Different stages of disease detection in squash plant based on machine learning.

To increase agriculture production, accurate and fast detection of plant disease is required. Expert advice is needed to detect disease in plants, nutrition deficiencies or any other abnormalities caused by extreme weather conditions. But this process is very tedious, costly, and takes more time. In this paper, hyperspectral imaging and machine learning were used to detect different stages (early, middle, and critical stage) of the powderly mildew disease (PMD) in squash plants. An unmanned aerial vehicle (UAV) was used to collect the data from the field and Locality Preserving Discriminative Broad Learning (LPDBL) was used to distinguish the diseased and healthy plants. In addition, the ability to detect the diseased plant by the proposed method was evaluated using 10 different spectral vegetation indices (VIs). The results show the proposed method detected the disease accurately in the early, middle, and critical stages of the squash plant. The proposed method's performance is compared with six different PMDs under indoor laboratory test and UAV-based field test conditions. The comparison's results show that the LPDBL provides better accuracy in detecting disease in the squash plant.

Biostimulant Substances for Sustainable Agriculture: Origin, Operating Mechanisms and Effects on Cucurbits, Leafy Greens, and Nightshade Vegetables Species.

Climate change is a pressing matter of anthropogenic nature to which agriculture contributes by abusing production inputs such as inorganic fertilizers and fertigation water, thus degrading land and water sources. Moreover, as the increase in the demand of food in 2050 is estimated to be 25 to 70% more than what is currently produced today, a sustainable intensification of agriculture is needed. Biostimulant substances are products that the EU states work by promoting growth, resistance to plant abiotic stress, and increasing produce quality, and may be a valid strategy to enhance sustainable agricultural practice. Presented in this review is a comprehensive look at the scientific literature regarding the widely used and EU-sanctioned biostimulant substances categories of silicon, seaweed extracts, protein hydrolysates, and humic substances. Starting from their origin, the modulation of plants' hormonal networks, physiology, and stress defense systems, their in vivo effects are discussed on some of the most prominent vegetable species of the popular plant groupings of cucurbits, leafy greens, and nightshades. The review concludes by identifying several research areas relevant to biostimulant substances to exploit and enhance the biostimulant action of these substances and signaling molecules in horticulture.

Identification and mapping of CpPM10.1, a major gene involved in powdery mildew (race 2 France of Podosphaera xanthii) resistance in zucchini (Cucurbita pepo L.).

KEY MESSAGE: Powdery mildew resistance in zucchini is controlled by one major dominant locus, CpPM10.1. CpPM10.1 was fine mapped. The expression of candidate gene Cp4.1LG10g02780 in resistant individuals was significantly upregulated after inoculation with the powdery mildew. Powdery mildew (PM) is one of the most destructive fungal diseases, reducing the productivity of Cucurbita crops globally. PM influences the photosynthesis, growth and development of infected zucchini and seriously reduces fruit yield and quality. In the present study, the zucchini inbred line 'X10' had highly stable PM resistance, and the inbred line 'Jin234' was highly susceptible to PM in the seedling stage and adult stages. Genetic analysis revealed that PM resistance in 'X10' is controlled by one major dominant locus. Based on the strategy of QTL-seq combined with linkage analysis and developed molecular markers, the major locus was found to be located in a 382.9-kb candidate region on chromosome 10; therefore, the major locus was named CpPM10.1. Using 1,400 F2 individuals derived from a cross between 'X10' and 'JIN234' and F2:3 offspring of the recombinants, the CpPM10.1 locus was defined in a region of approximately 20.9 kb that contained 5 coding genes. Among them, Cp4.1LG10g02780 contained a conserved domain (RPW8), which controls resistance to a broad range of PM pathogens. Cp4.1LG10g02780 also had nonsynonymous SNPs between the resistant 'X10' and susceptible 'Jin234.' Furthermore, the expression of Cp4.1LG10g02780 was strongly positively involved in PM resistance in the key period of inoculation. Further allelic diversity analysis in zucchini germplasm resources indicated that PM resistance was associated with two SNPs in the Cp4.1LG10g02780 RPW8 domain. This study not only provides highly stable PM resistance gene resources for cucurbit crops but also lays the foundation for the functional analysis of PM resistance and resistance breeding in zucchini.

