Ramie fibers, one of the most important natural fibers in China, are mainly composed of lignin, cellulose, and hemicellulose. As the high lignin content in the fibers results in a prickly texture, the lignin content is deemed to be an important trait of the fiber quality. In this study, the genetic basis of the fiber lignin content was evaluated, resulting in the identification of five quantitative trait loci (QTLs). Three genes, whole_GLEAN_10021050, whole_GLEAN_10026962, and whole_GLEAN_10009464 that were identified on the QTL regions of qLC7, qLC10, and qLC13, respectively, were found to be homologs of the Arabidopsis lignin biosynthetic genes. Moreover, all three genes displayed differential expression in the barks located in the top and middle parts of the stem, where lignin was not being synthesized and where it was being biosynthesized, respectively. Sequence comparison found that these three genes had wide variations in their coding sequences (CDSs) and putative promoter regions between the two parents, especially the MYB gene whole_GLEAN_10021050, whose protein had insertions/deletions of five amino acids and substitutions of two amino acids in the conserved domain. This evidence indicates that these three genes are potentially involved in lignin biosynthesis in ramie fibers. The QTLs identified from this study provide a basis for the improvement of lignin content and fiber quality in ramie breeding. The characterization of the three candidate genes here will be helpful for the future clarification of their functions in ramie.

The KNOX transcription factor plays crucial roles in regulating fiber growth in plants. Although the genome of ramie, an important fiber crop in China, is available, knotted related homeobox (KNOX) genes have not been systematically explored in this crop. In this study, seven members of the KNOX gene from the ramie genome were identified and assigned to two groups, Class I and II. The intron–exon structure, conserved domain architecture, cis-regulating elements, and expression pattern showed distinct differences among the seven KNOX regulators. One of the genes, Bnt07G011994, encodes an ortholog of Arabidopsis fiber-growth-related KNAT7, and is differentially expressed among barks undergoing different stages of fiber growth. The overexpression of Bnt07G011994 dramatically decreases the fiber number in transgenic Arabidopsis, indicating a negative role played by this gene in modulating fiber growth. Further transcriptome analysis of transgenic Arabidopsis revealed that the overexpression of Bnt07G011994 resulted in an expression change in 14 pectin biosynthesis-/metabolism-related genes. These findings provide a useful foundation for further investigating the function of KNOX genes in ramie, and provide an important insight into the involvement of the ramie KNOX gene in fiber growth.

NAM, ATAF, and CUC (NAC) proteins are plant-specific transcription factors that play crucial roles in fiber growth by regulating the secondary wall thickening. In this study, a systematical investigation of NAC genes was performed in the genome of ramie, an important fiber crop, resulting in a total of 60 ramie NAC genes identified. Phylogenetic analysis of these 60 NAC members in conjunction with 111 Arabidopsis NAC proteins identified 11 subfamilies, three of which showed considerable contraction in the ramie genome. Ten ramie NAC genes were identified to encode the orthologs of Arabidopsis NAC regulators involved in the control of secondary wall biosynthesis. Of these ten genes, most showed relatively high expression in the stems, and eight displayed a differential expression between the barks from the top and middle section of the stems where fiber growth is under different stages. Furthermore, the overexpression of three of these ten NAC genes significantly promoted fiber growth in transgenic Arabidopsis. These results indicated that these ten NAC genes were associated with the fiber growth of ramie. This study provided an important basis for researching the regulatory mechanism of fiber growth.