作物学报 ›› 2018, Vol. 44 ›› Issue (01): 53-62.doi: 10.3724/SP.J.1006.2018.00053
赵立娜1,2,**,刘子会1,**,段硕楠1,张园园1,2,李国良1,*,郭秀林1,*
ZHAO Li-Na1,2,**,LIU Zi-Hui1,**,DUAN Shuo-Nan1,ZHANG Yuan-Yuan1,2,LI Guo-Liang1,*,GUO Xiu-Lin1,*
摘要:
植物热激转录因子(heat shock transcription factor, Hsf)是响应热胁迫的主要调节因子,通过调节热激蛋白基因表达进而增强植物耐热性。小麦Hsf家族至少含有56个成员,其中B族11个,含B2亚族5个。本研究采用同源克隆技术,从37°C热处理的两叶一心小麦幼叶中克隆获得TaHsfB2d (序列号:AK331994)cDNA序列,序列长1191 bp,编码396个氨基酸。蛋白序列包括DNA结合结构域DBD和核定位信号序列NLS。同源分析表明,TaHsfB2d蛋白与大麦未知蛋白的相似性最高,为92%。荧光定量分析表明,TaHsfB2d在小麦多个组织器官中组成型表达,其中在成熟植株根系中表达量较高。37°C热胁迫、外源水杨酸(SA)和H2O2处理均能不同程度上调TaHsfB2d的表达,热激能显著增强SA和H2O2对TaHsfB2d表达的诱导。H2O2合成抑制剂DPI和羟自由基清除剂DMTU联合处理显著抑制热激对TaHsfB2d表达的上调作用、完全抑制SA对TaHsfB2d表达的上调。通过在洋葱内表皮瞬时表达TaHsfB2d并观察GFP荧光发现,正常条件下,TaHsfB2d蛋白定位于细胞核。酵母中耐热性鉴定表明,正常条件下,转TaHsfB2d的酵母细胞与转空载体对照酵母细胞的长势没有明显差异,热激处理同时降低,但前者的长势相对更强,TaHsfB2d的导入不影响细胞的生长发育。推测TaHsfB2d通过水杨酸途径介导植株耐热性调控过程,该过程依赖于H2O2存在。
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