State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Rice Research Institute, Sichuan Agricultural University, 211 Huimin Road, Wenjiang District, Chengdu, 611130, China.
BMC Plant Biol. 2020 Jul 22;20(1):345. doi: 10.1186/s12870-020-02541-x.
Calvin cycle plays a crucial role in carbon fixation which provides the precursors of organic macromolecules for plant growth and development. Currently, no gene involved in Calvin cycle has been identified in monocotyledonous plants through mutant or/and map-based cloning approach.
Here, we isolated a low-tillering mutant, c6635, in rice (Oryza sativa). The mutant displayed light green leaves and intensely declined pigment contents and photosynthetic capacity at early growth stage. Moreover, its individual plant showed a much smaller size, and most individuals produced only two tillers. At mature stage, its productive panicles, grain number and seed setting rate were significantly decreased, which lead to a sharp reduction of the grain yield. We confirmed that a single nucleotide mutation in LOC_Os04g16680 gene encoding sedoheptulose 1,7-bisphosphatase (SBPase) involved in Calvin cycle was responsible for the mutant phenotype of c6635 through map-based cloning, MutMap analysis and complementation experiments. Sequence analysis suggested that the point mutation caused an amino acid change from Gly-364 to Asp at the C-terminal of SBPase. In addition, OsSBPase gene was mainly expressed in leaf, and the encoded protein was located in chloroplast. The mutation of OsSBPase could significantly affect expression levels of some key genes involved in Calvin cycle.
We successfully identified a SBPase gene in monocotyledonous plants. Meanwhile, we demonstrated that a single nucleotide substitution at the 3'-end of this gene severely affects plant growth and grain yield, implying that the Gly-364 at the C-terminal of SBPase could play an important role in SBPase function in rice.
卡尔文循环在碳固定中起着至关重要的作用,为植物的生长和发育提供了有机大分子的前体。目前,通过突变体或/和图谱克隆的方法,尚未在单子叶植物中鉴定出参与卡尔文循环的基因。
我们从水稻中分离到一个矮秆突变体 c6635。该突变体表现出淡绿叶色,色素含量和光合能力在早期生长阶段明显下降。此外,其单株植株较小,大多数植株只产生两个分蘖。在成熟阶段,其有效穗数、粒数和结实率显著降低,导致产量明显下降。我们通过图谱克隆、MutMap 分析和互补实验证实,LOC_Os04g16680 基因编码的 sedoheptulose 1,7-双磷酸酶(SBPase)中的一个单核苷酸突变导致了 c6635 的突变表型。序列分析表明,该点突变导致 C 末端的甘氨酸-364 突变为天冬氨酸。此外,OsSBPase 基因主要在叶片中表达,编码的蛋白位于叶绿体中。OsSBPase 基因的突变显著影响了一些参与卡尔文循环的关键基因的表达水平。
我们成功地在单子叶植物中鉴定出一个 SBPase 基因。同时,我们证明该基因 3'-末端的单个核苷酸取代严重影响植物的生长和籽粒产量,表明 SBPase 蛋白 C 末端的 Gly-364 在水稻 SBPase 功能中可能起着重要作用。