中国生态农业学报Issue(2)：210-217,8.DOI:10.13930/j.cnki.cjea.151113
白菜型冬油菜质外体抗冻蛋白研究
Study on apoplast anti-freeze proteins in winter turnip rape (Brassica rape L.)
摘要
Abstract
The objective of this paper was to lay the basis for studying cold resistance of winter rapeseed. The anti-freeze activities of apoplast proteins were determined in the ‘Longyou 6’ winter rape leaves and roots under cold vernalization. The apoplast proteins were separated by SDS-PAGE and high expression proteins identified in MALDI-TOF/TOF mass spectrometry under field and pot experiments. The results showed that apoplast protein content of ‘Longyou 6’ leaves increased significantly (P < 0.05) after cold acclimation in an artificial climate chamber, reaching 92.31 µg•g-1(FW) on the fifth day, which represented an increase of 246.12% over CK. Apoplast protein content after 10-15 days of cold acclimation dropped compared with that after 5 days, but was still significantly higher than that of CK (P < 0.05). Apoplast protein content continued to increase with increasing cold acclimation time from 20 to 25 days (P < 0.05). Apoplast protein content decreased significantly with after 10 days of de-acclimation. In the process of cold acclimation, apoplast protein content of ‘Longyou 6’ leaves significantly accumulated. However, it decreased significantly after de-acclimation. Obviously, apoplast proteins of‘Longyou 6’ winter rape belonged to low temperature induced proteins. Anti-freeze activity detection analysis suggested that apoplast proteins had re-crystallization inhibition activity. Mass spectrometry identification revealed a variety of proteins with unclear functions along with β-1-3-glucanase consistent anti-freeze proteins reported in winter rye. The class glucanase detected by mass spectrometry was found to have weaker ice crystal forms due to modification effect with reclamation and anti-freeze activity test. The test suggested that this class glucanase was a low activity anti-freeze protein. Many anti-freeze proteins were synthesized and secreted by winter rape in apoplast of leaves and roots under low temperature stress. The proteins likely played a positive role against outside low temperature. Furthermore, it was speculated that there were many undetected anti-freeze apoplast proteins responsible for the strong winter hardiness of winter rapeseed.关键词
冬油菜/质外体/抗冻蛋白/葡聚糖酶/SDS-PAGE/质谱技术Key words
Winter turnip rape/Apoplast/Anti-freeze protein/Glucanase/SDS-PAGE/Mass spectrometry分类
农业科技引用本文复制引用
杨刚,刘林波,杨建胜,方园,张娟,史鹏辉,孙万仓,刘自刚,曾秀存,武军艳,方彦,李学才,陈奇..白菜型冬油菜质外体抗冻蛋白研究[J].中国生态农业学报,2016,(2):210-217,8.基金项目
国家高技术研究发展计划(863计划)项目(2011AA10A104)、国家重点基础研究发展计划(973计划)项目(2015CB150206)、国家公益性行业(农业)科研专项(200903002-04)、现代农业产业技术体系建设专项资金(CARS-13)、国家自然基金项目(31460356)和国家农业科技成果转化项目(2014G10000317)资助* This study was supported by the National High-tech R&D Program of China (863 Program)(No.2011AA10A104), the National Basic Research Program of China (973 Program)(No.2015CB150206), the Special Fund for Agro-scientific Research in the Public Interest (No.200903002-04), Special Fund for the Industrial Technology System Construction of Modem Agriculture (No. CARS-13), the National Natural Science Foundation of China (No.31460356) and the National Agricultural Science and Technology Achievement Transformation Project (No.2014G10000317) （863计划）
