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The pericarp is the predominant tissue determining the structural characteristics of most fruits. However, the molecular and genetic mechanisms controlling pericarp development remain only partially understood. Previous studies have identified that CLASS-II KNOX genes regulate fruit size, shape, and maturation in Arabidopsis thaliana and Solanum lycopersicum. Here we characterized the roles of the S. lycopersicum CLASS-II KNOX (TKN-II) genes in pericarp development via a detailed histological, anatomical, and karyotypical analysis of TKN-II gene clade mRNA-knockdown (35S:amiR-TKN-II) fruits. We identify that 35S:amiR-TKN-II pericarps contain more cells around their equatorial perimeter and fewer cell layers than the control. In addition, the cell sizes but not the ploidy levels of these pericarps were dramatically reduced. Further, we demonstrate that fruit shape and pericarp layer number phenotypes of the 35S:amiR-TKN-II fruits can be overridden by the procera mutant, known to induce a constitutive response to the plant hormone gibberellin. However, neither the procera mutation nor exogenous gibberellin application can fully rescue the reduced pericarp width and cell size phenotype of 35S:amiR-TKN-II pericarps. Our findings establish that TKN-II genes regulate tomato fruit anatomy, acting via gibberellin to control fruit shape but utilizing a gibberellin-independent pathway to control the size of pericarp cells.

The pericarp is the predominant tissue determining the structural characteristics of most fruits. However, the molecular and genetic mechanisms controlling pericarp development remain only partially understood. Previous studies have identified that CLASS-II KNOX genes regulate fruit size, shape, and maturation inArabidopsis thalianaandSolanum lycopersicum. Here we characterized the roles of theSolanum lycopersicumCLASS-II KNOX (TKN-II) genes in pericarp development via a detailed histological, anatomical, and karyotype analysis of theTKN-IIknockdown (35S:amiR-TKN-II) fruits. We identify that35S:amiR-TKN-IIpericarps contain more cells around their equatorial perimeter and fewer cell layers than the control. In addition, the cell sizes but not the ploidy levels of these pericarps were dramatically reduced.Further, we demonstrate that fruit shape and pericarp layer number phenotypes of the35S:amiR-TKN-IIfruits can be overridden by theproceramutant, known to induce a constitutive response to the plant hormone gibberellin. However, neither theproceramutation nor exogenous gibberellin application can fully rescue the reduced pericarp width and cell size phenotype of35S:amiR-TKN-IIpericarps. Our findings establish that TKN-II genes regulate tomato fruit anatomy, acting via gibberellin to control fruit shape but utilizing a gibberellin-independent pathway to control the size of pericarp cells.HighlightTomatoCLASS-II KNOXgenes regulate fruit and pericarp anatomy via GA-dependent and independent pathways.

Fruits can be divided into dry and fleshy types. Dry fruits mature through senescence and fleshy fruits through ripening. Previous studies have indicated that partially common molecular networks could govern fruit maturation in these different fruit types. However, the nature of such networks remains obscure. CLASS-II KNOX genes were shown to regulate the senescence of the Arabidopsis (Arabidopsis thaliana) dry fruits, the siliques, but their roles in fleshy-fruit development are unknown. Here, we investigated the roles of the tomato (Solanum lycopersicum) CLASS-II KNOX (TKN-II) genes in fleshy fruit ripening using knockout alleles of individual genes and an artificial microRNA line (35S:amiR-TKN-II) simultaneously targeting all genes. 35S:amiR-TKN-II plants, as well as a subset of tkn-II single and double mutants, have smaller fruits. Strikingly, the 35S:amiR-TKN-II and tknII3 tknII7/+ fruits showed early ripening of the locular domain while their pericarp ripening was stalled. Further examination of the ripening marker-gene RIPENING INHIBITOR (RIN) expression and 35S:amiR-TKN-II rin-1 mutant fruits suggested that TKN-II genes arrest RIN activity at the locular domain and promote it in the pericarp. These findings imply that CLASS-II KNOX genes redundantly coordinate maturation in both dry and fleshy fruits. In tomato, these genes also control spatial patterns of fruit ripening, utilizing differential regulation of RIN activity at different fruit domains.

