Salinity inhibits growth and development of most plants. The response to salinity is complex and varies between plant organs and stages of development. It involves challenges of ion toxicities and deficiencies as well as osmotic and oxidative stresses. The range of functions affected by the stress is reflected in elaborate changes to the transcriptome. The mechanisms involved in the developmental-stage specificity of the inhibitory responses are not fully understood. The present study took advantage of the well characterized developmental progression that exists along the maize leaf, for identification of salinity induced, developmentally-associated changes to the transcriptome. Differential subtraction screening was conducted for cells of two developmental stages: from the center of the growth zone where the expansion rate is highest, and from older cells at a more distal location of the growing zone where the expansion rate is lower and the salinity restrictive effects are more pronounced. Real-Time PCR analysis was used for validation of the expression of selected genes.

Deeply conserved plant microRNAs (miRNAs) function as pivotal regulators of development. Nevertheless, in the model crop Solanum lycopersicum (tomato) several conserved miRNAs are still poorly annotated and knowledge about their functions is lacking. Here, the tomato miR171 family was functionally analyzed. We found that the tomato genome contains at least 11 SlMIR171 genes that are differentially expressed along tomato development. Downregulation of sly-miR171 in tomato was successfully achieved by transgenic expression of a short tandem target mimic construct (STTM171). Consequently, sly-miR171-targeted mRNAs were upregulated in the silenced plants. Target upregulation was associated with irregular compound leaf development and an increase in the number of axillary branches. A prominent phenotype of STTM171 expressing plants was their male sterility due to a production of a low number of malformed and nonviable pollen. We showed that sly-miR171 was expressed in anthers along microsporogenesis and significantly silenced upon STTM171 expression. Sly-miR171-silenced anthers showed delayed tapetum ontogenesis and reduced callose deposition around the tetrads, both of which together or separately can impair pollen development. Collectively, our results show that sly-miR171 is involved in the regulation of anther development as well as shoot branching and compound leaf morphogenesis.

Powdery mildew (PM) diseases may severely limit the production of various crops, including members of the family Cucurbitaceae. Successful PM infection relies on the Mildew Resistance Locus O ( MLO) plant gene family, which encodes susceptibility factors essential for fungus penetration into the host cell. In cucumber ( Cucumis sativus), natural mutations in CsaMLO8 confer resistance to the PM pathogen Podosphaera xanthii. Here, we used CRISPR/Cas9-mediated mutagenesis to generate PM resistance in the susceptible cucumber cultivar Ilan. Two transgene-free Csamlo8 CRISPR mutant lines ( Csamlo-cr-1 and Csamlo-cr-2) were isolated, the first with a 5-bp deletion in exon 1, and the second harboring a 1,280-bp deletion and 10-bp insertion between exons 1 and 5. Both lines showed high resistance to PM under semicommercial growth conditions in the summer growing seasons of 2019 and 2021. These results provide the basis for generating transgene-free powdery mildew resistance in cucumber in any genetic background. This method can directly be employed on commercial cultivars and hybrid parental lines, and thereby substantially shorten and simplify the breeding process for PM resistance in cucumber.