Detection of Invisible Damages in ‘Rojo Brillante’ Persimmon Fruit at Different Stages Using Hyperspectral Imaging and Chemometrics

The main cause of flesh browning in ‘Rojo Brillante’ persimmon fruit is mechanical damage caused during harvesting and packing. Innovation and research on nondestructive techniques to detect this phenomenon in the packing lines are necessary because this type of alteration is often only seen when the final consumer peels the fruit. In this work, we have studied the application of hyperspectral imaging in the range of 450–1040 nm to detect mechanical damage without any external symptoms. The fruit was damaged in a controlled manner. Later, images were acquired before and at 0, 1, 2 and 3 days after damage induction. First, the spectral data captured from the images were analysed through an algorithm based on principal component analysis (PCA). The aim was to automatically separate intact and damaged fruit, and to detect the damage in the PC images when present. With this algorithm, 90.0% of intact fruit and 90.8% of damaged fruit were correctly detected. A model based on partial least squares—discriminant analysis (PLS-DA), was later calibrated using the mean spectrum of the pixels detected as damaged, to determine the moment when the fruit was damaged. The model differentiated fruit corresponding correctly to 0, 1, 2 and 3 days after damage induction, achieving a total accuracy of 99.4%.

A High-throughput Color Measurement System for Evaluating Flesh Browning in Apples

The development of new high-quality apple (Malus ×domestica) cultivars that are resistant to flesh browning is needed to expand the use of apples in the food service and catering industry. However, conventional methods for evaluating apple flesh browning can be both time-consuming and costly, thereby rendering such methods unsuitable for breeding programs that must characterize a large number of product samples. Therefore, it is necessary to develop new, simple, and inexpensive methods. The aim was to develop a method for simultaneously measuring the color values of 42 apple samples using a digital camera. The processing time per sample was reduced to less than one-tenth of that of the conventional method. The measurement dispersion [sd of the color difference between two colors ] of this system was less than 0.08, equivalent to the nominal value of a general colorimeter. Time-series analysis of six apple cultivars using this method showed that the calculated browning index values correlated well with the degree of browning judged by human perception. Further, the measurement data showed that the CIE L* a* b* value trends associated with browning in reddish- and watercored-flesh samples, was different from the corresponding trends in yellowish-flesh samples. This work reports the development of a high-throughput analytical system of apple browning and provides cautionary notes for evaluating reddish- and watercored-flesh browning, which should be measured on a different basis from that used for normal-flesh browning.

Simultaneous CRISPR/Cas9 Editing of Three PPO Genes Reduces Fruit Flesh Browning in Solanum melongena L.

Polyphenol oxidases (PPOs) catalyze the oxidization of polyphenols, which in turn causes the browning of the eggplant berry flesh after cutting. This has a negative impact on fruit quality for both industrial transformation and fresh consumption. Ten PPO genes (named SmelPPO1-10) were identified in eggplant thanks to the recent availability of a high-quality genome sequence. A CRISPR/Cas9-based mutagenesis approach was applied to knock-out three target PPO genes (SmelPPO4, SmelPPO5, and SmelPPO6), which showed high transcript levels in the fruit after cutting. An optimized transformation protocol for eggplant cotyledons was used to obtain plants in which Cas9 is directed to a conserved region shared by the three PPO genes. The successful editing of the SmelPPO4, SmelPPO5, and SmelPPO6 loci of in vitro regenerated plantlets was confirmed by Illumina deep sequencing of amplicons of the target sites. Besides, deep sequencing of amplicons of the potential off-target loci identified in silico proved the absence of detectable non-specific mutations. The induced mutations were stably inherited in the T1 and T2 progeny and were associated with a reduced PPO activity and browning of the berry flesh after cutting. Our results provide the first example of the use of the CRISPR/Cas9 system in eggplant for biotechnological applications and open the way to the development of eggplant genotypes with low flesh browning which maintain a high polyphenol content in the berries.

Browning of Early and Late-Harvested ‘Empire’ Apples Affected by Cold Storage and 1-MCP

The effects of harvest time, 1-methylcyclopropene (1-MCP) and air storage time on the susceptibility of flesh browning in ‘Empire’ apples were studied during a seven-day shelf life period after air storage at 0.5 °C for seven months. Early- or late-harvested ‘Empire’ apples without 1-MCP increased production of ethylene, respiration rates and internal ethylene concentration during the shelf life. Respiration rates increased in the late-harvested fruit for the shelf life period with/without 1-MCP treatment. The 1-MCP-treated fruit was approximately 10 N firmer than fruit not treated with 1-MCP at an early harvest for the entire shelf life duration. Peroxidase activity and percent change in electrical conductivity in the flesh were elevated in late-harvested fruit for the shelf life duration, whereas polyphenol oxidase activities were found to be stimulated by 1-MCP treatment, regardless of harvest time. Late-harvested fruit treated with 1-MCP exhibited increased susceptibility to flesh browning during shelf life, mostly due to reduction of the antioxidant defense mechanism of the fruits to stress in extending storage life, increasing polyphenol oxidase (PPO) activity and electrolyte leakage rate.