EVALUACIÓN DE ESPECIES DE MOSCAS DE LA FRUTA Y SUS HOSPEDEROS EN LA ZONA DE SATIPO

Las plantas hospederas de mosca de la fruta fueron evaluados de los fundos de la zona de Satipo, en campo y laboratorio. Los resultados muestran que los cítricos están infestadas por Anastrepha fraterculus; mientras que los frutales nativos con otras especies de Anastrepha, como son A. striata, A. leptozona, y A. distincta. Se ha encontrado un díptero amarillo en el caimito que no pertenece a los tephritidae. Se tiene 19 frutales distintos hospederas, siendo los cítricos C. reticulata x C. aurantifolia, Citrus limettioides, Citrus aurantifolia, Citrus reticulata, Citrus reshni, Citrus sinensis, C. reticulata x C. sinensis, C. reticulata x C. paradisi; y los frutales nativos y exóticos, Chrysophillum caimito, Averrhoa carambola, Spondias sp., Psidium guajaba, Manguifera indica, Inga edulis Mart, Anacardiaceae y Matisia cordata. El uvo de monte, guayaba y caimito sufren un daño de entre 83 Y 100%. El zapote y pacae muestran daños de entre 66 a 53%. El mango y carambola muestran daños de 31 a 26%. En los cítricos, la naranja Washington muestra los niveles más altos de infestación con 45%,; las mandarinas entre 27 y 32%, las naranjas entre 14 y 22%, los híbridos entre 20 y 22%, los portainjertos entre 26 y 27%. Las pérdidas económicas provinciales del 14% por moscas de la fruta sólo en naranja Valencia se estima en U.S.$ 2,025,000,00. Palabras clave: Anastrepha, fraterculus, striata, cítricos, Satipo,

Genetic diversity and population structure of sweet orange [Citrus sinensis (L.) Osbeck] germplasm of India revealed by SSR and InDel markers

Sweet orange (Citrus sinensis (L.) Osbeck) is an important commercial citrus fruit crop, cultivated in India and across the world. In India most of the cultivated sweet orange species were introduced varieties. In this study, we used two molecular markers, SSR and InDels, to understand the genetic diversity and population structure of seventy-two sweet orange genotypes. Genetic parameters consisted of a total number of alleles, a number of polymorphic alleles (effective alleles); genetic diversity (G.D.), expected heterozygosity (He), and the polymorphic information content (PIC) were calculated based on molecular data. Two dendrograms were constructed based on the InDels and SSR. In both the cases, they formed three major clusters showing various degrees of variations with respect to members of the clusters. Population structure analysis revealed the presence of two distinct subpopulations. Therefore, in order to address various challenges and develop sweet orange varieties with desirable traits, there is a need to broaden the genetic base of sweet orange through the intensive collection in the northeastern region. These results of intraspecific genetic variability of the collections will dictate the path for the sweet orange breeding and conservation programs in India.

Highly Efficient Generation of Canker-Resistant Sweet Orange Enabled by an Improved CRISPR/Cas9 System

Sweet orange (Citrus sinensis) is the most economically important species for the citrus industry. However, it is susceptible to many diseases including citrus bacterial canker caused by Xanthomonas citri subsp. citri (Xcc) that triggers devastating effects on citrus production. Conventional breeding has not met the challenge to improve disease resistance of sweet orange due to the long juvenility and other limitations. CRISPR-mediated genome editing has shown promising potentials for genetic improvements of plants. Generation of biallelic/homozygous mutants remains difficult for sweet orange due to low transformation rate, existence of heterozygous alleles for target genes, and low biallelic editing efficacy using the CRISPR technology. Here, we report improvements in the CRISPR/Cas9 system for citrus gene editing. Based on the improvements we made previously [dicot codon optimized Cas9, tRNA for multiplexing, a modified sgRNA scaffold with high efficiency, citrus U6 (CsU6) to drive sgRNA expression], we further improved our CRISPR/Cas9 system by choosing superior promoters [Cestrum yellow leaf curling virus (CmYLCV) or Citrus sinensis ubiquitin (CsUbi) promoter] to drive Cas9 and optimizing culture temperature. This system was able to generate a biallelic mutation rate of up to 89% for Carrizo citrange and 79% for Hamlin sweet orange. Consequently, this system was used to generate canker-resistant Hamlin sweet orange by mutating the effector binding element (EBE) of canker susceptibility gene CsLOB1, which is required for causing canker symptoms by Xcc. Six biallelic Hamlin sweet orange mutant lines in the EBE were generated. The biallelic mutants are resistant to Xcc. Biallelic mutation of the EBE region abolishes the induction of CsLOB1 by Xcc. This study represents a significant improvement in sweet orange gene editing efficacy and generating disease-resistant varieties via CRISPR-mediated genome editing. This improvement in citrus genome editing makes genetic studies and manipulations of sweet orange more feasible.

