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Manjul Dutt
Citrus Research and Education Center, University of Florida, Lake Alfred, FL, United States of America

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Genetics
Published: 10 August 2021 in PLOS ONE
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The Australian finger lime is a unique citrus species that has gained importance due to its unique fruit characteristics and perceived tolerance to Huanglongbing (HLB), an often-fatal disease of citrus trees. In this study, we developed allotetraploid finger lime hybrids and cybrids by utilizing somatic cell fusion techniques to fuse diploid ‘OLL8’ sweet orange or ‘Page’ tangelo callus-derived protoplasts with finger lime (FL) mesophyll-derived protoplasts. Six somatic fusions were regenerated from the ‘OLL8’ + FL fusion, while three putative cybrids were regenerated from the ‘Page’ + FL fusion. Ploidy levels and nuclear-expressed sequence tag derived simple sequence repeat (EST-SSR) markers confirmed the somatic hybrid production, and mitochondrial DNA primer sets confirmed the cybrid nature. Several trees produced by the somatic fusion remained HLB negative even after 6 years of growth in an HLB-endemic environment. Pathogenesis related (PR) and other genes that are often upregulated in HLB-tolerant trees were also upregulated in our somatic fusions. These newly developed somatic fusions and cybrids could potentially be used as breeding parents to develop the next generation of improved HLB-tolerant rootstocks and scions.

ACS Style

Manjul Dutt; Lamiaa M. Mahmoud; Karen Chamusco; Daniel Stanton; Christine D. Chase; Ethan Nielsen; Maria Quirico; Qibin Yu; Frederick G. Gmitter; Jude W. Grosser. Utilization of somatic fusion techniques for the development of HLB tolerant breeding resources employing the Australian finger lime (Citrus australasica). PLOS ONE 2021, 16, e0255842 .

AMA Style

Manjul Dutt, Lamiaa M. Mahmoud, Karen Chamusco, Daniel Stanton, Christine D. Chase, Ethan Nielsen, Maria Quirico, Qibin Yu, Frederick G. Gmitter, Jude W. Grosser. Utilization of somatic fusion techniques for the development of HLB tolerant breeding resources employing the Australian finger lime (Citrus australasica). PLOS ONE. 2021; 16 (8):e0255842.

Chicago/Turabian Style

Manjul Dutt; Lamiaa M. Mahmoud; Karen Chamusco; Daniel Stanton; Christine D. Chase; Ethan Nielsen; Maria Quirico; Qibin Yu; Frederick G. Gmitter; Jude W. Grosser. 2021. "Utilization of somatic fusion techniques for the development of HLB tolerant breeding resources employing the Australian finger lime (Citrus australasica)." PLOS ONE 16, no. 8: e0255842.

Journal article
Published: 14 July 2021 in Plants
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Huanglongbing (HLB), caused by the phloem-limited bacterium Candidatus Liberibacter asiaticus (CaLas), is the primary biotic stress causing significant economic damage to the global citrus industry. Among the abiotic stresses, salinity affects citrus production worldwide, especially in arid and coastal regions. In this study, we evaluated open-pollinated seedlings of the S10 (a diploid rootstock produced from a cross between two siblings of the Hirado Buntan Pink pummelo (Citrus maxima (Burm.) Merr.) with the Shekwasha mandarin (Citrus reticulata Blanco)) for their ability to tolerate HLB and salinity stresses. In a greenhouse study, ‘Valencia’ sweet orange (either HLB-positive or negative) was grafted onto six clonally propagated lines generated from the screened seedlings in the greenhouse and the trees were irrigated with 150 mM NaCl after eight months of successful grafting and detection of CaLas in the leaf petioles. Cleopatra mandarin was used as a salt-tolerant and HLB-sensitive rootstock control. CaLas infection was monitored using a quantitative polymerase chain reaction before and after NaCl treatments. Following three months of NaCl treatment, ‘Valencia’ leaves on the S10 rootstock seedlings recorded lower levels of chlorophyll content compared to Cleopatra under similar conditions. Malondialdehyde content was higher in HLB-infected ‘Valencia’ grafted onto Cleopatra than in the S10 lines. Several plant defense-related genes were significantly upregulated in the S10 lines. Antioxidant and Na+ co-transporter genes were differentially regulated in these lines. Based on our results, selected S10 lines have potential as salt-tolerant rootstocks of ‘Valencia’ sweet orange under endemic HLB conditions. However, it is necessary to propagate selected lines through tissue culture or cuttings because of the high percentage of zygotic seedlings derived from S10.

ACS Style

Lamiaa Mahmoud; Patrick Huyck; Christopher Vincent; Frederick Gmitter; Jude Grosser; Manjul Dutt. Physiological Responses and Gene Expression Patterns in Open-Pollinated Seedlings of a Pummelo-Mandarin Hybrid Rootstock Exposed to Salt Stress and Huanglongbing. Plants 2021, 10, 1439 .

AMA Style

Lamiaa Mahmoud, Patrick Huyck, Christopher Vincent, Frederick Gmitter, Jude Grosser, Manjul Dutt. Physiological Responses and Gene Expression Patterns in Open-Pollinated Seedlings of a Pummelo-Mandarin Hybrid Rootstock Exposed to Salt Stress and Huanglongbing. Plants. 2021; 10 (7):1439.

Chicago/Turabian Style

Lamiaa Mahmoud; Patrick Huyck; Christopher Vincent; Frederick Gmitter; Jude Grosser; Manjul Dutt. 2021. "Physiological Responses and Gene Expression Patterns in Open-Pollinated Seedlings of a Pummelo-Mandarin Hybrid Rootstock Exposed to Salt Stress and Huanglongbing." Plants 10, no. 7: 1439.

