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Citrus (Citrus reticulata) is one of the world’s most widely planted and highest-yielding fruit trees. Citrus fruits are rich in a variety of nutrients. The content of citric acid plays a decisive role in the flavor quality of the fruit. There is a high organic acid content in early-maturing and extra-precocious citrus varieties. Reducing the amount of organic acid after fruit ripening is significant to the citrus industry. In this study, we selected a low-acid variety, “DF4”, and a high-acid variety, “WZ”, as research materials. Through WGCNA analysis, two differentially expressed genes, citrate synthase (CS) and ATP citrate-pro-S-lyase (ACL), were screened out, which related to the changing citric acid. The two differentially expressed genes were preliminarily verified by constructing a virus-induced gene-silencing (VIGS) vector. The VIGS results showed that the citric acid content was negatively correlated with CS expression and positively correlated with ACL expression, while CS and ACL oppositely control citric acid and inversely regulate each other. These results provide a theoretical basis for promoting the breeding of early-maturing and low-acid citrus varieties.

【目的】本研究では、鹿児島県産柑橘3種類（紅甘夏、河内晩柑、吹上小夏）を用いて果皮量10％及び20％の2種の果実酒を調製し、調製後の変化を経時的に探ることを目的とした。 【方法】果実酒は果実、果皮、氷砂糖、ホワイトリカーを用いて調製した。果実酒の漬け込み後4週間目に果皮と果実を取り出し、その後も20週目まで色差、酸度、糖度、pHの測定を行った。果実酒から取り出した果実および果皮は凍結乾燥後に35％、70％エタノールで抽出し、果実酒とともに総ポリフェノール量及びDPPHラジカル捕捉能の測定を行った。 【結果】果実酒の色差、酸度、糖度、pHを測定したところ、果皮20％使用果実酒では10％果実酒と比べL*、a*、b*値が高くなったが、酸度、糖度、pHは果皮の量による差はほとんどみられなかった。漬け込み後4週目まで、L*、a*、b*値及び酸度の上昇と糖度の減少が見られたが、4週目で果実及び果皮を取り出した後は、ほぼ横ばいとなった。果実酒中の総ポリフェノール濃度はいずれの柑橘においても果皮20％使用果実酒では10％果実酒と比べ多く、4週目以降は横ばいとなった。DPPHラジカル捕捉能については、総ポリフェノール濃度と同様に果皮20％使用果実酒では10％果実酒と比べ高くなり、4週目以降は横ばいまたは減少が見られた。, Purpose : The purpose of this study was to prepare two types of fruit wine with 10% and 20% peel using three types of citrus from Kagoshima Prefecture (Beni-amatsu, Kawachi-bankan, and Fukiage-konatsu), and to explore changes over time after preparation. Methods：Fruit wine was prepared using fruit, fruit peel, glacial sugar, and white liquor. The peels and fruit were removed from the fruit wine at 4 weeks after pickling, and the color difference, acidity, sugar content, and pH were measured thereafter until the 20th week. Fruits and peels removed from fruit wine were freeze-dried and extracted with 35% and 70% ethanol, and the total polyphenol content and DPPH radical scavenging capacity were determined along with the fruit wine. Results：The fruit wine with 20% fruit peel showed higher L*, a*, and b* values than the wine with 10% fruit peel, but the acidity, sugar content, and pH showed little difference due to the amount of fruit peel. L*, a*, and b* values and acidity increased and sugar content decreased until the fourth week after pickling, but remained almost unchanged after the fruit and peel were removed at the fourth week. The total polyphenol concentration in the fruit wine was higher in fruit wine with 20% fruit peel than in fruit wine with 10% fruit peel for all citrus, and remained unchanged after the fourth week. The DPPH radical scavenging capacity of fruit wine with 20% fruit peel was higher than that of wine with 10% fruit peel and remained flat or decreased after the 4th week.

Pectin is a polysaccharide with wider applications in foods, pharmaceuticals, and several of other industries. The extraction and determination of vitamins from pectin obtained from citrus fruit peels were studied. The pectin from citrus peels was extracted using ethanol. It was observed that pectin from orange peels gave the highest yield (28.31±2.85%) followed by red grape (14.72±1.83%), lime (12.10±0.97%), and the least in lime (10.15±0.61%).The level of vitamin A (mg/g) was high in pectin from red grape (1483.637±10.420) followed by orange peels (1156.301±8.840), lime peels (475.099±5.510), and the least in white grape (283.228±3.770). There was a significant difference (p≤0.05) in the level of vitamin A content in all the extracted pectin. The vitamin B2(mg/g) content was highest in white grape (0.183±0.050) followed by orange (0.150±0.040), red grape (0.110±0.030), and the least value in lime (0.083±0.020) respectively. There was no significant difference (p≤0.05) in the level of vitamin B2pectin samples. The vitamin B12(mg/g) content was highest in white grape (3.300±1.130) followed by lime (2.931±1.100), orange (1.900±0.930), and the least value with Red grape (0.888±0.870). There was no significant difference in the level of vitamin B12between pectin from lime and white grape. The increasing order of vitamin C (mg/g) content in the extracted pectins was as follows: white grape (11.706±1.150)

Colletotrichum species are well-known plant pathogens, saprobes, endophytes, human pathogens and entomopathogens. However, little is known about Colletotrichum as endophytes of plants and cultivars including Citrus grandis cv. “Tomentosa”. In the present study, 12 endophytic Colletotrichum isolates were obtained from this host in Huazhou, Guangdong Province (China) in 2019. Based on morphology and combined multigene phylogeny [nuclear ribosomal internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (gapdh), chitin synthase 1 (chs-1), histone H3 (his3) actin (act), beta-tubulin (β-tubulin) and glutamine synthetase (gs)], six Colletotrichum species were identified, including two new species, namely Colletotrichum guangdongense and C. tomentosae. Colletotrichum asianum, C. plurivorum, C. siamense and C. tainanense are identified as being the first reports on C. grandis cv. “Tomentosa” worldwide. This study is the first comprehensive study on endophytic Colletotrichum species on C. grandis cv. “Tomentosa” in China.

The role of agroforest systems in pathogen regulation through structural characteristics such as shade and tree diversity is recognized. However, few studies have examined the importance of these factors on the spread of diseases of associated fruit trees in agroforestry systems, especially cocoa-based agroforestry systems (CBAS). The aim of this study was to evaluate the effect of associated tree diversity and shade rate into CBAS on citrus Phytophthora foot rot disease (PFRD) intensity. The study was carried out in five citrus production basins of Cameroon, contrasted by ecology and rainfall regimes. A set of 33 CBAS plots was mapped and their species composition and vertical structure were determined. Field data were used to reconstruct CBAS in 3D with Shademotion 5.1.47 software. Using static simulations with shadow overlays, shade rate received by each citrus tree was calculated. Subsequently, citrus trees were grouped into three categories according to the shade rate they received: (i) full sun, with shade rate 70%. PFRD intensity was determined by measuring the canker extent relatively to the citrus crown circumference. A significant difference in PFRD intensity was observed between the different shade rates. Citrus trees receiving no shade were significantly more attacked by PFRD than those receiving little shade or dense shade. The antagonistic effect of shade was demonstrated. At the expense of factors related to the specific composition of the CBAS, shade, temperature, relative humidity and rainfall were found to be the main factors explaining PFRD intensity in CBAS. This study shows that microclimatic alterations due to the biodiversity in tropical agroforestry systems contribute to sustainable pest management.

Hypothesis: This review article represents a brief layout of the risk factors and pathophysiology responsible for obesity, customary treatment strategies, and nanotechnology-based nutraceutical for therapeutics of obesity. Experiments: An exhaustive search of the literature was done for this purpose, using Google Scholar, PubMed, and ScienceDirect databases. A study of the literature was conducted using publications published in peer-reviewed journals between the years 2000 and 2022. Findings: This was revealed that risk factors responsible for obesity were genetic abnormalities, environmental, and socio-economic factors. Number of research articles published between 2000 and 2022 were based on phytoconstituents based nanoformulation for obesity therapeutics and therefore, have been systematically compiled in this review. Various nutraceuticals like Garcinia cambogia, quercetin, resveratrol, capsaicin, Capsicum, Curcuma longa, Camella Sinensis, Zingiber officinalis, Citrus aurantium, Aegle marmelos, Coffea canephora, Asparagus officinalis, Gardenia jasminoides, Catha edulis, Clusia nemroisa, Rosmarinus officinalis, Cirsium setidens, Betula platyphylla, Tripterygium wilfordi possessing anti-obesity actions are discussed in this review along with their patents, clinical trials as well as their nanoformulation available. Conclusion: This review illustrates that the nanotechnology have a great propensity to impart promising role in the delivery of phytochemicals and nutraceuticals in management of obesity conditions and other related disorders.

Machine learning is a branch of computer science concerned with developing algorithms & models capable of ‘learning through data and iterations’. Deep learning simulates the structure and function of human organs and diseases using artificial neural networks with more than one hidden layer. The primary purpose of this work is to develop and test computer vision and machine learning algorithms for classifying Huanglongbing (HLB)-infected, healthy, and unhealthy leaves and fruits of the citrus plant. The images were segmented using a normalized graph cut, and texture information was extracted using a co-occurrence matrix. The collected attributes were used for classification and support vector machine (SVM), and deep learning methods were employed. When rating the classification outcomes, the accuracy of the classification and the number of false positives and false negatives were considered. The result shows that Deep Learning could create categories up to 96.8% of HLB-infected leaves and fruits. Despite a broad variance in intensity from leaves collected in North India, this method suggests it could be beneficial in diagnosing HLB.

