摘要
FCS样锌指蛋白(FLZ)与植物的生长发育和逆境胁迫反应相关.水稻的FLZ基因家族分析和功能研究较少.本研究利用 TBtools 对水稻基因组 Blast,鉴定到 29 个 OsFLZ 家族基因成员,并分析了相关基因位置、基因结构、motif和启动子顺式作用元件等特征.随后,通过水稻CREP数据库研究了FLZ家族成员的全生育期组织表达模式,并发现其中的OsFLZ13 基因在开花前的花药中特异高水平表达.随后β-D-葡萄糖苷酸酶(GUS)染色显示,OsFLZ13 在花药发育的第 8 阶段开始表达,并在开花前的第 14 阶段表达量最高.用CRISPR/Cas9 基因编辑获得的突变体植株结实率显著下降.相比野生型中花 11 的 94%结实率,Osflz13-1 和Osflz13-2 的结实率分别只有 44%和 36%.本研究表明 OsFLZ13 参与花药发育以及花粉育性的调控,为进一步研究该基因及其家族基因的功能提供参考,同时对水稻雄性不育利用具有潜在的价值.
Abstract
FCS-like zinc finger(FLZ)is a protein associated with plant growth and stress.At present,there are few reports on FLZ gene family analysis and functional studies in rice.In this study,TBtools were used to blast rice genome,and a total of 29 OsFLZ genes were identified.Their gene location,gene structure,motif and promoter sequences were analyzed.The relative ex-pression level of FLZ genes in rice from CREP database showed that,OsFLZ13,a member of this family,was predominantly ex-pressed in anthers before flowering.β-D-glucuronidase(GUS)staining assays exhibited that OsFLZ13 began to express at stage 9 and gradually peaked at stage 14 of stamen development before flowering.Furthermore,two independent mutant lines,namely Osflz13-1 and Osflz13-2,were obtained with CRISPR/Cas9 gene editing system.Compared with the 94%seed-setting rate of wild type Zhonghua 11,the setting rates of Osflz13-1 and Osflz13-2 were reduced to 44%and 36%,respectively.This study throws light on the evolution of FLZ in planta and indicates the roles of OsFLZ13 in anthers development and pollen fertility,which will be beneficial further studies of its functions.Additionally,it provides a reference for exploring the function of the FLZ family and highlights its potential value for the utilization in male sterility systems in rice.
基金项目
国家自然科学基金地区科学基金项目(31860296)
广西自然科学基金重点项目(31860296)
维普
万方数据