CAJ | 학술논문

为进一步了解紫花苜蓿在高温胁迫下的生理变化和相关差异蛋白表达情况,本研究以淮阴苜蓿(Medicago sativa L. cv. Huaiyin)为材料,以25℃为对照,对30℃、35℃和40℃不同高温胁迫下,淮阴苜蓿不同时期(0 h、0.5 h、1 h、4 h、12 h和24 h)幼苗叶片的电导率、丙二醛(MDA)、细胞活力的动态变化进行监测,并进一步对35℃和40℃高温处理24 h的幼苗叶片差异表达蛋白进行筛选、分析和鉴定研究。结果发现,淮阴苜蓿在35℃和40℃高温下,生长0.5 h,叶片的电导率含量开始急剧上升,1 h后电导率下降,并随着处理时间的延长,电导率变化趋缓；生长1 h时,叶片的MDA含量较高,4 h后MDA含量平缓下降；细胞活力随着时间的延长总体表现出下降的趋势。差异蛋白点筛选分析结果显示,在35℃处理24 h时,有27个点存在显著差异,其中19个点是上调的,与对照图谱相比,新增了5个特殊的蛋白点。在40℃处理24 h时,有51个点存在显著差异,其中40个点是上调的,与对照图谱相比,新增了11个特殊的蛋白点。对40℃时51个蛋白点进行二级质谱鉴定发现,20 kD chaperonin、Hsp70、Hsp23和Hsp17这4个蛋白与防御机制有关,说明40℃诱导产生更多的防御蛋白,对膜系统损伤起到一定的修复功能。
위진일보료해자화목숙재고온협박하적생리변화화상관차이단백표체정황,본연구이회음목숙(Medicago sativa L. cv. Huaiyin)위재료,이25℃위대조,대30℃、35℃화40℃불동고온협박하,회음목숙불동시기(0 h、0.5 h、1 h、4 h、12 h화24 h)유묘협편적전도솔、병이철(MDA)、세포활력적동태변화진행감측,병진일보대35℃화40℃고온처리24 h적유묘협편차이표체단백진행사선、분석화감정연구。결과발현,회음목숙재35℃화40℃고온하,생장0.5 h,협편적전도솔함량개시급극상승,1 h후전도솔하강,병수착처리시간적연장,전도솔변화추완；생장1 h시,협편적MDA함량교고,4 h후MDA함량평완하강；세포활력수착시간적연장총체표현출하강적추세。차이단백점사선분석결과현시,재35℃처리24 h시,유27개점존재현저차이,기중19개점시상조적,여대조도보상비,신증료5개특수적단백점。재40℃처리24 h시,유51개점존재현저차이,기중40개점시상조적,여대조도보상비,신증료11개특수적단백점。대40℃시51개단백점진행이급질보감정발현,20 kD chaperonin、Hsp70、Hsp23화Hsp17저4개단백여방어궤제유관,설명40℃유도산생경다적방어단백,대막계통손상기도일정적수복공능。
The Medicago sativa L. cv. Huaiyin variety of alfalfa was used to determine the physical changes and related differences in protein expressions of alfalfa in response to heat stress. Electrical conductivity, malonaldehyde content and cell activity were measured in alfalfa seedlings at 25 ℃, 30 ℃, 35 ℃ and 40 ℃ in combination with different durations of heat stress (0 h, 0.5 h, 1 h, 4 h, 12 h and 24 h). In addition, alfalfa seedlings treated at 35 ℃ and 40 ℃ for 24 h were used to analyze differentially expressed protein. The results showed that electrical conductivity of the test samples increased sharply after treatment at 35 ℃ and 40 ℃ for 0.5 h, but declined for treatment durations exceeding 1 h. However, malonaldehyde content of the samples increased for treatment durations exceeding 1 h but gradually declined after 4 h. Cell activity of test samples declined with increasing treatment duration. Screening analysis of the differences in proteins showed 27 significantly different spots after treatment at 35 ℃ for 24 h, 19 of which spots were raised and 5 were new. After treatment at 40 ℃ for 24 h, 51 significantly different spots were observed—40 out of the 51 spots were raised and 11 were new. Secondary mass spectrum analyses indicated that 4 proteins — 20 kD chaperonin, Hsp70, Hsp23 and Hsp17, were only observed in samples at 40 ℃ for 24 hours. This suggested that the 4 proteins correlated with heat stress defense mechanisms, the samples produced more defense proteins at 40 ℃ to repair damage to alfalfa seedling membranes.