A promising method for preparing metaphase spread for counting the number of chromosomes from the emerging shoot tissue is described in this report. In the present study, we adopted enzymatic digestion of shoot tips to analyse the chromosome number. The chromosomes in metaphase stage of cell division are highly condensed and easy to count in routine cytological technique. Even the morphological features like position of centromere can be seen in metaphase. In prophase it may not be clear as the chromosomes are getting ready for cell division. In enzymatic digestion even the prophase chromosomes are visible, which can be counted. Hence enzymatic digestion technique is more efficient in citrus as compared to acid digestion method as the citrus crop is a perennial crop with small-sized chromosomes. Furthermore, the sample collection in the field was easy and actively growing vegetative flush was available throughout the year. This technique was attempted in the tissue culture lab of ICAR- CCRI in various in vito and in vivo ploidy induction experiments in Citrus sinensis Osbeck (Sweet orange cv. mosambi), C. reticulata Blanco (Nagpur mandarin) and C. jambhiri Lush (Rough lemon), for confirmation of diploidy (2n=2x=18), triploidy (2n=3x=27), tetraploid (2n=4x=36), hexaploid (2n=6x=54).

Asian citrus psyllid (ACP), also known as Diaphorina citri, is the natural vector of Candidatus Liberibacter asiaticus (CLas), which is responsible for Huanglongbing (HLB), a devastating citrus disease. Previously, the pathogen was successfully excluded from diseased citrus plants by using the indigenous endophyte Bacillus subtilis L1-21. However, the pathogen elimination and colonization potential of B. subtilis L1-21 in the carrier vector ACP, as well as the recruitment of native microbial communities of psyllid in the presence of endophytes, are still unknown.Initially, we suggested that endophyte L1-21 reduced the CLas copies in ACP from 6.58 × 10We propose that B. subtilis L1-21 is a native endophyte in citrus and psyllid, which efficiently reduces the CLas pathogen in both citrus and psyllids, provides a more protective effect by increasing the number of cultivable endophytes, and successfully colonizes the midgut of ACP. © 2022 Society of Chemical Industry.

The particle film technology has been reported as a promising tool in pest control. The objective of this work was to evaluate the efficiency of kaolin-based products against the oviposition of South American fruit fly, Anastrepha fraterculus (Dip.:Tephritidae), and its effect on the quality of citrus fruits. The experiment was conducted in orchards of ‘Okitsu’ tangerine and ‘Valencia’ orange trees in the 2017, 2018 and 2019 harvests. The treatments were as follows: 1) kaolin 10% + 0.1% Break-Thru® adjuvant; 2) Surround® 5% WP; 3) 0.15% phosmet (Imidan® 500 WP), 75 g. a.i.; 4) without application (control). The sprays were performed every 21 days. At harvest, fruits were individually packed in a greenhouse for inspection after 25 days and infestation was recorded. Fruit samples were evaluated for average diameter, average mass, soluble solids, titratable acidity and peel colorimetry. Infestation of A. fraterculus in tangerines was reduced in plants treated with the two kaolin-based products in the 2017 harvest. In the 2017 and 2019 crops, Surround® WP reduced the infestation and the number of puparium/fruits in oranges. The mineral films did not alter the physicochemical characteristics of the fruits, representing a promising alternative for the management of A. fraterculus.

Objectives This study investigates whether a citrus and pomegranate complex (CPC) improves physical fitness, mental well-being, and blood biomarkers for oxidative stress and endothelial function in healthy elderly. Design A randomized placebo-controlled cross-over trial. Participants The study included 36 healthy elderly aged 60–75 years old. Intervention and Measurements Participants received four weeks of CPC supplementation and performed the handgrip strength and senior fitness test. Quality of life (QOL) was assessed and blood samples were analyzed for oxidative stress and endothelial function markers. Results After four weeks of CPC supplementation, handgrip strength significantly improved (p=0.019), compared to placebo. Moreover, the thinking, memory, learning, and concentration facets were improved (p=0.042), compared to placebo, and plasma malondialdehyde decreased, compared to placebo (p=0.033). The intervention did not affect senior fitness and the other QOL domains and blood parameters. Conclusion Four weeks of daily CPC supplementation significantly improves handgrip strength and self-evaluated measures of psychological function in healthy older adults. Further research should focus on mechanisms associated with physical performance.

Resumen Introducción. El psílido asiático (Diaphorina citri Kuwayama) es el principal vector de la bacteria Candidatus Liberibacter asiaticus que causa la enfermedad de Huanglongbing (HLB) en los cítricos en Panamá. Objetivo. Determinar la distribución geográfica y las plantas hospederas del psílido asiático dentro de las zonas citrícolas de Panamá. Materiales y métodos. El estudio se desarrolló de junio a octubre de 2021, a través de un muestreo sistemático dentro de 46 localidades asociadas a cítricos comerciales y huertos familiares en siete provincias de Panamá. En cada planta se procedió a embolsar con una bolsa plástica de cierre mágico dos brotes nuevos de ≈10 cm de largo para capturar los especímenes e introducirlos dentro de viales plásticos de 2 mm con alcohol al 95 % para su posterior identificación. La variable de estudio incluyó el conteo del número total de especímenes y los datos fueron analizados de manera porcentual. Resultados. Del total de 73 muestras colectadas, la presencia del psílido asiático se confirmó en un 21,92 % (dieciséis muestras), resultó positiva su presencia en las provincias de Chiriquí, Coclé, Herrera, Panamá y Panamá Oeste, en alturas desde los 17 hasta 1080 m s. n. m. No hubo presencia del psílido en las provincias de Los Santos y Veraguas. De las dieciséis muestras que resultaron positivas, el 50 % pertenecieron a la planta de mirto, seguida por la naranja (18,75 %) y el limón criollo (12,50 %). Conclusiones. La distribución del psílido asiático vector del HLB en parcelas de cítricos comerciales y de huertos familiares en las principales zonas citrícolas en Panamá, se presentó diferentes alturas sobre el nivel del mar, asociado a la presencia de plantas hospederas como mirto, naranja y limón criollo. Abstract Introduction. The asian psyllid (Diaphorina citri Kuwayama) is the main vector of the bacterium Candidatus Liberibacter asiaticus that causes Huanglongbing (HLB) disease in citrus in Panama. Objective. To determine the geographic distribution and host plants of the asian psyllid within the citrus growing areas in Panama. Materials and methods. The study was carried out from June to October 2021 through systematic sampling within 46 localities associated with commercial citrus and home gardens in seven provinces of Panama. In each plant, two new shoots of 10 cm long were bagged with a magic-seal plastic bag to capture the specimens and introduce them into 2 mm plastic vials with 95 % alcohol for later identification. The study variable included the count of the total number of specimens, and the data was analyzed as a percentage. Results. Of the total of 73 collected samples, the presence of the Asian psyllid was confirmed in 21.92 % (16 samples). Its presence was positive in the provinces of Chiriquí, Coclé, Herrera, Panama, and West Panama, at altitudes of 17 to 1080 m.a.s.l. There was no presence of the psyllid in the provinces of Los Santos and Veraguas. Of the 16 samples that were positive, 50 % belonged to the myrtle plant, followed by orange (18.75%), and creole lemon (12.50 %). Conclusion. The distribution of the asian psyllid vector of HLB in commercial citrus and home gardens in the main citrus growing areas in Panama was presented at different heights above sea level, mainly associated with the presence of the host plants such as myrtle, orange, and creole lemon.

Resumen Introducción. El cultivo de naranja (Citrus sinensis (L.) Osb.) es uno de los de mayor importancia a nivel mundial. Dentro de las principales limitantes en su producción está la caída de flores ocasionada por Colletotrichum. Objetivo. Determinar las especies de Colletotrichum asociadas a la antracnosis en naranja y estimar la efectividad in vitro de cuatro fungicidas sobre su control. Materiales y métodos. Este estudio se llevó a cabo entre junio del 2021 y febrero del 2022. Seis aislamientos de hongos, obtenidos de tejidos florales de naranja, fueron identificados vía reacción en cadena de la polimerasa (PCR). Se evaluó la eficacia de los fungicidas tebuconazol, pyraclostrobin, difenoconazol y ferbam sobre dos aislamientos de Colletotrichum. Se realizaron dos ensayos para determinar el grado de inhibición del crecimiento micelial y el porcentaje de efectividad de cada fungicida. Resultados. Colletotrichum abscissum se identificó en pétalos con síntomas y Colletotrichum gloeosporioides en cálices. Se verificó la efectividad de la dosis empleada de los fungicidas tebuconazol y difenoconazol, al presentar inhibición del crecimiento micelial a partir de la décima parte de la dosis comercial evaluada. El pyraclostrobin demostró poco control sobre ambos aislados. El ferbam no presentó un control eficiente en ninguna de las dosis evaluadas en ambas cepas. Conclusión. Los hongos asociados a la antracnosis en naranja, en la finca estudiada, fueron C. abscissum, presente en pétalos con síntomas iniciales y avanzados, y C. gloeosporioides presente en cálices persistentes. La efectividad de los fungicidas evaluados fue mayor para los triazoles, tebuconazol y difenoconazol, en ese orden. El pyraclostrobin evidenció un control deficiente sobre ambos aislados. El ferbam resultó poco eficaz en las pruebas in vitro. Abstract Introduction. Orange (Citrus sinensis (L.) Osb.) cultivation is one of the most important worldwide. Among the main limitations in its production is the fall of flowers caused by Colletotrichum. Objective. To determine the species of Colletotrichum associated with anthracnose in orange and to estimate the in vitro effectiveness of four fungicides on its control. Materials and methods. This study was carried out between June 2021 and February 2022. Six fungal isolates, obtained from orange floral tissues, were identified via polymerase chain reaction (PCR). The efficacy of the fungicides tebuconazole, pyraclostrobin, difenoconazole, and ferbam on two Colletotrichum isolates was evaluated. Two tests were carried out to determine the degree of inhibition of mycelial growth and the percentage of effectiveness of each fungicide. Results. Colletotrichum abscissum was identified on petals with symptoms and Colletotrichum gloeosporioides on calyxes. The effectiveness of the doses used of the fungicides tebuconazole and difenoconazole was verified by presenting inhibition of mycelial growth from a tenth of the commercial dose evaluated. Pyraclostrobin showed little control over both isolates. Ferbam did not present an efficient control in any of the doses evaluated in both strains. Conclusion. The fungi associated with anthracnose in orange, on the farm studied, were C. abscissum, present on petals with initial and advanced symptoms, and C. gloeosporioides present on persistent calyxes. The effectiveness of the fungicides evaluated was higher for triazoles, tebuconazole, and difenoconazole, in that order. Pyraclostrobin showed a poor control on both isolates. Ferbam was not effective in in vitro tests.

