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Fruit and leaf possess distinctly different metabolites. Here, metabolites and transcriptome were compared between mature leaves (ML) and juice sacs (JS) of Citrus grandis "Hirado Buntan" to investigate the possible reasons. Results indicated that the remarkable difference in starch, total flavonoids and carotenoids, l-ascorbate, and jasmonic acid between ML and JS was tightly related to the expression levels of their biosynthesis-related genes, while the significant difference in abscisic acid and citrate was mainly related to the gene expression level(s) of 9-cis-epoxycarotenoid dioxygenase and proton pump genes, respectively. In addition, ATP citrate lyase probably plays a key role in the levels of flavonoids between ML and JS via providing different levels of acetyl-CoA. Taken together, these results identified some key candidate genes responsible for the content of a given metabolite and will contribute to research in regulating such metabolite content in citrus fruits.

The productivity of agricultural produce is fairly dependent on the availability of nutrients and efficient use. Magnesium (Mg2+) is an essential macronutrient of living cells and is the second most prevalent free divalent cation in plants. Mg2+ plays a role in several physiological processes that support plant growth and development. However, it has been largely forgotten in fertilization management strategies to increase crop production, which leads to severe reductions in plant growth and yield. In this review, we discuss how the Mg2+ shortage induces several responses in plants at different levels: morphological, physiological, biochemical and molecular. Additionally, the Mg2+ uptake and transport mechanisms in different cellular organelles and the role of Mg2+ transporters in regulating Mg2+ homeostasis are also discussed. Overall, in this review, we critically summarize the available information about the responses of Mg deficiency on plant growth and development, which would facilitate plant scientists to create Mg2+-deficiency-resilient crops through agronomic and genetic biofortification.

Citrate is an important intermediate product for the biosynthesis of several metabolites in plants. As two important organs of the citrus plant, fruits and leaves have their own metabolites characteristics; among them, citrate is normally high in fruit juice sacs (JS) and low in leaves. In this study, citrate content and transcript levels of citrate synthesis, transport, storage, and utilization related genes were compared between leaves and fruit JS of Citrus reticulata cv. 'Huagan No. 2', C. grandis cv. 'Hirado Buntan', and C. sinensis cv. 'Anliu'. Results indicated that the citrate content in fruit JS was significantly higher than in leaves of each cultivar. Only the relative mRNA levels of a P-type proton pump gene, CsPH8, was significantly lower in leaves than in fruit JS of three citrus cultivars, while other genes related to citrate biosynthesis, transport, storage, and utilization were highly expressed in leaves as compared to fruit JS. Furthermore, CsPH8 transient and stable transformation in leaves indicated that the change in citrate content is highly consistent with the change of CsPH8 transcript levels. Taken together, our results strongly suggest that the low accumulation of citrate in citrus leaves is mainly due to the low expression level of CsPH8; additionally, the high level of expression of citrate-utilizing genes would prevent citrate accumulation in the leaf organ.

Although foliar boron (B) fertilization is regarded as an efficient way to remedy B deficiency, the mechanisms of foliar B transport from leaves to roots are still unclear. In this study, performed with 1-year-old “Newhall” navel orange (Citrus sinensis) grafted on trifoliate orange (Poncirus trifoliata) plants, we analyzed the B concentration in leaves and roots, B-sucrose complex in the phloem sap after foliar application of 10B, girdling, and/or shading treatments. Results indicated that 10B concentration was significantly increased in roots after foliar 10B treatment. On the other hand, both girdling the scion stem and shading over the plants with a black plastic net significantly reduced the B and 10B concentration in roots. LC-MS analysis revealed that foliar 10B-treated plants had higher concentration of sucrose and some sugar alcohols in the phloem sap as compared to foliar water-treated plants. Combining with the analysis in the artificial mixture of B and sucrose, a higher peak intensity of the 10B-sucrose complex was found in the phloem sap of foliar 10B-treated plants compared to the control plants. Taken together, it is concluded that foliar B can be long distance transported from leaves to roots via phloem, at least by forming a B-sucrose complex in citrus plants.

