Local Citrus Research Articles

Valorization of hydro-distillate of fruit peels of Citrus paradisi macfad. Cultivar. Foster: Chemical profiling, antioxidant evaluation and in vitro and in silico enzyme inhibition studies

The major commercial crops in Pakistan are citrus fruit trees, which are farmed extensively and serve as the country's principal source of foreign exchange. A local citrus plant, Citrus paradisi, variety Foster is famous for its valuable fruit and fruit juice, however, tons of peels of this fruit are thrown as waste, which otherwise can be utilized in formulating nutraceutical and cosmetics. In the present study, essential oil of fruit peels was obtained through hydro-distillation, which was then analyzed through GC-MS analysis and studied for its antioxidant and enzyme inhibition potential. GCMS analysis revealed the presence of several components; major were found to be limonene, α-terpineol, caryophyllene, δ-amorphene, elemol, γ-eudesoml, nootkatone and di-isooctyl phthalate. Although, the oil showed weak free radical inhibition, it was potentially active in CUPRAC, FRAP, phosphomolybdenum and metal chelating antioxidant assays. The oil also exhibited anti-glucosidase, anti-amylase activities and also exhibited potent inhibition of the enzyme tyrosinase, which makes it strong candidate for nutraceuticals and skin care products. The docking studies also substantiate our results and caryophyllene, γ-eudesoml and nootkatone showed good binding affinity α-glucosidase and α-amylase and all tested compounds showed the higher binding affinity towards the enzyme tyrosinase.

The Study of Local Commodity-Based Farming Development: Case Jujun Village, Keliling Danau District, Kerinci Regency

Orange is one of the mainstays and potential local horticultural commodities in Jambi Province and is especially cultivated in Kerinci Regency. This research will be carried out in Jujun Village, Keliling Danau District, Kerinci Regency, with the consideration that this village is an Integrated Laboratory Village of the Faculty of Agriculture, University of Jambi. The research respondents were local citrus owners and cultivators. This study aims to describe the description of local citrus farming and feasibility analysis of local citrus farming in Jujun Village, Keliling Danau District, Kerinci Regency. The analytical method used is the R/C ratio and B/C ratio, to determine the feasibility of local citrus farming.

Carriage of Enterotoxin Genes by Methicillin-Resistant Staphylococcus aureus and Anti-Staphylococcal Activity of Local Citrus Fruits

Background: Staphylococcus aureus (S. aureus) is one of the leading causes of skin infections and food poisoning worldwide. Citrus fruits enriched with phytochemicals can exhibit anti-staphylococcal activity. Objective: The study aims to investigate in vitro susceptibility of citrus fruits against S. aureus harboured enterotoxin genes among food handlers, personnel, and the environment. Methods: S. aureus was isolated from food handlers, personnel, and the environment. Methicillin-Resistant Staphylococcus aureus (MRSA) screening was performed using cefoxitin antibiotic discs. The antimicrobial activity of citrus fruits, including Citrus limon (Lemon), Citrus pseudolimon (galgal), Citrus tangerine (fruiter), Citrus sinensis (malta), and Citrus paradisi (grapefruit), was analyzed by performing well-diffusion assay and broth microdilution method. Genes for toxins production, sea and seg were detected using a Polymerase Chain Reaction (PCR). Results: Out of 200 swab samples, 50 (25%) S. aureus were isolated using the biochemical battery. Antibiotic susceptibility testing (AST) and cefoxitin disc revealed 12 (24%) MRSA strains. Most strains were sensitive to Citrus limon, followed by Citrus pseudolimon, Citrus tangerine, Citrus sinensis, and Citrus paradisi. PCR results detected carriage of the sea gene in 4 (8%) isolates, and 20 (40%) isolates harboured the seg gene. Conclusion: The prevalence of S. aureus, mainly MRSA, was high in the environment. These resistant strains' carriage of sea and sea enterotoxin genes increases their virulence and leads to untreatable infections. The antibacterial activities of citrus fruits should be explored using adavance methods.