Original article
Published: 06 January 2021 in Plant Physiology Reports
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Salinity is an obstacle to citriculture worldwide, and is a concern in arid, semiarid, and coastal regions. In the current study, we irrigated one-year-old ‘Valencia’ trees budded onto Kuharske rootstock with 60 mM sodium chloride (NaCl) solution for ten weeks. Subsequently, these trees were sprayed with 50, 75, and 100 mM SA or MeJA to determine whether these phytohormones could alleviate the detrimental effects of salinity. Control trees were not sprayed, including a positive control with saline irrigation and a negative control without saline irrigation. A reduction in plant growth and chlorophyll content following the NaCl treatment was recorded. Subsequent SA and MeJA treatments promoted tree growth and enhanced chlorophyll content in these trees. Additionally, these treatments enhanced the expression of enzymatic antioxidants: POD, CSD, CAT, PAL, APX1, GSTs, aquaporin proteins, and Na+ co-transporters. SA and MeJA treatments also altered the expression of PR1, PR3, PR4, and PR5, while only MeJA altered the expression of PR2. Abscisic acid (ABA) expression was negatively affected by SA treatments, whereas the MeJA application increased ABA expression relative to the salt treated control. Furthermore, sodium (Na+) and chloride (Cl−) analysis indicated that leaves had higher levels of Na+ and Cl− in response to SA and MeJA treatments when compared with the controls. Our results provide perspectives at the possible function of SA and MeJA in ameliorating salt stress in citrus.

ACS Style

Lamiaa M. Mahmoud; Christopher I. Vincent; Jude W. Grosser; Manjul Dutt. The response of salt-stressed Valencia sweet orange (Citrus sinensis) to salicylic acid and methyl jasmonate treatments. Plant Physiology Reports 2021, 26, 137 -151.

AMA Style

Lamiaa M. Mahmoud, Christopher I. Vincent, Jude W. Grosser, Manjul Dutt. The response of salt-stressed Valencia sweet orange (Citrus sinensis) to salicylic acid and methyl jasmonate treatments. Plant Physiology Reports. 2021; 26 (1):137-151.

Chicago/Turabian Style

Lamiaa M. Mahmoud; Christopher I. Vincent; Jude W. Grosser; Manjul Dutt. 2021. "The response of salt-stressed Valencia sweet orange (Citrus sinensis) to salicylic acid and methyl jasmonate treatments." Plant Physiology Reports 26, no. 1: 137-151.

Journal article
Published: 08 December 2020 in Scientific Reports
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Shortening the juvenile stage in citrus and inducing early flowering has been the focus of several citrus genetic improvement programs. FLOWERING LOCUS T (FT) is a small phloem-translocated protein that regulates precocious flowering. In this study, two populations of transgenic Carrizo citrange rootstocks expressing either Citrus clementina FT1 or FT3 genes under the control of the Arabidopsis thaliana phloem specific SUCROSE SYNTHASE 2 (AtSUC2) promoter were developed. The transgenic plants were morphologically similar to the non-transgenic controls (non-transgenic Carrizo citrange), however, only AtSUC2-CcFT3 was capable of inducing precocious flowers. The transgenic lines produced flowers 16 months after transformation and flower buds appeared 30–40 days on juvenile immature scions grafted onto transgenic rootstock. Gene expression analysis revealed that the expression of SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) and APETALA1 (AP1) were enhanced in the transgenics. Transcriptome profiling of a selected transgenic line showed the induction of genes in different groups including: genes from the flowering induction pathway, APETALA2/ETHYLENE RESPONSE FACTOR (AP2/ERF) family genes, and jasmonic acid (JA) pathway genes. Altogether, our results suggested that ectopic expression of CcFT3 in phloem tissues of Carrizo citrange triggered the expression of several genes to mediate early flowering.

ACS Style

Juliana M. Soares; Kyle C. Weber; Wenming Qiu; Daniel Stanton; Lamiaa M. Mahmoud; Hao Wu; Patrick Huyck; Janice Zale; Kawther Al Jasim; Jude W. Grosser; Manjul Dutt. The vascular targeted citrus FLOWERING LOCUS T3 gene promotes non-inductive early flowering in transgenic Carrizo rootstocks and grafted juvenile scions. Scientific Reports 2020, 10, 1 -18.

AMA Style

Juliana M. Soares, Kyle C. Weber, Wenming Qiu, Daniel Stanton, Lamiaa M. Mahmoud, Hao Wu, Patrick Huyck, Janice Zale, Kawther Al Jasim, Jude W. Grosser, Manjul Dutt. The vascular targeted citrus FLOWERING LOCUS T3 gene promotes non-inductive early flowering in transgenic Carrizo rootstocks and grafted juvenile scions. Scientific Reports. 2020; 10 (1):1-18.

Chicago/Turabian Style

Juliana M. Soares; Kyle C. Weber; Wenming Qiu; Daniel Stanton; Lamiaa M. Mahmoud; Hao Wu; Patrick Huyck; Janice Zale; Kawther Al Jasim; Jude W. Grosser; Manjul Dutt. 2020. "The vascular targeted citrus FLOWERING LOCUS T3 gene promotes non-inductive early flowering in transgenic Carrizo rootstocks and grafted juvenile scions." Scientific Reports 10, no. 1: 1-18.

Journal article
Published: 25 November 2020 in Horticulturae
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Our study aimed to evaluate the physiological responses following salinity treatment of three putatively salt-tolerant Citrus rootstocks recently developed by the University of Florida’s Citrus breeding program. Four-month-old seedlings from each of the three rootstocks (HS1, HS17, and HC15) were irrigated with 0, 60, 80, and 100 mm NaCl solution. The seedlings were evaluated together with the salt-tolerant Cleopatra mandarin as a positive control, Volkamer lemon as a moderately salt-tolerant rootstock, and the salt-sensitive Carrizo rootstock as a negative control. Our results demonstrated that chlorophyll content, net CO2 assimilation rate (A), transpiration rate (E), and stomatal conductance (gsw ) significantly decreased in response to salinity. Na+ and Cl− levels were higher in leaf tissues than in the roots. Relatively little damage to the cellular membrane was recorded in HC15 and Cleopatra rootstocks under the 100 mm NaCl treatment, along with high accumulation of total phenolic content (TPC), while HS17 had the highest proline levels. Our results indicate that HC15 and HS17 rootstocks exhibited salt tolerance capacity via different strategies under salt stress and could be suitable replacements to the commercially available, salt-tolerant Cleopatra rootstock.

