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  • 1.

    【2hr】Inhibition of apricot polyphenol oxidase by combinations of plant proteases and ascorbic acid

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    机译 结合植物蛋白酶和抗坏血酸抑制杏多酚氧化酶
    摘要：The present research investigates the long term inhibition of enzymatic browning by inactivating the polyphenol oxidase (PPO) of apricot, using combinations of plant proteases and ascorbic acid (AA). The selected proteases were able to inactivate PPO at pH 4.5, with the degree of inactivation proportional to incubation time and protease concentration. Papain was the most effective protease, with 50 μg completely inactivating PPO in less than one hour. AA prevented browning reactions that occur before or during PPO inactivation by protease. The combinations of AA/proteases were highly effective in vitro, where 2 mM AA/500 μg proteases inhibited PPO activity completely over 24 h. The combination of AA/proteases was also effective in vivo, as treated apricot purees preserved their color (p < 0.0001, compared to untreated samples after 10 days of storage). The results demonstrate that AA/proteases combinations constitute a promising practical anti-browning method with feasible application in the food industry.
  • 2.

    【2hr】In vitro and in vivo antioxidant properties of bioactive compounds from green propolis obtained by ultrasound-assisted extraction

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    机译 超声辅助提取获得的绿色蜂胶中生物活性化合物的体外和体内抗氧化性能
    摘要：Green propolis presents a potential source of bioactive compounds, responsible for its antioxidant capacity. The effects of ethanol concentration, solid-solvent ratio, and extraction time were evaluated in regard to the total phenolic content (TPC) and antioxidant capacity of the extracts by the use of central composite rotatable designs. Optimum extraction conditions lead to significant reduction of extraction time compared to conventional extraction methods. Under optimum conditions, extracts were composed of 1614.80 mg GAE. g−1 and 807 mg artepillin C. g−1. Extracts were effective in retarding the oxidation in oil-in-water emulsions subjected to accelerated tests. Green propolis extracts (up to 200 mg.kg−1) did not increase Saccharomyces cerevisiae cell damage after 4 h of exposure, indicating its antioxidant effect and potential innocuity. Results demonstrated the antioxidant properties of the propolis extract was similar or better than the ones from synthetic antioxidants and warrant further investigation to determine its potential industrial application.
  • 3.

    【2hr】Monitoring of selective androgen receptor modulators in bovine muscle tissue by ultra-high performance liquid chromatography-tandem mass spectrometry

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    机译 超高效液相色谱-串联质谱法监测牛肌肉组织中选择性雄激素受体调节剂
    摘要：Selective androgen receptor modulators (SARMs) are non-steroidal compounds widely reported as drugs of abuse in human and animal sports, with potential for misuse as growth promoters in animal-based food production. In this study, a first analytical methodology to simultaneous screen for a panel of emerging SARMs in bovine muscle was developed, validated (CCβ values from 0.5–5 ng g−1), and applied to detect 15 structurally diverse compounds from nine SARM families. Muscle samples (200 mg) were homogenised in extraction solvent (MeCN:H2O, 4:1, v/v) before clean-up (end-capped C18 dSPE), defatting (n-hexane pre-saturated with MeCN partitioning) and concentration prior to UHPLC-MS/MS analysis. In the absence of incurred bovine muscle, method applicability was demonstrated by the analysis of rodent muscle tissue. The developed screening assay serves as a rapid, simple and cost-effective tool for surveillance monitoring of SARM abuse in livestock production systems as a pre-emptive measure ensuring food safety.
  • 4.

    【2hr】Alterations in physicochemical and functional properties of buckwheat straw insoluble dietary fiber by alkaline hydrogen peroxide treatment

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    机译 碱性过氧化氢处理荞麦秸秆不溶性膳食纤维的理化和功能性质变化
    摘要：To enhance the physicochemical and functional properties of insoluble dietary fiber (IDF) from buckwheat straw, we investigated the effects of alkaline hydrogen peroxide (AHP) treatment. Electron microscopy showed that the IDF had regular and compact tubes that turned into wrinkled lamellar products. After AHP treatment, X-ray diffraction indicated that the crystalline structure of the IDF was perturbed. And an undesirable decrease was observed in the content of hydroxybenzoic acid derivatives, hydroxycinnamic acid derivatives, flavonoids and the antioxidant capacity of IDF modified by AHP; however, the hydration properties (such as water holding capacity), α-amylase inhibition activity and glucose adsorption capacity of IDF were significantly enhanced by AHP. Furthermore, AHP led to a redistribution of monosaccharides in soluble dietary fiber and IDF, an interesting finding hinting at the mechanism and potential applications of AHP modification of IDF. In this study, AHP enhanced the physiological and functional properties of buckwheat straw IDF.
  • 5.

