Stomata are two-celled valves that control epidermal pores whose spacing optimizes shoot-atmosphere gas exchange. They develop from protodermal cells after unequal divisions followed by an equal division and differentiation. The concentration of the hormone auxin, a master plant developmental regulator, is tightly controlled in time and space, but its role, if any, in stomatal formation is obscure. Here dynamic changes of auxin activity during stomatal development are monitored using auxin input (DII-VENUS) and output (DR5:VENUS) markers by time-lapse imaging. A decrease in auxin levels in the smaller daughter cell after unequal division presages the acquisition of a guard mother cell fate whose equal division produces the two guard cells. Thus, stomatal patterning requires auxin pathway control of stem cell compartment size, as well as auxin depletion that triggers a developmental switch from unequal to equal division.

SummaryIn most flowering plant species, pollination and fertilization occur during the hot summer, so plants must have evolved a mechanism that ensures normal growth of their pollen tubes at high temperatures. Despite its importance to plant reproduction, little is known about the molecular basis of thermotolerance in pollen tubes. Here we report the identification and characterization of a novel Arabidopsis gene, THERMOSENSITIVE MALE STERILE 1 (TMS1), which plays an important role in thermotolerance of pollen tubes. TMS1 encodes a Hsp40-homologous protein with a DnaJ domain and an a_ERdj5_C domain found in protein disulfide isomerases (PDI). Purified TMS1 expressed in Escherichia coli (BL21 DE3) had the reductive activity of PDI. TMS1 was expressed in pollen grains, pollen tubes and other vegetative tissues, including leaves, stems and roots. Heat shock treatment at 37°C increased its expression levels in growing pollen tubes as well as in vegetative tissues. A knockout mutation in TMS1 grown at 30°C had greatly retarded pollen tube growth in the transmitting tract, resulting in a significant reduction in male fertility. Our study suggests that TMS1 is required for thermotolerance of pollen tubes in Arabidopsis, possibly by functioning as a co-molecular chaperone.

No abstract

The cell wall is important for pollen tube growth, but little is known about the molecular mechanism that controls cell wall deposition in pollen tubes. Here, the functional characterization of the pollen-expressed Arabidopsis cellulose synthase-like D genes CSLD1 and CSLD4 that are required for pollen tube growth is reported. Both CSLD1 and CSLD4 are highly expressed in mature pollen grains and pollen tubes. The CSLD1 and CSLD4 proteins are located in the Golgi apparatus and transported to the plasma membrane of the tip region of growing pollen tubes, where cellulose is actively synthesized. Mutations in CSLD1 and CSLD4 caused a significant reduction in cellulose deposition in the pollen tube wall and a remarkable disorganization of the pollen tube wall layers, which disrupted the genetic transmission of the male gametophyte. In csld1 and csld4 single mutants and in the csld1 csld4 double mutant, all the mutant pollen tubes exhibited similar phenotypes: the pollen tubes grew extremely abnormally both in vitro and in vivo, which indicates that CSLD1 and CSLD4 are not functionally redundant. Taken together, these results suggest that CSLD1 and CSLD4 play important roles in pollen tube growth, probably through participation in cellulose synthesis of the pollen tube wall.