The potential of local orange peel-derived eco-enzymes in producing indole acetic acid

Background: Ecoenzymes, created from the fermentation of organic citrus waste, offer a sustainable method to produce Indole Acetic Acid (IAA), a phytohormone vital for plant growth. This study investigates the potential of these ecoenzymes in promoting sustainable agriculture. Objective: This study aims to evaluate the capacity of ecoenzymes derived from local citrus organic waste to synthesize IAA hormones. Methods: The ecoenzyme was extracted from fruit powders and centrifuged to separate the supernatant. One ml of ecoenzyme supernatant was then mixed with 2 ml of Salkowski reagent and incubated for 12 hours at room temperature in the dark to facilitate reaction. The presence and concentration of IAA were determined using spectrophotometry at a wavelength of 530 nm, while total protein levels were measured using the Warburg-Christian method. Results: ecoenzymes from local citrus sources contain IAA, with the highest concentration observed in sample 7A (30.26 µg/ml). The ecoenzyme exhibited favorable characteristics, including an average degree of acidity of 3.55, and the highest total protein content was found in sample 2A (144.277 mg/mL). Conclusion: Ecoenzymes from local orange peels successfully produce IAA, supported by fermentation-induced microbial activity and acidic conditions. This highlights their potential in sustainable agriculture.

The potential of local orange peel-derived eco-enzymes in producing indole acetic acid

Background: Ecoenzymes, created from the fermentation of organic citrus waste, offer a sustainable method to produce Indole Acetic Acid (IAA), a phytohormone vital for plant growth. This study investigates the potential of these ecoenzymes in promoting sustainable agriculture. Objective: This study aims to evaluate the capacity of ecoenzymes derived from local citrus organic waste to synthesize IAA hormones. Methods: The ecoenzyme was extracted from fruit powders and centrifuged to separate the supernatant. One ml of ecoenzyme supernatant was then mixed with 2 ml of Salkowski reagent and incubated for 12 hours at room temperature in the dark to facilitate reaction. The presence and concentration of IAA were determined using spectrophotometry at a wavelength of 530 nm, while total protein levels were measured using the Warburg-Christian method. Results: ecoenzymes from local citrus sources contain IAA, with the highest concentration observed in sample 7A (30.26 µg/ml). The ecoenzyme exhibited favorable characteristics, including an average degree of acidity of 3.55, and the highest total protein content was found in sample 2A (144.277 mg/mL). Conclusion: Ecoenzymes from local orange peels successfully produce IAA, supported by fermentation-induced microbial activity and acidic conditions. This highlights their potential in sustainable agriculture.

Comparative analysis of chloroplast genomes of two Chinese local citrus varieties and haplotype analysis with other citrus species

Citrus plants, comprising some of the most widely cultivated fruit crops worldwide, have great research value. We assembled and annotated the chloroplast genomes of two local important citrus varieties of China, ‘Tuju’ and ‘Zangju’, and comparatively analyzed their sequences. The full lengths chloroplast genomes of ‘Tuju’ and ‘Zangju’ were 160,121 and 160,694 bp, respectively. Both featured a quadripartite structure with two copies of a large inverted repeat (IR) separated by small (SSC) and large (LSC) single-copy regions. ‘Tuju’ chloroplast genome contained 134 genes. ‘Zangju’ had one less protein-coding gene (rpl22) than ‘Tuju’. We also found 27 and 21 optimal codons, all of which ended with A or U, for chloroplast protein-coding genes of ‘Tuju’ and ‘Zangju’, respectively. There were 32 haplotypes obtained from 43 citrus chloroplast genomes, some different species or cultivars could share one haplotype. Of them, ‘Tuju’, one individual of Citrus limon, one individual of Citrus maxima and Citrus platymamma shared one haplotype, which implied they had the same maternal origin. The phylogenetic trees supported that all haplotypes could be divided five groups, while haplotype network analysis tended to get three groups. Our study reveals the complex lineage relationship of citrus plants at a certain level, and provides a valuable reference tool for future research on citrus plants.

Comparative study on secondary metabolites from different citrus varieties in the production area of Zhejiang.

