Comparative study on comprehensive quality of Xinhui chenpi by two main plant propagation techniques.

Xinhui chenpi (XHCP), the sun-dried peel of the mandarin orange, Citrus reticulata "Chachi," is the most famous crude drug, as well as a traditional seasoning in Chinese cooking. The main cultivation methods of XHCP are cutting and grafting, but it is generally considered that the quality of XHCP after cutting is superior to that obtained from plants propagated by graftings, which had a negative impact on the marketing of the finished product. In our study, a total of 25 samples of XHCP obtained from plants cultivated by either traditional methods (i.e., from cuttings) or by grafting were collected to compare the contents of four types of metabolites (essential oils, flavonoids, synephrine, and total polysaccharides) as well as antioxidant activity. The results revealed that the quality of XHCP did not decline after cutting, and marked individual differences between XHCP samples, even when prepared from plants grown in the same way. In general, grafting had no significant effect on the most essential oils components, total polysaccharides, synephrine, total flavonoids, total polymethoxylated flavones, hesperidin, nobiletin, tangeretin content, and antioxidant activity. Nevertheless, five volatile compounds can be used as potential chemical markers (p < 0.05) to distinguish between cutting XHCP and grafted XHCP, while four volatile compounds showed high content in grafted XHCP. Our study is expected to provide a theoretical basis for XHCP breeding and cultivation, and thereby further standardize the market of XHCP.

Anti-inflammatory and anti-aggregating effects of rangpur in the first trimester of growth: ultra-performance liquid chromatography-electrospray mass spectrometry profile and quantification of hesperidin.

BACKGROUND: Citrus fruits are a rich source of valuable molecules, and their industrial processing produces bagasses, little explored to generate important by-products. These Citrus residues, including seeds and peels, also contain numerous pharmacologically important substances. To reduce the impact of these Citrus by-products, young, harvested fruits could be used as a functional supplemental food while another part is grown until maturity for industrial production. This study therefore aims to valorize rangpur (Citrus limonia) in the first 3 months of its growth by investigating and comparing its monthly chemical profiles using ultra-performance liquid chromatography-electrospray mass spectrometry (UPLC-ESI-MS) and its anti-inflammatory and antiplatelet activity. RESULTS: Extracts obtained from the fruits harvested in November, December, and January, 2017 and 2018 (L221117, L161217, and L160118) showed different UPLC-ESI-MS profiles. Twenty-five of the 26 detected metabolites were identified as cyclitol, pyrrolidine betaine, aryl propanoyl esters, chlorogenic acids, flavonoids, coumarins, and limonoids. Quantification studies indicated an increased concentration of hesperidin from the younger fruits to the older fruits of the series. L160118 reduced nitrogen oxide (NOx), tumor necrosis factor alpha (TNF-α), and interleukin 6 (IL-6) levels more than other extracts. Their activity followed the same trends as the hesperidin concentration in each fruit. In contrast, the most promising antiplatelet activity was observed with the extracts from the two youngest fruits. This suggests combined effects of the chemical components found in these fruits' extracts. CONCLUSION: The extracts obtained from these young fruits showed considerable anti-inflammatory and antiplatelet activity. Overall, young rangpur could be used as raw material to produce functional foods without producing any waste. © 2022 Society of Chemical Industry.

Bactericidal Effect of Entomopathogenic Bacterium Pseudomonas entomophila Against Xanthomonas citri Reduces Citrus Canker Disease Severity.

The bacterium Pseudomonas entomophila has been recognized as an exceptional species within the Pseudomonas genus, capable of naturally infecting and killing insects from at least three different orders. P. entomophila ingestion leads to irreversible gut damage resulting from a global blockage of translation, which impairs both immune and tissue repair systems in the insect intestine. In this study we isolated a P. entomophila bacterial strain from soil samples which displayed a strong activity against Xanthomonas citri subsp, citri (Xcc), the etiological agent of citrus canker disease. The antagonism potential of isolated bacteria against Xcc and its ability to reduce citrus canker severity was assessed both ex planta and in planta. Our findings show that pathogenicity assays in Citrus x limonia by pressure infiltration and spray with a mixture of P. entomophila and Xcc leaded to a significant reduction in the number of canker lesions in high susceptible citrus leaves, at 21 days post-infection. To the best of our knowledge this is the first report of antibacterial activity of P. entomophila against a phytopathogenic bacterium. Collective action of P. entomophila factors such as diketopiperazine production and the type 6 secretion system (T6SS) may be involved in this type of biological control of citrus canker. The results suggest that the P. entomophila strain could be a promising biocontrol agent acting directly against Xcc.

