Cultivated citrus are selections from, or hybrids of, wild progenitor species whose identities and contributions to citrus domestication remain controversial. Here we sequence and compare citrus genomes-a high-quality reference haploid clementine genome and mandarin, pummelo, sweet-orange and sour-orange genomes-and show that cultivated types derive from two progenitor species. Although cultivated pummelos represent selections from one progenitor species, Citrus maxima, cultivated mandarins are introgressions of C. maxima into the ancestral mandarin species Citrus reticulata. The most widely cultivated citrus, sweet orange, is the offspring of previously admixed individuals, but sour orange is an F1 hybrid of pure C. maxima and C. reticulata parents, thus implying that wild mandarins were part of the early breeding germplasm. A Chinese wild ‘mandarin’ diverges substantially from C. reticulata, thus suggesting the possibility of other unrecognized wild citrus species. Understanding citrus phylogeny through genome analysis clarifies taxonomic relationships and facilitates sequence-directed genetic improvement.
Leprosis is one of the most serious citrus plant diseases. Leprosis-affected plants, especially sweet orange [Citrus sinensis (L.) Osbeck], which is the most widely cultivated citrus fruit worldwide, show reduced photosynthetic capacity and severe defoliation. The aim was to evaluate the relationship between the Brevipalpus phoenicis (Geijskes) vector mite and citrus leprosis disease in Pera sweet orange plants grafted on different rootstocks. Data were analysed using numerical classification and conventional statistical analysis (ANOVA).
Mandarin orange (MO) is an important fruit crop of tropical and subtropical regions of the world. A total of 217 morphologically distinct rhizobacteria from MO orchards in 3 states of northeastern India were isolated and analyzed for 4 plant-growth-promoting (PGP) attributes: nitrogen fixation, production of indole acetic acid like substances, solubilization of phosphate, and ability to antagonize pathogenic fungi. Isolates were ranked based on in-vitro-assayed PGP attributes, and 10 superior isolates were selected to test their effect on seedling emergence and seedling growth in a completely randomized pot experiment. These 10 isolates increased seedling emergence over a noninoculated control within 45 days after sowing. Five isolates, namely RCE1, RCE2, RCE3, RCE5, and RCE7, significantly increased shoot length, shoot dry biomass, and root dry biomass of 120-day-old seedlings over the noninoculated control. The beneficial effects of 4 selected strains, namely Enterobacter hormaechei RCE-1, Enterobacter asburiae RCE-2, Enterobacter ludwigii RCE-5, and Klebsiella pneumoniae RCE-7, on growth of the seedlings were visible up to 1 year after their transfer to 8 kg capacity pots. These strains were superior both in terms of in-vitro-assayed PGP attributes and of their beneficial effect in low phosphorus soil and, thus, may be promising bioinoculants for promoting early emergence and growth of MO seedlings.
Alternaria brown spot is one of the most important diseases of tangerines and their hybrids worldwide. Recently, outbreaks in Mediterranean areas related to susceptible cultivars, refocused attention on the disease. Twenty representatives were selected from a collection of 180 isolates of Alternaria spp. from citrus leaves and fruit. They were characterized along with reference strains of Alternaria spp. Micro- and macroscopic characteristics separated most Alternaria isolates into six morphotypes referable to A. alternata (5) and A. arborescens (1). Phylogenetic analyses, based on endopolygalacturonase (endopg) and internal transcribed spacer (ITS), confirmed this finding. Moreover, a five-gene phylogeny including two anonymous genomics regions (OPA 1-3 and OPA 2-1), and the beta-tubulin gene (ß-tub), produced a further clustering of A. alternata into three clades. This analysis suggested the existence of intra-species molecular variability. Investigated isolates showed different levels of virulence on leaves and fruit. In particular, the pathogenicity on fruit seemed to be correlated with the tissue of isolation and the clade. The toxigenic behavior of Alternaria isolates was also investigated, with tenuazonic acid (TeA) being the most abundant mycotoxin (0.2-20 mg/L). Isolates also synthesized the mycotoxins alternariol (AOH), its derivate alternariol monomethyl ether (AME), and altenuene (ALT), although to a lesser extent. AME production significantly varied among the six morphotypes. The expression of pksJ/pksH, biosynthetic genes of AOH/AME, was not correlated with actual toxin production, but it was significantly different between the two genotypes and among the four clades. Finally, ten isolates proved to express the biosynthetic genes of ACTT1 phytotoxin, and thus to be included in the Alternaria pathotype tangerine. A significant correlation between pathogenicity on leaves and ACTT1 gene expression was recorded. The latter was significantly dependent on geographical origin. The widespread occurrence of Alternaria spp. on citrus fruit and their ability to produce mycotoxins might represent a serious concern for producers and consumers.
Oranges are an important nutritional source for human health and have immense economic value. Here we present a comprehensive analysis of the draft genome of sweet orange (Citrus sinensis). The assembled sequence covers 87.3% of the estimated orange genome, which is relatively compact, as 20% is composed of repetitive elements. We predicted 29,445 protein-coding genes, half of which are in the heterozygous state. With additional sequencing of two more citrus species and comparative analyses of seven citrus genomes, we present evidence to suggest that sweet orange originated from a backcross hybrid between pummelo and mandarin. Focused analysis on genes involved in vitamin C metabolism showed that GalUR, encoding the rate-limiting enzyme of the galacturonate pathway, is significantly upregulated in orange fruit, and the recent expansion of this gene family may provide a genomic basis. This draft genome represents a valuable resource for understanding and improving many important citrus traits in the future.