The vacuolar H+-ATPase (V-ATPase) proton pump plays an important role in the acidification of vacuoles; however, genes encoding V-ATPase in the citrus genome and their roles in citric acid accumulation remain unclear in citrus fruit. In this study, we found at least one gene encoding subunit A, B, C, D, G, c'', d or e; two genes encoding the subunit E, F, H or a; and four genes encoding subunit c in the citrus genome. Spatial expression analysis showed that most genes were predominantly expressed in the mature leaves and/or flowers but were less expressed in root and juice cells. Two sweet orange (Citrus sinensis) cultivars, 'Anliu' (AL) and 'Hong Anliu' (HAL), which differ in terms of fruit acidity, were used in this study. The citric acid content was significantly higher in 'AL' fruits than in 'HAL' fruits over the entire experimental period (82 days-236 days after full blossom, DAFB). Transcript analysis showed that the transcript levels of most subunit genes, including V1-A, V1-B, V1-C, V1-E1, V1-G, V1-H2 and V0-a2, V0-c", V0-c4, and V0-d, were significantly lower in 'HAL' than in 'AL' fruits during fruit development and ripening. Moreover, ABA injection significantly increased the citric acid content, simultaneously accompanied by the obvious induction of V1-A, V1-C, V1-E1, V1-F1, V1-H2, V0-a1, V0-a2, V0-c1, V0-c2, V0-c4, and V0-d transcription levels. In conclusion, the results demonstrated that V1-A, V1-C, V1-E1, V1-H2, V0-a2, V0-c4, and V0-d may play more roles than other subunit genes in the vacuole acidification of citrus fruits. The lower activity of V-ATPase caused by the transcript reduction of some subunit genes may be one reason for the low citrate accumulation in 'HAL' juice sacs.

The regulation of citric acid metabolism during fruit ripening has a major impact on the production of high-quality fruit. The impact of citric acid on organoleptic fruit quality attributes, fruit storage performance and the synthesis of several secondary metabolites has led to an exponential increase in research efforts during the last two decades. Recent research has focused on the relationship among citric acid biosynthesis, transportation, storage, and utilization. Among citrate metabolic processes, activities of a proton pump, especially the plasma membrane H+-ATPase on tonoplast and citrate catabolism in cytosol play important roles in the regulation of citrate accumulation in cell vacuoles. Moreover, we highlight recent advances and provide an overview of citrate metabolism, postharvest physiology of citrate metabolism, and the influence of agro-climatic factors on citrus fruits. It is the first review that provides a comprehensive model for citrate metabolism in citrus fruit juice sacs. We...

Sweet oranges are produced on large scale all over the world and demanded due to less number of seeds and are better juice quality. A study was carried out with the objective to characterize promising sweet orange cultivars for better quality fruits production under different agro-climatic conditions of Punjab (Pakistan). Fruits of three promising sweet orange (Citrus sinensis L. Osbeck) cultivars viz. ‘Blood Red’, ‘Musambi’ and were harvested from commercial citrus groves at five different locations. Starting from Multan, in the direction of increasing altitude, four more sites viz. Sahiwal, Faisalabad, Sargodha and Chakwal were selected after every 100–150 km distance. Fruits were evaluated in terms of different physical and biochemical characters. Results showed significant influence of climatic conditions on physico-chemical properties of the cultivars at all the locations. Fruits harvested from Multan and Sahiwal showed higher fruit weight, diameter and peel weight. More juicy fruits with higher vitamin C content were produced under mild temperature environmental conditions of Chakwal. Overall, ‘Musambi’ fruits had more total soluble solids and reducing and total sugars. ‘Musambi’ was found to be good source of bioactive compounds i. e. vitamin C and total phenolic content, had the maximum antioxidant activity as compared to ‘Blood Red’ and ‘Succari’ and may be explored as potential cultivar in Sargodha and Chakwal.