    【2hr】Ca2+ involved in GABA signal transduction for phenolics accumulation in germinated hulless barley under NaCl stress

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    机译 Ca2 +参与GABA信号转导，促进NaCl胁迫下萌发的大麦中酚类物质的积累
    摘要：Keywords: GABA, Ca2+, Phenolics, NaCl stress, Germinated hulless barley

    Abstract

    In this study, in order to investigate the role of Ca2+ in GABA signal transduction involved in phenolics accumulation in barley seedlings under NaCl stress, the seedlings were treated with exogenous GABA and its synthesis inhibitor, 3-mercaplopropionic acid (3-MP), as well as Ca2+ channel blockers La3+, Ca2+ chelator EGTA, and Ca2+ release channel inhibitor 2-aminoethoxydiphenyl borate (2-APB). The results showed that GABA significantly enhanced phenolics, calcium and calmodulin content. It also induced Ca2+ influx in barley root tips cells, and altered the distribution of Ca2+, making calcium precipitates more uniform and intensive. While, 3-MP treatment led to opposite changes, which suggested that GABA was essential for calcium content increase. In addition, accumulation of phenolics was inhibited by LaCl3, EGTA and 2-APB treatments, and this inhibition could be alleviated partly by exogenous GABA. Taken together, Ca2+ was involved in GABA signal transduction for phenolics accumulation in barley seedlings under NaCl stress.
  • 6.

    【2hr】Non-Saccharomyces yeast and lactic acid bacteria in Co-inoculated fermentations with two Saccharomyces cerevisiae yeast strains: A strategy to improve the phenolic content of Syrah wine

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    机译 与两种酿酒酵母酵母菌菌株共接种的发酵中的非酿酒酵母和乳酸菌：提高西拉葡萄酒中酚含量的策略
    摘要：Syrah must was co-inoculated with mixed cultures of Saccharomyces + O. oeni/Lb. plantarum and Saccharomyces + non-Saccharomyces + O. oeni/Lb. plantarum to evaluate the effect on phenolics and sensory attributes. Reference wines were produced by S. cerevisiae. Malvidin-3-O-glucoside, flavan-3-ols, flavonols and phenolic acids were quantified using a RP-HPLC technique. Physicochemical characteristics and sensory attributes were measured. Total acidity and alcohol in mixed co-inoculations were different from reference wines. The concentration of l-malic acid was 7-times less in mixed co-inoculations. Mixed co-inoculations had ca. 1.3-times more malvidin-3-O-glucoside and phenolic acids than reference wines. Flavan-3-ols and flavonols were not different between mixed co-inoculations and reference wines. Acidity and astringency were least in mixed co-inoculations. Mouthfeel and bitterness least in S. cerevisiae wines. Tasters preferred mixed co-inoculated wines. Mixed co-inoculation is a strategy to contemplate for Syrah vinification but the modalities of inoculation need further investigation. Success depends on a suitable combination of yeast/bacteria and consideration of strain variation.
  • 7.

    【2hr】Streamlined approach for careful and exhaustive aroma characterization of aged distilled liquors

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    机译 精简而详尽的陈年蒸馏酒香气表征方法
    摘要：Solvent-assisted flavor evaporation (SAFE) is considered to be the best overall method to produce a “clean” aroma extract to avoid the loss of labile aroma compounds or the formation of thermally generated artifacts during gas chromatographic (GC) analysis. However, SAFE is both time consuming and labor intensive, especially when applied repeatedly for quantitation by stable isotope dilution analysis (SIDA), which requires the addition of isotopes within specific mass ratio ranges relative to target analytes. The streamlined approach described herein allows for accurate quantitation of odor-active components in liquor products with a single SAFE operation. The quantitative results achieved by this method are nearly identical for most odor-active components, except for specific semi-volatile constituents not recovered well by SAFE (e.g., vanillin and syringaldehyde in oak-aged liquors). The streamlined approach provides a simple and convenient way to expedite the careful and exhaustive study of the flavor chemistry of aged liquors.
  • 8.