Analysis of the molecular and biological variability of Zucchini yellow mosaic virus isolates from Iran and Iraq.

During the growing season of 2018, several field-grown cucurbit plants in different parts of Iraq and Iran were surveyed for the presence of zucchini yellow mosaic virus (ZYMV), using two degenerate primer pairs (CIF/Rev and NIb2F/3R) targeting the two separated partial regions of the potyvirus genome (CI and NIb respectively). 7 out of 20 samples were confirmed to be infected with ZYMV. Phylogenetic analyses based on the CI gene grouped all Iranian and two Iraqi (ZYMV1 and ZYMV2) isolates together with isolates from the Middle East in the subgroup (AI), whereas the other Iraqi (ZYMV3 and ZYMV4) isolates were clustered in the subgroup (DI), which was only consisted of American isolates. The highest and lowest identity between the studied isolates and the GenBank isolates showed that the two genes (CI, NIb) of each isolate particularly the Iraqi isolates were more similar to a specific and geographically scattered mosaic of worldwide isolates, suggestive of mixed infection might have occurred between different worldwide isolates in Iraq. Furthermore, the first complete nucleotide sequence of an Iraqi ZYMV (ZYMV-Iq) isolate was done, using the Illumina sequencing technique. The complete nucleotide sequence of ZYMV-Iq isolate was 9650 nt, excluding the 3'poly (A) tail. ZYMV-Iq isolate shared the highest nt identity of 98.8% with an American (KC665630) isolate. Phylogenetic analysis based on the full genome sequence placed ZYMV-Iq in subgroup A of group I alongside 18 isolates from the US and two isolates from Australia. In addition, recombination analysis detected lone significant recombination between ZYMV-Iq and South Korean (AY279000) isolate. Moreover, the results showed that symptom intensity was varied across experimental host plants.

Fine mapping identified the gibberellin 2-oxidase gene CpDw leading to a dwarf phenotype in squash (Cucurbita pepo L.).

Dwarfism is an important agronomic trait in pumpkin that can increase yield. In this study, the dwarf Cucurbita pepo L. line X10 exhibited significantly longitudinally shorter cell length in the stem than did the normal-vine line JIN234. The dwarf stature of X10 was recovered with exogenous gibberellin (GA3) application, suggesting that X10 might be sensitive to GA biosynthesis. Genetic analysis revealed that this dwarf trait is controlled by a single completely dominant locus: CpDw (Cucurbita pepo L. Dwarf). Using 1,300 F2 individuals derived from a cross between X10 and JIN234, we mapped the CpDw locus to a region of approximately 24.6 kb on chromosome 10 that contain 5 annotated genes. The high expression level of Cp4.1LG10g05910.1 and high GA2ox enzyme activity in X10 revealed that the GA 2-oxidase gene Cp4.1LG10g05910.1 is a candidate gene for CpDw. Alignment of the Cp4.1LG10g05910.1 gene revealed two nonsynonymous single nucleotide polymorphism (SNP) mutations in the two exons, as well as several SNPs and InDels in the important functional elements of promoter between parental lines. Further allelic diversity analysis of the Cucurbita spp. germplasm resources indicated that Cp4.1LG10g05910.1 may be involved in vine growth during the early developmental stage in C. pepo but not in C. maxima or C. moschata. This study provides an important theoretical basis for the genetic regulation of vine length and crop breeding in pumpkin.

Does scion-rootstock compatibility modulate photoassimilate and hormone trafficking through the graft junction in melon-pumpkin graft combinations?

The effect of the rootstock on the acropetal and basipetal transport of photoassimilates and hormones was studied in the 'Kiran' (Ki) melon cultivar grafted onto pumpkin rootstocks with different degrees of compatibility. A complementary experiment was performed to compare the incompatible combination (as evidenced by plant collapse at the fruit ripening stage), designated Ki/r53, with self-grafted r53/r53 as a model compatible combination. Both experiments showed the accumulation of a number of amino acids, sugars, and sugar alcohols in the scion of the incompatible Ki/r53 grafts. Additionally, they showed a marked reduction of trans-zeatin-type cytokinins and an elevated content of cis-zeatin-type cytokinins in the rootstock, and the opposite pattern in the scion, hinting at the possible involvement of a hormonal signal for graft compatibility. There was no direct evidence of a blockage at the graft union, since hormone acropetal and basipetal trafficking was demonstrated for all combinations. Dye uptake experiments did not show xylem flow impairment. A possibly significant finding in the incompatible combination was the deposition of undifferentiated cells in the hollow space that replaces the pith region in melon and pumpkin. The link between the above findings and the collapse of the plants of the incompatible combination remains unclear.

