Wayne W. Fish

The Influence of Rootstock Selection on Fruit Quality Attributes of Watermelon

Grafting watermelon (Citrullus lanatus) to control Fusarium wilt has been practiced in Europe, the Middle East, and the Far East for decades. Until recently, grafting watermelon has not been practiced in the United States due to labor costs and land availability. There is some disagreement in the literature as to the effects that grafting has on water- melon fruit quality. This study was designed to determine the effects of grafted watermelon on fruit firmness, lycopene content, and total soluble solids (TSS) using five different rootstocks. When using Cucurbita ficifolia or Cucurbita maxima x Cucurbita moschata hybrid as the rootstock, watermelon fruit consistently had higher fruit firmness values. Other C. maxima x C. moschata hybrids or Lagenaria siceraria rootstocks generally produced lower or more varied fruit firmness values. Grafting increased fruit firmness by as much as 25% in some cases, but field and year effects were ob- served. In addition, grafting had no effect on lycopene content or TSS. Furthermore, no off-flavors were detected in fruit from grafted plants, but there was a 5- to 7 day delay in fruit maturity compared to their non-grafted counterpart. Although environment can have a major influence on fruit quality attributes, rootstock selection may be equally important in achieving the desired outcome.

Watermelon juice: a promising feedstock supplement, diluent, and nitrogen supplement for ethanol biofuel production

BackgroundTwo economic factors make watermelon worthy of consideration as a feedstock for ethanol biofuel production. First, about 20% of each annual watermelon crop is left in the field because of surface blemishes or because they are misshapen; currently these are lost to growers as a source of revenue. Second, the neutraceutical value of lycopene and L-citrulline obtained from watermelon is at a threshold whereby watermelon could serve as starting material to extract and manufacture these products. Processing of watermelons to produce lycopene and L-citrulline, yields a waste stream of watermelon juice at the rate of over 500 L/t of watermelons. Since watermelon juice contains 7 to 10% (w/v) directly fermentable sugars and 15 to 35 μmol/ml of free amino acids, its potential as feedstock, diluent, and nitrogen supplement was investigated in fermentations to produce bioethanol.ResultsComplete watermelon juice and that which did not contain the chromoplasts (lycopene), but did contain free amino acids, were readily fermentable as the sole feedstock or as diluent, feedstock supplement, and nitrogen supplement to granulated sugar or molasses. A minimum level of ~400 mg N/L (~15 μmol/ml amino nitrogen) in watermelon juice was required to achieve maximal fermentation rates when it was employed as the sole nitrogen source for the fermentation. Fermentation at pH 5 produced the highest rate of fermentation for the yeast system that was employed. Utilizing watermelon juice as diluent, supplemental feedstock, and nitrogen source for fermentation of processed sugar or molasses allowed complete fermentation of up to 25% (w/v) sugar concentration at pH 3 (0.41 to 0.46 g ethanol per g sugar) or up to 35% (w/v) sugar concentration at pH 5 with a conversion to 0.36 to 0.41 g ethanol per g sugar.ConclusionAlthough watermelon juice would have to be concentrated 2.5- to 3-fold to serve as the sole feedstock for ethanol biofuel production, the results of this investigation indicate that watermelon juice, either as whole juice fermented on-site or as a waste stream from neutraceutical production, could easily integrate with other more concentrated feedstocks where it could serve as diluent, supplemental feedstock, and nitrogen supplement.

The Purification, Physical/Chemical Characterization, and cDNA Sequence of Cantaloupe Fruit Polygalacturonase-Inhibiting Protein

ABSTRACT Polygalacturonase-inhibiting proteins (PGIPs) are believed to aid in plant defense against fungal pathogens by inhibiting polygalacturonases (PGs) secreted by the invading organism. In an effort to better understand this type of plant-pathogen interaction in cucurbits, we have isolated a cantaloupe PGIP (CmPGIP) from 5 to 15 day postanthesis cantaloupe fruit. CmPGIP inhibited crude extracts of PG from two of four fungal pathogens of cantaloupe that were tested. Results from assays for PG activity that utilized rate of substrate viscosity reduction or rate of reducing group formation were consistent with CmPGIP inhibition of endo-PG activity. The M(r) of CmPGIP by sedimentation equilibrium or MALDITOF MS was 38,500. The pI of CmPGIP was approximately 8.2, and its absorptivity at 280 nm was 0.93 ml/mg. The circular dichroism spectrum of native CmPGIP exhibited strong negative ellipticity in the near UV and possessed a far UV spectrum indicative of beta-sheet periodic structure. Amino acid sequences of the N terminus and a cyanogen bromide peptide were used to construct oligonucleotide primers for polymerase chain reaction sequencing. The sequenced open reading frame predicts a mature protein of 307 amino acids with up to 68% identity to other PGIP molecules. Northern blot analysis revealed differential expression during fruit development. The isolation and structural information obtained for CmPGIP by this investigation provide a foundation for the development of molecular strategies for pre- and postharvest crop protection.