    【2hr】Variability of carotenoids in a Musa germplasm collection and implications for provitamin A biofortification

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    机译 穆萨种质资源中类胡萝卜素的变异性及其对维生素原A生物强化的影响
    摘要：Bananas are important staples in tropical and sub-tropical regions and their potential as a source of provitamin A has recently attracted attention for biofortification. A collection of 189 banana genotypes (AAB-plantains, M. acuminata cultivars and bred hybrids) was screened to determine variability in fruit pulp provitamin A carotenoid (pVAC) content using high performance liquid chromatography. Total carotenoid content in tested genotypes varied from 1.45 µg/g for hybrid 25447-S7 R2P8 to 36.21 µg/g for M. acuminata cultivar ITC.0601 Hung Tu with a mean of 8.00 µg/g fresh weight. Predominant carotenoids identified were α-carotene (38.67%), trans-β-carotene (22.08%), lutein (22.08%), 13-cis-β-carotene (14.45%) and 9-cis-β-carotene (2.92%), indicating that about 78% of the carotenoids in bananas are pVAC. High pVAC genotypes were identified for integration into biofortification strategies to combat vitamin A deficiency in developing countries.
  • 9.

    【2hr】Detection and quantification of adulterations in aged wine using RGB digital images combined with multivariate chemometric techniques

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    机译 使用RGB数字图像结合多元化学计量学技术对陈年葡萄酒中的掺假进行检测和定量
    摘要：A method has been developed to authenticate aged high-quality wines and to quantify their potential adulterations through multivariate analysis and regression techniques applied to the obtained RGB digital images. Wines of pure Gran Reserva, Crianza, and Joven Rioja as well as synthetic adulterated Gran Reserva samples were studied. Digital images were obtained by a single and inexpensive lab-made device. Each sample was characterized by means of the 256 channels intensities from the RGB-colorgram. Multivariate image analysis revealed differences among the wine classes, and between genuine-aged and adulterated samples. Partial least squares regression was used to develop a model for estimating the adulteration degree of Gran Reserva wines. The model achieved good prediction (RMSEP = 1.6), appropriate precision (RSD = 2.5%) and suitable LOD (2.3%) to quantify cost-effective adulterations. The present method, due to simplicity and low cost, could provide an appropriate alternative to the traditional chemical authentication methods.
  • 10.

    【2hr】Comparative transcriptome analysis reveals fruit discoloration mechanisms in postharvest strawberries in response to high ambient temperature

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    机译 比较转录组分析揭示了采后草莓对高温环境的变色机理
    摘要：The aim of this study was to examine both physiological and molecular evidences related to fruit discoloration in postharvest strawberries under high ambient temperature. The results showed the total anthocyanin and their main components in the strawberries under 35 °C were greatly increased due to the significant up-regulations of anthocyanin biosynthetic genes and transportation genes. High ambient temperature greatly improved the activities of peroxidase (POD) and enhanced gene expressions of POD3, POD6 and POD63. At the same time, high storage temperature activated laccase genes expression including laccase-9 and laccase-14, which was closely related to anthocyanin degradation. Levels of reactive oxygen species (ROS) metabolism were also increased under high ambient temperature at transcript levels. Therefore, we concluded that high ambient temperature could enhance anthocyanin accumulation and degradation at the same time, which maybe the main reasons for the fruits discoloration of postharvest strawberries under high ambient temperature.
  • 11.

    【2hr】Composition and content of phenolic acids and avenanthramides in commercial oat products: Are oats an important polyphenol source for consumers?

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    机译 商业燕麦产品中酚酸和avenanthramides的组成和含量：燕麦对消费者而言是重要的多酚来源吗？
    摘要：Oats contain a range of phenolic acids and avenanthramides which may have health benefits. Analysis of 22 commercial oat products (oat bran concentrate, oat bran, flaked oats, rolled oats and oatcakes) using HPLC-DAD detected eleven bound and thirteen free + conjugated phenolic acids and avenanthramides. The oat products (excluding concentrate) provided between 15.79 and 25.05 mg total phenolic acids (9.9–19.33 mg bound, 4.96–5.72 mg free + conjugated) and between 1.1 and 2 mg of avenanthramides in a 40 g portion while an 11 g portion of oat concentrate provided 16.7 mg of total phenolic acids (15.17 mg bound, 1.53 mg free + conjugated) and 1.2 mg of avenanthramides. The compositions and concentrations of the components in the different products were broadly similar, with the major component being ferulic acid (58–78.1%). The results show that commercial oat products are a source of phenolic acids and avenanthramides for consumers.
  • 12.

