Functional Characterization of MdTAC1a Gene Related to Branch Angle in Apple (Malus x domestica Borkh.).

The Tiller Angle Control 1 (TAC1) gene belongs to the IGT family, which mainly controls plant branch angle, thereby affecting plant form. Two members of MdTAC1 are identified in apple; the regulation of apple branch angle by MdTAC1 is still unclear. In this study, a subcellular localization analysis detected MdTAC1a in the nucleus and cell membrane, but MdTAC1b was detected in the cell membrane. Transgenic tobacco by overexpression of MdTAC1a or MdTAC1b showed enlarged leaf angles, the upregulation of several genes, such as GA 2-oxidase (GA2ox), and a sensitive response to light and gravity. According to a qRT-PCR analysis, MdTAC1a and MdTAC1b were strongly expressed in shoot tips and vegetative buds of weeping cultivars but were weakly expressed in columnar cultivars. In the MdTAC1a promoter, there were losses of 2 bp in spur cultivars and 6 bp in weeping cultivar compared with standard and columnar cultivars. An InDel marker specific to the MdTAC1a promoter was developed to distinguish apple cultivars and F1 progeny. We identified a protein, MdSRC2, that interacts with MdTAC1a, whose encoding gene which was highly expressed in trees with large branch angles. Our results indicate that differences in the MdTAC1a promoter are major contributors to branch-angle variation in apple, and the MdTAC1a interacts with MdSRC2 to affect this trait.

Cyclanilide Induces Lateral Bud Outgrowth by Modulating Cytokinin Biosynthesis and Signalling Pathways in Apple Identified via Transcriptome Analysis.

Cyclanilide (CYC), a plant growth regulator, is a potent shoot branching agent in apple. However, its mechanism remains unclear. The current study revealed that CYC treatment resulted in massive reprogramming of the axillary bud transcriptome, implicating several hormones in the response. We observed a marked increase (approximately 2-fold) in the level of zeatin riboside and a significant decrease (approximately 2-fold) in the level of abscisic acid (ABA). Zeatin metabolism gene cytokinin (CTK) oxidase 1 (CKX 1) was down-regulated at 168 h after CYC treatment compared with the control. Weighted gene co-expression network analysis of differentially expressed genes demonstrated the turquoise module clusters exhibited the highest positive correlation with zeatin riboside (r = 0.92) and the highest negative correlation with ABA (r = -0.8). A total of 37 genes were significantly enriched in the plant hormone signal transduction pathway in the turquoise module. Among them, the expressions of CTK receptor genes WOODEN LEG and the CTK type-A response regulators genes ARR3 and ARR9 were up-regulated. ABA signal response genes protein phosphatase 2C genes ABI2 and ABI5 were down-regulated in lateral buds after CYC treatment at 168 h. In addition, exogenous application of 6-benzylaminopurine (6-BA, a synthetic type of CTK) and CYC enhanced the inducing effect of CYC, whereas exogenous application of lovastatin (a synthetic type of inhibitor of CTK biosynthesis) or ABA and CYC weakened the promoting effect of CYC. These results collectively revealed that the stimulation of bud growth by CYC might involve CTK biosynthesis and signalling, including genes CKX1 and ARR3/9, which provided a direction for further study of the branching promoting mechanism of CYC.

Transcriptome Analysis Reveals Multiple Genes and Complex Hormonal-Mediated Interactions with PEG during Adventitious Root Formation in Apple.

Adventitious root (AR) formation is a bottleneck for the mass propagation of apple rootstocks, and water stress severely restricts it. Different hormones and sugar signaling pathways in apple clones determine AR formation under water stress, but these are not entirely understood. To identify them, GL-3 stem cuttings were cultured on polyethylene glycol (PEG) treatment. The AR formation was dramatically decreased compared with the PEG-free control (CK) cuttings by increasing the endogenous contents of abscisic acid (ABA), zeatin riboside (ZR), and methyl jasmonate (JA-me) and reducing the indole-3-acetic acid (IAA) and gibberellic acid 3 (GA3) contents. We performed a transcriptomic analysis to identify the responses behind the phenotype. A total of 3204 differentially expressed genes (DEGs) were identified between CK and PEG, with 1702 upregulated and 1502 downregulated genes. Investigation revealed that approximately 312 DEGs were strongly enriched in hormone signaling, sugar metabolism, root development, and cell cycle-related pathways. Thus, they were selected for their possible involvement in adventitious rooting. However, the higher accumulation of ABA, ZR, and JA-me contents and the upregulation of their related genes, as well as the downregulation of sugar metabolism-related genes, lead to the inhibition of ARs. These results indicate that AR formation is a complicated biological process chiefly influenced by multiple hormonal signaling pathways and sugar metabolism. This is the first study to demonstrate how PEG inhibits AR formation in apple plants.

