Yang Feng, Debatosh Das, Song Tao, Zhang Jian-Hua
Co-Innovation Center for Sustainable Forestry in Southern China, College of Biology and the Environment, Nanjing Forestry University, Nanjing, 210037, China.
Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, 518057, Guangdong, China.
Rice (N Y). 2021 Mar 20;14(1):32. doi: 10.1186/s12284-021-00474-z.
Carotenoid biosynthesis is essential for the generation of photosynthetic pigments, phytohormone production, and flower color development. The light harvesting like 3 (LIL3) protein, which belongs to the light-harvesting complex protein family in photosystems, interacts with geranylgeranyl reductase (GGR) and protochlorophyllide oxidoreductase (POR) both of which are known to regulate terpenoid and chlorophyll biosynthesis, respectively, in both rice and Arabidopsis.
In our study, a CRISPR-Cas9 generated 4-bp deletion mutant oslil3 showed aberrant chloroplast development, growth defects, low fertility rates and reduced pigment contents. A comparative transcriptomic analysis of oslil3 suggested that differentially expressed genes (DEGs) involved in photosynthesis, cell wall modification, primary and secondary metabolism are differentially regulated in the mutant. Protein-protein interaction assays indicated that LIL3 interacts with phytoene synthase (PSY) and in addition the gene expression of PSY genes are regulated by LIL3. Subcellular localization of LIL3 and PSY suggested that both are thylakoid membrane anchored proteins in the chloroplast. We suggest that LIL3 directly interacts with PSY to regulate carotenoid biosynthesis.
This study reveals a new role of LIL3 in regulating pigment biosynthesis through interaction with the rate limiting enzyme PSY in carotenoid biosynthesis in rice presenting it as a putative target for genetic manipulation of pigment biosynthesis pathways in crop plants.
类胡萝卜素生物合成对于光合色素的生成、植物激素的产生以及花色发育至关重要。光捕获类3(LIL3)蛋白属于光系统中的光捕获复合体蛋白家族,它与香叶基香叶基还原酶(GGR)和原叶绿素酸氧化还原酶(POR)相互作用,已知这两种酶分别在水稻和拟南芥中调节萜类和叶绿素生物合成。
在我们的研究中,通过CRISPR-Cas9技术产生的4-bp缺失突变体oslil3表现出叶绿体发育异常、生长缺陷、低育性以及色素含量降低。对oslil3进行的比较转录组分析表明,参与光合作用、细胞壁修饰、初生和次生代谢的差异表达基因(DEGs)在该突变体中受到不同程度的调控。蛋白质-蛋白质相互作用分析表明,LIL3与八氢番茄红素合酶(PSY)相互作用,此外,PSY基因的表达受LIL3调控。LIL3和PSY的亚细胞定位表明,二者均为叶绿体类囊体膜锚定蛋白。我们认为LIL3直接与PSY相互作用以调节类胡萝卜素生物合成。
本研究揭示了LIL3在水稻类胡萝卜素生物合成中通过与限速酶PSY相互作用来调节色素生物合成的新作用,表明它是作物植物色素生物合成途径基因操作的一个潜在靶点。