    【2hr】Understanding the influence of processing conditions on the extraction of rhamnogalacturonan-I “hairy” pectin from sugar beet pulp

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    机译 了解加工条件对甜菜粕中鼠李糖半乳糖醛酸-I“毛”果胶的提取影响
    摘要：Sugar beet pectin is rich in rhamnogalacturonan-I (RG-I) region, which is a potential source of prebiotics. RG-I pectin cannot be extracted the same way as commercial homogalacturan-rich pectin using hot acid. Therefore, this study has explored several alternative methods, including microwave-assisted extraction (MAE) and conventional-solvent extraction (CSE) at atmospheric pressure using different solvents, and microwave-assisted hydrothermal extraction (MAHE) under pressure using water. No conclusive differences in microwave and conventional heating were found with heating rate controlled. The optimum treatment times of both MAE and CSE at 90 °C atmospheric pressure and regardless of the solvents used were 120 min; however, MAHE at 130 °C under pressure can dramatically reduce the time to 10 min. Alcohol-insoluble solids (AIS) extracted using pH13 solvent by MAE had both the highest RG-I yield at 25.3% and purity at 260.2 mg/g AIS, followed by AIS extracts using water by MAHE with 7.5% and 166.7 mg/g AIS respectively.
  • 13.

    【2hr】Effect of cold plasma on polyphenol oxidase inactivation in cloudy apple juice and on the quality parameters of the juice during storage

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    机译 冷血浆对阴天苹果汁中多酚氧化酶失活的影响及其对贮藏过程中果汁质量参数的影响
    摘要：Keywords: Cloudy apple juice, Cold plasma, Spark discharge, PPO inactivation, Quality parameters, Shelf life study

    Abstract

    Direct cold plasma treatment has been investigated as an alternative non-thermal technology as a means of maintaining and improving quality of fresh cloudy apple juice. Process variables studied included type of plasma discharge, input voltage and treatment time on polyphenol oxidase (PPO) inactivation. Spark discharge plasma at 10.5 kV for 5 min was the best treatment, with near total inactivation of PPO achieved, although good PPO inactivation was also recorded using shorter treatment times. Residual activity (RA) of PPO was 16 and 27.6% after 5 and 4 min of treatment respectively. This PPO inactivation was maintained throughout the storage trials, but decreased with samples treated for a shorter time. Plasma treatment improved key quality parameters of Golden delicious cloudy apple juice, with retention of critical quality parameters during extended storage trials. Color was the most noticeable change, which was enhanced with retention of a greener color. An increase of 69 and 64% was obtained in the total phenolic content after 4 and 5 min of treatment, respectively. Therefore, cold plasma was demonstrated to be a good alternative to traditional heat treatments for enhanced quality retention of fresh cloudy apple juice and over its storage.
  • 14.

    【2hr】Quality of lettuce Lactuca sativa (var. Tropicana M1) grown with two low-salinity shrimp effluents

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    机译 两种低盐虾废水生长的莴苣莴苣（变种Tropicana M1）的质量
    摘要：In a previous study, we investigated the use of shrimp effluents from well water (WW) and diluted seawater (DSW) (both with 2.7 dS m−1 electrical conductivity (EC)), and a hydroponic solution (HS) as the control treatment in greenhouse lettuce production. This new paper completes the previous one by focusing on the quality of lettuce harvested. Compared to the lettuce from the other treatments, WW-lettuce exhibited higher levels of phenolic compounds and a higher antioxidant capacity, mainly in the soluble fraction. The lettuce cultivated with DSW showed no significant difference in total phenolics and flavonoids with respect to the HS lettuce. These results reveal that the functional properties (antioxidant properties, polyphenols and flavonoid content) are even better in the lettuce produced with WW and DSW shrimp effluents. In contrast, agronomical properties (weight, number of leaves and yield) were found to be better in the case of lettuce grown with the hydroponic solution (control).
  • 15.

