高粱、玉米苗期抗旱生理与分子机制的比较研究
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摘要
干旱是农业生产中经常遇到的主要非生物因素逆境,严重影响着农作物的产量和分布,造成农业的减产和种植区域的限制,目前解决这一问题的方法之一是改善作物抗逆品质,培育或种植优良抗旱的作物和品种。由于抗旱性状是多基因作用的结果,目前,应用现代分子和遗传等生物工程方法对农作物品种进行的较单一的基因的改良还没有带来农业生产上实质性的突破,彻底解决粮食生产中存在的问题。所以,有必要进一步搞清植物的抗旱机理。高粱抗旱性较强,是研究抗旱机理的较好的实验材料,在大田生产和科研工作中,大家公认高粱的抗旱性状好于小麦、玉米等作物。本研究在阐述、总结前人对干旱机制研究进展和高粱抗旱研究的基础上,以高粱为研究对象,以较之抗旱性差的玉米为对照,开展了植物抗旱生理机制的比较研究。并在分子水平上,在遗传背景研究较多的玉米上首先阐明质膜水孔蛋白对植物抗旱作用,在此基础上,对遗传背景研究较少的高粱的质膜水孔蛋白进行了研究,进而在分子水平上进行了二者水孔蛋白抗旱机制的比较。得到了如下主要结果:
1.通过土壤自然干旱和复水实验,初步明确了渗透调节和抗氧化等耐旱途径是高粱和玉米所共有的,只是在主要渗透调节物质和抗氧化酶上存在物种间的差异。一般情况下,高粱的渗透调节物质主要是可溶性糖和可溶性蛋白,抗氧化酶主要是CAT,玉米的渗透调节物质则主要是脯氨酸、K+,抗氧化酶主要是SOD、POD。在抵御干旱胁迫和复水修复及生理功能恢复等方面,高粱表现出了较好的抗旱能力。另外,发现在同一物种不同抗旱性品种间也存在渗透调节和抗氧化上的差异,一般抗旱品种较抗旱性差的品种渗透调节能力和抗氧化能力更强;同时,发现在不同物种间的不同抗旱性品种上存在渗透调节物质和抗氧化酶在干旱或复水时变化趋势上的一致。表明植物抗旱是个复杂交织的网络体系。实验证实了高粱比玉米具有较高的水势,初步阐明了高粱在渗透调节和抗氧化上的耐旱机制。
2.在高粱上首次进行了水孔蛋白和抗旱关系的功能研究,并进行了和玉米间的物种比较,初步阐明了高粱在水孔蛋白分子水平上的抗旱机制。实验以遗传性背景研究较清楚的玉米为参照,结合玉米幼叶向基性生长,伸长区具有低水势下生长较快的特点,以玉米幼叶为实验材料,采用RT-PCR 和Western blotting 和in situ Western 等分子实验技术,初步明确了质膜水孔蛋白在玉米幼叶伸长生长和抗旱中的作用。之后结合运用到高粱水孔蛋白的研究上,说明了质膜水孔蛋白参与了高粱幼叶伸长生长和抗旱。并对高粱、玉米质膜水孔蛋白基因和蛋白水平上的差异进行了比较,在水孔蛋白抗旱机制上对高粱的抗旱机理得到了初步结论。
Drought is one major abiotic factor that adversely affects the crop production and distribution and account for the reduction in the yields of crops. At present, to improve the crop’s quality of water stress resistant ability through breeding anti-water stress plants or cultivars is one of the main methods to solve this problem. As the water stress resistant ability is controlled by multigenes, using modern molecular and gene technology to breed water stress resistant plant or genotype has not got the good result. So it is very important and necessary to give more attention to clear up the water stress resistant mechanism deeply. Sorghum is a model plant for researching the mechanism of water stress resistance. In this study, various sorghum cultivars were used as experimental materials to study the mechanism of water stress resistance and compared with maize, which water stress resistant ability is lower than sorghum while its geneology knowledge is more clear. In our study, we first compared the osmotic mechanism and antioxidant ability between sorghum and maize, and then on the base of revealing the function of aquaporin PIP1 in maize leaf growth and water stress resistant ability, we studied the role of the same type aquaporin in sorghum, and compared its change between sorghum and maize in the level of molecular mechanism. The main results are as follows:
1.Through the soil drought and recovered water experiment, we confirmed that the osmotic tolerance and antioxidant mechanism are all exit in sorghum and maize, while their main osmotic materials and antioxidant enzymes are different. In sorghum, the soluble sugar and protein are the main osmotic substance and CAT is the main antioxidant enzyme. In maize, it used the K+ and proline as the main osmotic materials and SOD and POD as the main antioxidant enzymes. We also found the difference in the osmotic tolerance and antioxidant ability between different genotype in sorghum or in maize, and the same mechanism change between sorghum and maize. So the anti-drought mechanism in the crop is a complex net.