To investigate the distribution pattern of bioactive components and their correlations between citrus varieties, we thoroughly analyzed secondary metabolites (including flavonoids, phenolic acids, carotenoids, and limonoids) in the peel and pulp of 11 citrus varieties from the production area of Zhejiang. Citrus peels accumulated metabolites far more than the pulp, and the accumulation varied significantly between species. Flavonoids were the most abundant compounds, followed by phenolic acids, with carotenoids and limonoids being far less abundant than the first two, but limonoids were more abundant than carotenoids. Hesperidin was the main flavonoid in most varieties, but cocktail grapefruit and Changshanhuyou contained naringin, with Ponkan having the most abundant polymethoxylated flavones (PMFs). The major components of phenolic acids, carotenoids, and limonoids were ferulic acid, β-cryptoxanthin, and limonin, respectively. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) indicated that these components were mostly correlated with each other, and these citrus varieties could be categorized into four groups by pulp and three groups by peel. The obtained results filled the data gap for secondary metabolites from local citrus and could provide data references for citrus resource utilization, selection and breeding of superior varieties, and other research.

THE INFLUENCE OF PRODUCT ATTRIBUTES, CONSUMER ATTITUDE AND CONSUMER INTEREST ON THE PURCHASE DECISION OF LOCAL ORANGE FRUIT IN MALANG CITY

This study aims to determine the effect of product attributes, consumer attitudes and consumer interest on purchasing decisions of local citrus fruits on consumers in Malang City. This research method is descriptive quantitative using Structural Equation Modeling (SEM) analysis with Warp Partial Least Square (WarpPLS) approach. Primary data was obtained through filling out a questionnaire as an online interview media. Meanwhile, secondary data was obtained through literature studies from various related sources. Sampling was done based on non-probability sampling technique and 124 samples were selected as respondents in this study. The results showed that product attributes had a positive and significant effect on consumer attitudes and interests, consumer attitudes had a positive and significant effect on purchasing decisions, and positive and significant consumer interest on purchasing decisions.

Comprehensive studies of biological characteristics, phytochemical profiling, and antioxidant activities of two local citrus varieties in China.

Citrus is widely grown all over the world, and citrus fruits have long been recognized for their nutritional and medical value for human health. However, some local citrus varieties with potentially important value are still elusive. In the current study, we elucidated the biological characteristics, phylogenetic and phytochemical profiling, antioxidants and antioxidant activities of the two local citrus varieties, namely Zangju and Tuju. The physiological and phylogenetic analysis showed that Zangju fruit has the characteristics of wrinkled skin, higher acidity, and phylogenetically closest to sour mandarin Citrus sunki, whereas, Tuju is a kind of red orange with vermilion peel, small fruit and high sugar content, and closely clustered with Citrus erythrosa. The phytochemical analysis showed that many nutrition and antioxidant related differentially accumulated metabolites (DAMs) were detected in the peel and pulp of Zangju and Tuju fruits. Furthermore, it was found that the relative abundance of some key flavonoids and phenolic acids, such as tangeritin, sinensetin, diosmetin, nobiletin, and sinapic acid in the peel and pulp of Zangju and Tuju were higher than that in sour range Daidai and satsuma mandarin. Additionally, Zangju pulp and Tuju peel showed the strongest ferric reducing/antioxidant power (FRAP) activity, whereas, Tuju peel and pulp showed the strongest DPPH and ABTS free radical scavenging activities, respectively. Moreover, both the antioxidant activities of peel and pulp were significantly correlated with the contents of total phenols, total flavonoids or ascorbic acid. These results indicate that the two local citrus varieties have certain nutritional and medicinal value and potential beneficial effects on human health. Our findings will also provide an important theoretical basis for further conservation, development and medicinal utilization of Zangju and Tuju.

Prospects of using peels of Siamese orange young from fruit thinning in waste minimization

Siamese orange (Citrus nobilis) is a local citrus that grows in Mount Omeh, Lima Puluh Kota Regency. The quality is improved through thinning which is the process of removing some of the young fruit. Therefore, this research aimed to use young oranges obtained from thinning fruit in the manufacture of essential oils to reduce waste from citrus plants. Orange peel is converted into essential oil through distillation for 6 hours at 90-1000 C. The results showed that the product obtained had a yield of 0.9% (V/V), pale yellow color, volatility with a specific gravity of 0.84%, refractive index of 1.46, 0.67 acid number, solubility in ethanol 1:19 clear, ester number 27, and 0.02 mg/kg Fe content. The socialization of the use of essential oil and hydrosol in soap making to citrus farmers and industrial craftsmen around the Lima Puluh Kota Regency was also performed. The outcome indicate a positive response from the participants and the local government, thereby increasing the commercial value of oranges.

