Polyphenol oxidase (PPO) plays a key role in the postharvest pericarp browning of litchi fruit, but its underlying mechanism remains unclear. In this study, we cloned the litchi PPO gene (LcPPO, JF926153), and described its expression patterns. The LcPPO cDNA sequence was 2120 bps in length with an open reading frame (ORF) of 1800 bps. The ORF encoded a polypeptide with 599 amino acid residues, sharing high similarities with other plant PPO. The DNA sequence of the ORF contained a 215-bp intron. After carrying out quantitative RT-PCR, we proved that the LcPPO expression was tissue-specific, exhibiting the highest level in the flower and leaf. In the pericarp of newly-harvested litchi fruits, the LcPPO expression level was relatively high compared with developing fruits. Regardless of the litchi cultivar and treatment conditions, the LcPPO expression level and the PPO activity in pericarp of postharvest fruits exhibited the similar variations. When the fruits were stored at room temperature without packaging, all the pericarp browning index, PPO activity and the LcPPO expression level of litchi pericarps were reaching the highest in Nandaowuhe (the most rapid browning cultivar), but the lowest in Ziniangxi (the slowest browning cultivar) within 2 d postharvest. Preserving the fruits of Feizixiao in 0.2-μm plastic bag at room temperature would decrease the rate of pericarp water loss, delay the pericarp browning, and also cause the reduction of the pericarp PPO activity and LcPPO expression level within 3 d postharvest. In addition, postharvest storage of Feizixiao fruit stored at 4°C delayed the pericarp browning while decreasing the pericarp PPO activity and LcPPO expression level within 2 d after harvest. Thus, we concluded that the up-regulation of LcPPO expression in pericarp at early stage of postharvest storage likely enhanced the PPO activity and further accelerated the postharvest pericarp browning of litchi fruit.

Litchi downy blight, caused by Peronophythora litchii, is one of the major diseases of litchi and has caused severe economic losses. P. litchii has the unique ability to produce downy mildew like sporangiophores under artificial culture. The pathogen had been placed in a new family Peronophytophthoraceae by some authors. In this study, the whole transcriptome of P. litchii from mycelia, sporangia, and zoospores was sequenced for the first time. A set of 23637 transcripts with an average length of 1284 bp was assembled. Using six open reading frame (ORF) predictors, 19267 representative ORFs were identified and were annotated by searching against several public databases. There were 4666 conserved gene families and various sets of lineage-specific genes among P. litchii and other four closely related oomycetes. In silico analyses revealed 490 pathogen-related proteins including 128 RXLR and 22 CRN effector candidates. Based on the phylogenetic analysis of 164 single copy orthologs from 22 species, it is validated that P. litchii is in the genus Phytophthora. Our work provides valuable data to elucidate the pathogenicity basis and ascertain the taxonomic status of P. litchii.

In the post-harvest phase, fruit is inexorably subjected to extrinsic stressors that expedite energy expenditure and truncate the storage lifespan. The present study endeavors to elucidate the response strategies of litchi to the alterations of energy state caused by 2,4-Dinitrophenol (DNP) treatment through energy metabolism and sugar metabolism. It was observed that the DNP treatment reduced the energy state of the fruit, exacerbated membrane damage and triggered rapid browning in the pericarp after 24 h of storage. Furthermore, the expression of genes germane to energy metabolism (LcAtpB, LcAOX1, LcUCP1, LcAAC1, and, LcSnRK2) reached their peak within the initial 24 h of storage, accompanied by an elevation in the respiratory rate, which effectively suppressed the rise in browning index of litchi pericarp. The study also posits that, to cope with the decrease of energy levels and membrane damage, litchi may augment the concentrations of fructose, glucose, inositol, galactose, and sorbose, thus safeguarding the canonical metabolic functions of the fruit. Collectively, these findings suggest that litchi can modulate energy and sugar metabolism to cope with fruit senescence under conditions of energy deficiency. This study significantly advances the understanding of the physiological responses exhibited by litchi fruit to post-harvest external stressors.

A simple efficient in vitro plant regeneration system was developed by indirect somatic embryogenesis of ‘Feizixiao’ litchi (Litchi chinensis Sonn.). Pollen in the anther of monocytes was used to induce callus. Two auxins (naphthalene acetic acid [NAA] and 2,4-dichloriphenoxyacetic acid [2,4-D]), and two cytokines (kinetin [KT] and 6-benzyladenine [BA]) were tested to explore their influence on callus induction. MS medium supplemented with 2.22 µM BA, 2.69 µM NAA, 13.57 µM 2,4-D, and 0.4 g/L LH (lactalbumin hydrolysate) showed the highest callus induction frequency. The callus obtained from anther was subcultured in MS medium containing 4.52 µM 2,4-D to obtain synchronized friable embryogenic callus. Different developmental stages of SEs were obtained from the callus on MS medium containing 6% (w/v) sucrose and different PGRs (plant growth regulators). On MS medium containing 6% (w/v) sucrose and supplemented with 0.54 µM NAA, 23.23 µM KT, 0.4 g/L LH, 0.56 µM inositol, and 10% (w/v) CW (coconut water), a higher number of SEs (globular, heart, torpedo and cotyledonary embryos) was achieved than on other media. Plantlets were established onto half-strength MS medium containing 1.44 µM GA3 (gibberellic acid) followed by successful acclimatization in the greenhouse. With flow cytometry and chromosome counting, ploidy analysis of regenerated plants revealed that the regenerated plantlets were all diploid. This study is the first report on somatic embryogenesis of ‘Feizixiao litchi’, providing an opportunity to improve the cultivar by biotechnology methods. Key words: litchi (Litchi chinensis Sonn.), anther culture, callus, regeneration, somatic embryogenesis.

Litchi downy blight, a destructive litchi disease caused by Peronophythora litchii, is controlled by intensive fungicide applying. Sources of resistance are used in conventional breeding approaches, but the mechanism is not well understood. Follow-up six years investigation, ‘Guiwei’ and ‘Heiye’ displayed stable susceptible and resistant against to P. litchii, respectively. After 72 hour inoculation, ‘Heiye’ showed few disease spots, while ‘Guiwei’ appeared brown and covered with white sporangia. Germination of sporangia and growth of mycelium in ‘Guiwei’ is more quickly than in ‘Heiye’. Transcript levels were measured at 6, 24, and 48 hour post-inoculation. ‘Oxidation-reduction process’ was dramatically enhanced in ‘Heiye’, which could promote its resistance to pathogen infection. A small ratio (3.78%) of common DEGs indicates that resistant and susceptible cultivars take different strategies to defense against P. litchii. At early infection stage, ‘Heiye’ induced a larger number of genes, including seven receptor-like kinases, which quickly recognized attack of pathogen and led to a rapidly resistance by regulation of degradation of proteasome, transcription factors, and cell wall remodeling. The early DGEs were exiguous in ‘Guiwei’, suggesting a weak response. Once the infection was successful, the resistance was repressed by down-regulated genes involved in phenylpropanoid metabolism, ET biosynthesis and signaling conduction in ‘Guiwei’. In conclusion, quickly recognition and early responses to pathogen, as well as minimal pathogen development and basal expression of resistance-related genes, were correlated with a high level of resistance in ‘Heiye’, while susceptible ‘Guiwei’ suffered massive infection due to lagging response and repressed signal transduction.