Effects of Indole-3-Butyric Acid (IBA) and rooting media on rooting and survival of air layered wax apple (Syzygium samarangense) CV Jambu Madu / Efeitos do Ácido Indol-3-Butírico (AIB) e meio de enraizamento no enraizamento e sobrevivência de maçã de cera em camada de ar (Syzygium samarangense) CV Jambu Madu

The wax apple or jambu madu, is a non-climacteric tropical fruit from Myrtaceae family and widely cultivated in South East Asia. The limited availability of good quality seedlings of wax apple is the main problem to development of flourish it's market share in the current fruit industry. Therefore, in order to produce good quality planting materials, a study aimed at optimizing propagation and adventitious rooting technique and survivability of wax apple air layer was conducted. In this study, four different levels of Indole-3-Butyric Acid (IBA) concentration (0, 1000, 1500 and 2000 mg L-1) and three rooting media (sphagnum moss, vermicompost and garden soil) were applied after removal of bark (phloem) on the shoot to determine the effect on rooting and survivability of the wax apple air layer under field conditions. The results showed that the wax apple shoots treated with 2000 mg L-1 IBA produced the significantly higher number of roots, increased length of root, diameter of branch, length of branch, number of leaf and leaf area of air layers. In addition, the highest chlorophyll content and stomatal aperture were recorded in 2000 mg L-1 IBA treatment compared to other treatments including control. Vermicompost medium was better than garden soil and sphagnum moss in respect of rooting and survivability of air layers. The results showed that the combination of 2000 mg L-1 IBA and vermicompost as rooting media give the best combination to root initiation, root number, root length and survival rate (100%) of wax apple air layers. From this study, it can be concluded that 2000 mg L-1 IBA and vermicompost treatment enhance the root initiation, early establishment and survivability of wax apple air layered under field conditions.

A maçã de cera, ou jambu madu, é uma fruta tropical não climatérica da família Myrtaceae e amplamente cultivada no Sudeste Asiático. A disponibilidade limitada de mudas de macieira de boa qualidade é o principal problema para o desenvolvimento de sua participação de mercado na fruticultura atual. Portanto, com o objetivo de produzir materiais de plantio de boa qualidade, foi realizado um estudo visando otimizar a técnica de propagação e enraizamento adventício e a sobrevivência da camada aérea da cera de macieira. Neste estudo, quatro diferentes níveis de concentração de ácido indol-3-butírico (AIB) (0, 1000, 1500 e 2000 mg L-1) e três meios de enraizamento (musgo esfagno, vermicomposto e solo de jardim) foram aplicados após a remoção da casca (floema) na parte aérea para determinar o efeito no enraizamento e capacidade de sobrevivência da camada de ar da macieira em condições de campo. Os resultados mostraram que os brotos de macieira tratados com 2000 mg L-1 de AIB produziram significativamente maior número de raízes, maior comprimento de raiz, diâmetro de galho, comprimento de galho, número de folhas e área foliar das camadas aéreas. Além disso, o maior teor de clorofila e abertura estomática foram registrados no tratamento 2000 mg L-1 IBA em comparação com outros tratamentos, incluindo o controle. O meio de vermicomposto foi melhor do que o solo de jardim e o musgo esfagno em relação ao enraizamento e capacidade de sobrevivência das camadas aéreas. Os resultados mostraram que a combinação de 2000 mg L-1 de AIB e vermicomposto como meio de enraizamento proporciona a melhor combinação para iniciação radicular, número de raízes, comprimento radicular e taxa de sobrevivência (100%) das camadas aéreas de macieira. A partir deste estudo, pode-se concluir que 2.000 mg L-1 de AIB e tratamento com vermicomposto melhoram a iniciação radicular, o estabelecimento precoce e a capacidade de sobrevivência de macieiras em camadas de ar em condições de campo.

