花器官分化期玉米干旱胁迫的蛋白质组学研究和170个地方种的遗传多样性分析
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摘要
玉米(Zea mays L.)是重要的粮、经、饲兼用作物,在国民经济发展中具有举足轻重的地位。干旱是严重影响玉米产量提高的一个主要限制因素,花器官分化期,即从减数分裂到四分体花粉母细胞的时期,对干旱胁迫最为敏感,该阶段的干旱胁迫将造成大幅度的减产。并且,我国玉米种质的遗传基础比较狭窄,但是我国具有丰富的玉米地方品种,这些地方种的环境适应性强、遗传基础丰富、含有优异的农艺性状。因此,为了寻找有用的耐旱相关基因、拓宽玉米遗传种质基础和保存种质资源,我们对花器官分化期不同耐旱性玉米的生理和蛋白质组变化,以及170个地方种的遗传多样性进行系统研究。主要结果如下：
(1)随着干旱程度加剧,旱21和吉81162的Fv/Fm和叶绿素含量都显著降低。
(2)对于不同干旱胁迫下的雄穗,APX活性先降低后增加；GR活性随着干旱胁迫时间的延长均显著性增加,旱21的活性高于吉81162；CAT活性随着干旱胁迫时间的延长均先增加后降低,旱21的增加幅度大于吉81162。旱21的SOD和DHAR的含量均降低不显著,而吉81162的却显著性降低。这些结果表明：APX、GR和CAT是雄穗干旱胁迫条件下活性氧清除的关键酶。
(3)对于不同干旱胁迫下的叶片,干旱胁迫下的SOD和APX的含量都高于对照,然而,旱21的增加幅度大于吉81162；DHAR活性变化不大；GR是显著性降低的；CAT先降低后增加。通过这些结果,我们发现SOD、APX和CAT则是叶片干旱胁迫条件下活性氧清除的关键酶。
(4)经双向凝胶电泳和质谱分析,旱21和吉81162雄穗中表达丰度达2倍差异的蛋白点分别为114和100个,分别鉴定出13种和9种相应干旱胁迫的蛋白质。其中,抗坏血酸过氧化物酶和肌动蛋白在旱21和吉81162中都差异表达；类果胶酯酶抑制蛋白、类豆球蛋白、前纤维蛋白、磷酸丙糖异构酶、NAD的表异构酶／脱水酶、GTP结合蛋白、14-3-3蛋白和真核翻译起始因子eIF-5A等仅在旱21中差异表达；铁氧还原蛋白、翻译起始因子eIF-4A、拟定蛋白、肌动蛋白解聚因子ADF和ADF-3等仅在吉81162中差异表达。
(5)我国西南地区具有丰富的遗传多样性,各省间有较强的基因流,能抵制遗传漂变的作用。基于遗传距离的聚类分析和基于数学模型的STRUCTURE混合分析表明,170个地方种归属于3个亚群。群Ⅰ主要含贵州和广西的地方种,群Ⅱ为混合群,群Ⅲ含有广西和云南的地方种。
Maize (Zea mays L.) is a very important feed, grain and industrial crop, which plays an important role in the development of national economy. Drought stress is one of the major environmental stresses that greatly affect plant growth and maize production. The period from meiosis to tetrad break-up in anthers is one of the most sensitive stages; crop yield will be caused serious reduction when the maize encounters drought stress at this stage. Meanwhile, the maize genetic background in China was narrow. However, maize landraces have strong compatibility in local environment, rich genetic background and a lot of useful characters. In order to find drought tolerance genes, widen maize hereditary basis and conserve germplasms, the physiological and proteomic analysis of maize inbred lines with different tolerance at meiotic stage and the genetic diversity of170landraces from southwest China were analyzed. The main results are as follows:
(1) As the drought stress intensified, the Fv/Fm and chlorophyll content of Han21and Ji81162were significantly decreased.
(2) As for the ear under control and drought stress, the activity of APX in both two inbred lines was decreased first and then increased. The activity of GR was significantly increased with the stress time prolonged. The GR activities of Han21were higher than Ji81162. The activity of CAT was first increased and then decreased with prolonged drought stress time. The SOD and DHAR activities of Han21were slightly decreased, while they were markedly declined. These results suggested that APX, GR and CAT were the key active oxygen scavenging enzymes in ears.
(2) As for the leaves under control and drought stress, the SOD and APX activities were higher than control, and the active of Han21were increased fasterly than Ji81162. The DHAR activities had no change between control and drought. The activity of GR was significantly decreased. The activity of CAT was decreased first and then increased for two inbred lines. These results indicated that SOD, APX and CAT were the key enzymes in leaves under drought stress.
(4) After proteomic analysis by two-dimensional gel electrophoresis and peptide mass fingerprinting,114and100proteins were found in Han21and Ji81162, respectively.13and9proteins, respectively, which were responed to drought stress, were identidied. APX and actin were found in two inbred lines. Pectinesterase inhibitor domain containing protein, legumin-like protein, profiling, triosephosphate isomerase, NAD dependent epimerase/dehydratase, GTP-binding protein,14-3-3-like protein and eukaryotic translation initiation factor5A (eIF-5A) were only found in Han21. While ferredoxin, eIF-4A, hypothetical protein, actin-depolymerizing factor (ADF) and ADF-3were only expressed in Ji81162.
