玉米ZmCBL-ZmCIPK參與鉀離子吸收轉(zhuǎn)運(yùn)的功能研究
發(fā)布時間：2019-06-03 00:56
【摘要】：鉀是植物生長發(fā)育所必需的礦質(zhì)元素。然而我國耕地土壤普遍缺鉀且鉀肥資源匱乏,這已經(jīng)成為制約農(nóng)作物生產(chǎn)的重要限制因素之一。因此,通過遺傳改良方法提高作物鉀吸收利用效率是解決我國農(nóng)作物生產(chǎn)中鉀供應(yīng)不足問題的重要途徑之一。本實(shí)驗(yàn)室前期研究發(fā)現(xiàn),AKT1是模式植物擬南芥根部介導(dǎo)鉀吸收的主要鉀離子通道,其通道活性受鈣結(jié)合蛋白CBL1/9和蛋白激酶CIPK23的調(diào)控。過量表達(dá)CBL1/9和CIPK23均可以提高擬南芥的低鉀耐受能力。ZMK1是AKT1在玉米中的同源蛋白,體外實(shí)驗(yàn)證明ZMK1的通道活性受玉米鈣結(jié)合蛋白ZmCBL1和蛋白激酶ZmCIPK3/9/23的調(diào)控。本論文工作主要研究這些ZmCBL和ZmmCIPK基因在玉米鉀吸收利用中的生理功能,進(jìn)而篩選鉀高效吸收利用的玉米轉(zhuǎn)基因材料。首先構(gòu)建了 ZmCBL1和ZmCIPK3/9/23的過量表達(dá)轉(zhuǎn)基因玉米材料,并對這些轉(zhuǎn)基因玉米自交系進(jìn)行了鉀吸收利用效率性狀的檢測。在低鉀水培條件下,ZmCBL1過表達(dá)植株的老葉出現(xiàn)提前發(fā)黃枯萎的表型。鉀含量測定結(jié)果顯示,低鉀條件下ZmCBL1過表達(dá)植株根部和冠部的鉀含量均顯著低于對照材料。這些結(jié)果表明過量表達(dá)ZmCBL1可能導(dǎo)致玉米對低鉀脅迫更為敏感。鉀營養(yǎng)耗竭實(shí)驗(yàn)結(jié)果顯示,ZmCIPK23過量表達(dá)植株的鉀離子吸收速率高于對照材料。然而ZmCIPK23過表達(dá)植株在低鉀下卻表現(xiàn)出老葉提前發(fā)黃枯萎的敏感表型。鉀含量測定結(jié)果顯示,在低鉀下ZmCIPK23過表達(dá)植株冠部鉀含量略低于對照材料。進(jìn)一步研究發(fā)現(xiàn)ZmCIPK23過表達(dá)植株的老葉和上部伸展葉中的鉀含量顯著低于對照材料,而莖節(jié)中的鉀含量卻顯著高于對照材料。基因表達(dá)檢測顯示,正常條件下ZmCIPK23主要在根中表達(dá),但低鉀處理后ZmCIPK23在老葉中的表達(dá)量顯著升高。推測,ZmCIPK23可能參與低鉀下玉米冠部鉀離子的再分配過程。過量表達(dá)ZmCIPK23可能會干擾鉀離子在冠部的再分配過程,從而導(dǎo)致轉(zhuǎn)基因植株的低鉀敏感表型。實(shí)驗(yàn)結(jié)果顯示ZmCIPK3和ZmCIPK9的過表達(dá)植株在鉀吸收利用效率上與對照材料相比并無顯著差異,推測ZmCIPK3/9在玉米體內(nèi)可能并不參與鉀吸收轉(zhuǎn)運(yùn)過程。本論文還對上述轉(zhuǎn)基因玉米自交系開展了田間表型檢測。結(jié)果顯示,減少鉀肥施用會影響玉米的生長狀況。在拔節(jié)期,部分轉(zhuǎn)基因自交系(ZmCBL1和ZmCIPK9過表達(dá)玉米材料)與對照材料植株比表現(xiàn)出生長優(yōu)勢。但在吐絲期,這些過表達(dá)自交系在株高、莖粗、穗位等方面與對照材料相比并沒有顯著差異。本論文的研究結(jié)果表明,ZmCBL1和ZmCIPK23有可能參與玉米的鉀吸收和再分配過程。但是在玉米中持續(xù)過量表達(dá)ZmCBL1和ZmCIPK3/9/23并不能提高水培條件下玉米的鉀吸收利用效率。推測,玉米中可能存在與擬南芥不同的鉀吸收調(diào)控機(jī)制。在低鉀條件下,ZmCBL1和ZmCIPK23等基因合理的時空表達(dá),以及正確的蛋白活性調(diào)控可能才是提高玉米鉀吸收利用效率的關(guān)鍵。本論文的研究結(jié)果對進(jìn)一步探討玉米鉀吸收和轉(zhuǎn)運(yùn)的調(diào)控機(jī)制以及未來玉米鉀吸收利用效率性狀的遺傳改良提供了一定的借鑒。
[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.
【學(xué)位授予單位】：中國農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：S513
本文編號：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.
【學(xué)位授予單位】：中國農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：S513
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 ;Identification and characterization of putative CIPK genes in maize[J];遺傳學(xué)報(bào);2011年02期
相關(guān)博士學(xué)位論文 前4條
1 李紅;擬南芥轉(zhuǎn)運(yùn)蛋白NRT1.5/NPF7.3調(diào)控K~+在木質(zhì)部裝載的分子機(jī)制研究[D];中國農(nóng)業(yè)大學(xué);2016年
2 韓敏;鉀轉(zhuǎn)運(yùn)體KUP7參與擬南芥響應(yīng)低鉀脅迫的功能研究[D];中國農(nóng)業(yè)大學(xué);2015年
3 李娟;水稻鉀離子通道OsAKT1及其調(diào)控因子參與水稻鉀吸收的實(shí)驗(yàn)證據(jù)[D];中國農(nóng)業(yè)大學(xué);2014年
4 龍雨;水稻鉀離子通道OsAKT1生理功能及其調(diào)控機(jī)制的電生理學(xué)研究[D];中國農(nóng)業(yè)大學(xué);2014年
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