Genotyping of Sumatera local variety of citrus using random amplified polymorphism DNA (RAPD) technique

Indonesia has local varieties of citrus that are no less than imported citrus, especially in terms of fruit freshness. However, people are more interested in the color of citrus peel so people prefer imported citrus to local citrus, especially in Sumatera. Therefore, it is necessary to carry out efforts to conserve and improve the characteristics of these citrus to improve their quality through plant breeding. This study aims to optimize DNA isolation methods for citrus fruit samples with Chelex-TE and to determine the genetic profile of local Sumatera citrus and imported citrus using the genotyping RAPD. The samples used were several local citrus in Sumatera (Citrus Siam Mountain Omeh, Citrus Madu, Citrus Keprok Maga, Citrus Keprok Brastepu and Citrus Pasaman) and imported Citrus (Citrus Sunkist, Citrus Clemengold, Citrus Murkot and Citrus Wokam). DNA was isolated using the 10% Chelex-TE method which was optimized for several parameters such as grain size, fruit skin and leaves. RAPD was performed using 10 RAPD primers. The results showed that the optimum 10% Chelex Chelex-TE isolation method was a sample size of 1 grain. The amplification of local Sumatran citrus and imported citrus using 10 single primers produced polymorphic bands. The value of jaccard's similarity indicates that the five samples of Sumatera local variety of citrus and imported citrus have high genetic variation. Indonesia has local varieties of citrus that are no less than imported citrus, especially in terms of fruit freshness. However, people are more interested in the color of citrus peel so people prefer imported citrus to local citrus, especially in Sumatera. Therefore, it is necessary to carry out efforts to conserve and improve the characteristics of these citrus to improve their quality through plant breeding. This study aims to optimize DNA isolation methods for citrus fruit samples with Chelex-TE and to determine the genetic profile of local Sumatera citrus and imported citrus using the genotyping RAPD. The samples used were several local citrus in Sumatera (Citrus Siam Mountain Omeh, Citrus Madu, Citrus Keprok Maga, Citrus Keprok Brastepu and Citrus Pasaman) and imported Citrus (Citrus Sunkist, Citrus Clemengold, Citrus Murkot and Citrus Wokam). DNA was isolated using the 10% Chelex-TE method which was optimized for several parameters such as grain size, fruit skin and leaves. RAPD was performed using 10 RAPD primers. The results showed that the optimum 10% Chelex Chelex-TE isolation method was a sample size of 1 grain. The amplification of local Sumatran citrus and imported citrus using 10 single primers produced polymorphic bands. The value of jaccard's similarity indicates that the five samples of Sumatera local variety of citrus and imported citrus have high genetic variation.

Physiological and Metabolic Changes in ‘Xinyu Mandarin’ Following Natural Tetraploidization

The mandarin is an important fruit crop worldwide, and ‘Xinyu mandarin’—a local Citrus reticulata variety—is widely cultivated in Jiangxi Province, in China. Autopolyploidy has frequently been used for the improvement of crop varieties. In a previous study, we identified a tetraploid (4X) material of ‘Xinyu mandarin’ generated from its diploid (2X) mother seedling via a spontaneous mutation. However, the physiological and metabolic variations after polyploidy are not clear enough, which is not conducive to the early screening. In the present study, we analyzed the morphological, physiological, and metabolic differences between the tetraploid and diploid plants. The tetraploids had larger and thicker leaves, and the activities of key enzymes in photosynthesis, the total chlorophyll, phenolic, and proline contents, were all higher in tetraploids. In the non-targeted metabolomic profile, a total of 522 metabolites were identified, of which 61 were significantly different between diploids and tetraploids. The differential metabolites included similar proportions of primary and secondary metabolites, and most of these were up-regulated in tetraploids, especially stress-related metabolites such as phenolic acids, flavonoids, alkaloids, and so on, indicating that tetraploids may possess better stress tolerance ability and vigor. Therefore, the tetraploids of ‘Xinyu mandarin’ may serve as an excellent female parent for the improvement of citrus varieties.

