玉米ZmCBL-ZmCIPK参与钾离子吸收转运的功能研究
发布时间：2019-06-03 00:56
【摘要】：钾是植物生长发育所必需的矿质元素。然而我国耕地土壤普遍缺钾且钾肥资源匮乏,这已经成为制约农作物生产的重要限制因素之一。因此,通过遗传改良方法提高作物钾吸收利用效率是解决我国农作物生产中钾供应不足问题的重要途径之一。本实验室前期研究发现,AKT1是模式植物拟南芥根部介导钾吸收的主要钾离子通道,其通道活性受钙结合蛋白CBL1/9和蛋白激酶CIPK23的调控。过量表达CBL1/9和CIPK23均可以提高拟南芥的低钾耐受能力。ZMK1是AKT1在玉米中的同源蛋白,体外实验证明ZMK1的通道活性受玉米钙结合蛋白ZmCBL1和蛋白激酶ZmCIPK3/9/23的调控。本论文工作主要研究这些ZmCBL和ZmmCIPK基因在玉米钾吸收利用中的生理功能,进而筛选钾高效吸收利用的玉米转基因材料。首先构建了 ZmCBL1和ZmCIPK3/9/23的过量表达转基因玉米材料,并对这些转基因玉米自交系进行了钾吸收利用效率性状的检测。在低钾水培条件下,ZmCBL1过表达植株的老叶出现提前发黄枯萎的表型。钾含量测定结果显示,低钾条件下ZmCBL1过表达植株根部和冠部的钾含量均显著低于对照材料。这些结果表明过量表达ZmCBL1可能导致玉米对低钾胁迫更为敏感。钾营养耗竭实验结果显示,ZmCIPK23过量表达植株的钾离子吸收速率高于对照材料。然而ZmCIPK23过表达植株在低钾下却表现出老叶提前发黄枯萎的敏感表型。钾含量测定结果显示,在低钾下ZmCIPK23过表达植株冠部钾含量略低于对照材料。进一步研究发现ZmCIPK23过表达植株的老叶和上部伸展叶中的钾含量显著低于对照材料,而茎节中的钾含量却显著高于对照材料。基因表达检测显示,正常条件下ZmCIPK23主要在根中表达,但低钾处理后ZmCIPK23在老叶中的表达量显著升高。推测,ZmCIPK23可能参与低钾下玉米冠部钾离子的再分配过程。过量表达ZmCIPK23可能会干扰钾离子在冠部的再分配过程,从而导致转基因植株的低钾敏感表型。实验结果显示ZmCIPK3和ZmCIPK9的过表达植株在钾吸收利用效率上与对照材料相比并无显著差异,推测ZmCIPK3/9在玉米体内可能并不参与钾吸收转运过程。本论文还对上述转基因玉米自交系开展了田间表型检测。结果显示,减少钾肥施用会影响玉米的生长状况。在拔节期,部分转基因自交系(ZmCBL1和ZmCIPK9过表达玉米材料)与对照材料植株比表现出生长优势。但在吐丝期,这些过表达自交系在株高、茎粗、穗位等方面与对照材料相比并没有显著差异。本论文的研究结果表明,ZmCBL1和ZmCIPK23有可能参与玉米的钾吸收和再分配过程。但是在玉米中持续过量表达ZmCBL1和ZmCIPK3/9/23并不能提高水培条件下玉米的钾吸收利用效率。推测,玉米中可能存在与拟南芥不同的钾吸收调控机制。在低钾条件下,ZmCBL1和ZmCIPK23等基因合理的时空表达,以及正确的蛋白活性调控可能才是提高玉米钾吸收利用效率的关键。本论文的研究结果对进一步探讨玉米钾吸收和转运的调控机制以及未来玉米钾吸收利用效率性状的遗传改良提供了一定的借鉴。
[Abstract]:K is a mineral element necessary for plant growth and development. However, there is a shortage of potassium and potassium in the cultivated land in China, which has become one of the important factors to restrict crop production. Therefore, the improvement of the utilization efficiency of the crop potassium by the genetic improvement method is one of the important ways to solve the problem of insufficient potassium supply in the production of crops in China. In this lab, AKT1 is the main potassium channel of the root-mediated potassium absorption in Arabidopsis thaliana, and its channel activity is regulated by the calcium-binding protein CBL1/9 and the protein kinase CIPK23. Overexpression of CBL1/9 and CIPK23 can improve the low-potassium tolerance of Arabidopsis. ZMK1 is the homologous protein of AKT1 in maize, and the in vitro experiments show that the channel activity of ZMK1 is regulated by the binding protein ZmCBL1 and protein kinase ZmCIPK3/9/23. The work of this thesis is to study the physiological functions of these ZmCBL and ZmCIPK genes in the absorption and utilization of K. The expression of ZmCBL1 and ZmCIP3/9/23 was first constructed, and the utilization efficiency of these transgenic maize inbred lines was tested. Under the condition of low potassium hydroponic culture, the leaves of ZmCBL1 over-expressed plants showed an early and withered phenotype. The results of the determination of potassium content showed that the potassium content of the root and crown of ZmCBL1 over-expressed plants was significantly lower than that of the control material under the condition of low potassium. These results indicate that overexpression of ZmCBL1 may