Qianyi Tao

Molecular isolation of the M gene suggests that a conserved-residue conversion induces the formation of bisexual flowers in cucumber plants.

Sex determination in plants involves a variety of mechanisms. Here, we report the map-based cloning and characterization of the unisexual-flower-controlling gene M. M was identified as a previously characterized putative 1-aminocyclopropane-1-carboxylic acid synthase gene, while the m allele that mutated at a conserved site (Gly33Cys) lost activity in the original enzymatically active allele.

A putative positive feedback regulation mechanism in CsACS2 expression suggests a modified model for sex determination in cucumber (Cucumis sativus L.)

It is well established that the plant hormone ethylene plays a key role in cucumber sex determination. Since the unisexual control gene M was cloned and shown to encode an ethylene synthase, instead of an ethylene receptor, the ‘one-hormone hypothesis’, which was used to explain the cucumber sex phenotype, has been challenged. Here, the physiological function of CsACS2 (the gene encoded by the M locus) was studied using the transgenic tobacco system. The results indicated that overexpression of CsACS2 increased ethylene production in the tobacco plant, and the native cucumber promoter had no activity in transgenic tobacco (PM). However, when PM plants were treated with exogenous ethylene, CsACS2 expression could be detected. In cucumber, ethylene treatment could also induce transcription of CsACS2, while inhibition of ethylene action reduced the expression level. These findings suggest a positive feedback regulation mechanism for CsACS2, and a modified ‘one-hormone hypothesis’ for sex determination in cucumber is proposed.

A mutant in the CsDET2 gene leads to a systemic brassinosteriod deficiency and super compact phenotype in cucumber (Cucumis sativus L.)

Key messageA novel dwarf cucumber mutant, scp-2, displays a typical BR biosynthesis-deficient phenotype, which is due to a mutation inCsDET2 for a steroid 5-alpha-reductase.AbstractBrassinosteroids (BRs) are a group of plant hormones that play important roles in the development of plant architecture, and extreme dwarfism is a typical outcome of BR-deficiency. Most cucumber (Cucumis sativus L.) varieties have an indeterminate growth habit, and dwarfism may have its value in manipulation of plant architecture and improve production in certain production systems. In this study, we identified a spontaneous dwarf mutant, super compact-2 (scp-2), that also has dark green, wrinkle leaves. Genetic analyses indicated that scp-2 was different from two previously reported dwarf mutants: compact (cp) and super compact-1 (scp-1). Map-based cloning revealed that the mutant phenotype was due to two single nucleotide polymorphism and a single-base insertion in the CsDET2 gene that resulted in a missense mutation in a conserved amino acid and thus a truncated protein lacking the conserved catalytic domains in the predicted steroid 5α-reductase protein. Measurement of endogenous hormone levels indicated a reduced level of brassinolide (BL, a bioactive BR) in scp-2, and the mutant phenotype could be partially rescued by the application of epibrassinolide (EBR). In addition, scp-2 mutant seedlings exhibited dark-grown de-etiolation, and defects in cell elongation and vascular development. These data support that scp-2 is a BR biosynthesis-deficient mutant, and that the CsDET2 gene plays a key role in BR biosynthesis in cucumber. We also described the systemic BR responses and discussed the specific BR-related phenotypes in cucumber plants.

Ethylene responsive factor ERF110 mediates ethylene-regulated transcription of a sex determination-related orthologous gene in two Cucumis species

In plants, unisexual flowers derived from developmental sex determination form separate stamens and pistils that facilitate cross pollination. In cucumber and melon, ethylene plays a key role in sex determination. Six sex determination-related genes have been identified in ethylene biosynthesis in these Cucumis species. The interactions among these genes are thought to involve ethylene signaling; however, the underlying mechanism of regulation remains unknown. In this study, hormone treatment and qPCR assays were used to confirm expression of these sex determination-related genes in cucumber and melon is ethylene sensitive. RNA-Seq analysis subsequently helped identify the ethylene responsive factor (ERF) gene, CsERF110, related to ethylene signaling and sex determination. CsERF110 and its melon ortholog, CmERF110, shared a conserved AP2/ERF domain and showed ethylene-sensitive expression. Yeast one-hybrid and ChIP-PCR assays further indicated that CsERF110 bound to at least two sites in the promoter fragment of CsACS11, while transient transformation analysis showed that CsERF110 and CmERF110 enhance CsACS11 and CmACS11 promoter activity, respectively. Taken together, these findings suggest that CsERF110 and CmERF110 respond to ethylene signaling, mediating ethylene-regulated transcription of CsACS11 and CmACS11 in cucumber and melon, respectively. Furthermore, the mechanism involved in its regulation is thought to be conserved in these two Cucumis species.

Expression analysis of the unisex-determine gene M in cu-cumber: Expression analysis of the unisex-determine gene M in cu-cumber

Sex determination in plants has various mechanism; however, a single gene locus controlling unisexual expression is unique in the Cucurbitaceae plants, particularly cucumber (Cucumis sativus L.) and melon (Cucumis melo L.). In this study, with quantitative RT-PCR method, two sets of near-isogenic lines (NILs) were used to analyze the expression pattern of gene CsACS2 (GenBank accession number FJ529216). Additionally, chemical applications (AgNO3 and AVG) were used to investigate the effect of the plant endogenous ethylene on CsACS2 expression. Expression analysis reveals that endogenous ethylene, which might be derived from F or M itself, could activate the expression of M gene.