ACS Style

Lamiaa Mahmoud; Manjul Dutt; Christopher Vincent; Jude Grosser. Salinity-Induced Physiological Responses of Three Putative Salt Tolerant Citrus Rootstocks. Horticulturae 2020, 6, 90 .

AMA Style

Lamiaa Mahmoud, Manjul Dutt, Christopher Vincent, Jude Grosser. Salinity-Induced Physiological Responses of Three Putative Salt Tolerant Citrus Rootstocks. Horticulturae. 2020; 6 (4):90.

Chicago/Turabian Style

Lamiaa Mahmoud; Manjul Dutt; Christopher Vincent; Jude Grosser. 2020. "Salinity-Induced Physiological Responses of Three Putative Salt Tolerant Citrus Rootstocks." Horticulturae 6, no. 4: 90.

Journal article
Published: 24 November 2020 in Metabolites
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The citrus industry at present is severely affected by huanglongbing disease (HLB). HLB is caused by the supposed bacterial pathogen “Candidatus Liberibacter asiaticus” and is transmitted by the insect vector, the Asian citrus psyllid, Diaphorina citri Kuwayama. Developing new citrus hybrids to improve HLB management is much needed. In this study, we investigated the metabolomic profiles of three new hybrids produced from the cross of C2-5-12 Pummelo (Citrus maxima (L.) Osbeck) × pollen from Citrus latipes. The hybrids were selected based on leaf morphology and seedling vigor. The selected hybrids exhibited compact and upright tree architecture as seen in C. latipes. Hybrids were verified by simple sequence repeat markers, and were subjected to metabolomic analysis using gas chromatography-mass spectrometry. The volatile organic compounds (VOCs) and polar metabolites profiling also showed that the new hybrids were different from their parents. Interestingly, the levels of stored VOCs in hybrid II were higher than those observed in its parents and other hybrids. The level of most VOCs released by hybrid II was also higher than that released from its parents. Additionally, the preference assay showed that hybrid II was more attractive to D. citri than its parents and other hybrids. The leaf morphology, compact and upright architecture of hybrid II, and its attraction to D. citri suggest that it could be used as a windbreak and trap tree for D. citri (double duty), once its tolerance to HLB disease is confirmed. Our results showed that metabolomic analysis could be successfully used to understand the biochemical mechanisms controlling the interaction of D. citri with its host plants.

ACS Style

Nabil Killiny; Shelley E. Jones; Faraj Hijaz; Abdelaziz Kishk; Yulica Santos-Ortega; Yasser Nehela; Ahmad A. Omar; Qibin Yu; Jr. Fred G. Gmitter; Jude W. Grosser; Manjul Dutt. Metabolic Profiling of Hybrids Generated from Pummelo and Citrus latipes in Relation to Their Attraction to Diaphorina citri, the Vector of Huanglongbing. Metabolites 2020, 10, 477 .

AMA Style

Nabil Killiny, Shelley E. Jones, Faraj Hijaz, Abdelaziz Kishk, Yulica Santos-Ortega, Yasser Nehela, Ahmad A. Omar, Qibin Yu, Jr. Fred G. Gmitter, Jude W. Grosser, Manjul Dutt. Metabolic Profiling of Hybrids Generated from Pummelo and Citrus latipes in Relation to Their Attraction to Diaphorina citri, the Vector of Huanglongbing. Metabolites. 2020; 10 (12):477.

Chicago/Turabian Style

Nabil Killiny; Shelley E. Jones; Faraj Hijaz; Abdelaziz Kishk; Yulica Santos-Ortega; Yasser Nehela; Ahmad A. Omar; Qibin Yu; Jr. Fred G. Gmitter; Jude W. Grosser; Manjul Dutt. 2020. "Metabolic Profiling of Hybrids Generated from Pummelo and Citrus latipes in Relation to Their Attraction to Diaphorina citri, the Vector of Huanglongbing." Metabolites 10, no. 12: 477.

Journal article
Published: 10 November 2020 in BMC Biotechnology
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Background Development of precise genome editing strategies is a prerequisite for producing edited plants that can aid in the study of gene function and help understand the genetic traits in a cultivar. Citrus embryogenic cell cultures can be used to rapidly produce a large population of genome edited transformed citrus lines. The ability to introduce specific mutations in the genome of these cells using two constructs (pC-PDS1 and pC-PDS2) was evaluated in this study. Results Citrus sinensis ‘EV2’ embryogenic cell cultures are amenable to Agrobacterium-mediated CRISPR/Cas9-based genome editing. Guide RNAs (gRNAs) targeting two locations in the phytoene desaturase (PDS) gene were either driven by the Arabidopsis U6–26 promoter (pC-PDS1) or assembled as a Csy4 array under the control of the CmYLCV promoter (pC-PDS2). All transgenic embryos were completely albino and no variegated phenotype was observed. We evaluated 12 lines from each construct in this study and the majority contain either insertion (1–2 bp), substitution (1 bp), or deletion (1–3 bp) mutations that occurred close to the protospacer adjacent motif. Conclusions Both the pC-PDS1 and pC-PDS2 could successfully edit the citrus embryogenic cell cultures. However, the editing efficiency was dependent on the gRNA, confirming that the selection of a proper gRNA is essential for successful genome editing using the CRISPR/Cas9 technique. Also, utilization of embryogenic cell cultures offers another option for successful genome editing in citrus.

ACS Style

Manjul Dutt; Zhonglin Mou; Xudong Zhang; Sameena E. Tanwir; Jude W. Grosser. Efficient CRISPR/Cas9 genome editing with Citrus embryogenic cell cultures. BMC Biotechnology 2020, 20, 1 -7.

