Creating Completely Both Male and Female Sterile Plants by Specifically Ablating Microspore and Megaspore Mother Cells

Abstract

Although genetically modified (GM) plants have improved commercially important traits, such as biomass and biofuel production, digestibility, bioremediation, ornamental value, and tolerance to biotic and abiotic stresses, there remain economic, political, or social concerns over potential ecological effects of transgene flow from GM plants. The current solution for preventing transgene flow from GM plants is genetically engineering sterility; however, approaches to generating both male and female sterility are limited. In addition, existing strategies for creating sterility lead to loss or modifications of entire flowers or floral organs. Here, we demonstrate that instead of the 1.5-kb promoter, the entire SOLO DANCERS (SDS) gene is required for its meiocyte-specific expression. We then developed an efficient method to specifically ablate microspore and megaspore mother cells using the SDS and BARNASE fusion gene, which resulted in complete sterility in both male and female reproductive organs in Arabidopsis (Arabidopsis thaliana) and tobacco (Nicotiana tabacum), but did not affect plant growth or development, including the formation of all flower organs. Therefore, our research provides a general and effective tool to prevent transgene flow in GM plants.

Carbonic Anhydrases Function in Anther Cell Differentiation Downstream of the Receptor-Like Kinase EMS1

“…To identify viable pollen grains, mature anthers prior to dehiscence were collected for Alexander staining of pollen (Zhao et al, 2002;Xin et al, 2017). To study anther cell differentiation, semithin sectioning was performed essentially as previously described (Zhao et al, 2002;Huang et al, 2016b). For semithin sectioning, dissected young buds were fixed in 2.5% (v/v) of glutaraldehyde in 0.1 M HEPES buffer (pH 7.2) and 0.02% of Triton X-100 overnight at 4°C.…”

“…In the later stage, tapetal cells are degenerated via programmed cell death and the resulting materials are deposited onto pollen to form the pollen coat (tryphine). Mutants lacking tapetum or plants with genetically ablated tapetum fail to produce pollen grains (Mariani et al, 1990;Zhao et al, 2002;Yang et al, 2003;Huang et al, 2016b). In addition, precocious or delayed tapetum degeneration causes abnormal pollen development (Zhang et al, 2006;Zhang and Yang, 2014).…”

Two SERK Receptor-Like Kinases Interact with EMS1 to Control Anther Cell Fate Determination

“…Alexander pollen staining and anther semithin sectioning were conducted as described in our previous studies (Huang et al, 2016a(Huang et al, , 2016c.…”

Morphological Characterization of a New and Easily Recognizable Nuclear Male Sterile Mutant of Sorghum (Sorghum bicolor)

“…With endoreduplication, polynucleate tapetal cells, which are highly active in metabolism, provide energy and materials (sugar, lipid, and protein) for pollen development. Lacking or abnormal tapetum causes pollen defects and consequently male sterility [18, 19, 38–41]. Moreover, stress-induced male sterility and yield loss are mainly ascribed to aberrant tapetum development [42, 43].…”

“…Alexander staining was used to determine pollen viability as described previously [18, 19]. Briefly, anthers just before anther dehiscence were dissected out and fixed 24 h in the fixative (methanol, 60mL; chloroform, 30mL; distilled water, 20 mL; picric acid, 1 g; and HgCl 2 , 1 g).…”