Horticultural performance of 'Fuji Suprema' and 'Maxi Gala' apple trees trained at Tall Spindle and Bi-Axis systems grafted on different rootstocks in Southern Brazil / Comportamento agronômico de macieiras 'Fuji Suprema' e 'Maxi Gala' conduzidas em Tall Spindle e Bi-Axis enxertadas em diferentes porta-enxertos no Sul do Brasil

The use of new rootstocks combined with different training systems have been studied to increase productivity, fruit quality, and reduce costs. Another important factor is the validation of studies under replanting conditions, due to the growing limitation of new areas for planting commercial apple orchards in Brazil. The present study aimed to evaluate the behavior of 'Fuji Suprema' and 'Maxi Gala' apple trees in two training systems grafted on two rootstocks. The study occurred during the seasons 2016/17, 2017/18 and 2018/19, in an orchard located in southern Brazil in an area previously cultivated with apple trees. A randomized block design consisting of a 2x2 factorial was used, with Tall Spindle and Bi-Axis as the training system and 'M.9' and 'G.213' as rootstocks of Fuji Suprema and Maxi Gala cultivars. Variables were evaluated to determine tree vigor, as well as yield and fruit quality characteristics. It was observed that 'G.213' promoted higher vigor for both cultivars as well as the Bi-Axis training system in the Maxi Gala cultivar. The highest yield was observed in Tall Spindle as well as the rootstock 'G.213' in both cultivars. The most significant changes on fruit quality occurred in the Bi-Axis training system, presenting higher intensity of red color as well as 'Maxi Gala' grafted on 'G.213' rootstock.

A utilização de novos porta-enxertos e diferentes sistemas de condução tem sido estudado para aumentar a produtividade, qualidade dos frutos e reduzir custos. Outro fator importante é a validação de estudos em condições de replantio, devido à crescente limitação de novas áreas para o plantio comercial de pomares de macieiras no Brasil. O presente estudo teve como objetivo avaliar o comportamento de macieiras 'Fuji Suprema' e 'Maxi Gala' em dois sistemas de condução enxertados sobre dois porta-enxertos. O estudo ocorreu durante as safras 2016/17, 2017/18 e 2018/19, em um pomar localizado no sul do Brasil, em área previamente cultivada com macieiras. Utilizou-se delineamento de blocos casualizados, consistindo em um fatorial 2x2, sendo Tall Spindle e Bi-Axis como sistemas de condução e 'M.9' e 'G.213' como porta-enxertos das cultivares Fuji Suprema e Maxi Gala. Foram avaliadas variáveis para determinar o comportamento vegetativo, bem como as características de produtividade e qualidade dos frutos. Observou-se que 'G.213' promoveu maior vigor para ambas cultivares, assim como o sistema de condução Bi-Axis na cultivar Maxi Gala. O maior rendimento foi observado nas macieiras conduzidas em Tall Spindle e enxertadas sobre 'G.213' em ambas cultivares. As mudanças mais significativas na qualidade dos frutos ocorreram nas macieiras conduzidas em Bi-Axis, apresentando maior intensidade de coloração vermelha, assim como na macieira 'Maxi Gala' enxertada sobre 'G.213'.

Morphological, sensory and biochemical characteristics of summer apple genotypes / Características morfológicas, sensoriais e bioquímicas de genótipos de maçã cultivada no verão

Summer apples are one of the most important plant community in Artvin province located Northeastern part of Turkey. In present study 22 local apple genotypes were characterized by phenological, morphological, biochemical and sensory properties. Harvest date was the main phenological data. Morphological measurements included fruit weight, fruit shape, fruit ground color, fruit over color, fruit over color coverage and fruit firmness, respectively. Sensory measurements were as juiciness and aroma and biochemical characteristics included organic acids, SSC (Soluble Solid Content), vitamin C, total phenolic content and antioxidant capacity. Genotypes exhibited variable harvest dates ranging from 11 July to 13 August and cv. Summered harvested 30 July 2017. The majority of genotypes were harvested before cv. Summered. Fruit weight were also quite variable among genotypes which found to be between 89 g and 132 g, and most of the genotypes had bigger fruits than cv. Summered. Pink, red, yellow and green fruit skin color was evident and main fruit shape were determined as round, conic and oblate among genotypes. ART08-9, ART08-4, ART08-21 and ART08-22 had distinct bigger fruits and ART08-1, ART08-2, ART08-5, ART08-12 and ART08-17 had higher total phenolic content and antioxidant capacity. The results of the study showed significant differences for most of the phenological, morphological, sensory and biochemical characteristics. Thus, the phonological, morphological, sensory and biochemical characteristics of summer apple genotypes were distinguishable and these results suggest that phonological, morphological, sensory and biochemical differences of the summer apple genotypes can be attributed to differences in genetic background of genotypes which placed different groups by PCoA analysis.

