不同小麦雄性不育类型AGPase活性及杂种光合特性的分析

Analysis of ADP-Glucose Pyrophosphorylase Activity of Male Sterile Lines and Photosynthetic Characteristics of Hybrid in Wheat

作者: 专业:作物遗传育种 导师:高庆荣 年度:2010 学位:硕士  院校: 山东农业大学

Keywords

Cytoplasmic male sterility (CMS), ADP-Glucose Pyrophosphorylase, Sterile mechanism, Photosynthetic rate, Physiological parameters

        为进一步探寻小麦不育系的不育机制和籽粒不饱满的生理机制,以冀5418核基因为遗传背景,对同核异质K、V、T型不育系叶片、幼穗和籽粒中的腺苷二磷酸葡萄糖焦磷酸化酶(ADP-glucose pyrophosphorylase,AGPase)活性和淀粉积累量进行了动态观测,并与各自的保持系进行了比较;为了探讨不同细胞质对其杂种光合生理特性的影响,以8个同核异质雄性不育系(K、V、T、CHA型不育系)及其恢复系758461-4配制的杂种F1为材料,在4个生育时期(抽穗期、开花期、灌浆中期、灌浆后期)测定了旗叶净光合速率(Pn)及相关生理参数(气孔导度Gs、蒸腾速率Tr、细胞间隙CO2浓度Ci、大气CO2浓度(Ca),并对水分利用率WUE和气孔限制值Ls进行了分析。主要研究结果如下:(1)雄性不育的可能原因是雌雄蕊原基分化期幼穗和叶片中AGPase活性高,幼穗发育所需能量供应不足;而四分体期幼穗AGPase活性低,影响了花粉中淀粉积累。在雌雄蕊原基分化期,不育系幼穗中AGPase活性较保持系高9.33~27.94μmol g?1 FW h?1,差异达极显著水平(F=133.81, P<0.0001);而在四分体期,不育系幼穗中该酶活性极显著低于保持系(F=13.97~75.20, P<0.0001),差异为4.27~7.44μmol g?1 FW h?1。雌雄蕊原基分化期至四分体时期,不育系叶片中AGPase活性较保持系高7.39~80.77μmol g?1 FW h?1,差异达极显著水平(F=135.76~5454.28, P<0.0001);(2)不育系对籽粒AGPase活性具有明显的不良胞质效应,降低了ADPG供应水平,影响淀粉的积累是籽粒不饱满的重要原因之一。不育系强、弱势粒中总淀粉、直链淀粉和支链淀粉积累量、AGPase平均活性、淀粉含量及直/支比均极显著低于保持系,且这些指标均表现为强势粒显著高于弱势粒。Logistic方程显示,不育系籽粒淀粉积累量的减少主要由淀粉积累速率降低引起;籽粒AGPase活性与淀粉积累速率显著或极显著正相关(r=0.4460~0.7150, P=0.0004~0.0487);灌浆期,叶片中AGPase活性与光合速率呈负相关(r=?0.28634, P=0.2823);(3)杂种F1的净光合速率和生理参数Ci、Gs、WUE的优势普遍存在,其中T型胞质杂种净光合速率的优势最强,超中亲(MP) 20.029%,K、V型杂种分别超MP 10.630%、7.099%;(4)杂种F1的净光合速率在一定程度上受气孔因素的限制。杂种F1及其亲本净光合速率、生理参数4个生育时期的变化趋势相同,从开花期到灌浆期,Pn与Ci、Gs的变化趋势相同,与Ls的相反;(5) K、V、T型细胞质对杂种F1的净光合速率、生理参数无不良影响。K型杂种的Pn较CHA型低0.077μmolCO2·m?2·s?1,V、T型杂种的Pn较CHA型分别高0.419μmolCO2·m?2·s?1、0.725μmolCO2·m?2·s?1,但差异不显著;(6)在配制杂种时,通过选择优良的基因型可以改善杂种F1的气孔导度。以太911289为母本配制的杂种的Gs比以冀5418为母本配制的杂种高29.127 mmolm?2·s?1,差异极显著。
    Cytoplasmic male-sterile (CMS) lines, especially K (Aegilops kotchyi), V (Aegilops ventricosa), and T (Triticum timopheevi) types of cytoplasms, are of high value in heterosis utilization in wheat (Triticum aestivum L.). The sterile mechanisms of these CMS types have been focused in hybrid wheat research. As a key and rate-limiting starch biosynthetic enzyme, ADP-glucose pyrophosphorylase (AGPase) is considered to play an important role in the sterility of CMS lines. For gaining an insight into the physiological basis of the sterility in CMS lines of wheat, the activities of AGPase in leaves, young spikes, and grains were measured in K, V, and T types of isogenic CMS lines (Ji 5418 background) at different growth stages, and compared with those of their maintainer line. The dynamic accumulations of starch, amylose, and amylopectin were also investigated in both superior (the first and the second grains in a spikelet) and inferior (grains except for the first and the second grains in a spikelet) grains; In present work, 2 nuclear background male sterile lines including K, V, T, CHA-cytoplasm male-sterile lines, and restorer line and their hybrids, were employed to study the effect of different cytoplasmic types on the photosynthesis and physiological parameters: stomatal conductance (Gs), transpiration rate (Tr), CO2 concertration (Ci), atmospheric CO2 concentration (Ca), water use efficiency (WUE), intercellar stomatal limiting value (Ls) of hybrids at heading stage, anthesis stage, mid-filling stage and later-filling stage. The major results are as follows:(1)the male sterility is probably in relation to the insufficiency of energy in leaves and young pikes at floret primordium stage resulting from the high AGPase activity, and the low accumulation of starch in pollens at tetrad stage caused by low AGPase activity in young spikes. In young spikes, the AGPase activities of the three CMS lines were significantly higher (F = 133.81, P < 0.0001) with the differences of 9.33 ? 27.94μmol g?1 FW h?1 at floret primordium stage, and significantly lower (F = 13.97 ? 75.20, P < 0.0001) by 4.27 ? 7.44μmol g?1 FW h?1 at tetrad stage as compared with the maintainer line. From floret primordium to tetrad stage, the CMS lines had continuously higher level of AGPase activities in leaves than the maintainer line, with the differences ranged from 7.39 to 80.77μmol g?1 FW h?1 (F = 135.76 ? 5454.28, P < 0.0001).(2)the sterile cytoplasm has a negative effect on AGPase activity, resulting in decreases of ADPG level and starch accumulation in grains which may result in unfilled grains in CMS lines. Compared with the maintainer line, the three CMS lines all presented lower accumulations (P < 0.05) of starch, amylose and amylopectin in grain. The starch content and AGPase activity in superior grains were significantly higher than those in inferior grains for CMS lines. The dynamic accumulations of starch in superior and inferior grains fitted Logistic equations, and positively correlated with the starch accumulation rate (r = 0.4460 ? 0.7150, P = 0.0004 ? 0.0487). The AGPase activity in leaves had an insignificantly negative correlation with net photosynthetic rate.(3) The heterosis on photosynthesis and physiological parameters Ci, Gs and WUE of hybrid F1 was common. The photosynthesis of T-cytoplasm hybrid had the highest heterosis with 20.029%, K, V-cytoplasm were 10.630%,7.099% higher than those of Mid-parent, respectively.(4) The dynamic changes of Pn were restricted by the stomatal factors to some extent. The dynamic changes of hybrid F1 is the same as their parents at four stages. The dynamic changes of Pn were the same as Ci and Gs, and were different from Ls. (5) K, V, T-type cytoplasm had no negative effect on hybrid F1. Compared with CHA-cytoplasm, the Pn value of K-cytoplasm hybrid was lower, and decreased by 0.077μmolCO2·m?2·s?1, while these of V, T-cytoplasm hybrid were higher with increasing of 0.419μmolCO2·m?2·s?1, 0.725μmolCO2·m?2·s?1, respectively, but the difference was not significant.(6) The physiological parameter stomatal conductance of hybrid F1 could be improved with excellent genotype. The Gs value of hybrid F1 with tai911289 as female parent was higher than that whose female parent was ji5418 with increasing of 29.127 mmolm?2·s?1, with significantly difference.
        