Sorghum has higher water potential than maize, and has more effective osmotic tolerance and antioxidant ability. It showed more tolerance to drought than maize. 2.We first report the role of aquaporin in sorghum, and compared its difference with maize. The sorghum water stress resistant mechanism was revealed in some extent in AQP. At first, using RT-PCR, Western blotting and in situ Western or so molecular technology, we studied the aquaporin PIP1 in maize leaf growth and its water stress resistance. In mRNA and protein level, We found the PIP1 play roles in maize leaf growth and maize water stress resistance, and showed different change in different maize genotype. Base on the maize experiment, we studied the function in sorghum and got the same conclusion. Then we compared the difference of PIP1 between sorghum and maize and found PIP1 gave much more function in maize than in sorghum at the protein level. The function of aquaporin is one of the water stress resistant mechanism in plant.
1.通过土壤自然干旱和复水实验,初步明确了渗透调节和抗氧化等耐旱途径是高粱和玉米所共有的,只是在主要渗透调节物质和抗氧化酶上存在物种间的差异。一般情况下,高粱的渗透调节物质主要是可溶性糖和可溶性蛋白,抗氧化酶主要是CAT,玉米的渗透调节物质则主要是脯氨酸、K+,抗氧化酶主要是SOD、POD。在抵御干旱胁迫和复水修复及生理功能恢复等方面,高粱表现出了较好的抗旱能力。另外,发现在同一物种不同抗旱性品种间也存在渗透调节和抗氧化上的差异,一般抗旱品种较抗旱性差的品种渗透调节能力和抗氧化能力更强;同时,发现在不同物种间的不同抗旱性品种上存在渗透调节物质和抗氧化酶在干旱或复水时变化趋势上的一致。表明植物抗旱是个复杂交织的网络体系。实验证实了高粱比玉米具有较高的水势,初步阐明了高粱在渗透调节和抗氧化上的耐旱机制。
2.在高粱上首次进行了水孔蛋白和抗旱关系的功能研究,并进行了和玉米间的物种比较,初步阐明了高粱在水孔蛋白分子水平上的抗旱机制。实验以遗传性背景研究较清楚的玉米为参照,结合玉米幼叶向基性生长,伸长区具有低水势下生长较快的特点,以玉米幼叶为实验材料,采用RT-PCR 和Western blotting 和in situ Western 等分子实验技术,初步明确了质膜水孔蛋白在玉米幼叶伸长生长和抗旱中的作用。之后结合运用到高粱水孔蛋白的研究上,说明了质膜水孔蛋白参与了高粱幼叶伸长生长和抗旱。并对高粱、玉米质膜水孔蛋白基因和蛋白水平上的差异进行了比较,在水孔蛋白抗旱机制上对高粱的抗旱机理得到了初步结论。
Drought is one major abiotic factor that adversely affects the crop production and distribution and account for the reduction in the yields of crops. At present, to improve the crop’s quality of water stress resistant ability through breeding anti-water stress plants or cultivars is one of the main methods to solve this problem. As the water stress resistant ability is controlled by multigenes, using modern molecular and gene technology to breed water stress resistant plant or genotype has not got the good result. So it is very important and necessary to give more attention to clear up the water stress resistant mechanism deeply. Sorghum is a model plant for researching the mechanism of water stress resistance. In this study, various sorghum cultivars were used as experimental materials to study the mechanism of water stress resistance and compared with maize, which water stress resistant ability is lower than sorghum while its geneology knowledge is more clear. In our study, we first compared the osmotic mechanism and antioxidant ability between sorghum and maize, and then on the base of revealing the function of aquaporin PIP1 in maize leaf growth and water stress resistant ability, we studied the role of the same type aquaporin in sorghum, and compared its change between sorghum and maize in the level of molecular mechanism. The main results are as follows:
1.Through the soil drought and recovered water experiment, we confirmed that the osmotic tolerance and antioxidant mechanism are all exit in sorghum and maize, while their main osmotic materials and antioxidant enzymes are different. In sorghum, the soluble sugar and protein are the main osmotic substance and CAT is the main antioxidant enzyme. In maize, it used the K+ and proline as the main osmotic materials and SOD and POD as the main antioxidant enzymes. We also found the difference in the osmotic tolerance and antioxidant ability between different genotype in sorghum or in maize, and the same mechanism change between sorghum and maize. So the anti-drought mechanism in the crop is a complex net.
Sorghum has higher water potential than maize, and has more effective osmotic tolerance and antioxidant ability. It showed more tolerance to drought than maize. 2.We first report the role of aquaporin in sorghum, and compared its difference with maize. The sorghum water stress resistant mechanism was revealed in some extent in AQP. At first, using RT-PCR, Western blotting and in situ Western or so molecular technology, we studied the aquaporin PIP1 in maize leaf growth and its water stress resistance. In mRNA and protein level, We found the PIP1 play roles in maize leaf growth and maize water stress resistance, and showed different change in different maize genotype. Base on the maize experiment, we studied the function in sorghum and got the same conclusion. Then we compared the difference of PIP1 between sorghum and maize and found PIP1 gave much more function in maize than in sorghum at the protein level. The function of aquaporin is one of the water stress resistant mechanism in plant.
引文
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