As maçãs cultivadas no verão são uma das culturas vegetais mais importantes da província de Artvin, localizada no nordeste da Turquia. No presente estudo, 22 genótipos locais de maçã foram caracterizados quanto às suas propriedades morfológicas, bioquímicas e sensoriais. As características analisadas foram peso do fruto, data da colheita, forma do fruto, coloração da casca, firmeza do fruto, suculência, aroma, teor de ácidos orgânicos e de sólidos solúveis, vitamina C, teor fenólico total e capacidade antioxidante. Os genótipos exibiram datas de colheita que variaram de 11 de julho a 13 de agosto, e a cultivar de verão foi colhida em 30 de julho 2017. A maioria dos genótipos foi colhida antes da cultivar de verão. O peso dos frutos também foi bastante variável entre os genótipos, apresentando entre 89 e 132 g, e a maioria dos genótipos apresentou frutos maiores que a cultivar de verão. As colorações rosa, vermelho, amarelo e verde da casca dos frutos foram as mais evidentes, e a forma principal dos frutos foi determinada como redonda, cônica e oblata entre os genótipos. ART08-9, ART08-4, ART08-21 e ART08-22apresentaram frutos maiores distintos, e ART08-1, ART08-2, ART08-5, ART08-12 e ART08-17 apresentaram maior conteúdo fenólico total e capacidade antioxidante. Os resultados do estudo mostraram diferenças significativas para a maioria das características morfológicas, sensoriais e bioquímicas. Assim, essas características dos genótipos da maçã cultivadas no verão foram distintos, e esses resultados podem ser atribuídos a diferenças no contexto genético dos genótipos.

MxRop1-MxrbohD1 interaction mediates ROS signaling in response to iron deficiency in the woody plant Malus xiaojinensis.

Iron (Fe) deficiency affects crop production and quality. Rho of plants (ROPs) involves in multiple physiological processes in plants. While it has not been well characterized under Fe deficiency, especially in perennial woody plants. In our study, we cloned ROP homologous gene MxRop1 from Malus xiaojinenesis, then overexpressed it in Arabidopsis, showing enhanced plant tolerance to Fe deficiency, which demonstrated its gene function during this stress. Overexpression of MxRop1 also increased reactive oxygen species (ROS) levels. Moreover, active state of MxRop1 (CA-MxRop1) interacted with N-terminal region of MxrbohD1, one ROS synthesis gene. When MxrbohD1 was overexpressed in apple calli, it showed significantly increased H2O2 content, fresh weight and FCR activity, while ROS inhibitor application dramatically inhibited FCR activity, demonstrating ROS produced by MxrbohD1 regulated Fe deficiency responses. Furthermore, using Agrobacterium rhizogenes transformation, MxrbohD1 was overexpressed in apple roots, with increased expression of Fe deficiency-induced genes and increased root FCR activity. Under Fe deficiency, it exhibited slight leaf yellowing phenotype. Co-expression of CA-MxRop1 and MxrbohD1 significantly induced ROS generation. Finally, we proposed that MxRop1 interacted with MxrbohD1 to modulate ROS mediated Fe deficiency adaptive responses in Malus xiaojinensis, which will provide a guidance of cultivation of Fe-deficiency tolerant apple plant.

Transcriptomics of Improved Fruit Retention by Hexanal in 'Honeycrisp' Reveals Hormonal Crosstalk and Reduced Cell Wall Degradation in the Fruit Abscission Zone.

Apples (Malus domestica Borkh) are prone to preharvest fruit drop, which is more pronounced in 'Honeycrisp'. Hexanal is known to improve fruit retention in several economically important crops. The effects of hexanal on the fruit retention of 'Honeycrisp' apples were assessed using physiological, biochemical, and transcriptomic approaches. Fruit retention and fruit firmness were significantly improved by hexanal, while sugars and fresh weight did not show a significant change in response to hexanal treatment. At commercial maturity, abscisic acid and melatonin levels were significantly lower in the treated fruit abscission zone (FAZ) compared to control. At this stage, a total of 726 differentially expressed genes (DEGs) were identified between treated and control FAZ. Functional classification of the DEGs showed that hexanal downregulated ethylene biosynthesis genes, such as S-adenosylmethionine synthase (SAM2) and 1-aminocyclopropane-1-carboxylic acid oxidases (ACO3, ACO4, and ACO4-like), while it upregulated the receptor genes ETR2 and ERS1. Genes related to ABA biosynthesis (FDPS and CLE25) were also downregulated. On the contrary, key genes involved in gibberellic acid biosynthesis (GA20OX-like and KO) were upregulated. Further, hexanal downregulated the expression of genes related to cell wall degrading enzymes, such as polygalacturonase (PG1), glucanases (endo-ß-1,4-glucanase), and expansins (EXPA1-like, EXPA6, EXPA8, EXPA10-like, EXPA16-like). Our findings reveal that hexanal reduced the sensitivity of FAZ cells to ethylene and ABA. Simultaneously, hexanal maintained the cell wall integrity of FAZ cells by regulating genes involved in cell wall modifications. Thus, delayed fruit abscission by hexanal is most likely achieved by minimizing ABA through an ethylene-dependent mechanism.

