苦荞FtRLP48基因克隆及其变异对粒形和粒重的影响OA

Leucine-rich repeat receptor like proteins(LRR-RLPs)play an important role in plant growth and development.However,the relationship between LRR-RLPs and grain development remains unclear.FtRLP48,a member of the LRR-RLPs gene family,screened out from our previous research,may be involved in the regulation of grain shape and weight.In this study,to further clarify the gene function of FtRLP48,it was cloned from'Xiaomiqiao'and'Jinqiaomai 2',the two parents of the primary mapping population,and named FtRLP48-M and FtRLP48-J,respectively.The structure,coding protein information,genetic relationship and conserved domain of FtRLP48-M and FtRLP48-J were predicted via bioinformatics software.Transient transformation of tobacco leaves was used to analyze subcellular localization.The pBI121-FtRLP48-M and pBI121-FtRLP48-J vectors were transformed into indica rice to verify its function.The results showed that the CDS of FtRLP48-M and FtRLP48-J were 2 577 bp and 1 917 bp,respectively.A frameshift variation"ATGGA/C"was occurred at 1 902 bp of FtRLP48-J,resulting in the early termination of protein translation and the loss of transmembrane structure.The sequence alignments of amino acids and phylogenetic analysis showed that FtRLP48-M and FtRLP48-J had the highest similarity in protein sequence,and they were most closely related to Arabidopsis AtRLP48.Subcellular localization showed that FtRLP48-J protein was located in the cell membrane and nuclear membrane.Compared with wild type,the FtRLP48-J transgenic rice exhibited significantly higher grain length and grain width,as well as extremely significantly higher 1 000-grain weight,1 000-grain kernel weight and 1 000-grain hull weight.The grain width,1 000-grain hull weight and hull-grain ratio of FtRLP48-M transgenic rice were significantly higher than those of wild type.Our results clarified that FtRLP48 may take an active role in the formation of grain shape and grain weigh,providing a theoretical basis to dissect the molecular mechanism of grain development and explore molecular marker for Tartary buckwheat breeding.