A L-type lectin gene is involved in the response to hormonal treatment and water deficit in Volkamer lemon.

Combination of biotic and abiotic stress is a major challenge for crop and fruit production. Thus, identification of genes involved in cross-response to abiotic and biotic stress is of great importance for breeding superior genotypes. Lectins are glycan-binding proteins with a functions in the developmental processes as well as in the response to biotic and abiotic stress. In this work, a lectin like gene, namely ClLectin1, was characterized in Volkamer lemon and its expression was studied in plants exposed to either water stress, hormonal elicitors (JA, SA, ABA) or wounding to understand whether this gene may have a function in the response to multiple stress combination. Results showed that ClLectin1 has 100% homology with a L-type lectin gene from C. sinensis and the in silico study of the 5'UTR region showed the presence of cis-responsive elements to SA, DRE2 and ABA. ClLectin1 was rapidly induced by hormonal treatments and wounding, at local and systemic levels, suggesting an involvement in defence signalling pathways and a possible role as fast detection biomarker of biotic stress. On the other hand, the induction of ClLectin1 by water stress pointed out a role of the gene in the response to drought. The simultaneous response of ClLectin1 expression to water stress and SA treatment could be further investigated to assess whether a moderate drought stress may be useful to improve citrus performance by stimulating the SA-dependent response to biotic stress.

Quantification and localization of hesperidin and rutin in Citrus sinensis grafted on C. limonia after Xylella fastidiosa infection by HPLC-UV and MALDI imaging mass spectrometry.

A high performance liquid chromatography-ultraviolet (HPLC-UV) method was developed for quantifying hesperidin and rutin levels in leaves and stems of Citrus limonia, with a good linearity over a range of 1.0-80.0 and 1.0-50.0 µg mL(-1) respectively, with r(2)>0.999 for all curves. The limits of detection (LOD) for both flavonoids were 0.6 and 0.5 µg mL(-1), respectively, with quantification (LOQ) being 2.0 and 1.0 µg mL(-1), respectively. The quantification method was applied to Citrus sinensis grafted onto C. limonia with and without CVC (citrus variegated chlorosis) symptoms after Xylella fastidiosa infection. The total content of rutin was low and practically constant in all analyses in comparison with hesperidin, which showed a significant increase in its amount in symptomatic leaves. Scanning electron microscopy studies on leaves with CVC symptoms showed vessel occlusion by biofilm, and a crystallized material was noted. Considering the difficulty in isolating these crystals for analysis, tissue sections were analyzed by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) to confirm the presence of hesperidin at the site of infection. The images constructed from MS/MS data with a specific diagnostic fragment ion (m/z 483) also showed higher ion intensities for it in infected plants than in healthy ones, mainly in the vessel regions. These data suggest that hesperidin plays a role in the plant-pathogen interaction, probably as a phytoanticipin. This method was also applied to C. sinensis and C. limonia seedlings, and comparison with the graft results showed that the rootstock had an increased hesperidin content ∼3.6 fold greater in the graft stem than in the stem of C. sinensis seedlings. Increase in hesperidin content by rootstock can be related to induced internal defense mechanisms.