An HD-ZIP transcription factor, MxHB13, integrates auxin-regulated and juvenility-determined control of adventitious rooting in Malus xiaojinensis.

Adventitious root (AR) formation is a critical factor in the vegetative propagation of forestry and horticultural plants. Competence for AR formation declines in many species during the miR156/SPL-mediated vegetative phase change. Auxin also plays a regulatory role in AR formation. In apple rootstock, both high miR156 expression and exogenous auxin application are prerequisites for AR formation. However, the mechanism by which the miR156/SPL module interacts with auxin in controlling AR formation is unclear. In this paper, leafy cuttings of juvenile (Mx-J) and adult (Mx-A) phase Malus xiaojinensis were used in an RNA-sequencing experiment. The results revealed that numerous genes involved in phytohormone signaling, carbohydrate metabolism, cell dedifferentiation, and reactivation were downregulated in Mx-A cuttings in response to indole butyric acid treatment. Among the differentially expressed genes, an HD-ZIP transcription factor gene, MxHB13, was found to be under negative regulation of MdSPL26 by directly binding to MxHB13 promoter. MxTIFY9 interacts with MxSPL26 and may play a role in co-repressing the expression of MxHB13. The expression of MxTIFY9 was induced by exogenous indole butyric acid. MxHB13 binds to the promoter of MxABCB19-2 and positively affects the expression. A model is proposed in which MxHB13 links juvenility-limited and auxin-limited AR recalcitrance mechanisms in Mx-A.

Molecular mechanism of MdWUS2-MdTCP12 interaction in mediating cytokinin signaling to control axillary bud outgrowth.

Shoot branching is an important factor that influences the architecture of apple trees and cytokinin is known to promote axillary bud outgrowth. The cultivar 'Fuji', which is grown on ~75% of the apple-producing area in China, exhibits poor natural branching. The TEOSINTE BRANCHED1/CYCLOIDEA/PCF (TCP) family genes BRANCHED1/2 (BRC1/2) are involved in integrating diverse factors that function locally to inhibit shoot branching; however, the molecular mechanism underlying the cytokinin-mediated promotion of branching that involves the repression of BRC1/2 remains unclear. In this study, we found that apple WUSCHEL2 (MdWUS2), which interacts with the co-repressor TOPLESS-RELATED9 (MdTPR9), is activated by cytokinin and regulates branching by inhibiting the activity of MdTCP12 (a BRC2 homolog). Overexpressing MdWUS2 in Arabidopsis or Nicotiana benthamiana resulted in enhanced branching. Overexpression of MdTCP12 inhibited axillary bud outgrowth in Arabidopsis, indicating that it contributes to the regulation of branching. In addition, we found that MdWUS2 interacted with MdTCP12 in vivo and in vitro and suppressed the ability of MdTCP12 to activate the transcription of its target gene, HOMEOBOX PROTEIN 53b (MdHB53b). Our results therefore suggest that MdWUS2 is involved in the cytokinin-mediated inhibition of MdTCP12 that controls bud outgrowth, and hence provide new insights into the regulation of shoot branching by cytokinin.

Mdm-MIR393b-mediated adventitious root formation by targeted regulation of MdTIR1A expression and weakened sensitivity to auxin in apple rootstock.

Adventitious root (AR) formation is of great significance for apple rootstock breeding. It is widely accepted that miR393 influences AR formation in many plant species; however, the molecular mechanism by which factors regulate AR formation remains insufficient. In this study, the evolutionary relationship of mdm-miR393 and candidate target genes MdTIR1/AFB was systematically identified, and the expression patterns were analysed. Multisequence alignment analysis of miR393 family members suggests that miR393 conservatively evolved between different species. The evolutionary relationship of the TIR1/AFBs can be divided into G1, G2 and G3 subgroups. During AR formation, the expression level of mdm-miR393a/b/c was significantly upregulated at 1 d and 7 d by exogenous auxin treatment. Furthermore, the expression levels of MdTIR1A, MdTIR1D, MdAFB1, MdAFB2, MdAFB3, MdAFB4 and MdAFB8 also appeared to be significantly changed by exogenous auxin induction. Subsequently, tissue-specific expression analysis showed that the expression levels of mdm-miR393 and MdTIR1/AFBs in different tissues exhibited significant differences. The promoter of mdm-miR393 contains multiple elements that respond to ABA, adversity and light signals; auxin treatment can activate the mdm-MIR393b promoter but is obviously inhibited by NPA treatment. The targeting relationship between mdm-MIR393b and MdTIR1A was verified by expression patterns, degradation group data, transient tobacco conversion results, and genes functions experiments. Heterologous overexpression of mdm-MIR393b (35S::mdm-MIR393b) decreased the number of ARs in the phenotype and reduced the expression level of the target gene NtTIR1 in tobacco. Compared to the wild type, the 35S::mdm-MIR393b transgenic plants demonstrated insensitivity to auxin. Furthermore, tir1 mutant exhibited reduced root system structure relative to the control. The above results illustrated that mdm-MIR393b is involved in mediating AR formation by targeted regulation of MdTIR1A expression in apple rootstock.