Ripening is a complex developmental change of a mature organ, the fruit. In plants like a tomato, it involves softening, pigmentation, and biosynthesis of metabolites beneficial for the human diet. Examination of the transcriptional changes towards ripening suggests that redundant uncharacterized factors may be involved in the coordination of the ripening switch. Previous studies have demonstrated that Arabidopsis CLASS-II KNOX genes play a significant role in controlling the maturation of siliques and their transition to senescence. Here we examined the combined role of all four tomato CLASS-II KNOX genes in the maturation and ripening of fleshy fruits using an artificial microRNA targeting them simultaneously. As expected, the knockdown plants (35S::amiR-TKN-CL-II) exhibited leaves with increased complexity, reminiscent of the leaf phenotype of plants overexpressing CLASS-I KNOX, which antagonize CLASS-II KNOX gene functions. The fruits of 35S::amiR-TKN-CL-II plants were notably smaller than the control. While their internal gel/placenta tissue softened and accumulated the typical pigmentation, the pericarp color break took place ten days later than control, and eventually, it turned yellow instead of red.Additionally, the pericarp of 35S::amiR-TKN-CL-II fruits remained significantly firmer than control even after three weeks of shelf storage. Strikingly, the 35S::amiR-TKN-CL-II fruits showed early ethylene release and respiration peak, but these were correlated only with liquefaction and pigmentation of the internal tissues. Our findings suggest that CLASS-II KNOX genes are required to coordinate the spatial and temporal patterns of tomato fruit ripening.One sentence summaryTomato CLASS-II KNOX genes play antagonistic roles in the regulation of ripening at the internal fruit domains and pericarp.

Molecular tools provide new insights into phylogenetic relationships of plant species, and by relating phylogenetic groups to their geographical distribution, we can cast light upon the evolution history of plant clades. In the current study, we evaluated the phylogenetic position of the Sinai endemic Brassica deserti (Brassicaceae), later renamed as Erucastrum deserti, based on morphological data and 5.8S rDNA and ITS (Internal Transcribed Spacer) regions. Our results indicate that B. deserti belongs to an East-Mediterranean – Saharo Arabian clade and was not assigned to the core Brassica and Erucastrum clades, respectively, which evolved in the West Mediterranean area. We tentatively conclude that Brassica deserti evolved independently of core Brassica and Erucastrum.

The winter annuals Brassica tournefortii and Raphanus raphanistrum (Brassicaceae) share similar habitats and life-history traits, but differ in their reproduction system (self-compatibility vs. self-incompatibility, respectively). The two phylogenetically close species offer means to assess the effect of reproductive biology on genetic diversity between and within populations. In general, genetic diversity between populations of B. tournefortii was higher than that found between populations of R. raphanistrum, while higher genetic diversity indices were evident within populations of R. raphanistrum. In addition, the results of pairwise genetic distances indicated that the genetic distances between populations can be associated to the species’ reproductive biology and not to the population’s distribution pattern. We discuss whether knowledge of reproductive and habitat characteristics can be used to predict genetic diversity when planning sampling scheme for ex situ conservation.

The protective peel of potato tuber consists of periderm tissue, the outmost cell layers of which contain corky cell walls and are termed "skin". The skin protects the tuber from water loss and pathogen invasion, and its visual appearance is a highly important marketing factor. Physiological skin blemishes are of great concern, mainly russeting disorder and skinning injuries. We previously showed that application of calcium (Ca) reduces the rate and severity of skin russeting. Here, polyhalite fertilization was tested as an alternative source of Ca. The polyhalite mineral is a hydrated sulfate of potassium (K), Ca, and magnesium (Mg), and thus contains additional important nutrients that may contribute to skin quality. Furthermore, in view of the direct interaction of soil mineral elements with the tuber skin, we tested application of polyhalite at the end of the growth period, assuming that providing the mineral at the last stages of skin development may enhance its quality. Accordingly, polyhalite was applied at three time points: preplanting, in-season at around 3-4 weeks prior to haulm desiccation, and 2 days post-haulm desiccation. The experiments included several cultivars and locations. Data indicated that late application of polyhalite, after haulm desiccation, results in reduced concentrations of Ca and Mg and increased concentration of K in the tuber peel of fertilized plants compared to controls. Tuber appearance was improved, and the expression of FHT and CYP86A33, indicator genes for skin suberization, was significantly upregulated. Earlier applications of the polyhalite mineral did not alter mineral elements concentrations in the tuber peel compared to control plants. Overall, polyhalite fertilization positively affected tuber skin appearance and skin-related gene expression. However, the effect was moderate, and the mineral did not fully mitigate skin imperfections. The effect of polyhalite may be dependent on local conditions and cultivar type.