Nutritional balance and yield for green manure orange trees / Balanço nutricional e produtividade em laranjeiras manejadas com adubação verde

Intercropping could efficiently prevent soil nutrient losses caused by extensive agriculture. The present study aimed to assess the effect of green manure on the nutritional status of orange trees cultivar 'Pera' (Citrus sinensis (L.) Osbeck). The plants were grafted on 'Cravo' lime trees and were then planted in a 7x4m space. Four different treatments corresponding to the evaluated green manures were employed: jack bean (JB) (Canavalia ensiformis DC), lablab (LL) (Dolichos lablab L.), pigeon pea (PP) (Cajanus cajan L. Millsp), and Brachiaria (BQ) (Brachiaria brizantha Hochst ex A. Rich. Stapf) as control. The experimental design was in randomized blocks, in split-plot time, with six replicates, with four treatments (green manures) and two plants per evaluation. The nutritional status was assessed by using the DRIS method (Diagnosis and Recommendation Integrated System); the yield and the macro and micronutrient levels contained in green manures and in the control was also determined. The nutritional diagnosis indicated that, in the two years of experiment, plants treated with green manure showed better nutritional balance index compared to Brachiaria. This suggests that, over time, green manure can lead to better nutritional balance. Pigeon pea treatment showed the highest yields, compared to control, in the two evaluated crop cycles (2004/05 and 2005/06).

O manejo de culturas intercalares poderia prevenir eficientemente as perdas de nutrientes do solo, causadas pelo cultivo extensivo. O trabalho teve como objetivo avaliar o efeito da utilização de adubos verdes no estado nutricional de plantas de laranjeira 'Pêra' (Citrus sinensis (L.) Osbeck). O solo da área de cultivo é denominado Neossolo Quartzarênico. As plantas estavam enxertadas em limoeiro 'Cravo' e foram plantadas num espaçamento de sete por quatro metros. Foram empregados quatro tratamentos correspondentes aos adubos verdes avaliados, sendo eles: feijão de porco (FP) (Canavalia ensiformis DC), labe-labe (LL) (Dolichus lablab L.), feijão guandu anão (GA) (Cajanus cajan L. Millsp) e braquiária (BQ) (Brachiaria brizantha Hochst ex A. Rich. Stapf). Conduziu-se o experimento em delineamento de blocos casualizados em esquema de parcelas subdivididas no tempo, com seis repetições, quatro tratamentos (adubos verdes) e duas plantas úteis para as avaliações. Avaliou-se o estado nutricional pelo método DRIS (Sistema Integrado de Diagnose e Recomendação), bem como a produção e o teor de macro e micronutrientes contidos nos adubos verdes e na testemunha. O diagnóstico nutricional demonstrou que, nos dois anos de experimentação, houve um melhor índice de balanço nutricional para os adubos verdes, quando comparados com a braquiária (testemunha). Esta constatação permite inferir sobre a disponibilidade de, ao longo do tempo, os adubos verdes permitirem um melhor equilíbrio nutricional para as plantas. O tratamento com feijão de porco apresentou as maiores produções, quando comparado com a testemunha, nos dois ciclos agrícolas de avaliação (2004/05 e 2005/06).

Continuous, high-resolution biospeckle imaging reveals a discrete zone of activity at the root apex that responds to contact with obstacles.

BACKGROUND AND AIMS: Shining a laser onto biological material produces light speckles termed biospeckles. Patterns of biospeckle activity reflect changes in cell biochemistry, developmental processes and responses to the environment. The aim of this work was to develop methods to investigate the biospeckle activity in roots and to characterize the distribution of its intensity and response to thigmostimuli. METHODS: Biospeckle activity in roots of Zea mays, and also Jatropha curcas and Citrus limonia, was imaged live and in situ using a portable laser and a digital microscope with a spatial resolution of 10 µm per pixel and the ability to capture images every 0.080 s. A procedure incorporating a Fujii algorithm, image restoration using median and Gaussian filters, image segmentation using maximum-entropy threshold methods and the extraction of features using a tracing algorithm followed by spline fitting were developed to obtain quantitative information from images of biospeckle activity. A wavelet transform algorithm was used for spectral decomposition of biospeckle activity and generalized additive models were used to attribute statistical significance to changes in patterns of biospeckle activity. KEY RESULTS: The intensity of biospeckle activity was greatest close to the root apex. Higher frequencies (3-6 Hz) contributed most to the total intensity of biospeckle activity. When a root encountered an obstacle, the intensity of biospeckle activity decreased abruptly throughout the root system. The response became attenuated with repeated thigmostimuli. CONCLUSIONS: The data suggest that at least one component of root biospeckle activity resulted from a biological process, which is located in the zone of cell division and responds to thigmostimuli. However, neither individual cell division events nor root elongation is likely to be responsible for the patterns of biospeckle activity.