AMA Style

Manjul Dutt, Zhonglin Mou, Xudong Zhang, Sameena E. Tanwir, Jude W. Grosser. Efficient CRISPR/Cas9 genome editing with Citrus embryogenic cell cultures. BMC Biotechnology. 2020; 20 (1):1-7.

Chicago/Turabian Style

Manjul Dutt; Zhonglin Mou; Xudong Zhang; Sameena E. Tanwir; Jude W. Grosser. 2020. "Efficient CRISPR/Cas9 genome editing with Citrus embryogenic cell cultures." BMC Biotechnology 20, no. 1: 1-7.

Chapter
Published: 09 October 2020 in Genetically Modified Crops
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Citrus is a very nutritious fruit that is consumed worldwide, making it one of the most economically important fruits. Like other crops, citrus must cope with multiple biotic and abiotic stressors in its growing environment. These stressors represent obstacles which must be overcome by horticultural improvement programs. Conventional breeding and genetic transformation are the most popular techniques for citrus improvement. Because conventional breeding has several limitations, biotechnology-mediated strategies are being increasingly used, allowing better management of genetic resources and faster development of superior genotypes. Gene transfer in citrus demands reproducibility as well as effective genetic transformation and plant regeneration protocols. Although Agrobacterium-mediated transformation is the main method of citrus genetic modification, other direct gene transfer techniques have also attracted attention recently. Together, these methods have produced promising results against abiotic and biotic stresses which affect citrus production and has also helped in enhancing yield and fruit quality. Nevertheless, novel techniques are constantly being developed for increasing the efficiency of citrus improvement efforts. Currently, CRISPR/Cas9 is the most efficient genome-editing tool with good potential for developing edited citrus plants. In this chapter, we present the tools and techniques used with citrus biotechnology. We also highlight the efforts of the scientific community toward the generation of genetically modified citrus plants.

ACS Style

Sameena E. Tanwir; Juliana M. Soares; Stacy Welker; Jude W. Grosser; Manjul Dutt. Genetically Modified Citrus: Current Status, Prospects, and Future Challenges. Genetically Modified Crops 2020, 161 -201.

AMA Style

Sameena E. Tanwir, Juliana M. Soares, Stacy Welker, Jude W. Grosser, Manjul Dutt. Genetically Modified Citrus: Current Status, Prospects, and Future Challenges. Genetically Modified Crops. 2020; ():161-201.

Chicago/Turabian Style

Sameena E. Tanwir; Juliana M. Soares; Stacy Welker; Jude W. Grosser; Manjul Dutt. 2020. "Genetically Modified Citrus: Current Status, Prospects, and Future Challenges." Genetically Modified Crops , no. : 161-201.

Journal article
Published: 01 June 2020 in South African Journal of Botany
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Climate change and rising sea levels increasingly threaten agriculture, especially in the coastal regions. Banana cultivars, especially the Cavendish clones, are sensitive to salinity stresses, which results in decreased production. In the current study, we evaluated the water deficit stress remediating effects of silicon nanoparticles (SiO2NPs) on micropropagated banana cv. “Grand Nain” - either under in vitro conditions in the laboratory or ex vitro in the greenhouse. In vitro water deficit was induced with polyethylene glycol (PEG-8000). Addition of in vitro SiO2NPs enhanced shoot growth and chlorophyll content. Malondialdehyde content (MDA) and electrolyte leakage (EL) were reduced in 3% PEG-stressed plants following the addition of the 150 mg/l SiO2NPs (38.73 nmole and 4.93%) compared with the control plants (51.67 nmole and 5.76%). In the greenhouse study, plants under salt and water deficit treatments were sprayed with SiO2NPs at (0, 200, 400 and 600 mg/l). Under salinity stress conditions, SiO2NPs improved photosynthesis as the stressed plants had similar chlorophyll content as control unstressed plants (23.33). SiO2NPs at 400 mg/l application increased K+ level (10.54%) and decreased Na+ levels (1.17%). Under simulated water deficit stress conditions, 600 mg/l SiO2NPs significantly increased K+ and K+/Na+ percent (10.37 and 14.40%) along with improvement in chlorophyll content index at all levels of SiO2NPs application. The overall results revealed that SiO2NPs application can improve chlorophyll content, induce K+ uptake, modulate Na+ levels and decrease cell wall damage in the treated plants comparing to the untreated plants under abiotic stress. Our results suggest that the application of SiO2NPs can help maintain plant integrity in the banana cultivar ‘Grand Nain’ under simulated abiotic stress.

ACS Style

Lamiaa M. Mahmoud; Manjul Dutt; Ameer M. Shalan; Mahmoud E. El-Kady; Mohamed S. El-Boray; Yasser Shabana; Jude W. Grosser. Silicon nanoparticles mitigate oxidative stress of in vitro-derived banana (Musa acuminata ‘Grand Nain’) under simulated water deficit or salinity stress. South African Journal of Botany 2020, 132, 155 -163.

AMA Style

Lamiaa M. Mahmoud, Manjul Dutt, Ameer M. Shalan, Mahmoud E. El-Kady, Mohamed S. El-Boray, Yasser Shabana, Jude W. Grosser. Silicon nanoparticles mitigate oxidative stress of in vitro-derived banana (Musa acuminata ‘Grand Nain’) under simulated water deficit or salinity stress. South African Journal of Botany. 2020; 132 ():155-163.

Chicago/Turabian Style

Lamiaa M. Mahmoud; Manjul Dutt; Ameer M. Shalan; Mahmoud E. El-Kady; Mohamed S. El-Boray; Yasser Shabana; Jude W. Grosser. 2020. "Silicon nanoparticles mitigate oxidative stress of in vitro-derived banana (Musa acuminata ‘Grand Nain’) under simulated water deficit or salinity stress." South African Journal of Botany 132, no. : 155-163.

