杨试管苗与10年生植株叶片下表皮气孔密度、气孔大小、气孔开度日变化、气孔开张率日变化进行了较为系统的研究,同时从离体和活体两个方面研究了萎蔫、H2O2、CaCl2、ABA、SA、乙烯利、黑暗、培养基中不同浓度ABA、培养基中不同浓度CaCl2、培养基中糖对试管苗气孔开度的影响。本研究对于进一步了解试管苗气孔运动机制及调控具有重要理论意义和实际应用价值。主要研究结果如下:1.转基因毛白杨试管苗叶片下表皮气孔密度为255.10个/mm2,10年生植株叶片下表皮气孔密度为374.15个/mm2,试管苗叶片下表皮气孔密度小于10年生植株,二者差异达0.01极显著水平。2.转基因毛白杨试管苗叶片下表皮气孔大小为610.86μm2,10年生植株叶片下表皮气孔大小为112.97μm2,试管苗叶片下表皮气孔明显比10年生植株气孔大,二者差异达0.01极显著水平。3.转基因毛白杨试管苗叶片下表皮气孔开度在60.53～92.17μm2之间变化,10年生植株则在16.70～18.74μm2之间变化,试管苗叶片下表皮气孔开度大于10年生植株。4.转基因毛白杨试管苗叶片下表皮气孔开度的变化明显,最小值与最大值之比约为0.66:1,日变化呈明显波峰状,两个高峰分别出现在9:00和15:00;10年生植株叶片下表皮气孔开度的变化很小,最小值与最大值之比约为0.89:1,日变化曲线平稳,无明显波动。5.转基因毛白杨试管苗叶片下表皮气孔开张率在一昼夜均为100%,10年生植株叶片下表皮气孔开张率在一昼夜的变化呈双曲线,气孔开张率在两高峰处低于100%,晚上21:00到第二天凌晨3:00气孔张开率为0%。6.转基因毛白杨试管苗在离体状态下,萎蔫、H2O2都能使转基因毛白杨试管苗叶片下表皮气孔开度降低,但是均不能促进其气孔关闭;Ca2+、ABA、SA、乙烯利在浓度分别达到1mol?L-1、200μmol?L-1、0.1mmol?L-1、0.4%时均可诱导转基因毛白杨试管苗叶片下表皮气孔关闭。7.转基因毛白杨试管苗在活体状态下,连续黑暗、0.5mg?L-1~3mg?L-1的ABA不能促进转基因毛白杨试管苗叶片下表皮气孔关闭;37.8g?L-1的CaCl2可诱导其气孔关闭;转基因毛白杨无糖试管苗与有糖试管苗相比,叶片下表皮气孔开度降低,气孔的关闭率为83%。更多还原

【Abstract】 Populus tomentosa Carr. is a native species of section populus China. It plays an important role in the urban and rural environmental afforestation and industrial and agricultural material as its fast growth, the adaptability with the straight trunk and beautiful shapes. Rapid propagation of improved varieties through tissue culture is a widely used technology, however, the long-term growth of tube seedlings in high humidity, weak light, temperature, no wind conditions make its stomatal morphology and physiology significantly different from that of plants grown under natural conditions, this affacts the survival rate of transplanations of tube seedlings. With nail polish, the change of stomatal density, stomatal size, the daily variation of stomatal aperture, the daily variation of stomatal open percentage of the leaves’lower epidermis of tube seedlings and the 10 years plant of transgenic Populus tomentosa Carr. were compared, and the effect of wilting, H2O2, Ca2+, ABA, SA, ethrel, darkness, different concentrations of ABA in medium, different concentrations of CaCl2 in medium, tube seedlings grown in medium without sugar on stomatal aperture were studied, and analyzed the reason of abnormal stomatal behavior. It is important for the further understanding of the guard cells signal transduction pathways and the mechanisms of tube seedlings. The main results were as follows:1. Stomatal density of the leaves’lower epidermis of tube seedlings of transgenic Populus tomentosa Carr. was 255.10/mm2, stomatal density of the leaves’lower epidermis of the 10 years plant was 374.15/mm2, tube seedlings’stomatal density was less than that of the 10 years plant, there was a significant difference in stomatal density between tube seedlings and the 10 years plant.2. Stomata of the leaves’lower epidermis of tube seedlings of transgenic Populus tomentosa Carr. was 610.86μm2, stomata of the leaves’lower epidermis of the 10 years plant was 112.97μm2, tube seedlings’stomata was obviously larger than that of the 10 years plant, there was a significant difference in stomata between tube seedlings and the 10 years plant.3. Stomatal aperture of the leaves’lower epidermis of tube seedlings of transgenic Populus tomentosa Carr. was 60.53～92.17μm2, stomatal aperture of the leaves’lower epidermis of the 10 years plant was 16.70～18.74μm2, stomatal aperture in tube seedlings was greater than that in the 10 years plant.4. The changes of stomatal aperture of the leaves’lower epidermis of tube seedlings of transgenic Populus tomentosa Carr. was evident, the ratio of minimum and maximum was about 0.66:1, changes of stomatal aperture during 24 hours appeared wave crest, the two peaks of stomatal aperture were 9:00 and 15:00; the changes of stomatal aperture of the leaves’lower epidermis of the 10 years plant was not evident, the ratio of minimum and maximum was about 0.89:1, changes of stomatal aperture during 24 hours was steady and no evident fluctuation.5. Stomatal open percentage reached 100% in tube seedlings of transgenic Populus tomentosa Carr. during a day, a bimodal curve change was observed in stomatal open percentage of the 10 years plant.6. In vitro, wilting , H2O2 can reduce stomatal aperture, but not promote stomatal closing; the Ca2+, ABA, SA, ethrel can induce stomatal closing when their concentration were respectively 1mol?L-1、200μmol?L-1、0.1mmol?L-1、0.4%.7. In vivo, continuous darkness, ABA in medium can not promote stomatal closing when their concentration were 0.5mg?L-1~3mg?L-1; CaCl2 in medium can induce stomatal closing when their concentration were 37.8g?L; tube seedlings grown in medium without sugar compared with tube seedlings grown in medium with sugar, stomatal aperture of tube seedlings grown in medium without sugar was reduced, stomatal close percentage of tube seedlings grown in medium without sugar were 83%.更多还原