(5) The genetic diversity of landraces from southwest China were rich; the gene flow among provinces was strong enough to prevent differentiation through drift among provinces. The genetic distance-based and the model-based clustering both grouped170landraces into3groups:group Ⅰ contained landraces mainly from Guizhou and Guangxi provinces; group Ⅱ was the mixed group; group Ⅲ consisted of landraces mainly from Yunnan and Guangxi provinces.
(1)随着干旱程度加剧,旱21和吉81162的Fv/Fm和叶绿素含量都显著降低。
(2)对于不同干旱胁迫下的雄穗,APX活性先降低后增加；GR活性随着干旱胁迫时间的延长均显著性增加,旱21的活性高于吉81162；CAT活性随着干旱胁迫时间的延长均先增加后降低,旱21的增加幅度大于吉81162。旱21的SOD和DHAR的含量均降低不显著,而吉81162的却显著性降低。这些结果表明：APX、GR和CAT是雄穗干旱胁迫条件下活性氧清除的关键酶。
(3)对于不同干旱胁迫下的叶片,干旱胁迫下的SOD和APX的含量都高于对照,然而,旱21的增加幅度大于吉81162；DHAR活性变化不大；GR是显著性降低的；CAT先降低后增加。通过这些结果,我们发现SOD、APX和CAT则是叶片干旱胁迫条件下活性氧清除的关键酶。
(4)经双向凝胶电泳和质谱分析,旱21和吉81162雄穗中表达丰度达2倍差异的蛋白点分别为114和100个,分别鉴定出13种和9种相应干旱胁迫的蛋白质。其中,抗坏血酸过氧化物酶和肌动蛋白在旱21和吉81162中都差异表达；类果胶酯酶抑制蛋白、类豆球蛋白、前纤维蛋白、磷酸丙糖异构酶、NAD的表异构酶／脱水酶、GTP结合蛋白、14-3-3蛋白和真核翻译起始因子eIF-5A等仅在旱21中差异表达；铁氧还原蛋白、翻译起始因子eIF-4A、拟定蛋白、肌动蛋白解聚因子ADF和ADF-3等仅在吉81162中差异表达。
(5)我国西南地区具有丰富的遗传多样性,各省间有较强的基因流,能抵制遗传漂变的作用。基于遗传距离的聚类分析和基于数学模型的STRUCTURE混合分析表明,170个地方种归属于3个亚群。群Ⅰ主要含贵州和广西的地方种,群Ⅱ为混合群,群Ⅲ含有广西和云南的地方种。
Maize (Zea mays L.) is a very important feed, grain and industrial crop, which plays an important role in the development of national economy. Drought stress is one of the major environmental stresses that greatly affect plant growth and maize production. The period from meiosis to tetrad break-up in anthers is one of the most sensitive stages; crop yield will be caused serious reduction when the maize encounters drought stress at this stage. Meanwhile, the maize genetic background in China was narrow. However, maize landraces have strong compatibility in local environment, rich genetic background and a lot of useful characters. In order to find drought tolerance genes, widen maize hereditary basis and conserve germplasms, the physiological and proteomic analysis of maize inbred lines with different tolerance at meiotic stage and the genetic diversity of170landraces from southwest China were analyzed. The main results are as follows:
(1) As the drought stress intensified, the Fv/Fm and chlorophyll content of Han21and Ji81162were significantly decreased.
(2) As for the ear under control and drought stress, the activity of APX in both two inbred lines was decreased first and then increased. The activity of GR was significantly increased with the stress time prolonged. The GR activities of Han21were higher than Ji81162. The activity of CAT was first increased and then decreased with prolonged drought stress time. The SOD and DHAR activities of Han21were slightly decreased, while they were markedly declined. These results suggested that APX, GR and CAT were the key active oxygen scavenging enzymes in ears.
(2) As for the leaves under control and drought stress, the SOD and APX activities were higher than control, and the active of Han21were increased fasterly than Ji81162. The DHAR activities had no change between control and drought. The activity of GR was significantly decreased. The activity of CAT was decreased first and then increased for two inbred lines. These results indicated that SOD, APX and CAT were the key enzymes in leaves under drought stress.
(4) After proteomic analysis by two-dimensional gel electrophoresis and peptide mass fingerprinting,114and100proteins were found in Han21and Ji81162, respectively.13and9proteins, respectively, which were responed to drought stress, were identidied. APX and actin were found in two inbred lines. Pectinesterase inhibitor domain containing protein, legumin-like protein, profiling, triosephosphate isomerase, NAD dependent epimerase/dehydratase, GTP-binding protein,14-3-3-like protein and eukaryotic translation initiation factor5A (eIF-5A) were only found in Han21. While ferredoxin, eIF-4A, hypothetical protein, actin-depolymerizing factor (ADF) and ADF-3were only expressed in Ji81162.
(5) The genetic diversity of landraces from southwest China were rich; the gene flow among provinces was strong enough to prevent differentiation through drift among provinces. The genetic distance-based and the model-based clustering both grouped170landraces into3groups:group Ⅰ contained landraces mainly from Guizhou and Guangxi provinces; group Ⅱ was the mixed group; group Ⅲ consisted of landraces mainly from Yunnan and Guangxi provinces.
引文
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