Review
Published: 25 May 2020 in Tropical Plant Pathology
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Citrus cultivation is challenging due to the plethora of abiotic and biotic stresses faced by the crop. In recent years, production has been severely affected by diseases such as citrus canker and huanglongbing (HLB). Disease management is hampered as there is no field resistance to these diseases in any of the important commercially planted varieties. Traditionally, conventional breeding approaches have been applied for the improvement of the susceptible cultivars; however, this technique is laborious and time consuming. Genetic transformation of citrus allows for the rapid integration of novel genes into the plant’s genome to develop disease-resistant transgenic plants. Therefore, efforts have been made to utilize genetic engineering tools to develop genetically modified citrus that are resistant to citrus canker and HLB. This review summarizes the major achievements made in the development of citrus canker and HLB tolerance using transgenic technologies.

ACS Style

Juliana M. Soares; Sameena E. Tanwir; Jude W. Grosser; Manjul Dutt. Development of genetically modified citrus plants for the control of citrus canker and huanglongbing. Tropical Plant Pathology 2020, 45, 237 -250.

AMA Style

Juliana M. Soares, Sameena E. Tanwir, Jude W. Grosser, Manjul Dutt. Development of genetically modified citrus plants for the control of citrus canker and huanglongbing. Tropical Plant Pathology. 2020; 45 (3):237-250.

Chicago/Turabian Style

Juliana M. Soares; Sameena E. Tanwir; Jude W. Grosser; Manjul Dutt. 2020. "Development of genetically modified citrus plants for the control of citrus canker and huanglongbing." Tropical Plant Pathology 45, no. 3: 237-250.

Journal article
Published: 16 March 2020 in International Journal of Molecular Sciences
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Huanglongbing (HLB), a bacterial disease caused by Candidatus Liberibacter asiaticus (CLas), is a major threat to the citrus industry. In a previous study conducted by our laboratory, several citrus transgenic trees expressing the Arabidopsis thaliana NPR1 (AtNPR1) gene remained HLB-free when grown in a field site under high HLB disease pressure. To determine the molecular mechanisms behind AtNPR1-mediated tolerance to HLB, a transcriptome analysis was performed using AtNPR1 overexpressing transgenic trees and non-transgenic trees as control, from which we identified 57 differentially expressed genes (DEGs). Data mining revealed the enhanced transcription of genes encoding pathogen-associated molecular patterns (PAMPs), transcription factors, leucine-rich repeat receptor kinases (LRR-RKs), and putative ankyrin repeat-containing proteins. These proteins were highly upregulated in the AtNPR1 transgenic line compared to the control plant. Furthermore, analysis of protein–protein interactions indicated that AtNPR1 interacts with CsNPR3 and CsTGA5 in the nucleus. Our results suggest that AtNPR1 positively regulates the innate defense mechanisms in citrus thereby boosting resistance and effectively protecting the plant against HLB.

ACS Style

Wenming Qiu; Juliana Soares; Zhiqian Pang; Yixiao Huang; Zhonghai Sun; Nian Wang; Jude Grosser; Manjul Dutt. Potential Mechanisms of AtNPR1 Mediated Resistance against Huanglongbing (HLB) in Citrus. International Journal of Molecular Sciences 2020, 21, 2009 .

AMA Style

Wenming Qiu, Juliana Soares, Zhiqian Pang, Yixiao Huang, Zhonghai Sun, Nian Wang, Jude Grosser, Manjul Dutt. Potential Mechanisms of AtNPR1 Mediated Resistance against Huanglongbing (HLB) in Citrus. International Journal of Molecular Sciences. 2020; 21 (6):2009.

Chicago/Turabian Style

Wenming Qiu; Juliana Soares; Zhiqian Pang; Yixiao Huang; Zhonghai Sun; Nian Wang; Jude Grosser; Manjul Dutt. 2020. "Potential Mechanisms of AtNPR1 Mediated Resistance against Huanglongbing (HLB) in Citrus." International Journal of Molecular Sciences 21, no. 6: 2009.

Original article
Published: 28 February 2020 in Plant Cell, Tissue and Organ Culture (PCTOC)
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Finger lime is a small-leaved citrus native to Australia that is becoming increasingly popular worldwide. Finger lime is monoembryonic and cannot be propagated true to type from seeds. Initial studies to develop a tissue culture protocol for the mass production of elite lines being developed in our breeding program were unsuccessful due to complete leaf abscission during the culture establishment phase. In this study, silver-containing compounds such as silver thiosulfate (STS), silver nitrate (AgNO3), and silver nanoparticles (AgNPs) were applied at three concentrations (20, 40, and 60 µM) to manage ethylene biosynthesis in finger lime tissue cultures. The Murashige and Skoog (MS) medium supplemented with 2.2 µM BAP and 60 µM silver thiosulfate was the best medium for culture establishment. The same level of STS with 0.70 µM GA3 produced the largest number of shoots and prevented leaf abscission. Micropropagated shoots were successfully rooted in half-strength MS medium supplemented with 0.10 µM NAA. The addition of 60 µM silver thiosulfate (STS) to the MS medium resulted in efficient micropropagation of the finger lime explants and controlled leaf abscission.

ACS Style

Lamiaa M. Mahmoud; Jude W. Grosser; Manjul Dutt. Silver compounds regulate leaf drop and improve in vitro regeneration from mature tissues of Australian finger lime (Citrus australasica). Plant Cell, Tissue and Organ Culture (PCTOC) 2020, 141, 455 -464.

AMA Style

Lamiaa M. Mahmoud, Jude W. Grosser, Manjul Dutt. Silver compounds regulate leaf drop and improve in vitro regeneration from mature tissues of Australian finger lime (Citrus australasica). Plant Cell, Tissue and Organ Culture (PCTOC). 2020; 141 (3):455-464.

Chicago/Turabian Style

Lamiaa M. Mahmoud; Jude W. Grosser; Manjul Dutt. 2020. "Silver compounds regulate leaf drop and improve in vitro regeneration from mature tissues of Australian finger lime (Citrus australasica)." Plant Cell, Tissue and Organ Culture (PCTOC) 141, no. 3: 455-464.