lead to more sensitive maize to low-potassium stress. The results showed that the potassium ion absorption rate of ZmCIPK23 over-expressed plants was higher than that of the control material. However, the expression of ZmCIPK23 over-expressed plants in low-potassium shows the sensitive phenotype of the leaves of the old leaves in advance. The results of the determination of potassium content showed that the potassium content of the plant crown was slightly lower than that of the control material under the low K. The results showed that the potassium content of the leaves and the upper extension leaves of ZmCIPK23 over-expressed plants was significantly lower than that of the control material, while the potassium content in the stem was significantly higher than that of the control material. The expression of ZmCIPK23 was mainly expressed in the roots under normal conditions, but the expression of ZmCIPK23 in the old leaves increased significantly after the low potassium treatment. It is assumed that ZmCIPK23 may be involved in the re-distribution of the potassium ion of the corn crown under low potassium. Overexpression of ZmCIPK23 may interfere with the redistribution of potassium ions in the crown, resulting in a low-potassium-sensitive phenotype of the transgenic plant. The results showed that the over-expression plants of ZmCIP3 and ZmCIPK9 had no significant difference in the utilization efficiency of K, and it was suggested that ZmCIP3/9 could not participate in the process of potassium absorption and transport in maize. The field phenotype of the above-mentioned transgenic maize inbred line was also studied in this paper. The results showed that the reduction of the application of potassium fertilizer would affect the growth of the corn. In the jointing stage, a part of the transgenic inbred lines (ZmCBL1 and ZmCIPK9 over-expressed corn material) and the control material plant ratio showed a growth advantage. However, in the silk-laying period, the over-expression inbred lines were not significantly different from the control materials in terms of plant height, stem size, ear position and so on. The results of this paper show that ZmCBL1 and ZmCIPK23 are likely to be involved in the process of potassium absorption and redistribution of maize. However, the sustained over-expression of ZmCBL1 and ZmCIPK3/9/23 in the corn does not improve the utilization efficiency of the potassium in the corn under water culture conditions. It is suggested that there may be different potassium absorption and control mechanisms in maize. Under the condition of low potassium, the expression of ZmCBL1 and ZmCIPK23 and other genes is reasonable, and the correct regulation of protein activity may be the key to improve the utilization efficiency of potassium in maize. The results of this study provide some reference for the further study of the regulation and control mechanism of the absorption and transport of potassium in corn and the genetic improvement of the utilization efficiency of potassium in the future.