Source:sink imbalance detected with leaf- and canopy-level spectroscopy in a field-grown crop.

The finely tuned balance between sources and sinks determines plant resource partitioning and regulates growth and development. Understanding and measuring metabolic indicators of source or sink limitation forms a vital part of global efforts to increase crop yield for future food security. We measured metabolic profiles of Cucurbita pepo (zucchini) grown in the field under carbon sink limitation and control conditions. We demonstrate that these profiles can be measured non-destructively using hyperspectral reflectance at both leaf and canopy scales. Total non-structural carbohydrates (TNC) increased 82% in sink-limited plants; leaf mass per unit area (LMA) increased 38% and free amino acids increased 22%. Partial least-squares regression (PLSR) models link these measured functional traits with reflectance data, enabling high-throughput estimation of traits comprising the sink limitation response. Leaf- and canopy-scale models for TNC had R2 values of 0.93 and 0.64 and %RMSE of 13 and 38%, respectively. For LMA, R2 values were 0.91 and 0.60 and %RMSE 7 and 14%; for free amino acids, R2 was 0.53 and 0.21 with %RMSE 20 and 26%. Remote sensing can enable accurate, rapid detection of sink limitation in the field at the leaf and canopy scale, greatly expanding our ability to understand and measure metabolic responses to stress.

Characterization of Starch in Cucurbita moschata Germplasms throughout Fruit Development.

Pumpkins (Cucurbita moschata; Cucurbitaceae) are the rich source of nutrients and valued for their biologically active substances to be used for the treatment of several diseases. The contents, composition, and conformation of starch are the significant quality traits of C. moschata. Two germplasms were targeted for analysis regarding the taste difference. Results indicated that the total starch contents and amylose/amylopectin ratio were high in CMO-X as compared to CMO-E during each fruit development stage. Scanning electron microscopy and transmission electron microscopy observations revealed that smooth surface starch granules fused together to enhance the starch accumulation. For a comparison of fruit development in CMO-E and CMO-X, the putative pathway for starch metabolism was developed and homologs were identified for each key gene involved in the pathway. GBSS and SBE were correlated with the difference in the amylose/amylopectin ratio of CMO-E and CMO-X. Conclusively, the developmental regulation of genes associated with starch accumulation can be considered as an important factor for the determination of fruit quality.

Potential of endophytic fungi collected from Cucurbita pepo roots grown under three different agricultural mulches as antagonistic endophytes to Verticillium dahliae in western Washington.

Verticillium dahliae is a significant pathogen in cucurbit cropping systems for which there are limited control options outside of soil fumigation. Endophytes, fungi and bacteria that live within plant hosts without impacting the host negatively, have exhibited antagonism to V. dahliae. The objectives of this study were to survey potential V. dahliae-antagonistic endophytes from roots of 'Cinnamon Girl' pumpkin (Cucurbita pepo) grown under either polyethylene (PE), an experimental polylactic acid/ poly(hydroxalkanoate) (PLA/PHA) mulch, Weed Guard Plus, or no mulch, as well as from 'Sugar Baby' watermelon (Citrullus lanatus), and 'Tetsukabuto' squash (C. maxima x C. moschata). Four selected endophytes were screened for antagonism against V. dahliae in the laboratory, greenhouse, and field. A total of 777 isolates of potential fungal endophytes were recovered from pumpkin, watermelon, and squash roots between 2015 and 2016 of which 198 isolates were identified down to the genus level. Of those isolates, frequency of isolation was greatest for Dichotomopilus/Chaetomium spp. (5%), Cladosporium spp. (15.2 %), Clonostachys spp. (5.6 %), Epicoccum spp. (22.2 %), and Fusarium spp. (24.7 %). All five genera only weakly associated with roots grown under a particular mulch treatment (Cramer's V = 0.22) or cucurbit host (Cramer's V = 0.1925). In a laboratory culture plate assay, V. dahliae isolate JAW-113 was plated against one of four prospective endophytes (Dichotomopilus sp., Epicoccum sp., Microdochium sp., or Schizothecium sp.). The area under the Verticillium culture growth curve (AUVGC) was significantly highest (P < 0.0001) when V. dahliae was by Schizothecium sp. or Dichotomopilus sp. In a greenhouse study using a Mason jar assay with V. dahliae amended potting mix, pumpkin plant vigor, plant fresh weight, root fresh weight, and root dry weight were significantly higher (P < 0.05) for plants inoculated with Dichotomopilus sp., Epicoccum sp., Microdochium sp., and Schizothecium sp. compared to plants without endophyte inoculation. Subsequent field trials in 2017 and 2018 showed no significant differences in foliar disease severity or fruit yield, regardless of whether plants were inoculated with an endophyte or not. However, recovery of V. dahliae colony forming units from pumpkin stem sap was significantly lower (P < 0.0001) for plants inoculated with either Dichotomopilus sp. or Schizothecium sp. in 2017 or Dichotomopilus sp. in 2018.