Genetic transformation
Published: 08 January 2020 in In Vitro Cellular & Developmental Biology - Animal
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Full-length and 5′ deletion fragments of three constitutively expressed gene promoters identified from the Citrus sinensis L. genome (cyclophilin (CsCYP), glyceraldehyde-3-phosphate dehydrogenase C2 (CsGAPC2), and elongation factor 1-alpha (CsEF1)) were evaluated for their ability to express the uidA gene in leaf, stem, and root tissues of transgenic N. benthamiana plants. According to the fluorometric GUS assays, the CsGAPC2 promoter activity was significantly reduced in the leaves when the sequence between position − 1090 and − 497 was removed, while the activity remained unchanged in the roots. The CsCYP promoter activity was not affected by the different deletions, and even the smallest evaluated fragment was sufficient to maintain the expression of the uidA gene in N. benthamiana leaves and roots. Truncated promoter fragments of CsEF1 conferred higher GUS activity in leaves compared with the full-length promoter, while GUS activity was not affected in the roots. The removal of the intron in the 5′ UTR of the CsEF1 promoter reduced gene expression in the roots, although it did not cause any reduction in the gene expression level in the leaves. These results indicate that any of the three deletions of the CsCYP promoter can be utilized for efficient gene expression. In addition, the full-length CsGAPC2 promoter and two of the truncated CsEF1 promoter fragments were sufficient for efficient uidA gene expression.

ACS Style

L. Erpen-Dalla Corte; B. M. J. Mendes; Francisco De Assis Alves Mourão Filho; J. W. Grosser; M. Dutt. Functional characterization of full-length and 5′ deletion fragments of Citrus sinensis-derived constitutive promoters in Nicotiana benthamiana. In Vitro Cellular & Developmental Biology - Animal 2020, 56, 280 -289.

AMA Style

L. Erpen-Dalla Corte, B. M. J. Mendes, Francisco De Assis Alves Mourão Filho, J. W. Grosser, M. Dutt. Functional characterization of full-length and 5′ deletion fragments of Citrus sinensis-derived constitutive promoters in Nicotiana benthamiana. In Vitro Cellular & Developmental Biology - Animal. 2020; 56 (3):280-289.

Chicago/Turabian Style

L. Erpen-Dalla Corte; B. M. J. Mendes; Francisco De Assis Alves Mourão Filho; J. W. Grosser; M. Dutt. 2020. "Functional characterization of full-length and 5′ deletion fragments of Citrus sinensis-derived constitutive promoters in Nicotiana benthamiana." In Vitro Cellular & Developmental Biology - Animal 56, no. 3: 280-289.

Review
Published: 13 December 2019 in Plants
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Horticultural crops, including fruit, vegetable, and ornamental plants are an important component of the agriculture production systems and play an important role in sustaining human life. With a steady growth in the world’s population and the consequent need for more food, sustainable and increased fruit and vegetable crop production is a major challenge to guarantee future food security. Although conventional breeding techniques have significantly contributed to the development of important varieties, new approaches are required to further improve horticultural crop production. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) has emerged as a valuable genome-editing tool able to change DNA sequences at precisely chosen loci. The CRISPR/Cas9 system was developed based on the bacterial adaptive immune system and comprises of an endonuclease guided by one or more single-guide RNAs to generate double-strand breaks. These breaks can then be repaired by the natural cellular repair mechanisms, during which genetic mutations are introduced. In a short time, the CRISPR/Cas9 system has become a popular genome-editing technique, with numerous examples of gene mutation and transcriptional regulation control in both model and crop plants. In this review, various aspects of the CRISPR/Cas9 system are explored, including a general presentation of the function of the CRISPR/Cas9 system in bacteria and its practical application as a biotechnological tool for editing plant genomes, particularly in horticultural crops.

ACS Style

Lígia Erpen-Dalla Corte; Lamiaa M. Mahmoud; Tatiana S. Moraes; Zhonglin Mou; Jude W. Grosser; Manjul Dutt. Development of Improved Fruit, Vegetable, and Ornamental Crops Using the CRISPR/Cas9 Genome Editing Technique. Plants 2019, 8, 601 .

AMA Style

Lígia Erpen-Dalla Corte, Lamiaa M. Mahmoud, Tatiana S. Moraes, Zhonglin Mou, Jude W. Grosser, Manjul Dutt. Development of Improved Fruit, Vegetable, and Ornamental Crops Using the CRISPR/Cas9 Genome Editing Technique. Plants. 2019; 8 (12):601.

Chicago/Turabian Style

Lígia Erpen-Dalla Corte; Lamiaa M. Mahmoud; Tatiana S. Moraes; Zhonglin Mou; Jude W. Grosser; Manjul Dutt. 2019. "Development of Improved Fruit, Vegetable, and Ornamental Crops Using the CRISPR/Cas9 Genome Editing Technique." Plants 8, no. 12: 601.

Journal article
Published: 09 December 2019 in Plant Physiology
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Citrus greening or Huanglongbing (HLB) is caused by the phloem-limited intracellular Gram-negative bacterium Candidatus Liberibacter asiaticus (CLas). HLB-infected citrus phloem cells undergo structural modifications that include cell wall thickening, callose and p-protein induction, and cellular plugging. However, very little is known about the intracellular mechanisms that take place during CLas cell-to-cell movement. Here, we show that CLas movement through phloem pores of sweet orange (Citrus sinensis) and grapefruit (Citrus paradisi) is carried out by the elongated form of the bacteria. The round form of CLas is too large to move, but can change its morphology to enable its movement. CLas cells adhere to the plasma membrane of the phloem cells specifically adjacent to the sieve pores. Remarkably, CLas was present in both mature sieve element cells and nucleated non-sieve element cells. The sieve plate plugging structures of host plants were shown to have different composition in different citrus tissues. Callose deposition was the main plugging mechanism in the HLB-infected flush, where it reduced the open space of the pores. In the roots, pores were surrounded by dark extracellular material, with very little accumulation of callose. The expression of CALLOSE SYNTHASE 7 and PHLOEM PROTEIN 2 genes was upregulated in the shoots, but downregulated in root tissues. In seed coats, no phloem occlusion was observed, and CLas accumulated to high levels. Our results provide insight into the cellular mechanisms of Gram-negative bacterial cell-to-cell movement in plant phloem.