Produção de porta-enxerto de limão cravo em resposta a adubação organomineral / Produced of rootstock of raugpur lime in response to organic-mineral fertilization

A adubação do substrato pode permitir a obtenção de porta-enxertos de alta qualidade de limão cravo. O presente trabalho teve como objetivo avaliar a produção de porta-enxerto de limão cravo (Citrus limonia Osbeck), em resposta a aplicação em cobertura, de doses de um adubo organomineral, a base de turfa. O experimento foi conduzido em casa de vegetação, sendo as sementes plantadas em tubetes de 50 mL, preenchidos por substrato comercial a base de casca de Pinus e turfa. O delineamento experimental utilizado foi o inteiramente casualizado com 5 tratamentos, 5 repetições e 8 plantas por unidade experimental. Os tratamentos utilizados foram doses (0; 0,5; 1,0; 1,5; 2,0 mL) do produto comercial diluído em 1 litro de água, sendo aplicado 5 mL por tubete semanalmente. As adubações iniciaram aos 70 dias após a semeadura e aos 115 dias foram avaliadas as alturas da planta, número de folhas, diâmetro do caule e massa seca da parte aérea e da raiz e relação entre estas. Para a produção de portas-enxerto vigorosos de limão cravo pode ser aplicado, em cobertura no substrato, uma solução de 2,0 mL do adubo organomineral.

The fertilization of substrate can provided high quality of raugpur lime tree rootstock. This work has the objective to evaluate the production of the raugpur lime tree rootstock (Citrus limonia Osbeck), in response to broadcasting application to organic-mineral fertilizer of substrate. The experiment was developed in a greenhouse conditions in pots filled with commercial substratum based on Pinus rind and turf. The completely randomized experimental design with 5 treatments and, 5 replications and 8 plants per pot was use. The treatments were doses (0; 0.5; 1.0; 1.5; 2.0 mL) of a dilute commercial fertilizer in 1 liter of water, being applied 5 mL in each pot weekly. Fertilizations started 70 days after sowing and 115 days evaluations were perfomed which consisted of plant height, leaf number, diameter stem, weight of the root and shoot dry matter and relationship between these. For the production of vigorous rangpur lime tree rootstock can be applied in brodcasting the substrate, a solution of 2.0 mL of organic mineral fertilizer

Nitrogênio, fósforo e potássio na nutrição e no crescimento de mudas de laranjeiravalência, enxertadas sobre limoeiro cravo / Effect of nitrogen, phosphorus, and potassium levels on nutrition and production of seedlings of 'Valencia' sweet orange grafted on 'cravo' lemon rootstock

O experimento foi conduzido em casa de vegetação telada na FCAV/Unesp campus Jaboticabal-SP, durante o período de novembro de 2005 a janeiro de 2007. Conduziu-se este estudo, com o objetivo de avaliar componentes do desenvolvimento e do estado nutricional de mudas de laranjeira Valência (Citrus sinensis Osbeck), enxertadas sobre limoeiro Cravo (Citrus limonia Osbeck), em função de doses de nitrogênio, fósforo e potássio. O delineamento experimental foi inteiramente casualizado, em esquema fatorial 3³ + 1, sendo 3 fatores (nitrogênio, fósforo e potássio - NPK), 3 doses e uma testemunha (sem adubação), com 3 repetições. A unidade experimental foi representada por uma muda de laranjeira por sacola com 5 dm³ com 2,5 kg de substrato casca de Pinus spp. e vermiculita. Os tratamentos foram constituídos pela metade, uma vez e duas vezes a dose padrão recomendada, de 4.590; 920 e 4.380 mg sacola-1, de N, P e K, respectivamente. As adubações com N e K foram realizadas via fertirrigações três vezes por semana e o P foi adicionado ao substrato antes do replantio das mudas. Aos 424 dias após o transplantio, as plantas foram subdivididas em raízes e parte aérea para determinação da massa da matéria seca, altura, área foliar, diâmetro do caule e conteúdo de nutrientes. A adubação com N, P e K proporcionou maior desenvolvimento e maior acúmulo desses macronutrientes na parte aérea e nas raízes das mudas de laranjeira Valência, sobre limoeiro Cravo. Houve adequado desenvolvimento das plantas com a metade da dose recomendada de N, P e K pela literatura, aproximadamente de 918, 184 e 876 mg dm-3, respectivamente.