Hypoglycemic effect and bioactive compounds associated with the ripening stages of the Cucurbita ficifolia Bouché fruit.

BACKGROUND: The fruit of Cucurbita ficifolia Bouché is known in Mexico as 'chilacayote'. The scientific interest that C. ficifolia Bouché has acquired is due to its important hypoglycemic effect. The present research aimed (i) to discover whether this hypoglycemic property is present at different stages of development of this fruit, and (ii) to characterize some bioactive compounds with antioxidant or anti-inflammatory properties. Ethylene production, respiration rate, and maturity indices were determined during fruit development. The chemical characterization of the aqueous extracts of each stage of maturity studied was determined and their hypoglycemic effects were bioassayed using groups of normal mice with diabetes induced by streptozotocin at a dose of 500 mg-1 kg-1 body weight. RESULTS: Respiration rate and ethylene production showed a typical pattern for non-climacteric fruit and the quality parameters did not show significant changes. Phenolic compounds such as gallic acid and chlorogenic acid were found to have the highest concentration at 15 days of development. Extracts at 15 days showed a hypoglycemic effect that was 11% greater than that of glibenclamide in diabetized mice. CONCLUSION: All stages of development of C. ficifolia fruit had a hypoglycemic effect; however, the aqueous extract from the fruit at 15 days of development showed a better effect than glibenclamide. This finding highlights the potential of this maturity stage, and shows that it is appropriate for inclusion in treatments of type 2 diabetes mellitus. The results also indicate that phenolic compounds are mainly responsible for this effect and not d-chiro-inositol as previously thought. © 2020 Society of Chemical Industry.

Evapotranspiration and crop coefficients of Italian zucchini cultivated with recycled paper as mulch.

Recycled paper has the potential to be used as a mulch for vegetable production and can be adopted for the cultivation of Italian zucchini. However, there have been no studies about the water savings or crop coefficient values used in irrigation management in this system; therefore, there is a need for more research. In view of the above, this study aimed to evaluate the effects of recycled paper mulch on evaporation and evapotranspiration in Italian zucchini and to determine the crop coefficients in its developmental stages. The study was carried out in two cultivation cycles conducted at the Lysimetric Station in Viçosa, MG, Brazil. The experiments were installed in a randomized block design with four replicates. Four lysimeter cultivation treatments were applied: without mulch (C); with recycled paper as mulch (CP); with only recycled paper (P); and with Bahia grass (G). Irrigation and drainage measurements were performed daily to calculate the crop and reference evapotranspiration, and thus the crop coefficient (Kc) values. The following characteristics were evaluated: fruit yield, NDVI and water productivity. For the cultivation of Italian zucchini using paper as mulch, Kc values of 0.54, 0.77 and 0.44 and Kcb values of 0.15, 0.45 and 0.18 are recommended for the initial, intermediate and final stages, respectively. NDVI can be used to estimate the Kc values for Italian zucchini. The use of recycled paper as mulch reduces the water consumption of Italian zucchini.

Early Holocene crop cultivation and landscape modification in Amazonia.