Gibberellins can promote the inhibition of floral induction in citrus in order to reduce alternate bearing. However, not much is known about the responsiveness of Citrus deliciosa to exogenous applications of gibberellic acid. Furthermore, the successive application of gibberellins is barely studied. The main objective of this work was to evaluate the effect of successive applications of gibberellic acid (GA3) on the reduction of spring flowering subsequent to low fruit load periods (off-year) in ‘Montenegrina’ and ‘Rainha’ mandarin trees, in southern Brazil. The treatments consisted of one to four successive applications of GA3 (40 mg L-1) with 21-day interval in off-year trees. Applications began in May and ended in July. There was a significant reduction in sprouting and flowering of ‘Montenegrina’ mandarin with more than two GA3 successive applications compared to the control. To ‘Rainha’ mandarin, only two GA3 applications reduced the sprouting and solely one GA3 application was enough to reduce sprouting in relation to the control. GA3 use reduced sprouting and flowering of Montenegrina and Rainha cultivars. The orchard age may be related to its level of alternate bearing and, therefore, to the greater or lesser sensitivity of gibberellin applications. Resumo As giberelinas podem promover a inibição da indução floral em citros com intuito de reduzir a alternância de produção. Contudo, pouco se sabe sobre a responsividade de Citrus deliciosa às aplicações exógenas de ácido giberélico. Além disso, a aplicação sucessiva de giberelinas é pouco estudada. Desta forma, objetivou-se avaliar o efeito de aplicações sucessivas de ácido giberélico (AG3) sobre a redução do florescimento primaveril subsequente a safras de baixa carga de frutos (ano off) em tangerineiras ‘Montenegrina’ e ‘Rainha’, no Sul do Brasil. Os tratamentos consistiram em uma a quatro aplicações sucessivas de 40 mg L-1 de AG3, com intervalos de 21 dias, em plantas em ano off. O início das aplicações foi em maio, e o término, em julho. Houve redução significativa de brotação e de florescimento com mais de duas aplicações sucessivas de AG3 para a tangerineira ‘Montenegrina’ em relação ao controle. Para a tangerineira ‘Rainha’, apenas com duas aplicações de AG3, o florescimento foi reduzido, enquanto apenas uma aplicação já foi suficiente para reduzir a brotação em relação ao controle. O uso de AG3 reduz brotação e florescimento das cultivares Montenegrina e Rainha. A idade do pomar pode estar relacionada ao seu nível de alternância produtiva e, desta forma, à maior ou à menor sensibilidade a aplicações de giberelinas.

Citrus species have a “wet”-type stigma with abundant exudate, and their style contains numerous canals. For successful seed formation, the process includes pollen grain germination on the stigma, pollen tube development and growth through the stigma and style and, ultimately, successful ovule fertilization. However, preventing the fertilization process can be useful for many agronomic and plant-breeding purposes, such as seedless fruits or for developing new varieties. Several studies have recently shown the inhibition effect of sulfur on pollen tube development inside the Nadorcott mandarin stigma and its effective application to obtain seedless mandarins. However, when applied to the stigma, how can sulfur inhibit pollen tube growth? Moreover, does sulfur have the same effect on other species? The main objective of the present study is to clarify the mode of action of sulfur on the ultrastructure of the Nadorcott mandarin stigma and style. To fulfill this goal, untreated flowers and flowers treated with sulfur were pollinated 24 h later. The treated and untreated stigmas were analyzed and compared with several microscopy techniques. The main results showed that sulfur specifically caused an alteration to the outer layer of stigma papillary cells. This marked alteration resulted in papillary cells losing their functionality due to the deterioration and degradation of their cellular structure. Basal papillae, the stigmatic tissue and stylar canals also underwent major alteration. Sulfur also modified the quantity and uniformity distribution of the stigmatic exudate. All these alterations collectively prevented pollen tube development inside the stigma. These effects have been observed in several Citrus species and varieties, and in some other horticultural species, which suggests a generic (non species-specific) action.

Interstocks have been used in fruit tree cultivation to regulate tree size and improve fruit production and quality. In this study, several Huanglongbing (HLB)-tolerant open-pollinated pummelo interstock candidates were evaluated as interstocks between the Swingle rootstock and the ‘Valencia’ scion, with Swingle serving as the control interstock. After 5 years in the field, most trees did not exhibit visual HLB symptoms, although the trees were infected with HLB, and the CaLas Ct values in the ‘Valencia’ leaves of the different interstock treatments ranged between 25.88 and 27.82. Although the foliar chlorophyll content among the interstock treatments was not highly significant (p-value = 0.0313), the foliar starch content was significantly different (p-value = 0.0018). ‘Valencia’ grafted onto 5-1-99-3 and HBJL-4 interstocks (both open pollinated seedlings of the Hirado Buntan pummelo) exhibited the highest total phenolic compound (TPC) levels (46.44 and 46.36 mg gallic acid g−1 FW). Transcripts of CsPR1 and CsPR2, two pathogenesis-related (PR) genes, were upregulated in ‘Valencia’ grafted onto open pollinated seedling selections of the red shaddock pummelo, Liang Ping Yau pummelo, and Hirado Buntan pummelo compared with ‘Valencia’ grafted onto Swingle. All interstocks influenced the tree growth rate and improved canopy volume in the field compared to the control trees without any interstocks (p-value = 0.0085). The 5-4-99-7 (red shaddock pummelo) and 8-1-99-1B (Liang Ping Yau pummelo) interstock trees had the highest canopy volume among all the treatments. We propose, based on our current results, that HLB-tolerant citrus accessions, when judiciously used as interstocks, may enhance plant defense and provide increased HLB tolerance to susceptible scions.

In this study, the DEA–Malmquist index method was used to measure the total factor productivity of citrus in seven major mandarin-producing provinces and seven major tangerine-producing provinces in China from 2006 to 2020. Moran’s I index was used to test the spatial correlation of total factor productivity of mandarin and tangerine, and its σ convergence and β convergence characteristics were explored using coefficient of variation and spatial panel models. The results show that from the perspective of time series evolution, the growth rate of total factor productivity of mandarin and tangerine in China slowed down year by year after reaching the maximum value in 2008. Technological progress was the main factor affecting the total factor productivity of citrus. The total factor productivity growth of tangerine was more stable than that of mandarin, and the pure technical efficiency index and scale efficiency change index of mandarin and tangerine were not stable. From the perspective of regional differences, the total factor productivity of China’s main citrus-producing provinces all indicated positive growth, showing an increasing trend from east to west. The drivers of growth were mainly technological progress and scale efficiency. The regional differences in total factor productivity growth for mandarin were more obvious than for tangerine. The total factor productivity of mandarin and tangerine showed obvious spatial correlation characteristics; the positive spatial spillover effect was significant; and there were σ convergence, absolute β convergence, and conditional β convergence. Regional disparities in citrus industry development can be more objectively reflected by convergence analysis that takes spatial factors, economic and social factors, and other factors into account.

A precise estimation of plant water use is crucial for efficient irrigation management and understanding of plant–water interactions. Metalized-polyethylene mulch as a ground cover, combined with irrigation management, can improve water and fertilizer use, efficiency growth, and yield of young citrus trees, compared to bare ground. Thus, a study was undertaken to (i) compare daily water use, using the stem heat balance (SHB) method on ≤4-year-old citrus trees using three irrigation methods, and (ii) to determine soil moisture content, as well as total available soil water (TASW) in the irrigated zone. Three irrigation methods, regulated deficit irrigation (RDI), conventional drip irrigation (CD), and microsprinkler irrigation (MS), were applied on two sandy soils in central and southwest Florida with treatments further split between trees grown on 2.4 m mulched beds (M) and bare ground (NM) to form a factorial design. Measurements included sap flow, canopy size, leaf area index (LAI) and soil moisture. The result of this study shows greater values for the trunk cross-sectional area, canopy volume and LAI for both mulch treatments, in comparison to bare ground treatments. The sap flow data suggest that RDI resulted in higher water use than both CD and MS. The hourly sap flow was 120, 99 and 163% greater in M-RDI than M-CD, NM-CD and NM-MS, respectively. Mulched ground cover showed a higher soil moisture average at all soil layers (8-, 15- and 45-cm depth). For instance, the mulched ground cover showed higher soil moisture (37%) at a 15 cm depth at the Entisol site, followed by 30% at a 45 cm depth and 25.3% at depth 45 cm for the Spodosol site. The TASW values varied between 100 to 136% for both mulch and bare ground treatments. The irrigation systems used showed water contents close to field capacity, indicating that water was not limiting despite the different irrigation schedules. The high-water uptake is ascribed to frequent irrigation applications and pulses, and improved water distribution in the irrigated zone as a result of the mulch. These findings demonstrate the potential of metalized-polyethylene mulch and regulated drip irrigation to enhance water use efficiency.