Application of RAPD Molecular Technique to Study the Genetic Variations of Citrus in South Sulawesi, Indonesia

Indonesia is a tropical country with superior local citrus species and varieties. Citrus fruits were very useful for cultivation, especially to meet the nutritional needs of the wider community. A province in Indonesia, namely South Sulawesi is one of the centers for citrus development. A preliminary study showed various cultivated citrus in South Sulawesi namely Mandarin Orange cultivar Selayar, Mandarin Orange cultivar Batu, JC lime, Pummelo Pangkep, tangerine, Santang Madu, Dekopon, lime, and Kaffir lime. This study aims to evaluate genetic diversity at several citrus plantation centers in South Sulawesi using the RAPD technique. The analysis was carried out using five primers. In this study, RAPD primers could be used to characterize the genetic diversity and similarity of thirteen citrus cultivars in South Sulawesi. One informative RAPD primer based on its PIC value was OPC-09. The results of the genetic similarity analysis are presented in the form of a dendrogram. The first cluster consisted of Mandarin Orange cultivar Selayar (Seeded Selayar, Selayar-Selayar, JC-Selayar), JC lime, Mandarin Orange cultivar Batu, Santang Madu, and Pummelo Pangkep (cultivar Pangkep Merah, Pangkep Putih, Pangkep Golla-golla). The second cluster consisted of Mandarin Orange cultivar Selayar (selayar-selayar, JC-selayar), Santang Madu, tangerine, Mandarin Orange cultivar Batu, Dekopon, lime, and Kaffir lime. The clusters with the most distant genetic relationship are cluster A with cluster B, with a genetic similarity of 62%. Meanwhile, clusters with the closest genetic relationship are clusters I and II, with 79% genetic similarity.

LIQUEFIED WATER AND ANTIOXIDANTS INFLUENCE ON VEGETATIVE GROWTH AND PHYSIOLOGICAL TRAITS OF SWEET ORANGE

Citrus production faces many problems; a major one consists of irrigating citrus seedlings grown at Agricultural Research Stations and various nurseries with saline well water, which determines the growth, reproduction, and spread of citrus in Iraq. The said problem needs addressing to reduce its effects on the growth and reproduction of citrus fruits. Therefore, the latest research aimed to determine the effects of regular liquefied water and saline well water, in addition to a foliar spray of aspartic and ascorbic acids, on the growth and physiological properties of the grafted orange seedlings. The local citrus seedlings of the same age and size were selected and used as the original and grafted with local orange buds. They were shifted and tested for the above three factors at the Horticulture and Forestry Division, Najaf Agriculture Directorate, Iraq. The findings showed that irrigation with regular liquefied water had improved the growth traits, i.e., plant height, leaves plant-1, leaf area plant-1, and dry biomass weight of sweet orange seedlings, compared with saline well water. Results further revealed that foliar application of aspartic acid (100 mg L-1) positively affected the leaves plant-1 and dry biomass weight compared with the control. The ascorbic acid (4000 mg L-1) application gave highly superior and well-responsive reactions for most of the traits, i.e., leaves plant-1 (69.3 leaves plant-1), leaf area plant-1 (3888 cm2), dry biomass weight (0.3629 g), and total chlorophyll content (7.03 mg 100 g-1 fresh weight) in orange seedlings.

Keragaman Buah Pamelo Nambangan Hasil Induksi Sinar Gamma

Citrus has high commercial values both in local and global markets. Consumption of citrus as a fresh and functional fruit continues to increase along with the increase in population and the value of its benefits. The diversity of Indonesian local citrus with their specific value could be found in almost all regions. Nevertheless, the application of technology is necessary to improve the quality of local fruit performance and production quantity. Improvement of local fruit quality can be done through various methods, one of which is mutation by gamma ray induction. The aim of the study was to determine the fruit diversity of Nambangan Pummelo resulted from gamma ray induction. The research was conducted at the Experimental Field of Kraton Pasuruan and the Integrated Laboratory of Balitjestro in 2014. The results showed that the induced mutation contributed significantly to the increase in fruit diversity. Induced mutation has an effect on decreasing the number of seeds of the mutant fruit of Nambangan compared to control plant, increased the diversity of fruit flesh color, the level of bitterness and the shape of the fruit.