The onset of plant cultivation is one of the most important cultural transitions in human history1-4. Southwestern Amazonia has previously been proposed as an early centre of plant domestication, on the basis of molecular markers that show genetic similarities between domesticated plants and wild relatives4-6. However, the nature of the early human occupation of southwestern Amazonia, and the history of plant cultivation in this region, are poorly understood. Here we document the cultivation of squash (Cucurbita sp.) at about 10,250 calibrated years before present (cal. yr BP), manioc (Manihot sp.) at about 10,350 cal. yr BP and maize (Zea mays) at about 6,850 cal. yr BP, in the Llanos de Moxos (Bolivia). We show that, starting at around 10,850 cal. yr BP, inhabitants of this region began to create a landscape that ultimately comprised approximately 4,700 artificial forest islands within a treeless, seasonally flooded savannah. Our results confirm that the Llanos de Moxos is a hotspot for early plant cultivation and demonstrate that-ever since their arrival in Amazonia-humans have markedly altered the landscape, with lasting repercussions for habitat heterogeneity and species conservation.

Two androecious mutations reveal the crucial role of ethylene receptors in the initiation of female flower development in Cucurbita pepo.

Ethylene is the key regulator of sex determination in monoecious species of the family Cucurbitaceae. This hormone determines which individual floral meristems develop as female or male flowers and the female flowering transition. The sex determination genes discovered so far code for ethylene biosynthesis enzymes, but little is known about the importance of ethylene signaling components. In this paper we characterize two novel ethylene-insensitive mutations (etr1a-1 and etr1b) which block the female flowering transition of Cucurbita pepo; this makes plants produce male flowers indefinitely (androecy). Two missense mutations in the ethylene-binding domain of the ethylene receptors CpETR1A or CpETR1B were identified as the causal mutations of these phenotypes by using whole-genome resequencing. The distinctive phenotypes of single and double mutants for four etr mutations have demonstrated that the final level of ethylene insensitivity depends upon the strength and dosage of mutant alleles for at least three cooperating ETR genes, and that the level of ethylene insensitivity determines the final sex phenotype of the plant. The sex phenotype ranges from monoecy in ethylene-sensitive wild-type plants to androecy in the strongest ethylene-insensitive ones, via andromonoecy in partially ethylene-insensitive plants. The induction of female flowering transition was found to be associated with upregulation of CpACS11, CpACO2 and CpACS27, three ethylene biosynthesis genes required for female flower development. A model is proposed herein, integrating both ethylene biosynthesis and receptor genes into the genetic network which regulates sex determination in C. pepo.

Metabolic and transcriptomic analysis of two Cucurbita moschata germplasms throughout fruit development.

BACKGROUND: Pumpkins (Cucurbita moschata; Cucurbitaceae) are valued for their fruits and seeds and are rich in nutrients. Carotenoids and sugar contents, as main feature of pumpkin pulp, are used to determine the fruit quality. RESULTS: Two pumpkin germplasms, CMO-X and CMO-E, were analyzed regarding the essential quality traits such as dry weight, soluble solids, organic acids, carotenoids and sugar contents. For the comparison of fruit development in these two germplasms, fruit transcriptome was analyzed at 5 different developmental stages from 0 d to 40 d in a time course manner. Putative pathways for carotenoids biosynthesis and sucrose metabolism were developed in C. moschata fruit and homologs were identified for each key gene involved in the pathways. Gene expression data was found consistent with the accumulation of metabolites across developmental stages and also between two germplasms. PSY, PDS, ZEP, CRTISO and SUS, SPS, HK, FK were found highly correlated with the accumulation of carotenoids and sucrose metabolites, respectively, at different growth stages of C. moschata as shown by whole transcriptomic analysis. The results of qRT-PCR analysis further confirmed the association of these genes. CONCLUSION: Developmental regulation of the genes associated with the metabolite accumulation can be considered as an important factor for the determination of C. moschata fruit quality. This research will facilitate the investigation of metabolic profiles in other cultivars.

Identification of Fruit-Associated QTLs in Winter Squash (Cucurbita maxima Duchesne) Using Recombinant Inbred Lines.