The ex situ collection of the Israel Plant Gene Bank (IGB) aims to encompass the rich local flora and its genetic diversity with an emphasis on crop wild relatives. However, to properly establish a core collection, collection efforts must be prioritized and strategized. We previously classified local plant genetic resources into four priority groups that assisted in strategizing the collection activities. The following years of intensive collection activity yielded over 4200 banked accessions. However, these do not necessarily represent the distribution range of the target species for collection (TSC) and consequently, their genetic diversity. To best cover the latter, the collecting area was divided into botanical districts and the magnitude of the collection was determined according to prioritization group, e.g., a wild relative of an agricultural crop with a vast distribution range should be represented by a larger number of banked accessions than one with a smaller range. Continuous evaluation of specific needs shapes the collection scheme of the IGB to maximize collection efforts, better represent the presumed genetic diversity of TSC, and establish its core collection.

Background Naturally growing populations of olive trees are found in the Mediterranean garrigue and maquis in Israel. Here, we used the Simple Sequence Repeat (SSR) genetic marker technique to investigate whether these represent wild var. sylvestris. Leaf samples were collected from a total of 205 trees at six sites of naturally growing olive populations in Israel. The genetic analysis included a multi-locus lineage (MLL) analysis, Rousset’s genetic distances, Fst values, private alleles, other diversity values and a Structure analysis. The analyses also included scions and suckers of old cultivated olive trees, for which the dominance of one clone in scions (MLL1) and a second in suckers (MLL7) had been shown earlier. Results The majority of trees from a Judean Mts. population and from one population from the Galilee showed close genetic similarity to scions of old cultivated trees. Different from that, site-specific and a high number of single occurrence MLLs were found in four olive populations from the Galilee and Carmel which also were genetically more distant from old cultivated trees, had relatively high genetic diversity values and higher numbers of private alleles. Whereas in two of these populations MLL7 (and partly MLL1) were found in low frequency, the two other populations did not contain these MLLs and were very similar in their genetic structure to suckers of old cultivated olive trees that originated from sexual reproduction. Conclusions The genetic distinctness from old cultivated olive trees, particularly of one population from Galilee and one from Carmel, suggests that trees at these sites might represent wild var. sylvestris. The similarity in genetic structure of these two populations with the suckers of old cultivated trees implies that wild trees were used as rootstocks. Alternatively, trees at these two sites may be remnants of old cultivated trees in which the scion-derived trunk died and was replaced by suckers. However, considering landscape and topographic environment at the two sites this second interpretation is less likely. Electronic supplementary material The online version of this article (doi:10.1186/s12870-016-0947-5) contains supplementary material, which is available to authorized users.

The rind of muskmelon (Cucumis melo L. var. reticulatus) fruits contains a network of suberized periderm tissue, referred as the 'net', which originates in response to cracking of the fruit surface during its enlargement. Shallow cuts were made on the surface of melons to mimic naturally occurring cracking and induce net-like periderm development. Histological analysis of wounded fruits of the climacteric netted variety Krimka, and of two smooth melon varieties: the climacteric Momordica and the non-climacteric Tamdew, indicated that smooth melon varieties can undergo netting when their rind is fissured. Furthermore, the results implied that the climacteric character is not essential for net-tissue development, even though most netted varieties are climacteric. The involvement of ethylene in net- like periderm development was studied by analyzing the expression pattern of the ethylene-biosynthesis genes 1-aminocyclopropane-1-carboxylate synthase 1 and 1-aminocyclopropane-1-carboxylate oxidase 1 following wounding and during periderm development, and by applying the ethylene-generating chemical, Ethrel, as a lanolin paste, on the fresh cuts. Results suggested ethylene involvement in periderm initiation, but continuous exposure may interfere with further tissue development and organization. General implications of the current study on periderm development are further discussed.