    【2hr】Effects of emulsifiers on the moisture sorption and crystallization of amorphous sucrose lyophiles

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    机译 乳化剂对无定形蔗糖冻干物水分吸收和结晶的影响
    摘要：The crystallization of amorphous sucrose can be problematic in food products. This study explored how emulsifiers (a range of sucrose esters, polysorbates, and soy lecithin) impact the moisture sorption and crystallization of amorphous sucrose lyophiles. Solutions containing sucrose with and without emulsifiers were lyophilized, stored in desiccators, and analyzed by X-ray diffraction, infrared spectroscopy, and polarized light microscopy over time. Moisture sorption techniques, Karl Fischer titration, and differential scanning calorimetry were also used. Different emulsifiers had varying impacts on sucrose crystallization tendencies. Polysorbates enhanced sucrose crystallization, decreasing both the RH and time at which sucrose crystallized. These lyophiles did not collapse upon crystallization, unlike all other samples, indicating the likelihood of variations in nucleation sites and crystal growth. All other emulsifiers stabilized amorphous sucrose by up to a factor of 7x, even in the presence of increased water absorbed and independent of glass transition temperatures, indicating emulsifier structure governed sucrose crystallization tendencies.
  • 16.

    【2hr】Arsenic speciation analysis in rice milk using LC-ICP-MS

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    机译 使用LC-ICP-MS分析米浆中的砷形态
    摘要：The consumption of rice milk has increased, mainly by individuals intolerant to lactose or allergic to cow milk. However, rice milk contains As. In this sense, the concentration of As in rice milk should be controlled. In the present study it is proposed a methodology for determination of As(III), dimethylarsenic (DMA), monomethylarsenic (MMA) and As(V) species in rice milk using LC-ICP-MS. The main features of the methodology are fast analysis, easy and simple sample preparation, where the sample is 3-fold diluted in the mobile phase and then filtered. The four arsenic species investigated were detected in the analysed samples, being As(V) the main species. The limit of quantification of the method ranges from 0.25 to 0.43 μg L−1 As. The analyte recovery ranged from 81 to 116% for samples spiked to 1.00 μg L−1 or 5.00 μg L−1 As and the relative standard deviation was better than 5%.
  • 17.

    【2hr】Application of two-dimensional gel electrophoresis technique for protein profiling of Indian black gram varieties and detection of adulteration in black gram-based food products using comparative proteomics

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    机译 二维凝胶电泳技术在印度黑克品种蛋白质谱分析中的应用以及使用比较蛋白质组学检测黑克类食品中的掺假
    摘要：In the present study, protein fingerprints from six novel Indian black gram varieties were obtained using 2D-GE. The results revealed the presence of thirteen well-resolved protein spots in all six varieties. Analysis of the fingerprints using PD Quest™ revealed differential protein expression. In addition, six proteins were found to be uniquely expressed in varieties PDKV Black Gold and TAU-1. Further, analysis of 2D patterns of black gram and potential adulterants such as refined wheat flour and white pea using PD Quest™ revealed the presence of proteins with MW 15.0 kDa, pI 4.89 (refined flour) and MW 21.5 kDa, pI 5.70 (white pea), which can be considered as biomarkers for their presence in black gram food products. The method was sensitive enough to detect adulteration at 5% level and could successfully detect the potential presence of refined flour in one of the black gram papad samples analysed during the study.
  • 18.

    【2hr】Physicochemical, functional and morphological characterization of starches isolated from three native potatoes of the Andean region

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    机译 从安第斯地区的三个原生马铃薯中分离的淀粉的理化，功能和形态特征
    摘要：Three varieties of native potato (Imilla blanca, Imilla negra and Loc’ka) that grow in the Andean region at more than 3800 m.a.s.l. were selected fot the extraction and characterization or their starch. Instrumental techniques such as scanning electron microsocopic (SEM), differential scanning calorimetry (DSC), Fourier transformed infrarred spectroscopy (FTIR), X-ray diffraction, colorimetry and polarized light microscopy were used. The results showed that only Loc’kás starch had a unimodal granule size distribution, whereas Imilla negra and Imilla blanca starches showed two and three granule size populations, respectively. The starch from Imilla negra showed higher apparent amylose content, peak viscosity, phosphorous content and paste clarity. The starch from Imilla blanca showed high relative crystallinity, while Imilla blanca and Imilla negra had higher intensity ratios than that from Loc’ka, suggesting high molecular order. Cooked starch from Imilla negra showed higher resistant starch (RS) fraction than the other starches studied.
  • 19.