Morpho-chemical characterization of Huanglongbing in mandarin (Citrus reticulata) and orange (Citrus sinensis) varieties from Pakistan

Huanglongbing (HLB) is one of the world’s worst diseases of citrus trees. The research was conducted to characterize positive citrus samples of two local varieties, ‘Kinnow’ mandarin (Citrus reticulata Blanco) and ‘Mosambi’ orange (C. sinensis (L.) Osbeck), based on morphological, physical and biochemical characters. Physical and biochemical parameters of greening infected and healthy samples of both the cultivars were significantly different. Citrus fruits were lopsided and half to full green in the HLB infected trees compared to healthy. The HLB infected leaves samples were small and narrow. Fruit diameter, leaf area, and juice weight of HLB infected samples were significantly less than healthy samples. The rind thickness of infected fruits (70% and 80%) was significantly higher than healthy ones (20% and 30%). Starch content of leaves from mandarin (5.54 μg mL-1) and orange (6.12 μg mL-1) were significantly higher in HLB infected plants, while juice percentage of HLB infected fruit samples was lesser in mandarin and orange (22.73%, and 14.70%, respectively) than healthy fruits (38.22% and 31.30%, respectively). Acidity was significantly increased in juice of HLB infected mandarin and orange fruits (0.57%, and 0.53%, respectively). Similarly, there was significant reduction in biochemical parameters; total sugars in mandarin and orange (5.16% and 4.79%), total soluble solids (8.54% and 8.28%), total chlorophyll (13.50 and 14.92 mg mL-1) and ascorbic acid (34.28% and 34.12%) in HLB infected samples of both cultivars. This study determined differences of physical and biochemical parameters of two local citrus varieties, commonly cultivated in the Asian region.

Leaf Volatiles and Relevant Gene Expression as the Specific Characteristics in Citrus depressa Accession Discrimination

Citrus depressa Hayata is the native and widespread citrus species in Taiwan. The notable character is that C. depressa has a distinct aroma different from local citrus. The ex situ germplasm of scions from different collection regions has variant leaf shapes and different odor characteristics. Establishing volatile biomarkers for classifying the local C. depressa is beneficial to commercial development. Volatile organic compounds (VOCs) of fresh leaves from seven C. depressa accessions which were collected from different locations in Taiwan were extracted by headspace solid-phase microextraction and analyzed by GC-MS. The volatile compositions from each season showed the diversity, and linalool, of which the average relative content is 52.7%, was the most volatile component in any season. The other main VOCs of leaves of C. depressa were γ-terpinene, limonene, β-ocimene, and α-terpineol. The result of linear discriminant analysis by VOC markers shows that there are two main different types which are (1) accessions from the central and the east of Taiwan and (2) accessions which are closer to C. depressa in Okinawa, Japan. Five major VOC-related synthase genes were selected and the gene expression was used to classify the varieties. The clustering result is the same with VOC-based discrimination. Our results reveal leaf volatile profiling is capable of being the discrimination markers, and the possibility for constructing molecular markers is directly related to characteristics from secondary metabolites phenotyping.

First report of post-harvest Fusarium rot of mandarin Citrus reticulata cv. 'Kinnow' caused by Fusarium equiseti in Pakistan.