ACS Style

Diann Achor; Stacy Welker; Sulley K Ben-Mahmoud; Chunxia Wang; Svetlana Yuryevna Folimonova; Manjul Dutt; Siddarame Gowda; Amit Levy. Dynamics of Candidatus Liberibacter asiaticus Movement and Sieve-Pore Plugging in Citrus Sink Cells. Plant Physiology 2019, 182, 882 -891.

AMA Style

Diann Achor, Stacy Welker, Sulley K Ben-Mahmoud, Chunxia Wang, Svetlana Yuryevna Folimonova, Manjul Dutt, Siddarame Gowda, Amit Levy. Dynamics of Candidatus Liberibacter asiaticus Movement and Sieve-Pore Plugging in Citrus Sink Cells. Plant Physiology. 2019; 182 (2):882-891.

Chicago/Turabian Style

Diann Achor; Stacy Welker; Sulley K Ben-Mahmoud; Chunxia Wang; Svetlana Yuryevna Folimonova; Manjul Dutt; Siddarame Gowda; Amit Levy. 2019. "Dynamics of Candidatus Liberibacter asiaticus Movement and Sieve-Pore Plugging in Citrus Sink Cells." Plant Physiology 182, no. 2: 882-891.

Journal article
Published: 13 September 2019 in EDIS
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The large number of different rootstock varieties currently available for citrus production in Florida is unprecedented. This new 4-page article, chapter 4 of the forthcoming UF/IFAS Citrus Nursery Production Guide, provides information on rootstock uses in Florida, rootstock propagation, and the impact of tissue culture, as well as factors to consider when deciding which rootstock to plant. Written by Ute Albrecht, Manjul Dutt, and Jude Grosser and published by the UF/IFAS Horticultural Sciences Department. https://edis.ifas.ufl.edu/hs1340

ACS Style

Ute Albrecht; Manjul Dutt; Jude Grosser. Citrus Nursery Production Guide, Chapter 4: Rootstock Selection. EDIS 2019, 2019, 1 .

AMA Style

Ute Albrecht, Manjul Dutt, Jude Grosser. Citrus Nursery Production Guide, Chapter 4: Rootstock Selection. EDIS. 2019; 2019 (5):1.

Chicago/Turabian Style

Ute Albrecht; Manjul Dutt; Jude Grosser. 2019. "Citrus Nursery Production Guide, Chapter 4: Rootstock Selection." EDIS 2019, no. 5: 1.

Journal article
Published: 21 October 2018 in Plant Science
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In citrus, genetic improvement via biotechnology is challenging due to insufficient understanding of molecular barriers that prevent regeneration by somatic embryogenesis (SE). Our previous study indicated that LEC genes were involved in SE in citrus, but their regulatory roles remain to be elucidated. Here, we cloned one of the LEC genes, CsFUS3, and show that it is preferentially expressed during SE and in the embryogenic callus (EC) derived from citrus varieties with strong embryogenic competence. The overexpression of CsFUS3 in recalcitrant citrus callus restored embryogenic competence. Complementation of the loss-of-function Arabidopsis fus3 mutant with the CsFUS3 gene restored normal late embryogenesis, which is consistent with the CsFUS3 and AtFUS3 proteins contributing to the same regulatory network in Arabidopsis. Transcriptome profiling revealed that the expression of particular TFs that promote SE was up-regulated in the citrus overexpression (OE) line. The 104 differentially expressed genes associated with hormone biosynthesis, catabolism, and signaling are particularly noteworthy. The dynamic change in the ratio of ABA to GA during SE in wild-type callus mirrored the expression pattern of CsFUS3. In contrast, in the OE line, the ratio of ABA to GA was higher and the capacity for SE was greater when the OE line was separately treated with ABA and GA biosynthesis inhibitors. Taken together, our results demonstrate that the overexpression of CsFUS3 appears to establish a cellular environment favorable to SE, at least in part by promoting a high ABA to GA ratio and by regulating the expression of TFs that promote SE.

ACS Style

Zheng Liu; Xiao-Xia Ge; Wen-Ming Qiu; Jian-Mei Long; Hui-Hui Jia; Wei Yang; Manjul Dutt; Xiao-Meng Wu; Wen-Wu Guo. Overexpression of the CsFUS3 gene encoding a B3 transcription factor promotes somatic embryogenesis in Citrus. Plant Science 2018, 277, 121 -131.

AMA Style

Zheng Liu, Xiao-Xia Ge, Wen-Ming Qiu, Jian-Mei Long, Hui-Hui Jia, Wei Yang, Manjul Dutt, Xiao-Meng Wu, Wen-Wu Guo. Overexpression of the CsFUS3 gene encoding a B3 transcription factor promotes somatic embryogenesis in Citrus. Plant Science. 2018; 277 ():121-131.

Chicago/Turabian Style

Zheng Liu; Xiao-Xia Ge; Wen-Ming Qiu; Jian-Mei Long; Hui-Hui Jia; Wei Yang; Manjul Dutt; Xiao-Meng Wu; Wen-Wu Guo. 2018. "Overexpression of the CsFUS3 gene encoding a B3 transcription factor promotes somatic embryogenesis in Citrus." Plant Science 277, no. : 121-131.