The experiment was carried out in greenhouse at the FCAV/Unesp, Jaboticabal-SP during the period of November 2005 to January 2007. The objective of this study was to evaluate development components and nutritional status of seedlings of 'Valencia' (Citrus sinensis Osbeck) sweet orange grafted on 'Cravo' lemon rootstock (Citrus limonia Osbeck), in response to Nitrogen, Phosphorus, and Potassium levels. The experimental design was entirely randomized, in factorial scheme 3³ + 1, being 3 factors (Nitrogen, Phosphorus, and Potassium -NPK), 3 levels and a control (without fertilization) with 3 replications. The experimental unit constituted of a 'Valencia' sweet orange grafted on 'Cravo' lemon rootstock in a plastic bag with 2,5 kg of substratum. The treatments consisted of half, one, and two times the standard recommended dose for production of orange seedlings (4,590, 920, and 4,380 mg bag-1 of N, P, and K, respectively). Fertilization with N and K was carried out by three weekly fertirrigations, and the P was added to the substratum of Pinus spp. bark and vermiculite before sowing. After 424 days, the plants were subdivided in roots and aerial parts for determination of dry matter, height, leaf area, diameter of stem, and nutrient content. Fertilization with N, P, and K provided for greater development and greater accumulation of these macronutrients in the aerial part and root of the 'Valencia' sweet orange grafted on 'Cravo' lemon rootstock. There was adequate development of the plants with half the recommended dose of N, P, and K according to literature, approximately of 918, 184, and 876 mg dm-3 of N, P, and K, respectively. Satisfactory development was obtained with half the recommended dose of N, P and K, in general. Slightly greater development occurred in the recommended dose of 4,590, 920, and 4,380 mg bag-1 or 918, 184, and 876 mg dm-3 of substratum, respectively.

Proteção da gema e épocas de forçamento da brotação na enxertia da lima ácida 'Tahiti' / Bud protection and timing of bud break treatments on scion, in grafting of Tahiti lime

A produção de mudas de citros em recipientes é uma tecnologia recente, portanto o sistema de produção encontra-se em aperfeiçoamento, havendo necessidade de informações em várias etapas do sistema produtivo, entre elas os métodos de forçamento da brotação pós-enxertia. Objetivou-se, neste trabalho, estudar o efeito da cobertura das gemas com fitas plásticas e da época de forçamento da brotação sobre o pegamento e desenvolvimento de enxertos da lima ácida 'Tahiti'. O porta-enxerto utilizado foi o limoeiro 'Cravo' (Citrus limonia Osbeck), oriundo da germinação de sementes, e a variedade copa a limeira ácida 'Tahiti' (Citrus latifolia Tanaka). Foram testadas épocas de forçamento de brotação da enxertia (10, 15, 20, 25, 30 e 35 dias após a realização da enxertia) e formas de proteção do enxerto (gema coberta e gema exposta). Após a enxertia as plantas foram mantidas em casa de vegetação com esquema de irrigação por gotejamento. O delineamento experimental foi em blocos casualizados, com cinco repetições e 3 plantas por parcela. Não houve interação entre os fatores estudados, sendo que o índice de pegamento dos enxertos não foi influenciado pelos tratamentos e a cobertura total da borbulha pela fita plástica favoreceu a dormência da gema. O forçamento da brotação do enxerto de lima ácida 'Tahiti', utilizando o método de curvamento do porta-enxerto apresentou os melhores resultados quando esse foi realizado aos 10 dias após a enxertia.

