Kn1 Mutant Research Articles

Unequal Redundancy in Maizeknotted1 homeoboxGenes

The knotted1 (kn1) homeobox (knox) gene family was first identified through gain-of-function dominant mutants in maize (Zea mays). Class I knox members are expressed in meristems but excluded from leaves. In maize, a loss-of-function phenotype has only been characterized for kn1. To assess the function of another knox member, we characterized a loss-of-function mutation of rough sheath1 (rs1). rs1-mum1 has no phenotype alone but exacerbates several aspects of the kn1 phenotype. In permissive backgrounds in which kn1 mutants grow to maturity, loss of a single copy of rs1 enhances the tassel branch reduction phenotype, while loss of both copies results in limited shoots. In less introgressed lines, double mutants can grow to maturity but are shorter. Using a KNOX antibody, we demonstrate that RS1 binds in vivo to some of the KN1 target genes, which could partially explain why KN1 binds many genes but modulates few. Our results demonstrate an unequal redundancy between knox genes, with a role for rs1 only revealed in the complete absence of kn1.

The Interaction of knotted1 and thick tassel dwarf1 in Vegetative and Reproductive Meristems of Maize

In Arabidopsis, SHOOT MERISTEMLESS (STM) and CLAVATA1 (CLV1) competitively regulate meristem homeostasis. Here, we explore the interaction of their maize homologs knotted1 (kn1) and thick tassel dwarf1 (td1). kn1 mutants form fewer lateral organs and td1 inflorescences are fasciated with additional floral organs. Double mutants show kn1 epistatic to td1 in seedling and ear development but dose-sensitivity exists later to promote leaf initiation. Thus kn1 and td1 function in a pathway to maintain meristem homeostasis but their products may interact with different partners during development.

Loss-of-function mutations in the rice homeobox gene OSH15 affect the architecture of internodes resulting in dwarf plants.

The rice homeobox gene OSH15 (Oryza sativa homeobox) is a member of the knotted1-type homeobox gene family. We report here on the identification and characterization of a loss-of-function mutation in OSH15 from a library of retrotransposon-tagged lines of rice. Based on the phenotype and map position, we have identified three independent deletion alleles of the locus among conventional morphological mutants. All of these recessive mutations, which are considered to be null alleles, exhibit defects in internode elongation. Introduction of a 14 kbp genomic DNA fragment that includes all exons, introns and 5'- and 3'- flanking sequences of OSH15 complemented the defects in internode elongation, confirming that they were caused by the loss-of-function of OSH15. Internodes of the mutants had abnormal-shaped epidermal and hypodermal cells and showed an unusual arrangement of small vascular bundles. These mutations demonstrate a role for OSH15 in the development of rice internodes. This is the first evidence that the knotted1-type homeobox genes have roles other than shoot apical meristem formation and/or maintenance in plant development.

Molecular genetic approaches to leaf development: Knotted and beyond

Molecular genetics provides a promising alternative to other experimental approaches for furthering our understanding of the mechanisms controlling leaf development. We investigated the molecular basis of dominant Knotted (Kn1) mutations in maize, which cause cells associated with the lateral veins of the leaf blade to acquire characteristics of sheath or auricle and sporadically form outgrowths called knots. The kn1 gene encodes a homeodomain, a DNA-binding domain shared by many transcription factors that regulate developmental processes in animals and fungi. In normal plants, the expression of kn1 is confined to the shoot apex, but in Kn1 mutants, the gene is also expressed ectopically in the veins of developing leaves, apparently causing cells to change their developmental fates. The kn1 gene may function in the shoot apex of normal plants to promote indeterminate growth. Consistent with this hypothesis, when kn1 is expressed constitutively at high levels in the leaves of transgenic tobacco, shoots are formed on the leaf surface. Thus, our results indicate that while the kn1 gene may normally have no function in leaf development, it can alter the development of maize and tobacco leaves when it is expressed in the leaf inappropriately. Genes that normally play a role in leaf development are more likely to be defined by recessive mutations that alter leaf morphogenesis and histogenesis. Key words: leaf development, molecular genetics, Knotted.

DEVELOPMENTAL GENETICS OF MUTANTS THAT SPECIFY KNOTTED LEAVES IN MAIZE

Of seven dominant knotted-leaf mutants tested, six mapped at or near Kn1 on the long arm of chromosome 1, and one was not linked to Kn1. Comparisons of phenotypes among these mutants allowed us to focus on a systematic abnormality: the parenchyma cells associated with lateral veins do not fully differentiate into bundle sheath, mesophyll or upper sclerenchyma. The more dramatic expression of Kn1 mutants-knots, ligule alterations and twisting-is sporadic and dependent on the time when the mutant acts in leaf primordium development. Using lw to mark leaf sectors that lose Kn1 following X-irradiation, we show that the knotted-leaf phenotype encoded by chromosome 1L is autonomous. Analysis of sectors lacking a particular Kn1 gene ( Kn1-N2) suggests that Kn1 itself, rather than a linked modifier gene, is autonomous in the leaf primordium. Aneuploid studies using various translocations involving 1L and marked by Adh1 allozymes are compared. The Kn1 mutant appears to encode a "new" function or a considerable overproduction of an extant product in the leaf. Kn1/- 1L hypoploids either express knotted poorly or not at all; transvection is ruled out, but the cause for this modification of Kn1 expression is not yet known.-Our working hypothesis is that Kn1 mutants permit the expression of a product that is usually not produced in leaf primordial cells. We suggest that this product interferes with the early cell-type commitments of cells near lateral veins. Thus, relatively uncommitted cells are present in more mature blades, where they may divide unexpectedly into knots or may induce bits of ligule.