Although the phloem is a highly specialized tissue, certain pathogens, including phytoplasmas, spiroplasmas, and viruses, have evolved to access and live in this sequestered and protected environment, causing substantial economic harm. In particular, Candidatus Liberibacter spp. are devastating citrus in many parts of the world. Given that most phloem pathogens are vectored, they are not exposed to applied chemicals and are therefore difficult to control. Furthermore, pathogens use the phloem network to escape mounted defenses. Our review summarizes the current knowledge of phloem anatomy, physiology, and biochemistry relevant to phloem/pathogen interactions. We focus on aspects of anatomy specific to pathogen movement, including sieve plate structure and phloem-specific proteins. Phloem sampling techniques are discussed. Finally, pathogens that cause particular harm to the phloem of crop species are considered in detail.

Cottony cushion scale, Icerya purchasi Maskell, is an occasional pest of citrus, especially when insecticides disrupt vedalia beetle, Novius cardinalis (Mulsant) (Coleoptera: Coccinellidae). In two field tests conducted in successive years, navel orange trees were artificially infested with I. purchasi to determine the impact of I. purchasi on fruit yield and quality. In the first year, for which adult scale densities ranged between 0 and > 45 per branch and between 0 and > 500 on trunk counts, there was a highly significant negative linear relationship between the number of I. purchasi adults on branches or the trunk and its effect on fruit number, the percentage of large fruit, and the mass of fruit, and a highly significant positive relationship for the percentage of sooty mold-affected fruit and the percentage of juice grade fruit. In the second year, in which adult scale densities ranged between 0 and 10.5 per branch and between 0 and 35 on trunk counts, only the percentage of sooty mold-affected fruit showed a relationship. Damage to fruit was better predicted by population estimates from branch samples versus trunk counts. Analysis of the two years, estimated significant damage ranging from 2 adults per branch to no detectable effects of insect density, depending on the given metric of fruit number or quality, at the time that treatment decisions would be made. These results support May–June monitoring of branches for adult scales and application of treatments at an action threshold of

As a waste material, the amazing potential of cellulose nanocrystals (CNCs) isolated from Citrus medica L. pericarp in being a natural resource of lingo-cellulosic products has never been investigated before. In the present study, an alkaline pretreatment and a two-step bleaching procedure were applied to conduct the desired acid hydrolysis by the usage of 64% sulfuric acid at 50°C for 105 min. The extracted CNCs were distinguished through the means of transmission electron microscopy (TEM), field emission scanning electron microscopy (FESEM), X-ray diffraction (XRD), atomic force microscopy (AFM), Fourier-transform infrared (FTIR) spectroscopy, dynamic light scattering (DLS), zeta potential, and energy-dispersive X-ray spectroscopy (EDX). The elimination of peaks, which were accountable for the inducement of hemicelluloses and lignin, was confirmed by the FTIR results and were also validated by the outcomes of XRD that proved the gentle removal of non-cellulosic components. The morphology and size of CNCs were indicated through the FESEM and TEM results. In addition, the spherical forms of synthesized CNCs were observed with a diameter of 46 nm throughout the FESEM images, while displaying a value of 42.54 nm as well due to TEM micrographs. The obtained zeta potential displayed a reasonable negative surface charge for CNCs. Furthermore, the cytotoxicity assessment of this product on fibroblast cells was performed to study their susceptibility for bio-medical and cosmetic industrial applications, which resulted in the lack of exhibiting any cytotoxic effects. In conformity to the outcomes of TEM and FESEM, the results of AFM revealed a fine dispersion and spherical form of cellulose nanoparticles. The interaction between HMG-CoA reductase and CNC was studied by the usage of multi-spectroscopic methods and enzyme kinetics to explore the binding mechanism of HMG-CoA reductase-CNC system. Reduced catalytic activity of the occurrence of changes in the secondary structure of HMG-CoA reductase was as a result of interacting with CNC causing a reduction in its catalytic activity.

Citrus plants are prone to infection by different viroids which deteriorate their vigor and production. Citrus viroid V (CVd-V) is among the six citrus viroids, belongs to genus Apscaviroid (family Pospiviroidae) which induces symptoms of mild necrotic lesions on branches and cracks on trunk portion.A survey was conducted to evaluate the prevalence of CVd-V in core and non-core citrus cultivated areas of Punjab, Pakistan. A total of 154 samples from different citrus cultivars were tested for CVd-V infection by RT-PCR. The results revealed 66.66% disease incidence of CVd-V. Citrus cultivars Palestinia Sweet lime, Roy Ruby, Olinda Valencia, Kaghzi lime, and Dancy were identified as new citrus hosts of CVd-V for the first time from Pakistan. The viroid infection was confirmed by biological indexing on indicator host Etrog citron. The reported primers used for the detection of CVd-V did not amplify, rather showed non-specific amplification, which led to the designing of new primers. Whereas, new back-to-back designed primers (CVd-V AF1/CVd-V AR1) detected CVd-V successfully and obtained an expected amplified product of CVd-V with 294 bp. Sequencing analysis confirmed the new host of CVd-V showing 98-100% nucleotide sequence homology with those reported previously from other countries while 100% sequence homology to the isolates reported from Pakistan. Based on phylogenetic analysis using all CVd-V sequences in GenBank, two main CVd-V groups (I and II) were identified, and newly identified isolates during this study fall in the group I.The study revealed that there are some changes in the nucleotide sequences of CVd-V which made difficult for their detection using reported primers. All isolates of Pakistan showed high sequence homology with other isolates of CVd-V from Iran and USA whereas; the isolates from China, Japan, Tunisia, and Africa are distantly related. It is evident that CVd-V is spreading in all citrus cultivars in Pakistan.

Linarin (acacetin-7-O-rutinoside), isorhoifolin (apigenin-7-O-rutinoside), and diosmin (diosmetin-7-O-rutinoside) are chemically and structurally similar flavone rutinoside (FR) compounds found in Chrysanthemum L. (Anthemideae, Asteraceae) plants. However, their biosynthetic pathways remain largely unknown. In this study, we cloned and compared FRs and genes encoding rhamnosyltransferases (RhaTs) among eight accessions of Chrysanthemum polyploids. We also biochemically characterized RhaTs of Chrysanthemum plants and Citrus (Citrus sinensis and Citrus maxima). RhaTs from these two genera are substrate-promiscuous enzymes catalyzing the rhamnosylation of flavones, flavanones, and flavonols. Substrate specificity analysis revealed that Chrysanthemum 1,6RhaTs preferred flavone glucosides (e.g. acacetin-7-O-glucoside), whereas Cs1,6RhaT preferred flavanone glucosides. The nonsynonymous substitutions of RhaTs found in some cytotypes of diploids resulted in the loss of catalytic function. Phylogenetic analysis and specialized pathways responsible for the biosynthesis of major flavonoids in Chrysanthemum and Citrus revealed that rhamnosylation activity might share a common evolutionary origin. Overexpression of RhaT in hairy roots resulted in 13-, 2-, and 5-fold increases in linarin, isorhoifolin, and diosmin contents, respectively, indicating that RhaT is mainly involved in the biosynthesis of linarin. Our findings not only suggest that the substrate promiscuity of RhaTs contributes to the diversity of FRs in Chrysanthemum species but also shed light on the evolution of flavone and flavanone rutinosides in distant taxa.

This study aimed to investigate the effects of foliar application of different potassium (K) sources on yield and fruit nutrient uptake in 28-year-old Valencia oranges (Citrus sinensis, L.) grafted onto Trifoliata citrange rootstock. In the trial that was conducted with different foliar potassium sources in the region of Koycegiz (Turkey) in the years 2016, 2017, and 2018 with 5 replications according to the random blocks trial design for this purpose, in addition to the control group where no foliar treatment was applied, the treatment groups received Potassium Nitrate (KNO3, 46% K2O; 300 g/100L/da), Potassium Thiosulfate (25% K2O; 400 ml/100L/da) and Potassium Phosphite (25% K2O; 500 ml/100 L/da) treatments. For all examined parameters and for all 3 years, the highest results were obtained in the potassium phosphite treatment. This may be due to its higher mobility inside the plant compared to other K sources. The effects of all treatments on yield were found significant, where the yield values for 2016, 2017, and 2018 were found as 139, 143, and 140 kg/tree, respectively. This may be explained with the role of K on promoting the fruit reserves. Regarding the effects of potassium phosphite on all fruit nutrient uptake amounts, the effect of the treatment in all 3 years was found significant except for Fe, Cu, and Mn. In conclusion, among the potassium sources, the most effective fertilizer was determined as potassium phosphite.

Aluminum (Al) toxicity has conspicuous detrimental effects on citrus production whereas boron (B) has been shown to alleviate its toxicity. Lignin plays a critical role in the cell wall extensibility and root elongation under stressed conditions. Hence, the interaction between B and Al on cell wall structure and lignin-related metabolic pathway was investigated in root of trifoliate orange (Poncirus trifoliata (L.) Raf.) seedlings. The results showed B supply considerably decreased the Al content in root, particularly in cell wall, and reduced Al-induced damage on growth-related parameters and thickness of cell wall. Boron application decreased the hydrogen peroxide (H

We previously discovered that the SPC/Fyn/Rho-kinase (ROK) pathway mediates the Ca

Citrus rootstock breeding is important to improve their resistance to diseases, pests and adverse environmental conditions. The majority of citrus species have nuceller embryony. Although Clementine mandarin is monoembryonic, in some cases (intercrossing etc.) abortive seed formation can be seen frequently. In this study, Clementine mandarin × Carrizo citrange were crossed. In the study, interspecies hybridization (Citrus × Poncirus) was used. Embryo rescue technique was used to prevent the loss of hybrid individuals due to abortive seed formation. The seeds were gathered from the crossed plants 80, 100, and 120 days after pollination. The immature embryos were removed from the seeds and these embryos were transferred to control, 0.5, and 1.0 mg l-1 GA3 containing medium to germination. The highest germination rate with 100% was observed from 1.0 mg l-1 GA3 containing media harvested 120 days after pollination. Additionally, the highest rate of trifoliate seedlings was obtained from embryos gathered after 120 days of the pollination.

