Positional cloning and characterization reveal the role of a miRNA precursor gene ZmLRT in the regulation of lateral root number and drought tolerance in maize

Lateral roots play essential roles in drought tolerance in maize(Zea mays L.). However, the genetic basis for the variation in the number of lateral roots in maize remains elusive. Here, we identified a major quantitative trait locus(QTL),q LRT5-1, controlling lateral root number using a recombinant inbred population from a cross between the maize lines Zong3(with many lateral roots) and 87-1(with few lateral roots).Fine-mapping and functional analysis determined that the candidate gene for qLRT5-1,ZmLRT, expresses the primary transcript for the microRNA miR166a. ZmLRT was highly expressed in root tips and lateral root primordia,and knockout and overexpression of ZmLRT increased and decreased lateral root number,respectively. Compared with 87-1, the ZmLRT gene model of Zong3 lacked the second and third exons and contained a 14 bp deletion at the junction between the first exon and intron,which altered the splicing site. In addition,ZmLRT expression was significantly lower in Zong3 than in 87-1, which might be attributed to the insertions of a transposon and over large DNA fragments in the Zong3 ZmLRT promoter region. These mutations decreased the abundance of mature miR166a in Zong3, resulting in increased lateral roots at the seedling stage.Furthermore, miR166a post-transcriptionally repressed five development-related class-Ⅲ homeodomain-leucine zipper genes. Moreover,knockout of ZmLRT enhanced drought tolerance of maize seedlings. Our study furthers our understanding of the genetic basis of lateral root number variation in maize and highlights ZmLRT as a target for improving drought tolerance in maize.

The TaTCP4/10-B1 cascade regulates awn elongation in wheat(Triticum aestivum L.)

Awns are important morphological markers for wheat and exert a strong physiological effect on wheat yield.The awn elongation suppressor B1 has recently been cloned through association and linkage analysis in wheat.However,the mechanism of awn inhibition centered around B1 remains to be clarified.Here,we identified an allelic variant in the coding region of B1 through analysis of re-sequencing data;this variant causes an amino acid substitution and premature termination,resulting in a long-awn phenotype.Transcriptome analysis indicated that B1 inhibited awn elongation by impeding cytokinin-and auxinpromoted cell division.Moreover,B1 directly repressed the expression of TaRAE2 and TaLks2,whose orthologs have been reported to promote awn development in rice or barley.More importantly,we found that TaTCP4 and TaTCP10 synergistically inhibited the expression of B1,and a G-to-A mutation in the B1 promoter attenuated its inhibition by TaTCP4/10.Taken together,our results reveal novel mechanisms of awn development and provide genetic resources for trait improvement in wheat.

TaANR1-TaMADS25 module regulates lignin biosynthesis and root development in wheat(Triticum aestivum L.)

Common wheat is a staple food for 35%of the global population,therefore increasing wheat yield in an ever-changing environment is essential for food security in the present day(Peng et al.,2011).Root system is responsible for water and nutrient acquisition,thus crucial for competitive fitness and crop yield in challenging environments(Karlova et al.,2021;Liu et al.,2022).Over the past decades,extensive research has focused on identifying genes accountable for root growth and development in plants(Rogers and Benfey,2015).However,only a limited number of genes have been cloned in wheat(Li et al.,2021).

Fine mapping and characterization of a major QTL for grain length,QGl.cau-2D.1,that has pleiotropic effects in synthetic allohexaploid wheat

Grain size is one of the determinants of grain yield,and identifying the genetic loci that control grain size will be helpful for increasing grain yield.In our previous study,a quantitative trait locus(QTL)for grain length(GL),QGl.cau-2D.1,was identified from an F2 population developed from the cross between the natural(TAA10)and synthetic(XX329)allohexaploid wheat.In the present study,we mainly fine mapped and validated its genetic effects.To this end,multiple near-isogenic lines(NILs)were obtained through marker-assisted selection with TAA10 as the recurrent parent.The secondary populations derived from 25 heterozygous recombinants were used for fine mapping of QGl.cau-2D.1,and the allele from XX329 significantly increased GL,thousand-grain weight(TGW),total spikelet number per spike(TSN)and spike compactness(SC).Using NILs for XX329(2D+)and TAA10(2D−),we determined the genetic and pleiotropic effects of QGl.cau-2D.1.The target sequences were aligned with the wheat reference genome RefSeq v2.1 and spanned an~0.9 Mb genomic region.TraesCS2D03G0114900(ortholog of Os03g0594700)was predicted as the candidate gene based on whole-genome re-sequencing and expression analyses.In summary,the map-based cloning of QGl.cau-2D.1 will be useful for improving grain weight with enhanced GL and TSN.

Ectopic expression of VRT-A2 underlies the origin of Triticum polonicum and Triticum petropavlovskyi with long outer glumes and grains

Polish wheat (Triticum polonicum) is a unique tetraploid wheat species characterized by an elongated outer glume. The genetic control of the long-glume trait by a single semi-dominant locus, P1 (from Polish wheat), was established more than 100 years ago, but the underlying causal gene and molecular nature remain elusive. Here, we report the isolation of VRT-A2, encoding an SVP-clade MADS-box transcription factor, as the P1 candidate gene. Genetic evidence suggests that in T. polonicum, a naturally occurring sequence rearrangement in the intron-1 region of VRT-A2 leads to ectopic expression of VRT-A2 in floral organs where the long-glume phenotype appears. Interestingly, we found that the intron-1 region is a key ON/OFF molecular switch for VRT-A2 expression, not only because it recruits transcriptional repressors, but also because it confers intron-mediated transcriptional enhancement. Genotypic analyses using wheat accessions indicated that the P1 locus is likely derived from a single natural mutation in tetraploid wheat, which was subsequently inherited by hexaploid T. petropavlovskyi. Taken together, our findings highlight the promoter-proximal intron variation as a molecular basis for phenotypic differentiation, and thus species formation in Triticum plants.