The helper component-proteinase (HC-Pro) protein of potyviruses is a suppressor of gene silencing and has been shown to elicit plant developmental-defect-like symptoms. In Zucchini yellow mosaic virus (ZYMV), a mutation in the highly conserved FR ₁₈₀ NK box of HC-Pro to FI ₁₈₀ NK causes attenuation of these symptoms. At 5 days postinoculation and before symptoms appear, virus accumulation, HC-Pro protein levels, and viral short interfering RNA (siRNA) levels are similar for the severe (FRNK) and attenuated (FINK) strains. At this stage, ZYMV ^FRNK caused greater accumulation of most microRNAs (miRNAs), and especially of their complementary miRNA “passenger” strands (miRNA*s), in systemically infected leaves than the attenuated ZYMV ^FINK did. HC-Pro ^FRNK specifically bound artificial siRNA and miRNA/miRNA* duplexes with a much higher affinity than the mutated HC-Pro ^FINK . Further analysis of the mutant and wild-type HC-Pro proteins revealed that suppressor activity of the ZYMV HC ^FINK mutant was not diminished. However, the FINK mutation caused a loss of HC-Pro suppressor function in other potyviruses. Replacement of the second positively charged amino acid in the ZYMV FRNK box to result in FRNA also caused symptom attenuation and reduced small RNA duplex-binding affinity without loss of suppressor activity. Our data suggest that the highly conserved FRNK box in the HC-Pro of potyviruses is a probable point of contact with siRNA and miRNA duplexes. The interaction of the FRNK box with populations of miRNAs directly influences their accumulation levels and regulatory functions, resulting in symptom development.

Shoot branching is an important aspect of plant architecture because it substantially affects plant biology and agricultural performance. Sugars play an important role in the induction of shoot branching in several species, including potato (Solanum tuberosum L.). However, the mechanism by which sugars affect shoot branching remains mostly unknown. In the present study, we addressed this question using sugar-mediated induction of bud outgrowth in potato stems under etiolated conditions. Our results indicate that sucrose feeding to detached stems promotes the accumulation of cytokinin (CK), as well as the expression of vacuolar invertase (VInv), an enzyme that contributes to sugar sink strength. These effects of sucrose were suppressed by CK synthesis and perception inhibitors, while CK supplied to detached stems induced bud outgrowth and VInv activity in the absence of sucrose. CK-induced bud outgrowth was suppressed in vinv mutants, which we generated by genome editing. Altogether, our results identify a branching-promoting module, and suggest that sugar-induced lateral bud outgrowth is in part promoted by the induction of CK-mediated VInv activity.

SUMMARY: Taxonomy: Zucchini yellow mosaic virus (ZYMV) is a member of genus Potyvirus , family Potyviridae. ZYMV is a positive‐strand RNA virus.  Physical properties: Virions are flexuous filaments of 680–730 nm in length and 11–13 nm in diameter, composed of about 2000 subunits of a single 31‐kDa protein (calculated). The genome RNA size is 9.6 kb covalently linked to a viral‐encoded protein (the VPg) at the 5′ end, and with a 3′ poly A tail. The 5′ end of the sequence is AU‐rich (69%).  Viral proteins: The genome is expressed as a polyprotein cleaved by three viral proteases and processed into ten putative mature proteins. The structural coat protein is processed from the carboxyl terminus of the polyprotein and is highly immunogenic.  Host and symptoms: Natural and experimental infection has been reported mainly in the Cucurbitaceae. Experimental local lesion hosts include Chenopodium amaranticolour , C. quinoa and Gomphrena globosa . Some ZYMV strains cause symptomless infection as in Ranunculus sardous , Nicotiana benthamiana and Sesamum indicum . ZYMV causes stunting and major foliar deformation with dark green blisters and mosaics in cucurbit hosts, eventually developing a filamentous leaf phenotype. In general, symptoms are severe on cucurbit hosts and cause dramatic reductions in yields due to severe fruit deformation. The virus is present in all the plant tissues at relatively high concentrations ( c . 0.1 mg/mL of purified virus per 1 g fresh leaf tissue). The most suitable species for maintenance and purification is Cucurbita pepo.  Transmission: ZYMV is efficiently transmitted by aphids in a non‐persistent manner. The coat protein (CP) and the helper component‐protease (HC‐Pro) are required for aphid transmission, through the CP DAG motif and the HC‐Pro KLSC and PTK motifs. Mechanical transmission is efficient both in the laboratory and naturally.  Economic importance: ZYMV disease is a major constraint in the production of cucurbits world‐wide. The virus can cause massive damage (to total loss) to cucurbit crops, and prevents the growth of some cucurbit crops in certain areas. Control of ZYMV requires the integration of conventional resistance and transgenic breeding along with cross‐protection technologies.

The flavonoids are phenylpropanoid-derived metabolites that are ubiquitous in plants, playing many roles in growth and development. Recently, we observed that fruit rinds of yellow casaba muskmelons (Cucumis melo ‘Inodorous Group’) accumulate naringenin chalcone, a yellow flavonoid pigment. With RNA-sequencing analysis of bulked segregants representing the tails of a population segregating for naringenin chalcone accumulation followed by fine mapping and genetic transformation, we identified a Kelch domain-containing F-box protein coding (CmKFB) gene that, when expressed, negatively regulates naringenin chalcone accumulation. Additional metabolite analysis indicated that downstream flavonoids are accumulated together with naringenin chalcone, whereas CmKFB expression diverts the biochemical flux toward coumarins and general phenylpropanoids. These results show that CmKFB functions as a posttranscriptional regulator that diverts flavonoid metabolic flux.

The 2b protein encoded by Cucumber mosaic virus (CMV) has been shown to be a silencing suppressor and pathogenicity determinant in solanaceous hosts, but a movement determinant in cucumber. In addition, synergistic interactions between CMV and Zucchini yellow mosaic virus (ZYMV) have been described in several cucurbit species. Here, it was shown that deletion of the 2b gene from CMV prevented extensive systemic movement of the virus in zucchini squash, which could not be complemented by co-infection with ZYMV. Thus, ZYMV expressing a silencing suppressor with a different target could not complement the CMV 2b-specific movement function. Expression of the 2b protein from an attenuated ZYMV vector resulted in a synergistic response, largely restoring infection symptoms of wild-type ZYMV in several cucurbit species. Deletion or alteration of either of two nuclear localization signals (NLSs) did not affect nuclear localization in two assays, but did affect pathogenicity in several cucurbit species, whilst deletion of both NLSs led to loss of nuclear localization. The 2b protein interacted with an Arabidopsis thaliana karyopherin α protein (AtKAPα) in the yeast two-hybrid system, as did each of the two single NLS-deletion mutants. However, 2b protein containing a deletion of both NLSs was unable to interact with AtKAPα. These data suggest that the 2b protein localizes to the nucleus by using the karyopherin α-mediated system, but demonstrate that nuclear localization was insufficient for enhancement of the 2b-mediated pathogenic response in cucurbit hosts. Thus, the sequences corresponding to the two NLSs must have another role leading to pathogenicity enhancement.