The production of citrus nursery trees in portable containers is a new developing technology and requires investment in research to acquire information on various stages of their production, including the methods to force post grafting bud break. The aim of this work was to investigate the effect of the coverage of buds with plastic tapes as well as the time to force bud break on the establishment and development of 'Tahiti' lime scions. The rootstock used was the 'Rangpur' lime (Citrus limonia Osbeck), obtained from the germination of seeds, while the lime variety 'Tahiti' (Citrus latifolia Tanaka) was used as the scion. The treatments tested were buds covered with plastic tape and uncovered buds at 10, 15, 20, 25, 30, and 35 days after the completion of the budding. After budding, the plants were kept in a greenhouse with dripping irrigation. The experimental design was the randomized blocks, with five replicates and three plants per plot. There was no interaction among the factors studied, while the index of positive grafting was not influenced by the treatments and total coverage of the buds with plastic tape encouraged dormancy of the buds. The bud break of 'Tahiti' lime using the method of bending showed the best results when applied 10 days after grafting.

Aproveitamento de plântulas de porta-enxertos cítrico oriundas do desbaste e seu desenvolvimento vegetativo inicial / Utilization of rootstock citrus seedlings deriving of thinning and its initial vegetative development

Avaliou-se o desenvolvimento vegetativo inicial de porta-enxertos cítricos cultivados em casa de vegetação, por meio de semeadura e de repicagem de raiz nua. O experimento foi conduzido em casa de vegetação, na Estação Experimental Agronômica da UFRGS, em Eldorado do Sul, RS, a partir do mês de setembro de 2003, afim de testar a viabilidade de aproveitamento de plântulas oriundas do desbaste. O delineamento experimental adotado foi o de blocos casualizados, com quatro repetições, em esquema de parcela subdividida e cada subparcela constituída por 22 tubetes. Nas parcelas principais foram testados dois sistemas de propagação: semeadura e repicagem no momento do desbaste. Nas subparcelas foram testados três porta-enxertos cítricos: Trifoliata (Poncirus trifoliata [L.] Raf.), citrangeiro "C37" (P. trifoliata x Citrus sinensis [L.] Osbeck.) e limoeiro "Cravo" (C. limonia Osbeck.). A semeadura foi realizada em tubetes cônicos de polietileno preto (capacidade de 120cm³), com duas a três sementes por tubete. A repicagem foi feita no momento do desbaste da bandeja semeada quando as plântulas estavam, em média, com duas a quatro folhas. Os resultados demonstraram que há viabilidade de aproveitamento das plântulas resultantes do desbaste, sem que ocorra diferenças no desenvolvimento vegetativo. Além disso, foi observado que o porta-enxerto "C37" apresentou um vigor inicial maior que o Trifoliata e o limoeiro "Cravo".

The initial vegetative growth of citrus seedling rootstocks was evaluated by means of sowing and transplantation of bare-root seedling. The experiment was conduced in September 2003 in greenhouse conditions in the Estação Experimental Agronômica/UFRGS in Eldorado do Sul, Brazil. The objective of this study was to test the viability of use of seedlings deriving from thinning. The experiment was in randomized blocks with four replications in a split plot design and each sub-plot consisted of 22 tubs. In the main plot were tested two propagation systems: sowing and transplantation at the thinning moment. In sub-plots were tested three citrus rootstocks: Trifoliate orange (Poncirus trifoliata [ L. ] Raf.); 'C37' citrange (P. trifoliata x Citrus sinensis [ L. ] Osbeck.), and 'Rangpur' lime (C. limonia Osbeck.). The sowing was carried in conical tubs of black polyethylene (capacity of 120cm³) with two to three seeds in each tub. Transplantation was made at the thinning moment, when the seedlings had, in average, two to four leaves. The results demonstrate that it is possible to use seedlings resulting from thinning, without differences in the vegetative development comparatively to sowing. The 'C37' citrus rootstock presents a larger initial vigor when compared to the other citrus rootstocks tested.