    【2hr】Physico-elemental analysis of roasted organic coffee beans from Ethiopia, Colombia, Honduras, and Mexico using X-ray micro-computed tomography and external beam particle induced X-ray emission

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    机译 使用X射线微计算机断层扫描和外部束粒子诱导的X射线发射，对来自埃塞俄比亚，哥伦比亚，洪都拉斯和墨西哥的烘焙有机咖啡豆进行物理元素分析
    摘要：Abbreviations: PIXE, particle induced X-ray emission; LDA, linear discriminant analysis; micro-CT, X-ray micro-computed tomographyKeywords: Ethiopia, Colombia, Honduras, External beam PIXE, Mexico, Organic, Roasted coffee bean, X-ray micro-computed tomography

    Abstract

    The physico-elemental profiles of commercially attained and roasted organic coffee beans from Ethiopia, Colombia, Honduras, and Mexico were compared using light microscopy, X-ray micro-computed tomography, and external beam particle induced X-ray emission. External beam PIXE analysis detected P, S, Cl, K, Ca, Ti, Mn, Fe, Cu, Zn, Br, Rb, and Sr in samples. Linear discriminant analysis showed that there was no strong association between elemental data and production region, whilst a heatmap combined with hierarchical clustering showed that soil-plant physico-chemical properties may influence regional elemental signatures. Physical trait data showed that Mexican coffee beans weighed significantly more than beans from other regions, whilst Honduras beans had the highest width. X-ray micro-computed tomography qualitative data showed heterogeneous microstructural features within and between beans representing different regions. In conclusion, such multi-dimensional analysis may present a promising tool in assessing the nutritional content and qualitative characteristics of food products such as coffee.
  • 20.

    【2hr】Comprehensive characterization of hydroxycinnamoyl derivatives in green and roasted coffee beans: A new group of methyl hydroxycinnamoyl quinate

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    机译 生咖啡豆和烘焙咖啡豆中的羟基肉桂酰基衍生物的全面表征：一组新的羟基肉桂肉桂酸甲酯
    摘要：Chemical compounds studied in this article: 3-O-p-Coumaroylquinic acid (PubChem CID: 9945785), 5-O-p-Coumaroylquinic acid (PubChem CID: 6441280), 4-O-p-Coumaroylquinic acid (PubChem CID: 5281766), Caffeic acid (PubChem CID: 689043), Ferulic acid (PubChem CID: 445858), 1-O-Caffeoylquinic acid (PubChem CID: 10155076), 5-O-Caffeoylquinic acid (PubChem CID: 1794427), 4-O-Caffeoylquinic acid (PubChem CID: 9798666), 5-O-Feruloylquinic acid (PubChem CID: 15901362), 3,4-Di-O-Caffeoylquinic acid (PubChem CID: 6474309), 3,5-Di-O-Caffeoylquinic acid (PubChem CID: 6474310), 4,5-Di-O-Caffeoylquinic acid (PubChem CID: 5281780), Caffeoyl-N-tryptophan (PubChem CID: 15228042)Keywords: Green and roasted coffee beans, Methyl hydroxycinnamoyl quinates, UPLC-DAD-QToF-MS

    Abstract

    The aim of this study was to quantitatively characterize 19 green and roasted coffee beans by ultra-performance liquid chromatography coupled with diode array detector and quadrupole time-of-flight mass spectrometry. A total of 57 phenolic acids including nine methyl ester of mono-, di-caffeoylquinic acid, and feruloylquinic acid were identified. The methyl hydroxycinnamoyl quinates are reported for the first time from Coffea arabica and Coffea robusta. The total phenolic content ranged from 5628 ± 227 to 8581 ± 109 mg/100 g dry weight (DW) in green, and from 791 ± 63 to 1891 ± 37 mg/100 g DW roasted beans. The methyl caffeoylquinates accounted for 2.1% of the total phenolic acids. The result suggested that the phenolic composition was affected by the type of species, cultivars, and roasting process. Hence, to retain the balance between health beneficial phenolics and sensory attributes, optimization of roasting condition specific to the cultivar type substantially required.
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