Male sterility derived from Satsuma mandarin (Citrus unshiu) has been used in Japanese citrus breeding programs to obtain seedless cultivars, which is a desirable trait for consumers. Male sterility has often been evaluated by anther development or pollen fertility; however, the inheritance and heritability of male sterility derived from Satsuma is poorly understood. In this study, we investigated the mode of inheritance and broad-sense heritability of male sterility derived from Satsuma. Initially, we evaluated the total number of pollen grains per anther and apparent pollen fertility, as indicated by lactophenol blue staining, in 15 citrus cultivars and selections to understand the male sterility of Satsuma. The results indicated that male sterility was primarily caused by decreased number of pollen grains per anther in progeny of Satsuma. We also evaluated these traits in three F1 populations (hyuganatsu × ‘Okitsu No. 56’, ‘Okitsu No. 46’ × ‘Okitsu No. 56’ and ‘Okitsu No. 46’ × ‘Kara’), of which the parents are derived from Satsuma. Individuals in these populations showed strong segregation for number of pollen grains per anther. The apparent fertility of pollen also showed segregation but was almost constant at 70%-90%. The estimated broad-sense heritability for the number of pollen grains per anther was as high as 0.898 in the ‘Okitsu No. 46’ × ‘Okitsu No. 56’ and ‘Okitsu No. 46’ × ‘Kara’ populations. These results indicated that the number of pollen grains per anther primarily determined male sterility among progeny of Satsuma, and this trait was inherited by the progeny. Development of DNA markers closely linked to male sterility using the F1 populations of ‘Okitsu No. 46’ × ‘Okitsu No. 56’ and ‘Okitsu No. 46’ × ‘Kara’ is expected to contribute to the breeding of novel seedless citrus cultivars.
Flowering dates and the timing of late season frost are both driven by local ambient temperatures. However, under climatic warming observed over the past century, it remains uncertain how such impacts affect frost risk associated with plant phenophase shifts. Any increase in frost frequency or severity has the potential to damage flowers and their resultant yields and, in more extreme cases, the survival of the plant. An accurate assessment of the relationship between the timing of last frost events and phenological shifts associated with warmer climate is thus imperative. We investigate spring advances in citrus flowering dates (orange, tangerine, sweet lemon, sour lemon and sour orange) for Kerman and Shiraz, Iran from 1960 to 2010. These cities have experienced increases in both T max and T min, advances in peak flowering dates and changes in last frost dates over the study period. Based on daily instrumental climate records, the last frost dates for each year are compared with the peak flowering dates. For both cities, the rate of last frost advance lags behind the phenological advance, thus increasing frost risk. Increased frost risk will likely have considerable direct impacts on crop yields and on the associated capacity to adapt, given future climatic uncertainty.
Three essential oils (EOs) were isolated from the peel of citrus fruits Citrus reticulata L., Citrus reticulata chinase Blanco and Citrus sinensis (L.) Osbeck (Family: Rutaceae) and evaluated against Culex pipiens L.1758 (Family: Culicidae). Chemical composition indicated that the EOs were rich in essential phytochemicals including hydrocarbons, monoterpenes and sesquiterpenes. These constituents revealed some variability among the oils displaying interesting chemotypes limonene (35-51%), 1R-α-pinene (1.04-2.5%), γ-terpinene (0.46-5.65%) and sabinene (0.51-5.42%). The toxicity proved that C. sinensis oil had more effect than C. reticulata chinase and C. reticulata oils against larvae (LC50 = 15.35, 16.11 and 32.84 mg/L, respectively). However, C. reticulate was the most active as fumigant against adults (LC50 2.74 μL/L air). The in vivo effect on acetylcholine esterase (AChE), carboxyl esterase (CbE), acid phosphatase (ACP), alkaline phosphatase (ALP), adenosine triphosphatase (ATPase) and glutathione-S-transferase (GST) were also demonstrated. To the best of our knowledge, this is the ﬁrst report about the chemical composition and mosquitocidal activity of C. reticulata chinase essential oils. Conclusively, the tested essential oils could be used as eco-friendly alternatives in mosquitoes control programme.
The freshness of citrus fruits commonly available in the market was non-destructively assessed by Raman spectroscopy. Intact clementine, mandarin and tangerine species were characterised concerning their carotenoids skin Raman signalling in a time course from the moment they were acquired as fresh stock, supplying the market, to the physical degradation, when they were no longer attractive to consumers. The freshness was found to strongly correlate to the peel Raman signal collected from the same area of the intact fruits in a time course of a maximum of 20days. We have shown that the intensity of the carotenoid Raman signal is indeed a good indicator of fruit freshness and introduced a Raman coefficient of freshness (CFresh), whose time course is linearly decreasing, with different slope for different citrus groups. Additionally, we demonstrated that the freshness assessment could be achieved using a portable Raman instrument. The results could have a strong impact for consumer satisfaction and the food industry.
Composition and changes in free volatiles have been extensively studied in citrus fruit such as mandarin. However, components of glycosidically bound volatiles and changes during fruit ripening have been rarely investigated. A total of 56 glycosidically-bound volatiles were identified in fruit peel at four ripening stages. The highest concentrations in glycosidically-bound volatiles were observed for methyl salicylate in ripening fruit. Concentration of total bound volatiles increased from color conversion stage at 150days after bloom (DAB), peaked at yellow stage (190DAB), followed by a decrease at orange stage (210DAB). Satsuma mandarin fruit at different ripening stages could be separated in a partial least squares-discriminant analysis (PLS-DA) plot using glycosidically bound volatiles as variables. In total 35 glycosidically bound volatiles were identified with variable importance in projection (VIP) score exceeding 1, which may be potential markers for separating fruit at different ripening stages.