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The development of trichomes (spines) on cucumber fruits is an important agronomic trait. It has been reported that two MYB family members, CsMYB6 (Csa3G824850) and CsTRY (Csa5G139610) act as negative regulators of trichome or fruit spine initiation. To further study the functions of these two genes, we overexpressed them in tobacco, and found that the flowers and seed coats of transformants overexpressing CsTRY displayed an unexpected defect in pigmentation that was not observed in plants overexpressing CsMYB6. Moreover, the expression of key genes in the flavonoid synthesis pathway was repressed in CsTRY overexpressing plants, which resulted in the decrease of several important flavonoid secondary metabolites. In addition, CsTRY could interact with the AN1 homologous gene CsAN1 (Csa7G044190) in cucumber, which further confirmed that CsTRY not only regulates the development of fruit spines, but also functions in the synthesis of flavonoids, acting as the repressor of anthocyanin synthesis.

Powdery mildew is a common disease in plants, and it is also one of the main diseases in the middle and final stages of cucumber (Cucumis sativus). Powdery mildew on plant leaves affects the photosynthesis, which may reduce the plant yield. Therefore, it is of great significance to automatically identify powdery mildew. Currently, most image-based models commonly regard the powdery mildew identification problem as a dichotomy case, yielding a true or false classification assertion. However, quantitative assessment of disease resistance traits plays an important role in the screening of breeders for plant varieties. Therefore, there is an urgent need to exploit the extent to which leaves are infected which can be obtained by the area of diseases regions. In order to tackle these challenges, we propose a semantic segmentation model based on convolutional neural networks (CNN) to segment the powdery mildew on cucumber leaf images at pixel level, achieving an average pixel accuracy of 96.08%, intersection over union of 72.11% and Dice accuracy of 83.45% on twenty test samples. This outperforms the existing segmentation methods, K-means, Random forest, and GBDT methods. In conclusion, the proposed model is capable of segmenting the powdery mildew on cucumber leaves at pixel level, which makes a valuable tool for cucumber breeders to assess the severity of powdery mildew.

In cucumber (Cucumis sativus L.), the differentiation and development of female flowers are important processes that directly affect the fruit yield and quality. Sex differentiation is mainly controlled by three ethylene synthase genes, F (CsACS1G), M (CsACS2), and A (CsACS11). Thus, ethylene plays a key role in the sex differentiation in cucumber. The “one-hormone hypothesis” posits that F and M regulate the ethylene levels and initiate female flower development in cucumber. Nonetheless, the precise molecular mechanism of this process remains elusive. To investigate the mechanism by which F and M regulate the sex phenotype, three cucumber near-isogenic lines, namely H34 (FFmmAA, hermaphroditic), G12 (FFMMAA, gynoecious), and M12 (ffMMAA, monoecious), with different F and M loci were generated. The transcriptomic analysis of the apical shoots revealed that the expression of the B-class floral homeotic genes, CsPI (Csa4G358770) and CsAP3 (Csa3G865440), was immensely suppressed in G12 (100% female flowers) but highly expressed in M12 (∼90% male flowers). In contrast, CAG2 (Csa1G467100), which is an AG-like C-class floral homeotic gene, was specifically highly expressed in G12. Thus, the initiation of female flowers is likely to be caused by the downregulation of B-class and upregulation of C-class genes by ethylene production in the floral primordium. Additionally, CsERF31, which was highly expressed in G12, showed temporal and spatial expression patterns similar to those of M and responded to the ethylene-related chemical treatments. The biochemical experiments further demonstrated that CsERF31 could directly bind the promoter of M and promote its expression. Thus, CsERF31 responded to the ethylene signal derived from F and mediated the positive feedback regulation of ethylene by activating M expression, which offers an extended “one-hormone hypothesis” of sex differentiation in cucumber.

Powdery mildew (PM) is a serious fungal disease of cucumber worldwide. The identification of resistance genes is very important for resistance breeding to ensure cucumber production. Here, natural loss-of-function mutations at an MLO homologous locus, CsMLO1, were found to confer durable PM resistance in cucumber. CsMLO1 encoded a cell membrane protein, was mainly expressed in leaves and cotyledons, and was up-regulated by PM at the early stage of host-pathogen interaction. Ectopic expression of CsMLO1 rescued the phenotype of the PM resistant Atmlo2 Atmlo12 double mutant to PM susceptible in Arabidopsis. Domesticated and wild resistant cucumbers originating from various geographical regions of the world were found to harbor three independent natural mutations that resulted in CsMLO1 loss of function. In addition, between the near-isogenic lines (NILs) of PM resistant and susceptible, S1003 and NIL(Pm5.1), quantitative RT-PCR revealed that there is no difference at expression levels of several genes in the pathways of ethylene, jasmonic acid or salicylic acid. Moreover, the two NILs were used for transcriptome profiling to explore the mechanism underlying the resistance. Several genes correlated with plant cell wall thickening are possibly involved in the PM resistance. This study revealed that loss of function of CsMLO1 conferred durable PM resistance, and that this loss of function is necessary but alone may not be sufficient for PM resistance in cucumber. These findings will facilitate the molecular breeding of PM resistant varieties to control this destructive disease in cucumber. [ABSTRACT FROM AUTHOR]