不同小麦雄性不育类型AGPase活性及杂种光合特性的分析

摘要8-10
Abstract10-12
1 引言13-33
    1.1 研究背景13-16
    1.2 CMS 不育机理的细胞生物学研究16-18
        1.2.1 败育的主要时期16-17
        1.2.2 花丝和花隔维管束与CMS17
        1.2.3 胼胝质与CMS17-18
        1.2.4 绒毡层与CMS18
        1.2.5 细胞质与CMS18
    1.3 CMS 不育机理的生理生化研究18-26
        1.3.1 能量代谢与CMS19
        1.3.2 呼吸速率与CMS19
        1.3.3 酶活性与CMS19-22
        1.3.4 核酸、氨基酸与CMS22-23
        1.3.5 Ca2+与CMS23-24
        1.3.6 细胞骨架与CMS24-25
        1.3.7 内源激素与CMS25
        1.3.8 蛋白质与CMS25-26
        1.3.9 糖类与CMS26
    1.4 CMS 不育机理的分子生物学研究26-31
        1.4.1 线粒体与CMS27-30
            1.4.1.1 线粒体结构和数量与CMS27
            1.4.1.2 线粒体多肽差异与CMS27
            1.4.1.3 线粒体DNA 与CMS27-30
        1.4.2 叶绿体与CMS30-31
    1.5 目的意义31-33
2 材料与方法33-38
    2.1 试验材料33
    2.2 试验设计33
    2.3 取样方法33-34
    2.4 测定项目34-37
        2.4.1 AGPase 活性的测定34
        2.4.2 淀粉含量的测定34-36
        2.4.3 淀粉积累过程拟合36-37
        2.4.4 旗叶净光合速率的测定37
    2.5 数据统计与分析37-38
3 结果与分析38-54
    3.1 AGPase 活性对育性的影响38-39
    3.2 AGPase 活性对籽粒形成的影响39-46
        3.2.1 淀粉积累39
        3.2.2 淀粉积累过程的Logistic 拟合39-40
        3.2.3 强弱势粒淀粉积累量差异分析40-41
        3.2.4 籽粒AGPase 活性41-42
        3.2.5 强弱势粒AGPase 活性比较42-43
        3.2.6 籽粒AGPase 活性对直链淀粉和支链淀粉的影响43-45
        3.2.7 旗叶AGPase 活性对净光合速率的影响45-46
    3.3 杂种F1 光合生理特性的分析46-54
        3.3.1 杂种F1 净光合速率、生理参数的优势表现46-47
        3.3.2 不同生育时期杂种 F1 及其亲本净光合速率、生理参数的变化趋势47-50
        3.3.3 不同细胞质对杂种F1 净光合速率、生理参数的影响50-51
        3.3.4 核质互作效应51-52
        3.3.5 母本基因型对杂种F1 净光合速率、生理参数的影响52-54
4 讨论54-57
    4.1 AGPase 活性与育性的关系54
    4.2 AGPase 活性对籽粒形成的影响54-55
    4.3 K、V、T 型细胞质对杂种F1 光合特性的影响55-57
5 结论57-58
参考文献58-68
致谢68-69
攻读硕士学位期间发表的论文69
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