The authors would like to thank the Deanship of Scientific Research at Majmaah University for supporting this work under Project Number No. R-1441-22. Quality of service (QoS)-aware data gathering in static-channel based underwater wireless sensor networks (UWSNs) is severely limited due to location and time-dependent acoustic channel communication characteristics. This paper proposes a novel cross-layer QoS-aware multichannel routing protocol called QoSRP for the internet of UWSNs-based time-critical marine monitoring applications. The proposed QoSRP scheme considers the unique characteristics of the acoustic communication in highly dynamic network topology during gathering and relaying events data towards the sink. The proposed QoSRP scheme during the time-critical events data-gathering process employs three basic mechanisms, namely underwater channel detection (UWCD), underwater channel assignment (UWCA) and underwater packets forwarding (UWPF). The UWCD mechanism finds the vacant channels with a high probability of detection and low probability of missed detection and false alarms. The UWCA scheme assigns high data rates channels to acoustic sensor nodes (ASNs) with longer idle probability in a robust manner. Lastly, the UWPF mechanism during conveying information avoids congestion, data path loops and balances the data traffic load in UWSNs. The QoSRP scheme is validated through extensive simulations conducted by NS2 and AquaSim 2.0 in underwater environments (UWEs). The simulation results reveal that the QoSRP protocol performs better compared to existing routing schemes in UWSNs. Deanship of Scientific Research at Majmaah University R-1441-22

Citric acid homeostasis patterns and its content are diversified among the fruits of citrus cultivars, but the cause remained unclear. In this study we showed that changes of citric acid content were highly associated with the expression profiles of a P-type proton pump gene (CsPH8) in the fruits of six citrus cultivars; moreover, analysis of 21 different fruit samples indicated that the correlation coefficient between titratable acid content and CsPH8 transcript level was 0.5837 with a significant level (P 0.05). Overexpression of CsPH8 in acidless pumelo juice sacs, strawberry fruit, and tomato fruit significantly increased the titratable acid or citric acid content besides the gene transcript level. On another hand, RNA interference of CsPH8 in acidic pumelo juice sacs significantly decreased the CsPH8 transcript level and the titratable acid or citric acid content as well. In addition, severe drought significantly increased the CsPH8 transcript level besides the titratable acid content. Taken together, these findings address the function of CsPH8 in citrus vacuolar acidification, confirm that CsPH8 plays a key role in the variation of citric acid content, and supported that the acid fluctuation influenced by drought, is at least partly due to the change of CsPH8 transcript level.

The moist temperate forests in Northern Pakistan are home to a variety of flora and fauna that are pivotal in sustaining the livelihoods of the local communities. In these forests, distribution and richness of vegetation, especially that of medicinal plants, is rarely reported. In this study, we carried out a vegetation survey in District Balakot, located in Northeastern Pakistan, to characterize the diversity of medicinal plants under different canopies of coniferous forest. The experimental site was divided into three major categories (viz., closed canopy, open spaces, and partial tree cover). A sampling plot of 100 m2 was established on each site to measure species diversity, dominance, and evenness. To observe richness and abundance, the rarefaction and rank abundance curves were plotted. Results revealed that a total of 45 species representing 34 families were available in the study site. Medicinal plants were the most abundant (45%) followed by edible plants (26%). Tree canopy cover affected the overall growth of medicinal plants on the basis of abundance and richness. The site with partial canopy exhibited the highest diversity, dominance, and abundance compared to open spaces and closed canopy. These findings are instrumental in identifying the wealth of the medicinal floral diversity in the northeastern temperate forest of Balakot and the opportunity to sustain the livelihoods of local communities with the help of public/private partnership.

