16Ovules contain the female gametophytes which are fertilized during pollination to initiate 17 seed development. Thus, the number of ovules that are produced during flower 18 development is an important determinant of seed crop yield and plant fitness. Mutants 19 with pleiotropic effects on development often alter the number of ovules, but specific 20 regulators of ovule number have been difficult to identify in traditional mutant screens. 21We used natural variation in Arabidopsis accessions to identify new genes involved in 22 the regulation of ovule number. The ovule numbers per flower of 189 Arabidopsis 23 accessions were determined and found to have broad phenotypic variation that ranged 24 from 39 ovules to 84 ovules per pistil. Genome-Wide Association tests revealed several 25 genomic regions that are associated with ovule number. T-DNA insertion lines in 26 candidate genes from the most significantly associated loci were screened for ovule 27 48
Key Words 49Ovule number, NERD1, Arabidopsis, Branching, Carpel Margin Meristem, GWAS 50 52During plant reproduction, pollen tubes deliver two sperm cells to female gametophytes 53 contained within ovules. This allows double fertilization to occur in order to produce the 54 embryo and endosperm in the developing seed. Angiosperms with all kinds of 55 pollination syndromes (insect-, wind-, and self-pollinated) produce much more pollen 56 than ovules in order to ensure successful pollination. For example, most soybean 57 varieties produce only 2 ovules per flower, but more than 3,000 pollen grains, a 1,500-58 fold difference(1). Wind-pollinated plants such as maize have an even more extreme 59 difference in pollen production vs. ovule production per plant, with more than 1 million 60 pollen grains versus an average of 250 ovules per plant (a 4000-fold difference (2)). 61Arabidopsis thaliana, which is a self-pollinating plant, also produces an excess of pollen, 62 with at least 2000 pollen grains per flower compared to an average of 60 ovules per 63 flower(3). Since pollen is produced in excess, in self-pollinated plants the number of 64 ovules (i.e. female gametes) sets the maximum seed number per flower. 65 66The ability to manipulate ovule number to increase the reproductive potential of plants 67requires an understanding of the molecular pathways that control ovule initiation. 68The model plant Arabidopsis thaliana produces flowers with four whorls of organs: 69 sepals, petals, stamens, and carpels. The inner whorls (3 and 4) are responsible for 70 sexual reproduction, with pollen (the male gametophytes) produced in the whorl 3 71 stamens and the female gametophytes (also known as the embryo sacs), produced in 72 ovules contained within the whorl 4 carpels. Specification of the 4 whorls is controlled by 73 phenotype is exacerbated when ant is combined with mutations in other carpel 97 development transcriptional regulators, such as SEUSS (SEU), LEUNIG (LUG), 98 SHATTERPROOF1 and 2 (SHP1 and SHP2), CRABSCLAW (CRC), FILAMENTOUS 99 FLOWER (FIL), and YABBY3 (YAB3). ...