Interaction of Sodium Dodecyl Sulfate with Watermelon Chromoplasts and Examination of the Organization of Lycopene within the Chromoplasts

The properties of plant-derived precipitates of watermelon lycopene were examined in aqueous sodium dodecyl sulfate (SDS) as part of an ongoing effort to develop simpler, more economical ways to quantify carotenoids in melon fruit. Levels of SDS >0.2% were found to increase the water solubility of lycopene in the state in which it was isolated from watermelon. Electron microscopy and chemical analyses suggested that the watermelon lycopene as isolated is packaged inside a membrane to form a chromoplast. Spectral peaks in the visible region of the watermelon chromoplasts in SDS exhibited a bathochromic shift from those in organic solvent. Watermelon chromoplasts in SDS exhibited pronounced circular dichroic activity in the visible region. Binding measurements indicated that about 120 molecules of SDS were bound per molecule of lycopene inside the chromoplast; likely, the detergent molecules are bound to the chromoplast membrane. Around 80% of the chromoplast-SDS complexes were retained on a 0.45 mum membrane filter. Together, these observations are consistent with lycopene in a J-type chiral arrangement inside a membrane to form a chromoplast. The binding of SDS molecules to the chromoplast membrane form a complex that is extensively more water-soluble than the chromoplast alone.

Polygalacturonase-inhibiting protein activity in cantaloupe fruit as a function of fruit maturation and tissue origin

Netted cantaloupe (Cucumis melo var. cantalupensis cv. Magnum 45) were harvested from 5 to 35 days postanthesis. The fruit of each age group were divided into exocarp, outer mesocarp, mid mesocarp, inner mesocarp, placenta, and seed. Each tissue was extracted and assayed for polygalacturonase-inhibiting protein (PGIP) activity against polygalacturonases (PGs) from three fungal pathogens of cantaloupe fruit. The PGIP activity of all tissues except placenta was high from the flower stage through the first week of fruit development but decreased markedly between 5 and 10 days postanthesis. PGIP activity against Phomopsis cucurbitae PG remained high and nearly constant in placental tissue throughout fruit development. However in this same tissue, PGIP activity against Fusarium solani PG decreased during fruit development to about 25% of its level in the 5-day-old fruit. This differential change in PGIP activity toward the two PGs suggests that different forms of the inhibitor are expressed between early and late stages of cantaloupe fruit development. The results also illustrate the importance of using multiple pathogen enzyme systems that can provide an opportunity for more accurate elucidation of mechanisms involved in the host–pathogen interaction.

Modeling the Inhibitor Activity and Relative Binding Affinities in Enzyme-Inhibitor-Protein Systems: Application to Developmental Regulation in a PG-PGIP System

Within a number of classes of hydrolytic enzymes are certain enzymes whose activity is modulated by a specific inhibitor‐protein that binds to the enzyme and forms an inactive complex. One unit of a specific inhibitor‐protein activity is often defined as the amount necessary to inhibit one unit of its target enzyme by 50 %. No objective quantitative means is available to determine this point of 50 % inhibition in crude systems such as those encountered during purification. Two models were derived: the first model is based on an irreversible binding approximation, and the second, or equilibrium, model is based on reversible binding. The two models were validated using the inhibition data for the polygalacturonase‐polygalacturonase‐inhibiting protein (PG‐PGIP) system. Theory and experimental results indicate that the first model can be used for inhibitor protein activity determination and the second model can be used for inhibitor protein activity determination as well as for comparison of association constants among enzymes and their inhibitor‐proteins from multiple sources. The models were used to identify and further clarify the nature of a differential regulation of expression of polygalacturonase‐inhibiting protein in developing cantaloupe fruit. These are the first relations that provide for an objective and quantitative determination of inhibitor‐protein activity in both pure and crude systems. Application of these models should prove valuable in gaining insights into regulatory mechanisms and enzyme‐inhibitor‐protein interactions.