Geranyl diphosphate synthase (GPPS) is a plastid localized enzyme that catalyzes the biosynthesis of Geranyl diphosphate (GPP), which is a universal precursor of monoterpenes. Wintersweet (Chimonanthus praecox L.), a famous deciduous flowering shrub with a strong floral scent character, could have GPPS-like homologs that are involved in monoterpenes biosynthesis, but it remains unclear. In the present study, five full-length GPPS and geranylgeranyl diphosphate synthases (GGPPS) genes were identified in the wintersweet transcriptome database. The isolated cDNAs showed high protein sequence similarity with the other plants GPPS and GGPPS. The phylogenetic analysis further classified these cDNAs into four distinct clades, representing heterodimeric GPPS small subunits (SSU1 and SSU2), homodimeric GPPS, and GGPPS. Analysis of temporal expression revealed that all genes have the highest transcript level at the full-open flower stage. From tissue-specific expression analysis, CpGPPS.SSU1 and CpGGPPS1 were predominantly expressed in petal and flower, whereas CpGPPS.SSU2, GPPS, and GGPPS2 showed a constitutive expression. Additionally, the subcellular localization assay identified the chloroplast localization of SSUs and GGPPSs proteins, and the yeast two-hybrid assay showed that both CpGPPS.SSU1 and CpGPPS.SSU2 can interact with the GGPPS proteins. Taken together, these preliminary results suggest that the heterodimeric GPPS can regulate floral scent biosynthesis in wintersweet flower.

Citric acid metabolism is considered to be the central cellular process of metabolite conversions. ATP-citrate lyase (ACL) and cytosolic aconitase (cyt-ACO) are the two citrate-degrading enzymes that decide the carbon flux towards different metabolite biosynthesis pathways. However, the correlation of their activities with metabolite concentrations in citrus fruits is still unclear. Here, the concentrations of soluble sugars, organic acids, acetyl-CoA, flavonoids, carotenoids, and &gamma, aminobutyric acid, as well as the activities of ACL, cyt-ACO, acetyl-CoA C-acetyltransferase, and acetyl-CoA carboxylase, were compared among the fruits of six citrus cultivars during fruit development and ripening. The results showed that the correlation between citrate concentration and cyt-ACO or ACL activity varied greatly among cultivars, while the activities of cyt-ACO and ACL had a significantly negative correlation (r = &minus, 0.4431). Moreover, ACL overexpression and RNA interference in the Citrus callus indicated that increasing and decreasing the ACL activity could reduce and induce cyt-ACO activity, respectively. In addition, significant correlation was only observed between the ACL activity and the concentration of acetyl-CoA (r = 0.4333). Taken together, the present study suggested that ACL and cyt-ACO synergistically control the citrate fate for the biosynthesis of other metabolites, but they are not the key determinants for the accumulation of citrate, as well as other metabolites in citrus fruits.

The persistent consumption of citrus fruit mainly depends on its sweet–sour taste as well as pulp texture. To date, the simple facility by covering plastic film just over trees is commonly used in different citrus species for harvest time extension and quality improvement. In this study, the effect of covering trees of Citrus reticulata cv. Kinokuni with plastic films was investigated. Results indicated that simply covering plastic film over trees significantly decreased the soil water content, the light intensity, and then the photosynthetic rate, however, moderately increased the internal average temperature of 0–3 °C. Moreover, it significantly facilitated soluble sugar accumulation, increased water soluble pectin content, and decreased protopectin content. Investigating transcript profile showed that only 351 gene transcripts were significantly influenced by covering plastic film. Interestingly, most differently expressed genes (DEGs) were involved in carbohydrate and cell wall metabolisms except some DEGs were related to signal transduction and environmental adaption. Taken together, covering plastic film over trees comprehensively modified the internal environmental condition and improved the fruit sweetness and mastication trait by promoting soluble sugar accumulation, increasing water soluble pectin content, and decreasing protopectin content through the alteration of transcript level.

The underwater wireless sensor networks have been receiving a great attention towards the ocean monitoring systems in the past couple of years. They have different characteristics comparing with the territorial WSN, including the constrained bandwidth and limited power supply. Though, more work has been completed for developing protocols and models based on the terrestrial networks. These applications can rarely be applied for the applications and the products based on the underwater wireless sensor networks. Major works have been placed on the designing of the standard protocol with considerations of the underwater communication characteristics. With the use of the sensor nodes, more energy can be saved. The proposed system is based on the process of depth based routing protocol and it will be the basic idea of the proposed paper. The depth routing protocol is adapted into the process of checking the depth of sensor nodes. The cluster based routing protocol, is used in this paper for minimizing the consumption ratio of the energy and distributing the nodes equally to all the functions. The nodes may be damaged due to high load in the current system. The main issue will be faced is, constructing a standard clustering algorithm. In this paper, the underwater wireless sensor networks context term will be clustering and it will contribute the purpose of efficient use of the energy resources. The energy that is consumed by each node will be processed with equal probability for the selection of cluster head. Based on this type of processing, the stable time period of the network will be improved in the depth base routing (DBR). The cluster depth routing protocol will be used for forwarding the packets. By using this, it will maximize the overall performance of the network. The comparison is done on the various types of protocols used in it.