Infuence of kaolin application on most important fruit and leaf characteristics of two apple cultivars under sustained defcit irrigation

BACKGROUND: Apple is one of the oldest and most valuable fruits. Water restriction is one of the major problems in the production of this fruit in some planting areas. METHODS: Effects of kaolin spray treatments were studied on two early apple cultivars of Golab and Shafi-Abadi under sustained deficit irrigation (SDI) in Alborz province, Iran during 2017 and 2018. Irrigation treatments were 100%, 85%, and 70% ETc and kaolin application were concentrations of 0, 3 and 6% in 2017 and 0, 1.5 and 3% in 2018. RESULTS: Results showed that 85% ETc treatment compared to other irrigation treatments improved apple tree crown volume in 2017. Deficit irrigation treatments significantly reduced fruit weight in both years. Application with 6% kaolin resulted in 33.3% increase in apple fruit weight compared to non-kaolin treatment at 100% ETc irrigation in the first year. Severe deficit irrigation (70% ETc) significantly reduced apple fruit length in both years, but 6% kaolin increased fruit length in both apple cultivars in 2017. Severe deficit irrigation treatment increased the firmness of apple fruit compared to control and mild deficit irrigation (85% ETc) in the first year of experiment. There was no significant difference between irrigation treatments for apple fruit firmness in the second year of experiment. Kaolin treatments of 1.5% and 3% at full irrigation increased the soluble solids content of apple fruit by 36.6% and 44.1% in 2018, respectively. Deficit irrigation treatments significantly increased leaf proline content compared to control in both years. In the first year, kaolin treatments increased leaf proline but in the second year, leaf proline was not significant. Deficit irrigation treatment of 70% ETc and 6% kaolin had the highest amount of glycine betaine content, malondialdehyde and hydrogen peroxide in apple leaf in the first year of experiment. CONCLUSIONS: Severe deficit irrigation stress (70% ETc) increased the activity of nonenzymatic defense systems of apple trees. Kaolin as a drought stress reducing agent can be recommended in apple orchards of Golab and Shafi-Abadi cultivars as an effective and inexpensive method to improve tolerance to drought stress conditions.

ERF4 affects fruit firmness through TPL4 by reducing ethylene production.

The firmness of fleshy fruit crops has a significant effect on their quality, consumer preference, shelf life and transportability. In a combined quantitative trait locus and genome-wide association studies study of apple fruit texture, we identified a mutation (C-G) in the ethylene response factor-associated amphiphilic repression (EAR) motif in the coding region of the apple ETHYLENE RESPONSE FACTOR4 (ERF4) gene. Chromatin immunoprecipitation sequencing showed that ERF4 binds to the promoter of ERF3, which is involved in regulation of ethylene biosynthesis. The EAR mutation in ERF4 results in reduced repression of ERF3 expression, which is turn promotes ethylene production and loss of fruit firmness. ERF4 acts as a transcriptional repressor whose activity is modulated by a TOPLESS co-repressor 4 (TPL4)-binding EAR repression motif. Biolayer interferometry analysis showed that the mutation in the EAR motif causes a reduction in the interaction with TPL4. Suppression of ERF4 or TPL4 promoted fruit ripening and ethylene production. Taken together, our results provide insights into how ERF4 allelic variation underlies an important fruit quality trait.

Application of in vivo solid phase microextraction (SPME) in capturing metabolome of apple (Malus ×domestica Borkh.) fruit.