Invertase (INV)-mediated sucrose (Suc) hydrolysis, leading to the irreversible production of glucose (Glc) and fructose (Frc), plays an essential role in abiotic stress tolerance of plants. However, the regulatory network associated with the Suc catabolism in response to cold environment remains largely elusive. Herein, the cold-induced alkaline/neutral INV gene PtrA/NINV7 of trifoliate orange (Poncirus trifoliata (L.) Raf.) was shown to function in cold tolerance via mediating the Suc hydrolysis. Meanwhile, a nuclear matrix-associated region containing A/T-rich sequences within its promoter was indispensable for the cold induction of PtrA/NINV7. Two AT-Hook Motif Containing Nuclear Localized (AHL) proteins, PtrAHL14 and PtrAHL17, were identified as upstream transcriptional activators of PtrA/NINV7 by interacting with the A/T-rich motifs. PtrAHL14 and PtrAHL17 function positively in the cold tolerance by modulating PtrA/NINV7-mediated Suc catabolism. Furthermore, both PtrAHL14 and PtrAHL17 could form homo- and heterodimers between each other, and interacted with two histone acetyltransferases (HATs), GCN5 and TAF1, leading to elevated histone3 acetylation level under the cold stress. Taken together, our findings unraveled a new cold-responsive signaling module (AHL14/17-HATs-A/NINV7) for orchestration of Suc catabolism and cold tolerance, which shed light on the molecular mechanisms underlying Suc catabolism catalyzed by A/NINVs under cold stress.

The morbidity and mortality of diabetes have increased dramatically in recent decades. Novel strategies for treating diabetes and its complications with minimal side effects are in urgent need. New monomeric molecules extracted from herbal medicine, which is a form of alternative medicine, are being sought as drug candidates for the treatment of diabetes and its complications. Hesperetin (Hst), a citrus flavonoid, is of increasing interest in scientific studies recently due to its properties in combating diabetes and its complications, whereas existing studies are scattered and unsystematic. Here, we summarized the literature studies over the last 10 years to review the potential therapeutic role of Hst in the prevention and mitigation of diabetes and its complications, intending to provide promising strategies for the clinical management of diabetes and its complications.

During recent surveys on leafhoppers in urban and agroecosystem areas in Europe the Oriental Hishimonus diffractus Dai, Fletcher & Zhang, 2013 (Hemiptera, Cicadellidae) has been found. This species, described from China and Thailand, is recorded here for the first time from Europe in the following countries: France, Portugal (Madeira), Spain, Gibraltar, United Kingdom, and Italy (Sicily). H. diffractus has been found associated with Jasminum and Olea (Oleaceae), Citrus (Rutaceae), Pittosporum (Pittosporaceae), Rubus (Rosaceae), Nerium (Apocynaceae) and Vitis (Vitaceae). Presence of the adults, morphological features, and some biological notes are provided to H. diffractus. Furthermore, Hishimonus hamatus Kuoh, 1976 is here reported for the first time in Spain and France. All Hishimonus species introduced in Europe are considered, and their economic importance discussed. Details on the first record and country in Europe, origin area, current distribution and host plants are given for each species.

The Hindustan citrus mite, Schizotetranychus hindustanicus (Acari: Tetranychidae), is an invasive pest in South America and constitutes a threat to Brazilian citriculture. This study aimed to determine the contribution of weather variables to the seasonal abundance of S. hindustanicus and the best sampling scheme (sampling variable and unit) for this mite. Populations of S. hindustanicus were monitored monthly in an orange orchard for 31 months in Roraima state, Brazil. Eggs, mobile stages, and the symptoms caused by S. hindustanicus were sampled in different combinations of canopy quadrants, vertical tree strata, and branch sections. The optimal sampling variable and sampling unit for S. hindustanicus scouting were determined according to fidelity and precision criteria. Rainfall and high air temperature were the main factors reducing S. hindustanicus populations. The most suitable variable for S. hindustanicus sampling was egg count. The optimal sampling unit was a leaf collected in the central section of branches located in the middle tree stratum. In addition, the sample should be taken from the southwest quadrant. Collectively, this study adds to the understanding of S. hindustanicus population dynamics and provides a sampling scheme for better management of this pest.

Citri Reticulatae Pericarpium Viride (CRPV, Qing Pi in Chinese) has been widely used in traditional Chinese medicine. Polymethoxylated flavonoids (PMFs), which are a special group of flavonoids with strong antitumor activity, are broadly distributed in citrus peels. However, systematic investigation of antitumor PMFs in CRPV has received little attention to date.An MCF-7 cell biospecific extraction method integrated with neutral loss/diagnostic ion filtering-based HPLC-QTOF-MS/MS strategy was developed for rapid and specific profiling of antitumor PMFs and systematic identification of PMFs in CRPV.By incubating MCF-7 cells with CRPV extract, potential antitumor PMFs specifically bound to cells and were isolated. Then, by systematic investigation of fragmentation pathways, neutral loss and diagnostic ion filtering strategies were proposed to comprehensively and accurately identify PMFs.Sixteen antitumor PMFs were unambiguously or tentatively identified. Among them, minor compound 15 (5-hydroxy-6,7,8,3',4'-pentamethoxyflavone with a free hydroxyl group at C-5) exhibited excellent antitumor activity, with an ICTo the best of our knowledge, this is the first report on the systematic investigation of antitumor PMFs in CRPV. The study will lay a foundation for the quality control and clinical application of CRPV.

Pretreatment or treatment with anti-apoptotic, anti-inflammatory, or anti-oxidative approaches could be critical for attenuated the severity of myocardial ischemia/reperfusion (I/R) injury. Naringin, a natural flavonoid, plays important roles in inflammation-related diseases. Immature dry fruits of Citrus wilsonii Tanaka (Xiang Yuan) are rich in naringin that can be used as traditional Chinese medicine to treat inflammation-related symptoms. However, its roles in cardioprotective role remain unclear. This study aimed to isolate naringin from Citrus wilsonii Tanaka fruit and tested their cardioprotective effect. The dry fruits of Citrus wilsonii Tanaka were extracted with boiling water and then supernatants were freeze-dried to yield aqueous extract (ZQAE). The extract was chemoprofiled using UPLC-MS/MS to stand for major constituents, and then subjected to different chromatographic separation steps, and naringin was isolated in a high yield. The cardioprotective effects of the aqueous extract of ZQAE and naringin were investigated in a myocardial I/R rat model and to elucidate the mechanism underlying its cardioprotective effect. Our results indicated that 5-day ZQAE and naringin pretreatments both promoted histopathological changes and reduced myocardial enzymes (cTnl, CK-MB, CK and LDH) induced by I/R. Moreover, the 50 mg/kg and 100 mg/kg ZQAE dose pretreatments presented a significantly decreased infarct size as well as myocardial enzyme levels but also inhibited myocardial apoptosis (cleaved-caspase3 protein expression), the inflammatory response (IL-23, IL-6, and TNF-α) and oxidative stress (MDA and SOD). The cardioprotective effect of 5 mg/kg dose of naringin pretreatment is comparable with that of 5 mg/kg drug ditiazem pretreatment. Additionally, naringin pretreatment exhibited striking decreases in the apoptosis index and downregulation of the protein expression levels of cleaved-Caspase3, Bcl2 and Bax. Meanwhile, naringin downregulated HMGB1 expression and upregulated SIRT1 expression in the myocardium. These findings suggest that short-term pretreatments with ZQAE and naringin both protect against myocardial I/R injury by suppressing myocardial apoptosis, the inflammatory response, and oxidative stress. The cardioprotective effect of naringin involves SIRT1 activation and may interact with HMGB1 and inhibit the release of HMGB1.

Background: The full impact of the intake of citrus fruits on the risk of depression in individuals with chronic heart failure (HF) is unknown. Here, we examined the associations between the estimated habitual intakes of citrus fruits and depressive symptoms in patients with chronic HF. Methods: We enrolled 150 stable outpatients with chronic HF who had a history of worsening HF. To assess the patients' daily dietary patterns, we used a brief self-administered diet-history questionnaire to calculate the daily consumption of foods and nutrients. To assess the patients' mental state, we used a nine-item Patient Health Questionnaire (PHQ-9). Results: Twelve patients (8%) were identified as having moderate-to-severe depression (PHQ-9 score > 10). The patients with PHQ-9 > 10 had lower daily intakes of citrus fruits compared to those with no or mild depressive symptoms (PHQ-9 < 10). The daily intakes of various antioxidants, including vitamin C, beta- carotene, and beta-cryptoxanthin, all of which are abundant in citrus fruits, were reduced in the patients with PHQ-9 > 10, accompanied by higher serum levels of 8-isoprostane (an oxidative stress marker). A multivariate logistic regression analysis using forward selection showed that a lowered daily intake of citrus fruits was an independent predictor of the comorbidity of moderate-to-severe depression in patients with chronic HF, after adjustment for age, gender, and the hemoglobin value. Conclusions: A lower daily consumption of citrus fruits was associated with higher prevalence of depression in patients with chronic HF. Our findings support the hypothesis that a daily consumption of citrus fruits has a beneficial effect on the prevention and treatment of depression in chronic HF patients. (C)2021 Japanese College of Cardiology. Published by Elsevier Ltd. All rights reserved.