Original article
Published: 08 September 2018 in Plant Biotechnology Reports
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Anthocyanins are synthesized via the flavonoid pathway through a complex expression of several genes such as MYB transcription factors. Anthocyanins protect plants against biotic and abiotic stresses. Herein, we studied the effect of expression of MYB (VvmybA1 cloned from the red grape and Ruby cloned from ‘Moro’ blood orange) transcription factors in “Mexican” lime on juice quality and leaf pigments, leaf metabolites, and phytohormones. Anthocyanins, furanocoumarins, flavonoids, and hydroxycinnamates were analyzed with high-performance liquid chromatography–mass spectrometry, whereas chlorophylls, carotenoids, and xanthophylls were analyzed using HPLC coupled with photodiode array detector (PDA). The rest of metabolites were analyzed using gas chromatography–mass spectrometry. Overexpression of VvmybA1 and Ruby resulted in accumulation of anthocyanins in leaves, flowers, and fruits of the transgenic plants. However, the level of anthocyanins in Ruby plants was significantly lower than that in VvmybA1 plants. The level of anthocyanins and the gene expression of VvmybA1 and Ruby in young leaves were higher than mature leaves. On the other hand, the level of several furanocoumarins, and hydroxycinnamates decreased in mature VvmybA1 leaves, indicating a drainage of p-coumaric acid due to the induction of anthocyanins biosynthesis. The level of chlorophyll decreased in mature VvmybA1 leaves, whereas zeaxanthin level increased, indicated a photoprotection role for anthocyanins. Most of polar and volatile metabolites also decreased VvmybA1 leaves, indicating a decrease in the photosynthetic efficiency. Benzoic acid and salicylic acid increased, whereas auxins decreased. The level of abscisic acid was not affected by the overexpression of VvmybA1 and the plants showed normal growth and development. Overexpression of VvmybA1 highly increased the antioxidant activity of the transgenic juice and leaves, whereas overexpression of Ruby showed only a slight increase. The pH, °Brix value, and TA of the transgenic juice were not affected by the expression of VvmybA1 or Ruby.

ACS Style

Faraj Hijaz; Yasser Nehela; Shelley E. Jones; Manjul Dutt; Jude W. Grosser; John A. Manthey; Nabil Killiny. Metabolically engineered anthocyanin-producing lime provides additional nutritional value and antioxidant potential to juice. Plant Biotechnology Reports 2018, 12, 329 -346.

AMA Style

Faraj Hijaz, Yasser Nehela, Shelley E. Jones, Manjul Dutt, Jude W. Grosser, John A. Manthey, Nabil Killiny. Metabolically engineered anthocyanin-producing lime provides additional nutritional value and antioxidant potential to juice. Plant Biotechnology Reports. 2018; 12 (5):329-346.

Chicago/Turabian Style

Faraj Hijaz; Yasser Nehela; Shelley E. Jones; Manjul Dutt; Jude W. Grosser; John A. Manthey; Nabil Killiny. 2018. "Metabolically engineered anthocyanin-producing lime provides additional nutritional value and antioxidant potential to juice." Plant Biotechnology Reports 12, no. 5: 329-346.

Short communication
Published: 31 July 2018 in Scientia Horticulturae
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Genetic transformation of citrus has been mainly conducted via Agrobacterium tumefaciens using juvenile tissues of the epicotyl segments obtained from the in vitro germination of seeds. Transformation of the stem segments obtained from the mature material is an alternative to overcome or reduce the juvenile phase reported in perennial species such as citrus. However, some citrus species, including mandarins, are recalcitrant to these methods or have low genetic transformation efficiency. Thus, the alternative is the use of juvenile explants with high regenerative potential, such as cell suspensions obtained from citrus embryogenic callus or direct incorporation of DNA into protoplasts using polyethylene glycol (PEG). The present study aimed to compare these four genetic modification techniques for ‘W Murcott’, one of the main fresh market mandarin cultivars in the United States. The transgenic plants were obtained by the different techniques using a vector containing the EGFP gene under the control of the CaMV35S promoter. The genetic transformation efficiency was low, utilizing either juvenile epicotyl or mature stem segments, with an average of 1.23% and 0.33%, respectively. The genetic transformation efficiency using cell suspensions and cell suspension-derived protoplast explants were higher, with an average of 29% and 11%, respectively. Our results reveal the importance of utilizing cell suspension-derived cultures for the efficient transformation of ‘W Murcott'.

ACS Style

Manjul Dutt; Lígia Erpen; Jude W. Grosser. Genetic transformation of the ‘W Murcott’ tangor: comparison between different techniques. Scientia Horticulturae 2018, 242, 90 -94.

AMA Style

Manjul Dutt, Lígia Erpen, Jude W. Grosser. Genetic transformation of the ‘W Murcott’ tangor: comparison between different techniques. Scientia Horticulturae. 2018; 242 ():90-94.

Chicago/Turabian Style

Manjul Dutt; Lígia Erpen; Jude W. Grosser. 2018. "Genetic transformation of the ‘W Murcott’ tangor: comparison between different techniques." Scientia Horticulturae 242, no. : 90-94.

Journal article
Published: 04 January 2018 in EDIS
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Cylindrical Australian finger limes (Microcitrus australasica) taste like a combination of lemon, lime, and grapefruit, come in a rainbow of colors, and have a texture like caviar. Like other citrus fruits, finger limes are nutritious, low in calories, and vitamin-rich. So far in the United States only California grows finger limes commercially, but this 4-page fact sheet written by Aditya Singh, Edward Evans, Jeff Wasielewski, Manjul Dutt, and Jude Grosser and published by the UF/IFAS Food and Resource Economics Department makes the case that exotic, colorful finger limes would likely grow well in Florida, where they would appeal to hoteliers and restaurants and to adventurous, health-conscious consumers on the lookout for a delicious new fresh fruit snack to try. http://edis.ifas.ufl.edu/fe1033

ACS Style

Aditya Singh; Edward A. Evans; Jeff Wasielewski; Manjul Dutt; Jude Grosser. Finger Lime: An Alternative Crop with Great Potential in South Florida. EDIS 2018, 2018, 1 .

AMA Style

Aditya Singh, Edward A. Evans, Jeff Wasielewski, Manjul Dutt, Jude Grosser. Finger Lime: An Alternative Crop with Great Potential in South Florida. EDIS. 2018; 2018 (1):1.

Chicago/Turabian Style

Aditya Singh; Edward A. Evans; Jeff Wasielewski; Manjul Dutt; Jude Grosser. 2018. "Finger Lime: An Alternative Crop with Great Potential in South Florida." EDIS 2018, no. 1: 1.