Determination of Polyphenols Content and Antioxidant Activity of Citrus sinensis Fruit Peel

Citrus fruits have long been qualified as veritable foods in view of the many therapeutic benefits they bring to the body. Several researchers have stud-ied the relationship between the bioactive compounds of Citrus and the health benefits and reduction of the risk of disease. Citrus sinensis, used in the food industry and its extracts have also been used in traditional medicine to activate vital energy, circulation, and weight loss, and appetite control. However, limited efforts have been made on collecting data on antioxidant potential of peels orange from the northern region of Algeria. Our study, therefore, focuses on the evaluation of total polyphenols compounds and in vitro assessment of their antioxidant potential of peels orange from the northern region of Algeria. The ethyl acetate and n-butanolic fractions from peels orange have been tested for their antioxidant activities and their lipid peroxidation inhibiting effects. The total phenolic and flavonoids content showed high levels. The preliminary phytochemical screening of tannins, phenolic compounds, flavonoids, coumarin, and alkaloids was also used. DPPH assay possesses strong potency to scavenge free radicals. The NO. radi-cal scavenging test exerts a good inhibitory effect. Furthermore, orange peels have been shown to suppress the lipid peroxidation of linoleic acid. Results further revealed a strong correlation between antioxidant effects and polyphenolic compounds. The high antioxidant activity of peel orange suggests that it could serve as a good natural antioxidant additive or food dietary supplement.

Absorption and Distribution of Calcium (45Ca) Applied to the Surface of Orange (Citrus sinensis) Fruits at Different Developmental Stages

In this study, the rate of absorption and transport of calcium (Ca) in orange fruits (Citrus sinensis cv. Fukumoto) after surface treatment with 45Ca was evaluated by supplying treatments at different dates between fruit set and mid-phase II of growth (i.e., 30, 44, 66 and 99 days after full bloom, DAFB). 45Ca absorption was always detected, but 45Ca application at the fruit set was associated with the highest rates of Ca transport into the pulp (39%) compared to the other treatments (17–19%). Scanning electron microscopy SEM) images of the fruit surface showed the occurrence of stomata along with the entire rind at all the developmental stages evaluated. However, from the beginning of stage I, stomata began to collapse and develop plugs, and this became frequent as the fruit age increased. From 44 DAFB, oil gland density increased, and Ca oxalates (CaOx) were found in the fruit flavedo and albedo. Fruit Ca (not labeled) concentration increased from fruit set (30 DAFB) to 99 DAFB, although oil gland formation and rind growth and differentiation likely hindered Ca transport to internal fruit tissues (pulp). The total Ca concentration in the pedicel was always higher than that in the fruit, with no differences between the first three treatments. The information obtained in this study may be essential for improving Ca fertilizer efficacy in citrus by spray applications.

Genome Wide Identification of Respiratory Burst Oxidase Homolog (Rboh) Genes in Citrus sinensis and Functional Analysis of CsRbohD in Cold Tolerance

Respiratory burst oxidase homologs (Rbohs) are critical enzymes involved in the generation of reactive oxygen species (ROS) that play an important role in plant growth and development as well as various biotic and abiotic stresses in plants. Thus far, there have been few reports on the characterization of the Rboh gene family in Citrus. In this study, seven Rboh genes (CsRbohA~CsRbohG) were identified in the Citrus sinensis genome. The CsRboh proteins were predicted to localize to the cell membrane. Most CsRbohs contained four conserved domains, an EF-hand domain, and a transmembrane region. Phylogenetic analysis demonstrated that the CsRbohs were divided into five groups, suggesting potential distinct functions and evolution. The expression profiles revealed that these seven CsRboh genes displayed tissue-specific expression patterns, and five CsRboh genes were responsive to cold stress. Fourteen putative cis-acting elements related to stress response, hormone response, and development regulation were present within the promoters of CsRboh genes. The in-silico microRNA target transcript analyses indicated that CsRbohE might be targeted by csi-miR164. Further functional and physiological analyses showed that the knockdown of CsRbohD in trifoliate orange impaired resistance to cold stress. As a whole, our results provide valuable information for further functional studies of the CsRboh genes in response to cold stress.