【学位授予单位】：中国农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S513
本文编号：2491550
[Abstract]:K is a mineral element necessary for plant growth and development. However, there is a shortage of potassium and potassium in the cultivated land in China, which has become one of the important factors to restrict crop production. Therefore, the improvement of the utilization efficiency of the crop potassium by the genetic improvement method is one of the important ways to solve the problem of insufficient potassium supply in the production of crops in China. In this lab, AKT1 is the main potassium channel of the root-mediated potassium absorption in Arabidopsis thaliana, and its channel activity is regulated by the calcium-binding protein CBL1/9 and the protein kinase CIPK23. Overexpression of CBL1/9 and CIPK23 can improve the low-potassium tolerance of Arabidopsis. ZMK1 is the homologous protein of AKT1 in maize, and the in vitro experiments show that the channel activity of ZMK1 is regulated by the binding protein ZmCBL1 and protein kinase ZmCIPK3/9/23. The work of this thesis is to study the physiological functions of these ZmCBL and ZmCIPK genes in the absorption and utilization of K. The expression of ZmCBL1 and ZmCIP3/9/23 was first constructed, and the utilization efficiency of these transgenic maize inbred lines was tested. Under the condition of low potassium hydroponic culture, the leaves of ZmCBL1 over-expressed plants showed an early and withered phenotype. The results of the determination of potassium content showed that the potassium content of the root and crown of ZmCBL1 over-expressed plants was significantly lower than that of the control material under the condition of low potassium. These results indicate that overexpression of ZmCBL1 may lead to more sensitive maize to low-potassium stress. The results showed that the potassium ion absorption rate of ZmCIPK23 over-expressed plants was higher than that of the control material. However, the expression of ZmCIPK23 over-expressed plants in low-potassium shows the sensitive phenotype of the leaves of the old leaves in advance. The results of the determination of potassium content showed that the potassium content of the plant crown was slightly lower than that of the control material under the low K. The results showed that the potassium content of the leaves and the upper extension leaves of ZmCIPK23 over-expressed plants was significantly lower than that of the control material, while the potassium content in the stem was significantly higher than that of the control material. The expression of ZmCIPK23 was mainly expressed in the roots under normal conditions, but the expression of ZmCIPK23 in the old leaves increased significantly after the low potassium treatment. It is assumed that ZmCIPK23 may be involved in the re-distribution of the potassium ion of the corn crown under low potassium. Overexpression of ZmCIPK23 may interfere with the redistribution of potassium ions in the crown, resulting in a low-potassium-sensitive phenotype of the transgenic plant. The results showed that the over-expression plants of ZmCIP3 and ZmCIPK9 had no significant difference in the utilization efficiency of K, and it was suggested that ZmCIP3/9 could not participate in the process of potassium absorption and transport in maize. The field phenotype of the above-mentioned transgenic maize inbred line was also studied in this paper. The results showed that the reduction of the application of potassium fertilizer would affect the growth of the corn. In the jointing stage, a part of the transgenic inbred lines (ZmCBL1 and ZmCIPK9 over-expressed corn material) and the control material plant ratio showed a growth advantage. However, in the silk-laying period, the over-expression inbred lines were not significantly different from the control materials in terms of plant height, stem size, ear position and so on. The results of this paper show that ZmCBL1 and ZmCIPK23 are likely to be involved in the process of potassium absorption and redistribution of maize. However, the sustained over-expression of ZmCBL1 and ZmCIPK3/9/23 in the corn does not improve the utilization efficiency of the potassium in the corn under water culture conditions. It is suggested that there may be different potassium absorption and control mechanisms in maize. Under the condition of low potassium, the expression of ZmCBL1 and ZmCIPK23 and other genes is reasonable, and the correct regulation of protein activity may be the key to improve the utilization efficiency of potassium in maize. The results of this study provide some reference for the further study of the regulation and control mechanism of the absorption and transport of potassium in corn and the genetic improvement of the utilization efficiency of potassium in the future.
【学位授予单位】：中国农业大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：S513
【参考文献】
相关期刊论文 前1条
1 ;Identification and characterization of putative CIPK genes in maize[J];遗传学报;2011年02期
相关博士学位论文 前4条
1 李红;拟南芥转运蛋白NRT1.5/NPF7.3调控K~+在木质部装载的分子机制研究[D];中国农业大学;2016年
2 韩敏;钾转运体KUP7参与拟南芥响应低钾胁迫的功能研究[D];中国农业大学;2015年
3 李娟;水稻钾离子通道OsAKT1及其调控因子参与水稻钾吸收的实验证据[D];中国农业大学;2014年
4 龙雨;水稻钾离子通道OsAKT1生理功能及其调控机制的电生理学研究[D];中国农业大学;2014年
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