Analysis And Identification Of Alkali Induced Proteome In Cotton

At present, land salinization is serious with farmland was decreased very sharply. In order to expand cultivated land to cope with the huge pressure to survive brought by rapid population growth, so we need to overcome salinity damage to crops. The largest damage to crops on saline soil is alkali stress. Thus revealing the mechanism of alkali to cultivate anti-alkali crops and expanding the area of cultivation has become not only an inevitable trend but also in response to the national call for sustainable development. As the important industrial crops and have high resistance to the salt relatively, using molecular biological techniques to study the alkali resistance mechanism of cotton, then improving and transforming alkali resistance of cotton and develop more excellent salt-tolerant cotton varieties are very necessary for the development of agriculture. This study used Zhong H177 salt-tolerant cotton at four-leaf stage as material. Through the improvement method of TCA-acetone extraction get total protein of cotton leaf when it treatment with different concentration gradient NaHCO3 / Na2CO3 mixed alkali after 6 hours and 12 hours. After SDS gel electrophoresis, and POD enzyme activity test, the experiment conditions were determined for 60 mM NaHCO3 / Na2CO3 lye, and processing time is 12 hours. Using total protein extracted by cotton completed two-dimensional electrophoresis experiment, through research and analysis to get a experimental result. The result showed that we have got 22 protein points, but they had significant difference, seven up-regulated and 15 down-regulated. These proteins are related to photosynthesis, defense mechanism, energy metabolism, etc. Then we shorten the processing time as 6 hours to optimize the experiment condition, extract proteins from four leaf stages. Using the same method as two-dimensional electrophoresis, we found 29 significant difference proteins. These proteins associated with the photosynthesis, defense mechanism, energy metabolism and organic acids. The difference proteins in root were, 11 proteins were identified, including three up-regulation and eight down-regulation. These difference proteins were different from leaf significant difference proteins. After the test of 29 up-regulated proteins, in which five were expressions in high degree and with different transcription levels. These five were found only 60% of similarity between them.Some alkali stress related proteins of cotton were found from this study, this finding may shed new light for further understand of the mechanism of cotton alkali response.