An in vivo direct-immersion SPME sampling coupled to comprehensive two-dimensional gas chromatography - time-of-flight mass spectrometry (GCxGC-ToFMS) was employed to capture real-time changes in the metabolome of 'Honeycrisp' apples during ripening on the tree. This novel sampling approach was successful in acquiring a broad metabolic fingerprint, capturing unique metabolites and detecting changes in metabolic profiles associated with fruit maturation. Several metabolites and chemical classes, including volatile esters, phenylpropanoid metabolites, 1-octen-3-ol, hexanal, and (2E,4E)-2,4-hexadienal were found to be up-regulated in response to fruit maturation. For the first time, Amaryllidaceae alkaloids, metabolites with important biological activities, including anti-cancer, anti-viral, anti-parasitic, and acetylcholinesterase (AChE) inhibitory activity, were detected in apples. Considering the elimination of oxidative degradation mechanisms that adversely impact the representativeness of metabolome obtained ex vivo, and further evidence that lipoxygenase (LOX) pathway contributes to volatile production in intact fruit, in vivo DI-SPME represents an attractive approach for global plant metabolite studies.

Serological and Molecular Detection of Latent Viruses in the Apple Germplasm Bank of Santa Catarina

Abstract The Apple Germplasm Bank (AGB) of Santa Catarina Agricultural Research and Rural Extension Company - Epagri, AGB-Epagri, is the largest of the genus Malus in Brazil. Twenty-eight main accessions of this bank were virus screened through DAS-ELISA, RT-PCR and IC-RT-PCR during two consecutive reproductive cycles, and each accession showed latent mixed infection by at least two species, among ASGV, ASPV and ACLSV. The combined use of diagnostic methods helped overcome inconsistencies commonly found in apple virus detection and was shown essential for the AGB-Epagri can be safely used as a source of genetic variability and for the exchange of virus-free propagative material.

Ancient apple cultivars from Garfagnana (Tuscany, Italy): A potential source for 'nutrafruit' production.

Ancient apple cultivars are known for their organoleptic properties over a small geographic area, but little is known of their nutraceutical properties, which might be useful in large-scale breeding programmes. Nine ancient apple cultivars from Tuscany (Italy) were characterized for their organoleptic properties, phenolic profiles, and antioxidant activity. These cultivars had high polyphenol concentrations (principally flavanols and phenolic acids) and high total antioxidant capacity compared with most commercial apple cultivars. Fruits from the cultivars 'San Michele' and 'Del Debbio' showed a good compromise between fruit size and solid soluble content, and might be suitable for fresh consumption, while fruit from 'Benito', 'Della Piastra', 'Lugliese Grisanti', 'Del Sangue' and 'Ruggine' had high polyphenol contents and excellent antioxidant capacity, and may be suitable for breeding programmes. 'Ruggine' fruit could also be used for sweet juices with good nutraceutical properties due to their high soluble solid content and high flavanol concentration.

A comparison of the chemical composition and antioxidant activity of several new early- to mid-season apple cultivars for a warmer climate with traditional cultivars.

BACKGROUND: New early- to mid-season apple cultivars are being developed to help address warmer growing seasons due to climate change. Free sugars, organic acids, total phenolic content, total flavonoid content, antioxidant activity and phenolic composition were determined in the pulp and peel of six new and six traditional apple cultivars. In addition, the phenolic profiles of apple peels were characterized using high-resolution mass spectrometry. Forty-eight polyphenol compounds were identified, by accurate mass, in apple peel. RESULTS: Compared to Fuji apples, a new apple cultivar, Decobell, contained 2.6- and 1.4-fold higher levels of the sum of individual polyphenol levels in the peel and the pulp, respectively. Decobell apples showed similar sugar-to-acid ratio (0.27) to Fuji apples (0.25). CONCLUSIONS: The results indicate that the Decobell cultivar could have the best quality characteristics in terms of sugar-to-acid ratios and health-promoting activities due to the phenolic profiles. © 2019 Society of Chemical Industry.

Influence of the ethylene-related signal-inhibiting octapeptide NOP-1 on postharvest ripening and quality of 'Golden Delicious' apples.

BACKGROUND: Processes extending the shelf life of climacteric fruit play an important role in terms of a sustainable global food supply. In a previous study, a synthetic octapeptide (NOP-1) was shown to inhibit the interaction between ethylene receptor (ETR) and ethylene insensitive-2 (EIN2), and in consequence delay tomato ripening. We investigated for the first time the effect of NOP-1 on inhibiting the ripening of apples ('Golden Delicious') during postharvest. RESULTS: Using purified recombinant proteins from a bacterial expression system, we demonstrate here that EIN2 also interacts tightly (Kd = 136 ± 29 nmol L-1 ) with the corresponding apple ETR MdETR1. In line with previous binding studies on tomato ETRs, the ripening-delaying peptide NOP-1 clearly binds to the purified apple ETR. An NOP-1 solution (1000 µmol L-1 ) was applied with a brush or microdispenser and compared with apples treated with 1-methylcyclopropene (1-MCP) (SmartFresh™, Agrofresh) applied as gaseous treatment or untreated control fruits. NOP-1 inhibited colour development and chlorophyll degradation during shelf life. These effects were more pronounced with the brush application (surface film) than with microdroplets application (mimicking a sprayable formulation). NOP-1 did not alter ethylene release or respiration rate, whereas 1-MCP expectedly strongly suppressed both. There were no differences in quality parameters evaluated. CONCLUSION: Our study shows that NOP-1 binds to MdETR1 which results in delaying of ethylene-dependent ripening developments of skin colour and chlorophyll. Besides application methods, possible reasons for the weak effect of NOP-1 in comparison with previous tomato experiments could be different receptor affinity and penetration differences. © 2019 Society of Chemical Industry.