Polyploidy, frequently observed in citrus species, aids in achieving better adaptation to environmental stresses. In this context, the current work aims to develop stable tetraploids in citrus rootstock cultivars, viz., Rough lemon, Rangpur lime and Alemow, through in vitro colchicine treatments. Seed-derived explants were obtained by culturing sterile seeds on MS basal media. Seedlings with a size of 5–8 mm (hypocotyl) were exposed to colchicine treatment. After treatment, the surviving seedlings were minigrafted onto six-month-old rootstock for better growth. Colchicine concentrations of 0.1%, 0.2%, 0.3% and control for durations of 16 or 24 h were tested with respect to the induction of polyploidisation. Treatment with 0.1% colchicine for 24 h resulted in high rates of mutation for polyploidisation and showed the highest tetraploid induction percentage (18.3%) in all the rootstock cultivars. High colchicine concentration and long exposure time decreased the survival of the observed seedlings. Flow cytometry and cytological methods were used for confirmation of autotetraploidy in the analysed samples. The surviving seedlings were identified on the basis of morphological and cytological variables, such as leaf area and stomata size, which significantly increased with increasing ploidy level. The proposed method was found to be an effective way to induce the polyploidy in Rangpur lime, Rough lemon and Alemow rootstocks.

Citrus exhibits unique nutritional values. Most citrus cultivars are derived from mutations. However, the effect of these mutations on fruit quality is unclear. We have previously found a yellowish bud mutant in the citrus cultivar ‘Aiyuan 38’. Therefore, this study aimed to determine the effect of the mutation on fruit quality. ‘Aiyuan 38’ (WT) and a bud mutant variant (MT) were used to analyze variations in fruit color variation and flavor substances using colorimetric instruments, high-performance liquid chromatography (HPLC), headspace solid-phase microextraction-gas chromatography–mass spectrometry (HS-SPME-GC–MS), and odor activity values (OAVs). The mutation in MT conferred yellowish characteristics to its peel. Although the differences in total sugar and acid content of the pulp were not statistically significant between WT and MT, the MT glucose content was significantly lower and the malic acid level was significantly higher. HS-SPME-GC–MS analysis revealed that the MT pulp released more types and contents of volatile organic compounds (VOCs) than the WT, whereas the opposite trend was observed for the peel. Analysis of the OAV revealed that the MT pulp contains 6 unique VOCs, whereas the peel contains only 1. This study provides a useful reference for the study of flavor substances associated with citrus bud mutations.

Metabolic diseases have been related to the overdrinking of high-sugar content beverages. As a result, the demand for alternative formulations based on plant-based ingredients with health-promoting properties has increased during the last few years. Nonetheless, the design and production of effective formulations requires understanding the bioavailability of these compounds. For this purpose, a two-month longitudinal trial with 140 volunteers was conducted to measure the beneficial effects of a maqui–citrus beverage, rich in (poly)phenols. From data obtained by quantifying metabolites present in urine samples, biostatistical and machine learning (data imputation, feature selection, and clustering) methods were applied to assess whether a volunteer’s sex and the sweetener added to the beverage (sucrose, sucralose, or stevia) affected the bioavailability of (poly)phenol metabolites. Several metabolites have been described as being differentially influenced: 3,4-dihydroxyphenylacetic acid and naringenin with its derivatives were positively influenced by stevia and men, while eriodictyol sulfate and homoeridictyol glucunoride concentrations were enhanced with stevia and women. By examining groups of volunteers created by clustering analysis, patterns in metabolites’ bioavailability distribution as a function of sex and/or sweeteners (or even due to an uncontrolled factor) were also discovered. These results underline the potential of stevia as a (poly)phenol bioavailability enhancer. Furthermore, they also evidence sex affects the bioavailability of (poly)phenols, pointing at a sex-dependent metabolic pathway regulation.

In autumn and spring, citrus leaves with a Ponkan (Citrus reticulata Blanco cv. Ponkan) genetic background (Harumi, Daya, etc.) are prone to abnormal physiological chlorosis. The effects of different degrees of chlorosis (normal, mild, moderate and severe) on photosynthesis and the chlorophyll metabolism of leaves of Citrus cultivar (Harumi) were studied via field experiment. Compared with severe chlorotic leaves, the results showed that chlorosis could break leaf metabolism balance, including reduced chlorophyll content, photosynthetic parameters, antioxidant enzyme activity and enzyme activity related to chlorophyll synthesis, increased catalase and decreased enzyme activity. In addition, the content of chlorophyll synthesis precursors showed an overall downward trend expected for uroporphyrinogen III. Furthermore, the relative expression of genes for chlorophyll synthesis (HEMA1, HEME2, HEMG1 and CHLH) was down-regulated to some extent and chlorophyll degradation (CAO, CLH, PPH, PAO and SGR) showed the opposite trend with increased chlorosis. Changes in degradation were more significant. In general, the chlorosis of Harumi leaves might be related to the blocked transformation of uroporphyrinogen III (Urogen III) to coproporphyrinogen III (Coprogen III), the weakening of antioxidant enzyme system activity, the weakening of chlorophyll synthesis and the enhancement in degradation.

In Mali, citrus fruits contribute significantly to the nutritional balance of the population due to their high content of vitamin C, carotenoids and polyphenols. The citrus sector in Mali is rich in several species, but the current trend in citrus production is towards the production of Tangelo (orCitrus×tangelo). The objective of this study was to determine the effects of phytohormones 2,4,5-T, AG3and potassium nitrate (KNO3) on vegetative growth of Tangelo. The trial was conducted in Koulikoro district, especially on the experimental plot of the IPR/IFRA of Katibougou. The plant material used isTangelo orlandowith white pulp. The experimental design was a Fisher block with three replications. The three products and water (check) were applied by spraying the leaves of the young plants at 110 days after planting. Observations were made on the following growth parameters: graft diameter, graft length and number of vegetative branches at 90th, 180thand 270thdays after the application. The results of the analysis of variance of the different treatments revealed significant (p

This document aims to provide a framework for the efficient and effective global conservation of genetic resources of Citrus crops., The development of this Global Crop Conservation Strategy was funded by the German Federal Ministry of Food and Agriculture (BMEL) as part of the three-year project led by the Crop Trust: "Breathing new life into the Global Crop Conservation Strategies: Providing an Evidence Base for the Global System of Ex situ Conservation of Crop Diversity." The Crop Trust also cooperated with the Secretariat of The International Treaty on Plant Genetic Resources for Food and Agriculture (ITPGRFA) in the development of this document.

Although interactions between the cytoplasmic and nuclear genomes occurred during diversification of many plants, the evolutionary conflicts due to cytonuclear interactions are poorly understood in crop breeding. Here, we constructed a pan-mitogenome and identified chimeric open reading frames (ORFs) generated by extensive structural variations (SVs). Meanwhile, short reads from 184 accessions of citrus species were combined to construct three variation maps for the nuclear, mitochondrial, and chloroplast genomes. The population genomic data showed discordant topologies between the cytoplasmic and nuclear genomes because of differences in mutation rates and levels of heteroplasmy from paternal leakage. An analysis of species-specific SVs indicated that mitochondrial heteroplasmy was common and that chloroplast heteroplasmy was undetectable. Interestingly, we found a prominent divergence in the mitogenomes and the highest genetic load in the, which may provide the basis for cytoplasmic male sterility (CMS) and thus influence the reshuffling of the cytoplasmic and nuclear genomes during hybridization. Using cytoplasmic replacement experiments, we identified a type of species-specific CMS in mandarin related to two chimeric mitochondrial genes. Our analyses indicate that cytoplasmic genomes from mandarin have rarely been maintained in hybrids and that paternal leakage produced very low levels of mitochondrial heteroplasmy in mandarin. A genome-wide association study (GWAS) provided evidence for three nuclear genes that encode pentatricopeptide repeat (PPR) proteins contributing to the cytonuclear interactions in theCitrusgenus. Our study demonstrates the occurrence of evolutionary conflicts between cytoplasmic and nuclear genomes in citrus and has important implications for genetics and breeding.

[ES] A escala mundial, la Citricultura es uno de los sectores dentro de la Fruticultura con mayor producción, superando los 158 millones de toneladas en el año, pero es sensible de un gran número de estreses abióticos al ser un cultivo subtropical. La baja temperatura es uno de los estreses abióticos que más afecta a los cítricos produciendo grandes pérdidas anuales. El cambio climático está muy presente dando lugar cambios en los patrones de El tiempo originando un gran número de olas de frío y calor. Pero gracias a la técnica del injerto y al uso de patrones de cítricos los problemas causados por los estreses abióticos pueden mejorarse. El IVIA cuenta con un programa de mejora de patrones cuyo objetivo es la búsqueda de nuevo material vegetal con el fin de satisfacer los problemas de la citricultura. Tradicionalmente, el patrón más tolerante a la baja temperatura es Poncirus trifoliata, el cual se han realizado la mayoría de los trabajos en la bibliografía. Como patrón sensible, se empleó Citrus macrophylla ya que se trata de un patrón del tipo limonero que se sabe popularmente de su sensibilidad al frío. También se empleó el patrón citrange Carrizo, el cual se trata de patrón comercial más cultivado en España en la actualidad y la variedad Taroco Rosso, que se sabe de su pigmentación a causa de la baja temperatura. Así pues, los capítulos planteados en la presente tesis fueron: 1. Estrés a baja temperatura a corto plazo, 2. Estrés a baja temperatura a largo plazo y 3. El efecto del patrón en la variedad Tarocco Rosso., [CAT] A escala mundial, la Citricultura és un dels sectors dins de la Fruticultura amb més producció, superant els 158 milions de tones a l'any, però és sensible d'un gran nombre d'estressos abiòtics per ser un cultiu subtropical. La baixa temperatura és un dels estressos abiòtics que més afecta els cítrics produint grans pèrdues anuals. El canvi climàtic està molt present donant lloc a canvis en els patrons de El temps originant un gran nombre d'onades de fred i calor. Però gràcies a la tècnica de l'empelt i a l'ús de patrons de cítrics, els problemes causats pels estressos abiòtics es poden millorar. L'IVIA compta amb un programa de millora de patrons que te com a objectiu la recerca de nou material vegetal per tal de satisfer els problemes de la citricultura. Tradicionalment, el patró més tolerant a la baixa temperatura és Poncirus trifoliata, el qual s'han realitzat la majoria dels treballs a la bibliografia. Com a patró sensible, es va fer servir Citrus macrophylla ja que es tracta d'un patró del tipus llimoner que se sap popularment de la seva sensibilitat al fred. També es va emprar el patró citrange Carrizo, el qual es tracta del patró comercial més cultivat a Espanya actualment i la varietat Taroco Rosso, que se sap de la seva pigmentació a causa de la baixa temperatura. Així doncs, els capítols plantejats en aquesta tesi van ser: 1. Estrès a baixa temperatura a curt termini, 2. Estrès a baixa temperatura a llarg termini i 3. L'efecte del patró en la varietat Tarocco Rosso., [EN] On a global scale, Citriculture is one of the sectors within Fruit Growing with the highest production, exceeding 158 million tons per year, but it is sensitive to many abiotic stresses as it is a subtropical crop. Low temperature is one of the abiotic stresses that most affects citrus, producing large annual losses. Climate change is very present, giving rise to changes in weather patterns, causing many cold and heat waves. But thanks to the grafting technique and the use of citrus rootstocks, problems caused by abiotic stresses can be improved. IVIA institute has a rootstock improvement program whose objective is to search for new plant material to satisfy the problems of Citriculture. Traditionally, the most low-temperature tolerant rootstock is Poncirus trifoliata, on which most of the work in the literature has been done. As a sensitive rootstock, Citrus macrophylla was used since it is a lemon type rootstock that is popularly known for its sensitivity to cold. The citrange Carrizo rootstock was also used, which is currently the most cultivated commercial rootstock in Spain, and the Taroco Rosso variety, which is known for its pigmentation due to low temperature. Thus, the chapters proposed in this thesis were: 1. Short-term low-temperature stress, 2. Long-term low-temperature stress, and 3. The rootstock effect on the Tarocco Rosso variety.

