Combined QTL mapping and association study reveals candidate genes for leaf number and flowering time in maize

Twelve QTL for flowering and leaf number were detected. The ZmWRKY14^Hap4 could increase leaf number, flowering time and biomass yield which are promising for silage maize breeding. Silage maize, one of the most important feedstock for ruminants, is widely grown from temperate regions to the tropics. Flowering time and leaf number are two significantly correlated traits and important for the quality, adaptation and biomass yield of silage maize. In this study, a recombinant inbred line population consisting of 215 individuals and an association panel of 369 inbred lines were analysed in field conditions in three locations for 2 consecutive years, and five, four and three quantitative trait loci for the total leaf number, days to anthesis (DTA) and silking (DTS) were detected, which could explain 48.55, 35.37 and 34.22% of total phenotypic variation, respectively. Association analysis of qLN10 on chromosome 10 found that ZmWRKY14 was the candidate gene for leaf number, whose expression level was negatively correlated with the leaf number. There are five haplotypes for ZmWRKY14, and haplotype 4 could significantly increase flowering time, leaf number and biomass yield, but has no obvious influence on ear weight. The optimal allelic combination of ZmWRKY14 and ZCN8 could further increase leaf number and biomass yield. The results will provide important genetic information for silage maize breeding.