Artificial induction of grape bud dormancy release by hydrogen cyanamide (HC) serves as a reliable model system to explore the events occurring shortly after the induction of dormancy release. Recently, a group of genes with remarkable differences in expression level between HC-treated and control buds was identified. The identification of several calcium signalling-related genes within that group raised the hypothesis of the involvement of Ca(2+) signalling in grape bud dormancy release. Therefore, the effects of HC treatment on the expression profiles of several calcium sensors, the effect of the plasma membrane calcium channel blocker LaCl(3) and the calcium chelator EGTA on HC-induced and chilling-induced bud-break, and the effect of HC application on calcium-dependent protein phosphorylation activities in the bud tissue were studied. Here the HC-induced expression of Ca(2+)-ATPase is described, indicating that this treatment might evoke an increase in [Ca(2+)]cyt. Similar induction was confirmed for calmodulin, calmodulin-binding protein, and calcium-dependent protein kinase (CDPK). Both LaCl(3) and EGTA blocked the inducing effect of HC on bud-break, and their inhibitory effects were removed by supplying exogenous Ca(2+). Calcium-dependent histone phosphorylation was up to 70% higher in HC-treated buds. Endogenous protein phosphorylation assays detected four proteins exhibiting increased phosphorylation following HC treatment, of which two were phosphorylated in a calcium-dependent manner. One of these, a 47 kDa protein, presented strong and Ca(2+)-dependent phosphorylation only in HC-treated buds. The potential role of CDPK in the phosphorylation of this protein was supported by an immunoprecipitation assay. The data suggest, for the first time, that calcium signalling is involved in the mechanism of bud dormancy release.

The rind of fruits of muskmelon (Cucumis melo L. var. reticulatus) contains a network of suberized tissue referred to as the 'netting', and peroxidase (EC 1.11.1.7) activity is necessary to the polymerization of the aromatic domain of suberin. Peroxidase activity increased dramatically during the early stages of melon fruit netting, and in fruits exhibiting incomplete netting, peroxidase activity was significantly higher in netted than in non-netted regions of the same fruit. Moreover, analysis of peroxidase activity in three varieties of smooth-rind melons (Cucumis melo var. inodorous) indicated lower levels of the activity in rind samples, taken throughout fruit development, than in rinds of netted varieties. Netting-associated anionic peroxidase (NAPOD) was isolated from the melon rind at an early stage of netting development, partially purified, microsequenced and its cDNA was cloned. It was found to be a single-copy gene within the genome of netted and smooth melon varieties, and highly homologous to other Cucurbitaceous anionic peroxidases. A high transcript level was only detected in the rind of the netted variety. Monitoring the gene expression of netting-associated anionic peroxidase, together with other enzymes involved in the netting will shed light on the molecular control of the suberization processes in the melon rind and in plants in general.

SummaryThe suberized net tissue of the rind of muskmelon (Cucumis melo L.) is of agricultural importance as an indicator of fruit maturity and quality, and as a preventive factor against mechanical injury, both pre- and postharvest. The net originates from cracks that appear at the surface of the fruit. Therefore, a comparative study was conducted with two netted-rind and two smooth-rind melon varieties, to identify fruit characteristics associated with initiation of netting development. The developmental pattern of fruit expansion did not differ between smooth and netted melon varieties. Rather, our results indicated that inherited cuticular/epidermal characteristics are probably associated with the development of ®ssures in the fruit rind. The rind of the netted fruit was found to contain signi®cantly more cuticle than that of the smooth-rind variety during the period of maximal growth rate; moreover, in the rind of the smooth varieties the deposition of the cuticle was mainly on the outside of the epid...