Arabidopsis pollen-specific glycerophosphodiester phosphodiesterase-like genes are essential for pollen tube tip growth

In angiosperms,pollen tube growth is critical for double fertilization and seed formation.Many of the factors involved in pollen tube tip growth are unknown.Here,we report the roles of pollenspecific GLYCEROPHOSPHODIESTER PHOSPHO DIESTERASE-LIKE(GDPD-LIKE)genes in pollen tube tip growth.Arabidopsis thaliana GDPD-LIKE6(At GDPDL6)and At GDPDL7 were specifically expressed in mature pollen grains and pollen tubes and green fluorescent protein(GFP)-At GDPDL6 and GFP-At GDPDL7 fusion proteins were enriched at the plasma membrane at the apex of forming pollen tubes.Atgdpdl6 Atgdpdl7 double mutants displayed severe sterility that was rescued by genetic complementation with At GDPDL6 or At GDPDL7.This sterility was associated with defective male gametophytic transmission.Atgdpdl6 Atgdpdl7 pollen tubes burst immediately after initiation of pollen germination in vitro and in vivo,consistent with the thin and fragile walls in their tips.Cellulose deposition was greatly reduced along the mutant pollen tube tip walls,and the localization of pollen-specific CELLULOSE SYNTHASE-LIKE D1(CSLD1)and CSLD4 was impaired to the apex of mutant pollen tubes.A rice pollen-specific GDPD-LIKE protein also contributed to pollen tube tip growth,suggesting that members of this family have conserved functions in angiosperms.Thus,pollen-specific GDPDLIKEs mediate pollen tube tip growth,possibly by modulating cellulose deposition in pollen tube walls.

SLIDE, the Protein Interacting Domain of Imitation Switch Remodelers, Binds DDT-Domain Proteins of Different Subfamilies in Chromatin Remodeling Complexes

The Imitation Switch （ISWI） type adenosine triphosphate （ATP）-dependent chromatin remodeling factors are conserved proteins in eukaryotes, and some of them are known to form stable remodeling complexes with members from a family of proteins, termed DDT-domain proteins. Although it is well documented that ISWIs play important roles in different biological processes in many eukaryotic species, the molecular basis for protein interactions in ISWI complexes has not been fully addressed. Here, we report the identification of interaction domains for both ISWI and DDT-domain proteins. By analyzing CHROMATIN REMODELING11 （CH R11） and RINGLET1 （RLT1）, an Arabidopsis thaliana ISWI （AtlSWI） and AtDDT-domain protein, respectively, we show that the SLIDE domain of CHR11 and the DDT domain together with an adjacent sequence of RLT1 are responsible for their binding. The Arabidopsis genome contains at least 12 genes that encode DDT-domain proteins, which could be grouped into five subfamilies based on the sequence similarity. The SLIDE domain of AtlSWI is able to bind members from different AtDDT subfamilies. Moreover, a human ISWI protein SNF2H is capable of binding AtDDT-domain proteins through its SLIDE domain, suggesting that binding to DDT-domain proteins is a conserved biochemical function for the SLIDE domain of ISWIs in eukaryotes.

TAA family contributes to auxin production during de novo regeneration of adventitious roots from Arabidopsis leaf explants

Many differentiated plant organs have the ability to regenerate into a new plant after detachment via de novo organogenesis. During de novo root organogenesis from Arabidopsis thaliana leaf explants, wounding first induces endogenous auxin production in mesophyll cells. Auxin is then polar transported to, and accumulates in, regenerationcompetent cells near the wound to trigger the cell-fate transition. The TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS(TAA) family proteins and the YUCCA(YUC) family proteins catalyze two successive biochemical steps in auxin biogenesis, and YUCs have been shown to be involved in auxin production in mesophyll cells during de novo root organogenesis. In thisstudy, we show that the TAA family is also required for adventitious rooting. Inhibition of TAA blocked adventitious root formation from leaf explants. Intriguingly,whereas YUC1 and YUC4 have been shown to be highly induced by wounding, TAA genes retained consistent expression levels before and after leaf detachment.Therefore, we suggest that TAAs and YUCs are both required for auxin biogenesis in leaf explants, but they play different roles in regeneration. While YUC1 and YUC4 function in response to wounding to catalyze the ratelimiting step in auxin biosynthesis, TAAs probably serve as abiding and basal enzymes during de novo root organogenesis from leaf explants.

A Single Amino-Acid Substitution at Lysine 40 of an Arabidopsis thaliana α-tubulin Causes Extensive Cell Proliferation and Expansion Defects

Microtubules are highly dynamic cytoskeletal polymers of α/β-tubulin heterodimers that undergo multiple post-translational modifications essential for various cellular functions in eukaryotes. The lysine 40 （K40） is largely conserved in α-tubulins in many eukaryote species, and the post-translational modification by acetylation at K40 is critical for neuronal development in vertebrates. However, the biological function of K40 of α-tubulins in plants remains unexplored. In this study, we show in Arabidopsis thaliana that constitutive expression of mutated forms of α-tubulin6 （TUA6） at K40 （TUA6K40A or TUA6K40Q ）, in which K40 is replaced by alanine or glutamine, result in severely reduced plant size. Phenotypic characterization of the 35S：TUA6K40A transgenic plants revealed that both cell proliferation and cell expansion were affected. Cytological and biochemical analyses showed that the accumulation of α- and β-tubulin proteins was significantly reduced in the transgenic plants, and the cortical microtubule arrays were severely disrupted, indicating that K40 of the plant α-tubulin is critical in maintaining microtubule stability. We also constructed 35S：TUA6K40R transgenic plants in which K40 of the engineered TUA6 protein is replaced by an arginine, and found that the 35S：TUA6K40R plants were phenotypically indistinguishable from the wild-type. Since lysine and arginine are similar in biochemical nature but arginine cannot be acetylated, these results suggest a structural importance for K40 of α-tubulins in cell division and expansion.