Citrus reticulata cv. 'Kinnow' mandarin is the most popular and widely grown fruit crop in Pakistan. During 2017, a survey was conducted to the local citrus fruit markets of Faisalabad, Pakistan. Citrus fruits (n=50) exhibiting stem end rot and fruit rot were collected with 15% disease incidence. The stem end region showed light to dark brown lesions and white fungal growth was also observed in the severely infected fruit. Infected fruit were excised into 2mm2 segments, surface disinfected with 1% NaClO, rinsed with sterilized water and dried. Later, these tissues were placed on potato dextrose agar (PDA) medium and subsequently incubated at 25 °C. Purified isolates produced white colonies with beige pigmentation. The frequency of fungal isolation was 47%. Microscopic observations revealed that macroconidia (n=50) had 5 to 6 septations, with a prominent dorsiventral curvature, tapered and elongated apical cell, and a foot shape basal cell. The macroconidia were measuring 22 to 45 × 2.9 to 4.3 µm with an average of 31 × 3.6 µm. However, microconidia were not observed. Chlamydospores were globose, intercalary, solitary, or in pairs, appearing in chains (Leslie and Summerell 2006). For molecular identification, DNA was extracted from all isolates. The internal transcribed spacer region (ITS) ITS1/4 (White et al. 1990), translation elongation factor-1 alpha (TEF) EF1/2 (O'Donnell et al. 1998), and RNA polymerase II subunit 1 (RPB1) (O'Donnell et al. 2013) were amplified using PCR and the product was subsequently sequenced. Based on BLAST analysis, the isolate was identified as Fusarium equiseti (FUS-21). The sequences of the representative isolate FUS-21 were deposited in the GenBank with accession numbers (ITS, MH581300), (TEF, MK203749), and (RPB1, MW596599) showing more than 99% similarity with ITS accession GQ505683, TEF accession GQ505594, and 100% to RPB1 accession JX171481. To determine the pathogenicity, 40 healthy surface disinfested citrus fruit were taken. The fruit were inoculated by creating artificial wounds on the surface with a sterilized needle and 10 μL of 105 spores/mL was deposited in the wounds. In case of control fruit were inoculated with 10 μL sterilized distilled water only, and incubated at 25 °C. All fruit inoculated with the putative pathogen, developed symptoms like the original fruit from which they were isolated. The pathogenicity test was repeated twice. Visible white mycelium appeared at the stem end region and the fruits became dried as the infection progressed. However, the control fruit remained asymptomatic. The pathogen was re-isolated from infected fruit and identified based on morphometric and molecular analysis. Previously we have reported F. oxysporum causing citrus fruit rot in Pakistan (Moosa et al. 2020). This is the first report of F. equiseti causing post-harvest rot of citrus fruits in Pakistan. Kinnow is an important fruit crop of Pakistan with huge export value the management of Fusarium rot is quite important to save the loss of fresh produce.

Land suitability determination for citrus cultivation using a GIS-based multi-criteria analysis in Mersin, Turkey

Determination of suitable locations for agricultural products is an important subject to study, as it is essential to identify optimal areas for crop productivity and sustainable farming activities. This study proposes a site selection assessment model for citrus cultivation using Geographic Information System (GIS) Based Multi-Criteria Decision Analysis (MCDA) in the Mersin province of Turkey. The purpose of the study is to create a Multi-Criteria Assessment-Based Spatial Decision Support System which takes into account the opinions of local citrus farmers as well as those of experts. 14 preference factors and 10 exclusion factors were determined for the analysis of land suitability for citrus farming. The weighting of preference factors for citrus cultivation was determined using the Analytic Hierarchy Process, and a land suitability map was generated using the weighted overlay method in the GIS environment. Based on the opinions of farmers and experts, higher weights were assigned to the average temperature in the flowering period, the average temperature in the winter season, and elevation, as compared to all other preference factors. The results also demonstrate that WorldClim, a global climate dataset, is an essential source of data in such studies, as climate conditions play a crucial role in potential citrus expansion. The final land suitability map shows that the most suitable and moderately suitable areas for citrus cultivation have surface areas of 85 824 ha and 85 430 ha, respectively. The Receiver Operating Characteristic (ROC) curve was used to evaluate validation of the land suitability for citrus and the Area Under the Curve (AUC) value was calculated at 0.87, indicating substantial consistency. This study proposes a methodology for the determination of suitable sites for citrus cultivation which relies on realistic data. The land suitability map produced in the study will be essential for the creation of sustainable agriculture strategies in the eastern Mediterranean region.

Medicine and food with particular reference to chinpi, dried citrus peel, and a component of Ninjin'yoeito

Kampo medicines contain many kinds of herbal drugs. Chinpi and Kippi, dried citrus peels, are components of a substantial number of Kampo medicine. They contain abundant flavonoids and studies on hesperidin, narirutin, and nobiletin as active ingredient have been conducted. Conversely, in Kagoshima prefecture, located in the southwestern part of the Japanese Islands, various citrus products are cultivated. Among them, Tankan and Daimasaki are specialies. In this study, we conducted high- performance liquid chromatography to determine the difference in flavonoid contents among Tankan, Daimasaki, Tankan related product, Chinpi, and Kippi. As a result, several active components, such as hesperidin, narirutin, nobiletin, and tangeretin, in common with crude drug, Chinpi, were detected in local citrus fruits. In addition, some active components little or not found in Chinpi, for example hesperetin and rutin, were detected in the local products. A detailed analysis of active components considering their genetic origin, the time of fruit collection, and different parts of the fruit used (peel, albedo, edible parts, and the whole) will need to be discussed to get the most out of the citrus fruits or make best use of them for health and longevity.