Development and reprotuction of Spodoptera eridania on natural hosts and artificial diet / Desenvolvimento e reprodução de Spodoptera eridania em hospedeiros naturais e dieta artificial

Abstract The development and reproductive potential of Spodoptera eridania (Stoll, 1782) (Lepidoptera, Noctuidae) was compared on natural and artificial diets. Its biology was determined under laboratory conditions, providing an artificial diet, leaves of white clover and the peach cultivar 'Chimarrita' and fruits and leaves of the apple cultivar 'Gala'. Larvae of S. eridania could not complete their biological cycle when fed on apple fruits or peach leaves. The artificial diet provided the shortest development time, with a cycle of 28.5±0.14 days from egg to adult and 37.3% total viability. Apple leaves led to a longer development time from egg to adult (62.8±1.22 days) and lower total viability (23%) and white clover provided the highest total viability (54.3%). The results showed that S. eridania can use white clover and apple leaves to develop normally in apple orchards. The reduction of food sources such as white clover used by the larvae results in infestation and damage to fruits from "test bites".

Resumo O desenvolvimento e o potencial reprodutivo de Spodoptera eridania (Stoll, 1782) (Lepidoptera, Noctuidae) foi comparado em dietas naturais e artificial. A biologia de S. eridania foi determinada em condições de laboratório, comparando-se uma dieta artificial com folhas de trevo branco e de pêssego da cultivar 'Chimarrita' e frutos e folhas de maçã da cultivar 'Gala'. As larvas de S. eridania não puderam completar seu ciclo biológico quando alimentadas com frutas de maçã ou folhas de pêssego. A dieta artificial proporcionou o menor tempo de desenvolvimento, com um ciclo completo de 28,5 ± 0,14 dias de ovo a adulto e 37,3% de viabilidade total. As folhas de maçã levaram maior tempo de desenvolvimento de ovo a adulto (62,8 ± 1,22 dias) e menor viabilidade total (23%). O trevo branco apresentou a maior viabilidade total (54,3%). Os resultados mostraram que S. eridania pode usar trevo branco e folhas de maçã para se desenvolver normalmente em pomares de maçã. A redução de fontes alimentares, como o trevo branco usado pelas larvas, resulta em infestação e danos aos frutos através das "mordidas de prova".

Effects of deficit irrigation and kaolin application on vegetative growth and fruit traits of two early ripening apple cultivars

BACKGROUND: Drought is one of the main serious problems for agriculture production which its intensity is increasing in many parts of the world, hence, improving water use efficiency is a main goal for sustainable agriculture. RESULTS: Growth indices including relative shoot length growth (SL), relative stem diameter increase (SD) and relative trunk cross sectional area growth (TCSA) measured at the start and end of the season decreased by reducing the irrigation level. Chlorophyll index (CI) was decreased at 70% crop evapotranspiration, however water use efficiency (WUE), leaf and fruit total phenolic content (TPC), and fruit anthocyanin content (AC) were among the traits that showed increment by water deficit stress in both cultivars. Shafi-Abadi cultivar showed to be more sensitive to the water stress than 'Golab' Kaolin treatment improved SL, SD and CI traits, but this increase was statistically significant only for SD at 5% level. Kaolin had no significant effect on yield and water use efficiency (WUE), however, it had negative effect on yield efficiency (YE). Kaolin treatments also significantly increased fruit and leaf TPC (P< 0.01) but had no effect on leaf and fruit total antioxidant activity (AA), as well as fruit anthocyanin content (AC) and soluble proteins (SP). CONCLUSIONS: Irrigation at 85% ETc showed better results than 100% and 70% ETc levels for yield attributes. It seems that the more pronounced effect of kaolin on vegetative traits but not on the fruits, might be attributed to the early ripening and harvest time of the examined cultivars.