Cucurbita maxima Duchesne squash and pumpkins are cultivated world-wide. Cucurbita maxima fruits are produced for fresh market and are valuable for food processing. Therefore, fruit characteristics and yield are the traits of high economic importance for breeders. To date, the genetic basis of fruit-associated traits in C. maxima have been poorly understood. In the present study, we evaluated fruit-associated traits and conducted quantitative trait locus (QTL) analysis using recombinant inbred lines (RILs) derived from a cross of two inbred lines with different fruit morphotypes. Phenotypic data for nine fruit traits (earliness, weight, number per plant, yield per plant, length and diameter, shape index, flesh thickness, sucrose content and dry matter content) were collected for RILs in two open-field experiments. Pairwise analysis of the phenotypic data revealed correlations among the fruit and yield-associated traits. Using a previously developed genetic map, we identified 26 QTLs for eight traits. The QTLs were found in 10 locations on eight chromosomes of C. maxima. The QTLs were detected across experiments and explained up to 41.4% of the observed phenotypic variations. Major-effect QTLs for multiple fruit-associated traits were clustered on chromosome 4, suggesting that this genomic region has been under selection during diversification and/or domestication of C. maxima.

Effect of elevated levels of CO2 on powdery mildew development in five cucurbit species.

The environment is the key factor that influences the host-parasite relationship. Elevated CO2 levels resulting from various anthropogenic sources may directly affect the surroundings around pathogens and plants. It is hypothesized that plants may respond differently to pathogens in the environment containing an elevated concentration of CO2. To test the hypothesis an experiment was conducted to examine the effects of intermittent exposures of elevated levels of CO2 viz., 400, 500 and 600 ppm (5 hr/day on alternate days) on the development of Sphaerotheca fuliginea causing powdery mildew disease on five cucurbits species using open-top chambers. The elevated levels of CO2 acted as a growth promoter and significantly enhanced the plant growth of all five cucurbit species. Inoculation with the fungus incited specific mildew symptoms on the leaves and decreased the plant growth and biomass production of the cucurbits tested except bitter gourd. The intermittent exposures with elevated levels of CO2 aggravated the disease development. As a result, severe mildew developed on all five cucurbits, including bitter gourd, which expressed tolerance to the pathogen. Fungus colonization in terms of the number of conidia/cm2 leaf surface was significantly greater on the plants exposed to 500 or 600 ppm CO2. The stomata and trichome density and stomatal pore width were increased in the leaves of CO2 exposed plants. The CO2 exposures also accelerated the photosynthesis rate, but transpiration, stomatal conductance, salicylic acid and total phenols were decreased; fungus inoculation caused the effects just reverse of CO2. Interaction between S. fuliginea and CO2 was found synergistic at 500 ppm, whereas with rest of the concentrations it was near to additive.

Reynoutria sachalinensis extract elicits SA-dependent defense responses in courgette genotypes against powdery mildew caused by Podosphaera xanthii.

Powdery mildew (PM) caused by Podosphaera xanthii is one of the most important courgette diseases with high yield losses and is currently controlled by fungicides and sulphur applications in conventional and organic production. Plant derived elicitors/inducers of resistance are natural compounds that induce resistance to pathogen attack and promote a faster and/or more robust activation of plant defense responses. Giant knotweed (Reynoutria sachalinensis, RS) extract is a known elicitor of plant defenses but its mode of action remains elusive. The aim of this study was to investigate the mechanisms of foliar RS applications and how these affect PM severity and crop performance when used alone or in combination with genetic resistance. RS foliar treatments significantly reduced conidial germination and PM severity on both an intermediate resistance (IR) and a susceptible (S) genotype. RS application triggered plant defense responses, which induced the formation of callose papillae, hydrogen peroxide accumulation and the Salicylic acid (SA) - dependent pathway. Increased SA production was detected along with increased p-coumaric and caffeic acid concentrations. These findings clearly indicate that RS elicits plant defenses notably as a consequence of SA pathway induction.

The ethylene receptors CpETR1A and CpETR2B cooperate in the control of sex determination in Cucurbita pepo.