Iron (Fe) deficiency chlorosis is a yield-limiting problem in citrus production regions with calcareous soils. Physiological and transcriptional analyses of fragrant citrus (Citrus junos Sieb. ex Tanaka) leaves from Fe-sufficient (IS) and Fe-deficient (ID) plants were investigated in this study. The physiological results showed that Fe, potassium, and nitrogen levels decreased by 12, 15, and 41% in ID leaves, respectively. However, zinc and copper levels increased by 49 and 35% in ID leaves, respectively. The chlorophyll (Chl) content, photosynthesis rate, stomatal conductance, and transpiration rate in ID leaves decreased by 55, 33, 38, and 42%, respectively, compared with IS leaves. Moreover, transcriptional profiling analysis showed that genes associated with Chl metabolism, photosynthesis, and nitrogen metabolism were dramatically downregulated by Fe deficiency. The expression of glutamyl-tRNA reductase 1, chlorophyll(ide) b reductase, and geranylgeranyl diphosphate reductase in ID leaves was 0.26–0.37 times that in IS leaves. The expression levels of 16 photosynthesis-related genes were severely downregulated by Fe deficiency. In addition, the transcription levels of nitrate transporter, nitrate reductase, and ferredoxin-nitrite reductase genes in ID leaves were 0.38–0.45 times those in IS leaves. Taken together, these results indicated that the block of Chl biosynthesis, the reduction of photosynthesis, and the repression of nitrogen absorption resulted in the chlorosis symptoms observed in fragrant citrus leaves.

Boron (B) deficiency stress is frequently observed in citrus orchards and causes considerable loss of productivity and fruit quality. Carrizo citrange (Cc) has been reported as a rootstock more tolerant to B deficiency than Trifoliate orange (To). The ‘Newhall’ navel orange (Ns) performed better when grafted onto Cc (Ns/Cc) than when grafted onto To (Ns/To) under long-term B deficiency. The present study confirmed that Ns/Cc had higher boron content, leaf fresh weight, lower leaf chlorosis and stronger photosynthesis ability than Ns/To. Moreover, B-deficiency significantly reduced the chlorophyll and carotenoid content in Ns/To. The content of total soluble sugar and lignin were dramatically increased and the expression levels of photosynthesis-related genes were substantially down-regulated in Ns/To by B-deficient treatment. B-deficiency also strongly induced expression levels of chlorophyll decomposition-related genes, glucose synthesis-related genes and lignin synthesis-related genes, and significantly inhibited the expression of carotenoid synthesis-related genes in Ns/To. Overall, these findings suggested that the influence of To on the scion of Ns was worse than that of Cc due to differently regulating these metabolic pathways under the long term of B-deficiency. The transcriptome analysis provided further information for understanding the mechanism of the different responses of scion-rootstock combinations to B-deficiency stress.

In Wireless Sensors Networks (WSNs), researcher’s main focus is on energy preservation and prolonging network lifetime. More energy resources are required in case of remote applications of WSNs, where some of the nodes die early that shorten the lifetime and decrease the stability of the network. It is mainly caused due to the non-optimal Cluster Heads (CHs) selection based on single criterion and uneven distribution of energy. We propose a new clustering protocol for both homogeneous and heterogeneous environments, named as Optimized Path planning algorithm with Energy efficiency and Extending Network lifetime in WSN (OPEN). In the proposed protocol, timer value concept is used for CH selection based on multiple criteria. Simulation results prove that OPEN performs better than the existing protocols in terms of the network lifetime, throughput and stability. The results explicitly explain the cluster head selection of OPEN protocol and efficient solution of uneven energy distribution problem.