Mandarin production has increased in recent years, especially for fresh consumption, due to its ease of peeling, its aroma, and its content of bioactive compounds. In this sense, aromas play a fundamental role in the sensory quality of this fruit. The selection of the appropriate rootstock is crucial for the success of the crop and its quality. Therefore, the objective of this study was to identify the influence of 9 rootstocks (“Carrizo citrange”, “Swingle citrumelo CPB 4475”, “Macrophylla”, “Volkameriana”, “Forner-Alcaide 5”, “Forner-Alcaide V17”, “C-35”, “Forner-Alcaide 418”, and “Forner-Alcaide 517”) on the volatile composition of “Clemenules” mandarin. For this, the volatile compounds of mandarin juice were measured using headspace solid-phase micro-extraction in a gas chromatograph coupled to a mass spectrometer (GC-MS). Seventy-one volatile compounds were identified in the analyzed samples, with limonene being the main compound. The results obtained showed that the rootstock used in the cultivation of mandarins affects the volatile content of the juice, with “Carrizo citrange”, “Forner-Alcaide 5”, “Forner-Alcaide 418”, and “Forner-Alcaide 517” being those that presented the highest concentration.

Consumption of fast food and sedentary lifestyles has recently developed in society. This activity is one of the causes of various degenerative diseases, including cardiovascular disease. Therefore, efforts are needed to prevent it. Sweet orange (Citrus sinensis) is one of the most common types of citrus fruit, with total production reaching 70% of all types. This type of fruit is the most widely produced and consumed by the world’s population. Sweet orange is known for its bioactive compounds that have many benefits in cardiovascular health, such as alkaloids, flavonoids, tannins, phenols, vitamins C, B1, B2, and B3, glycosides, coumarin glycosides, folic acid, some organic acids, essential oils, and saponins. Flavonoids are components of secondary metabolites that play a role in the cardiovascular system. The main components of these compounds are hesperidin and naringenin. Various studies have shown that these compounds are active in reducing hyperlipidemia, improving endothelial function, acting as an anti-inflammatory and anti-atherosclerotic agent, preventing myocardial infarction, and preventing cardiac hypertrophy through various mechanisms with low toxicity.

This study aims to detect Clostridium botulinum and its control using natural leaf extracts of Citrus limon, Citrus sinensis, and Citrus unshiu in Egyptian fish products, e.g., canned tuna, canned sardine, canned mackerel, fesikh, moloha, and renga, as well the application of C. limon in tuna. Moreover, the antibacterial activity of the C. limon leaf extract was also estimated. In the water extract, ascorbic acid, total flavonoid content (TFC), and total phenolic content (TPC) were determined by volumetric, aluminum chloride, and Folin–Ciocalteu approaches, respectively. The antioxidant ability of the extract was analyzed in vitro via free radical scavenging (DPPH) and Ferric reducing assays. The results showed variability in the distribution of the total number of positive C. botulinum in fish samples from three different governorates under study, which were (24) Alexandria, (16) Beheira, and (17) Gharbia, out of the 120 tested samples in each governorate. Additionally, the findings revealed that all three Citrus extracts contain an appropriate number of secondary metabolites, with a sustainable presence of saponin and tannins in the C. limon extract. Furthermore, all Citrus extracts inhibited bacterial growth by increasing the inhibition zone, with C. limon being the best extract (25 mm) compared to C. sinensis and C. unshiu. The overall results showed the high antioxidant and anti-Clostridium powers (p < 0.05) of C. limon leaf extract, indicating its preservative activity in fishery products during storage. Finally, C. limon leaf extract can fight off C. botulinum and is considered a promising natural preservation candidate in ensuring safe and fresh fishery products.

‘Aliza’ is a new pomelo × mandarin hybrid (Citrus maxima, cv. Red Chandler × Citrus reticulata, cv. Ora) developed by the Israeli citrus breeding program at the Volcani Institute. Here, we aimed to characterize the quality and flavor of ‘Aliza’ fruit as compared to other commercial citrus fruit, specifically pomelo (C. maxima), grapefruit (Citrus paradisi), orange (Citrus sinensis) and mandarin (C. reticulata). ‘Aliza’ fruits have a similar size as grapefruits, but have a thinner peel and a unique yellowish/golden color. ‘Aliza’ fruits are completely seedless and have especially high juice contents. They also have a unique, highly preferred flavor, characterized by high sweetness and moderate bitterness and acidity, with strong citrusy and tropical fruity aromas. Sensory analyses conducted with the aid of a trained panel and an electronic tongue revealed that the flavor of ‘Aliza’ fruits is different from the flavors of other citrus species. Consumer acceptance and preference tests revealed that ‘Aliza’ fruit are highly appreciated and favored. The aroma volatile profile of ‘Aliza’ fruit was somewhat similar to those of pomelo and grapefruit, but very different from those of orange and mandarin. Overall, ‘Aliza’ fruits can be distinguished from other citrus fruits by their unique color, high juice content and exceptional, unique flavor.

[ES] En la actualidad se emplean fungicidas de síntesis para controlar las principales causas del desperdicio de los cítricos tras su recolección, pero su uso generalizado está empezando a ser cuestionado y restringido por muchos comercializadores en Europa y en el resto del mundo debido a sus posibles efectos adversos en la salud del consumidor y en el medio ambiente. El objetivo principal de este trabajo es la selección y validación industrial de un producto de química verde que sea compatible con los productos de agricultura ecológica y que aporte unos beneficios similares en el tratamiento postcosecha de cítricos a los obtenidos con los fungicidas de síntesis tradicionales. Para este fin se han realizado diversos ensayos semiindustriales de selección de producto y diferentes combinaciones con otras materias activas para ver sus efectos en la eficacia en control de podrido. Finalmente se ha realizado un ensayo a nivel industrial en condiciones reales de aplicación, con el cual se ha determinado que el producto escogido, cuyo nombre comercial es Greencide, cumple con los requisitos propuestos inicialmente y por lo tanto se da por válido, dada su alta eficacia en combinación con sorbato potásico y un desinfectante compuesto por ácido peracético y peróxido de hidrógeno. Gracias a los resultados obtenidos podemos concluir que con el uso de Greencide podemos reducir el impacto ambiental del procesado postcosecha de cítricos tanto los cultivados con métodos de agricultura convencional como ecológicos, manteniendo una alta eficacia en control de podrido que se mantiene en el tiempo, algo imprescindible para su uso en exportaciones de larga distancia., [EN] Synthetic fungicides are currently used to control the leading causes of citrus waste after harvesting. Still, their widespread use is beginning to be questioned and restricted by many marketers in Europe and the rest of the world due to their potential adverse effects on consumers¿ health and the environment. The main objective of this work is the selection and industrial validation of a green chemistry product compatible with organic farming products and that provides similar benefits in the postharvest treatment of citrus to those obtained with traditional synthetic fungicides. For this purpose, various semi-industrial tests have been carried out on the selection of the product and different combinations with other active ingredients to see its effects on effectiveness when it comes to decay control. Finally, an industrial-level test has been carried out under real application conditions, with which it has been determined that the chosen product, whose trade name is Greencide, meets the requirements initially proposed and it is therefore considered valid, given its high efficacy in combination with potassium sorbate and a disinfectant composed of peracetic acid and hydrogen peroxide. Considering the obtained results, we can conclude that with the use of Greencide we can reduce the environmental impact of postharvest processing of citrus fruits, both those grown with conventional and organic farming methods, maintaining a high efficiency in decay control that is maintained over time, something essential for its use in long-distance exports.