High-throughput screening of an ethyl methanesulfonate-generated mutant collection of Cucurbita pepo using the ethylene triple-response test resulted in the identification of two semi-dominant ethylene-insensitive mutants: etr1a and etr2b. Both mutations altered sex determination mechanisms, promoting conversion of female into bisexual or hermaphrodite flowers, and monoecy into andromonoecy, thereby delaying the transition to female flowering and reducing the number of pistillate flowers per plant. The mutations also altered the growth rate and maturity of petals and carpels in pistillate flowers, lengthening the time required for flowers to reach anthesis, as well as stimulating the growth rate of ovaries and the parthenocarpic development of fruits. Whole-genome sequencing allowed identification of the causal mutation of the phenotypes as two missense mutations in the coding region of CpETR1A and CpETR2B, each one corresponding to one of the duplicates of ethylene receptor genes highly homologous to Arabidopsis ETR1 and ETR2. The phenotypes of homozygous and heterozygous single- and double-mutant plants indicated that the two ethylene receptors cooperate in the control of the ethylene response. The level of ethylene insensitivity, which was determined by the strength of each mutant allele and the dose of wild-type and mutant etr1a and etr2b alleles, correlated with the degree of phenotypic changes in the mutants.

Morphological, Transcriptomic and Hormonal Characterization of Trimonoecious and Subandroecious Pumpkin (Cucurbita maxima) Suggests Important Roles of Ethylene in Sex Expression.

: Sex expression is a complex process, and in-depth knowledge of its mechanism in pumpkin is important. In this study, young shoot apices at the one-true-leaf stage and 10-leaf stage in Cucurbita maxima trimonoecious line '2013-12' and subandroecious line '9-6' were collected as materials, and transcriptome sequencing was performed using an Illumina HiSeqTM 2000 System. 496 up-regulated genes and 375 down-regulated genes were identified between shoot apices containing mostly male flower buds and only female flower buds. Based on gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, the differentially expressed genes were mainly enriched in the ethylene and auxin synthesis and signal transduction pathways. In addition, shoot apices at the 4-leaf stage were treated with the ethylene-releasing agent 2-chloroethylphosphonic acid (Ethrel), aminoethoxyvinyl glycine (AVG), AgNO3 and indoleacetic acid (IAA). The number of female flowers up to node 20 on the main stem of '2013-12' increased significantly after Ethrel and IAA treatment and decreased significantly after AVG and AgNO3 treatment. The female flowers in '9-6' showed slight changes after treatment with the exogenous chemicals. The expression of key genes in ethylene synthesis and signal transduction (CmaACS7, CmaACO1, CmaETR1 and CmaEIN3) was determined using quantitative RT-PCR, and the expression of these four genes was positively correlated with the number of female flowers in '2013-12'. The variations in gene expression, especially that of CmaACS7, after chemical treatment were small in '9-6'. From stage 1 (S1) to stage 7 (S7) of flower development, the expression of CmaACS7 in the stamen was much lower than that in the ovary, stigma and style. These transcriptome data and chemical treatment results indicated that IAA might affect pumpkin sex expression by inducing CmaACS7 expression and indirectly affecting ethylene production, and the ethylene synthesis and signal transduction pathways play crucial roles in pumpkin flower sex expression. A possible reason for the differences in sex expression between pumpkin lines '2013-12' and '9-6' was proposed based on the key gene expression. Overall, these transcriptome data and chemical treatment results suggest important roles for ethylene in pumpkin sex expression.

Safety and quality issues in summer squashes using handheld portable NIRS sensors for real-time decision making and for on-vine monitoring.

BACKGROUND: Portable handheld near infrared spectroscopy (NIRS) instruments currently present enormous advantages in terms of size, weight, and robustness. They also provide fast, precise information that can be obtained in situ, and they represent a viable option for controlling vegetable safety and quality during the growth period. The aim of this research was to evaluate three handheld portable NIRS instruments for in situ and real-time analysis of intact summer squashes. Traditional methods were used to analyze 221 summer squashes, and this work was used to develop calibration models for morphological, safety, and quality parameters. The longitudinal distribution of nitrate content in summer squashes weighing over 400 g was also studied, and the evolution of this parameter during the harvest period was tracked to determine which summer squashes and which zones of the vegetables (peduncle, equatorial, or stylar) could be earmarked for baby-food production. RESULTS: The robustness of the calibration models confirmed the expectations raised by NIRS technology for morphological, safety, and quality control of individual summer squashes, and the models developed with the MicroNIR-1700 instrument were those that provided more accuracy and precision, being the peduncle zone the part with higher nitrate content. CONCLUSIONS: It is in the peduncle zone, therefore, where measurements of this parameter must be carried out to decide on the destination of the harvested product. Summer squashes picked at the end of the harvest are those that must be used for baby-food production. © 2019 Society of Chemical Industry.