State Key Laboratory of Crop Genetics and Germplasm Enhancement, Jiangsu Plant Gene Engineering Research Center, Nanjing Agricultural University, Nanjing 210095, PR China.
State Key Laboratory of Crop Genetics and Germplasm Enhancement, Jiangsu Plant Gene Engineering Research Center, Nanjing Agricultural University, Nanjing 210095, PR China.
Plant Sci. 2018 Dec;277:89-99. doi: 10.1016/j.plantsci.2018.09.020. Epub 2018 Sep 26.
Mutations of stromal Hsp70 cause chloroplast developmental abnormalities and knockout mutants of stromal Hsp70 usually exhibit protein import deficiencies. However, their effects have not been studied in amyloplast development. Here, we identified an amyloplast abnormal development mutant, floury endosperm11 (flo11) that exhibited an opaque phenotype in the inner core and the periphery of grains. Semi-thin section revealed defective amyloplast development in the flo11 endosperm. Map-based cloning and subsequent complementation test demonstrated that FLO11 encoded a plastid-localized heat shock protein 70 (OsHsp70cp-2). OsHsp70cp-2 was abundantly expressed in developing endosperm, whereas its paralogous gene OsHsp70cp-1 was mainly expressed in photosynthetic tissues. Ectopic expression of OsHsp70cp-1 under the control of OsHsp70cp-2 promoter rescued the mutant phenotype of flo11. Moreover, simultaneous knockdown of both OsHsp70cp genes resulted in white stripe leaves and opaque endosperm. BiFC and Co-IP assays revealed that OsHsp70cp-2 was associated with Tic complex. Taken together, OsHsp70cp-2 may regulate protein import into amyloplasts, which is essential for amyloplast development in rice.
基质 Hsp70 的突变会导致叶绿体发育异常,基质 Hsp70 的敲除突变体通常表现出蛋白导入缺陷。然而,它们在淀粉体发育中的作用尚未得到研究。在这里,我们鉴定了一个淀粉体异常发育突变体,粉质胚乳 11(flo11),其在谷物的内芯和外周表现出不透明表型。半薄切片显示 flo11 胚乳中的淀粉体发育缺陷。基于图谱的克隆和随后的互补测试表明,FLO11 编码一种定位于质体的热休克蛋白 70(OsHsp70cp-2)。OsHsp70cp-2 在发育中的胚乳中大量表达,而其同源基因 OsHsp70cp-1 主要在光合组织中表达。在 OsHsp70cp-2 启动子的控制下异位表达 OsHsp70cp-1 挽救了 flo11 的突变表型。此外,同时敲低两个 OsHsp70cp 基因会导致白条纹叶片和不透明胚乳。BiFC 和 Co-IP 测定表明 OsHsp70cp-2 与 Tic 复合物相关。总之,OsHsp70cp-2 可能调节蛋白导入淀粉体,这对于水稻淀粉体的发育至关重要。