Intenzivan uzgoj agruma u Republici Hrvatskoj započinje u drugoj polovici 20. stoljeća, a danas su prema uzgojnim površinama i prinosima u domaćoj proizvodnji treća voćna kultura. Kao i sve kultivirane biljke, agrumi su podložni napadu entomofaune. Podredu Sternorrhyncha koji obuhvaća lisne buhe, štitaste moljce, lisne i štitaste uši pripadaju neki od najštetnijih kukaca koji su u prošlosti prouzročili propadanje čitavih plantaža agruma diljem svijeta. Razvojem svjetskih tržišta raste i međunarodna trgovina poljoprivrednim proizvodima. Kukci iz ovog podreda se lako prenose sadnim materijalom. S obzirom da je u posljednjih 20 godina i u Hrvatskoj otkriveno nekoliko novih štetnih organizama koji su regulirani EU zakonodavstvom, potpuno je jasno da niti naš teritorij nisu zaobišle ove negativne posljedice globalizacije. Stoga je cilj ovog rada bio pregledom literature istražiti koje su sve vrste kukaca iz podreda Sternorrhyncha štetnih na agrumima prisutne u Republici Hrvatskoj i usporediti ga sa zemljama s najvećom proizvodnjom agruma u svijetu, a nakon toga nadopuniti popis nalazima vlastitog faunističkog istraživanja u nasadima agruma u Republici Hrvatskoj, s posebnim osvrtom na novoutvrđene i karantenske vrste te izdvojiti potencijalno najštetnije. Faunističko istraživanje je provedeno u razdoblju od 2015. do 2020. godine u šest obalnih županija od Dubrovačko-neretvanske do Istarske županije. Ukupno je prikupljeno 1.495 uzoraka biljnog materijala iz kojih je izrađeno 1.416 trajnih mikroskopskih preparata. Determinirane su 22 vrste kukaca iz podreda Sternorrhyncha kojima domaćini mogu biti agrumi, od toga pet vrsta štitastih moljaca, sedam vrsta lisnih uši i deset vrsta štitastih uši. Utvrđena je prisutnost karantenske vrste štitastog moljca Aleurocanthus spiniferus Quaintance, 1903 i regulirane nekarantenske vrste Parabemisia myricae (Kuwana) 1927. U različitim tipovima nasada prvi put je na agrumima pronađeno pet vrsta lisnih uši i jedna vrsta štitaste uši. Aphis craccivora Koch, 1854, Aphis fabae Scopoli, 1763, Aphis gossypii Glover, 1877, Aphis neri Boyer de Fonscolombe, 1841, Aulacorthum solani Kaltenbach, 1843 i Pulvinaria floccifera (Westwood, 1870) su duže vremena prisutne u hrvatskoj entomofauni, ali su sada prvi put nađene na agrumima. Najveći broj vrsta (17) pronađen je u Dubrovačko-neretvanskoj i Splitsko-dalmatinskoj županiji (16) u kojima se nalazi glavnina proizvodnje agruma u Hrvatskoj, stoga je i statistička obrada podataka ograničena na ove dvije županije. Veći intenzitet šteta u proizvodnim nasadima agruma zabilježen je na tri lokacije u kojima je izostala primjena agrotehničkih mjera i mjera zaštite. U jednom od takvih nasada u Dubrovačko-neretvanskoj županiji prvi put je otkriven i štitasti moljac A. spiniferus. Za vrijeme istraživanja ovaj štitasti moljac se konstantno širio te je otkriven i u Splitsko-dalmatinskoj županiji. Provedeno istraživanje je prvi put objedinilo rezultate prisutnosti i raširenosti kukaca iz podreda Sternorrhyncha na agrumima u Hrvatskoj. Introduction: Citrus plants have been grown on the Croatian side of the Adriatic since the Middle Ages but intensive production started in the second half of 20th century. Satsuma mandarin is the 3rd important fruit culture in Croatia, following apple and olive with total growing areas of 2.100 hectares and annual fruit production from 20.000-65.000 tones in five year period. As all cultivated plants, species belonging to genus Citrus L. and Poncirus Raf. are susceptible to impact of entomofauna. Suborder Sternorrhyncha within the Hemiptera order comprises of four Superfamilies: Psylloidea, Aleyrodoidea, Aphidoidea and Coccomorpha. Many insects which may feed on citrus plants fall within the said superfamilies. Their direct damages on plants are manifested as vigour deteriorating and late tree dying. Indirect damage is pour commercial value of fruits due to sooty mold settled on honeydew excreted by these insects. On the other hand, more harmful to plants are insects which, apart from ability to cause direct and indirect damage, are vectors of plant diseases like Citrus tristeza virus, Citrus greening disease or Huanglongbing and virus like disease Citrus chlorotic dwarf. Former records show that vectors of the mentioned plant diseases like aphid Aphis (Toxoptera) citricidus (Kirkaldy) 1907, psyllid Trioza erytreae (Del Guercio 1918) and whitefly Parabemisia myricae (Kuwana) 1927 are already present in the Mediterranean part of EU in countries with considerable citrus production. It is important fact because planting material of host plants, although accompanied by plant passport, may freely circulate through EU territory and represent a potential source of infection. Up to now, several faunistic researches were conducted and check lists in Croatia were made for aphids, whiteflies and scale insects. There were no complete faunistic research for psyllids and its general status in Croatia is still unknown. Furthermore, previous faunistic research of insects from the suborder Sternorrhyncha only partially covered plants belonging to genus Citrus and Poncirus. Therefore the aim of this research was to make the inventory of insects from the suborder Sternorrhyncha in citrus production areas in Croatia in order to gain complete status of distribution of these insects and to provide scientific and sound support in case of accidentally imported quarantine pests. Materials and methods: Literature research was done to gain image about distribution of the 73 chosen important insects from the suborder Sternorrhyncha that may feed on citrus plants. These insects (11 psyllids, 14 whiteflies, 19 aphids i 29 scale insects), according to the literature data, are classified as a quarantine, economic or secondary pests on citrus plants. Research was conducted for 15 countries which, according to FAO, have the biggest production and export of citrus fruits in the Mediterranean, Asia, South Africa, North and South America. Research was also done for Croatia and neighbouring countries Slovenia and Montenegro. In addition, six year faunistic research (2015-2020) in citrus production areas in Croatia was done. During that period 266 sites in total were inspected, of which 181 orchards, 69 backyards and 16 nurseries and garden centres. Visual inspections and sampling were conducted from April till November in six counties in coastal part of Croatia on ten plant species belonging to genus Citrus and Poncirus. Targeted psyllids were investigated on 105 locations. Whiteflies were found on 112 locations where 289 samples were taken. Aphids were found on 228 locations where 639 samples were taken and scale insects were found on 185 locations where 518 samples were taken. Host plants in orchards, backyards and nurseries and garden centres were visually inspected with help of 10x hand magnifier. Vast majority of locations were inspected at least two times during the six year period. Due to presence of planting material of citrus and ornamental plants originating in other EU countries, nurseries and garden centres were inspected at least once every year. In case of suspected infestation, sample of infested plant material or insects were taken for laboratory slide preparation. Methods of sampling nymphs and adults of psyllids are described by Hodkinson and White (1979) and Ossiannilsson (1992). Infested plant material with puparium and pupal case of whiteflies were taken according to the methods described by Martin (1987, 1999) and Hodges and Evans (2005). Infested plant material and adults of aphids were taken according to the method described by Blackman and Eastop (2000). Infested plant material with adult females of scale insects were taken according to the methods described by McKenzie (1967), Kosztarab and Kozár (1988), Gill (1988, 1997) and Watson and Chandler (1999). For slide preparation of psyllids modified method by Watson and Chandler (1999) was used. Whiteflies were slide mounted according to modified method by Šimala (2008) with individual phases described in method by Watson and Chandler (1999). For slide preparation of aphids methods described by Martin (1983) and Blackman and Eastop (2000) were used. Scale insects were slide mounted according to modified method by Masten Milek (2007) with individual phases described in method by Watson and Chandler (1999). After that microscopic identification according to relevant morphological keys for every group of insects was done. In case of finding of Trioza species, identification key by Hollis (1984) would be used. Since at the moment there is no relevant key for Diaphorina species, identification is done according to description by Yang (1984). For identification of puparium and pupal case of whiteflies, identification keys by Martin et al. (2000), Suh (2010), Dubey and Ko (2012) and Wang et al. (2014) were used. Identification of aphids were done according to keys by Blackman and Eastop (2000). For identification of scale insects identification keys by Gill (1988, 1997), Camporese and Pellizzari (1994), Williams and Watson (1990), Williams and Granara de Willink (1992) were used. For all species found during faunistic research, dominance, constancy and ecological significance indices were calculated for orchards, backyards and nurseries and garden centres in every county. Biodiversity for every locality was calculated using biodiversity indices (Shannon-Wiener Index, Simpson Index, and Sörenson Index). In order to determine if there is a difference between type of growing site (orchards, backyards and nurseries and garden centres) and county (only for Dubrovačko-neretvanska and Splitsko-dalmatinska), number of species in every group of insects and number of individuals from every group of insects were subjected to analysis of variance. Mean values were ranked based on Duncan's Multiple Range test with the protection level of 95% (p

Turkey is one of the major exporters of mandarins in the Mediterranean region. Seedlessness in citrus, which is one of the most desired fruit quality traits, especially in fresh mandarin export markets, can be obtained via triploidy as in many fruit species. Triploid plants can be recovered by 2x × 2x hybridizations in citrus, as well as 2x × 4x and 4x × 2x crosses. Hence, we aimed to develop local triploid hybrids by using the embryo rescue technique in five crosses using eight different citrus varieties in the present study.Embryos isolated from abortive seeds derived by 135 days after pollinations were cultured on modified Murashige and Tucker (MT) medium by adding different levels of GAGenotyping of triploid plants confirmed using Simple Sequence Repeat (SSR) molecular markers and five SSRs were able to identify three alleles of triploid hybrids. Selected triploid mandarin hybrids have been grafted on several rootstocks for field trials and are in the process of yield and quality performances.