Screening Potential Citrus Rootstocks for Phytophthora nicotianae Tolerance

Seeds from four citrus rootstocks including sour orange, Bitters-C22 citrandarin, Sarawak pummelo 3 Rio Red grapefruit, and Sarawak pummelo 3Bower mandarin were exposed to high inoculum levels of Phytophthora nicotianae to screen for tolerance. Inoculation of pregerminated seeds (PGIS) and non-PGIS was carried out. The average P. nicotianae propagule counts from the soil samples where these seedlings were raised ranged from 424 to 1361 colony forming units/cm3. The proportion of live to dead plants was recorded at 11months postinoculation, which showed that Sarawak3Bower performed significantly better than other rootstocks. Evaluation of the rootstocks 18 months postinoculation resulted in only one surviving sour orange plant, which suggests potential rootstock resistance

Fungi Impacting Human Health

Among millions of fungal species, a vast majority are harmless and non-pathogenic. Only a few hundred are known to cause human disease. Pathogenic fungi cause diseases in humans leading to severe health issues, while several beneficial fungi provide nutrition, recycle nutrients in the ecosystem by decomposing complex organic materials, and offer commercial opportunities for pharmaceutical, food, and nutraceutical industries. Human diseases from fungal infections range from minor/mild diseases affecting close to a billion people to life-threatening diseases affecting hundreds of millions. Factors such as invasive surgical procedures, immunocompromised individuals, air quality, and pollution contribute to an elevated risk of invasive fungal infections resulting in hospitalizations and high mortalities globally. According to the Centers for Disease Control and Prevention (CDC), fungal diseases contribute several billion dollars in direct medical costs annually. An inadequate number of antifungal drugs are available to treat the multitude of fungal diseases, and antifungal resistance development in some fungi poses serious limitations on treatment options. Novel diagnostic methods provide opportunities for rapid, specific, sensitive, and cost-effective diagnosis of several fungal pathogens; however, diagnosing invasive fungal infections is still challenging, especially in immunocompromised patients. There is an urgent need for studies focused on fungal pathogens and infections in humans, improved diagnostic methods, and the development of vaccines and antifungal drugs with alternative modes of action. Enhanced funding support for research in fungal diseases and awareness of often neglected fungal diseases in people through public health programs will contribute to the reduced threat posed by these diseases. Advancements in industrial and commercial applications of benign fungi will result in improved human life

First report on citrus dry rot in sour orange rootstock in Texas

A grapefruit tree on sour orange rootstock in a residential property in Mission, TX was suspected to have citrus dry root rot disease based on symptoms. The causal organism was isolated from the infected root samples and based on fungal cultural and microscopic morphology and PCR, it was confirmed to be Fusarium solani (Martius) Appel & Wollenweber emend. Snyder & Hansen. A total of 10 healthy sour orange rootstock seedlings were inoculated using conidial suspension of the fungus by the standard root-dip method. After 9 month post inoculation, the inoculated fungus was re-isolated from root and stem sections of these plants. Plants were smaller in size and displayed the classical symptoms of dry rot. The fungal colonies were confirmed to be F. solani based on fungal morphology and PCR. This is the first report of F. solani infecting sour orange rootstock plants in Texas

First report on citrus dry rot in sour orange rootstock in Texas

A grapefruit tree on sour orange rootstock in a residential property in Mission, TX was suspected to have citrus dry root rot disease based on symptoms. The causal organism was isolated from the infected root samples and based on fungal cultural and microscopic morphology and PCR, it was confirmed to be Fusarium solani (Martius) Appel & Wollenweber emend. Snyder & Hansen. A total of 10 healthy sour orange rootstock seedlings were inoculated using conidial suspension of the fungus by the standard root-dip method. After 9 month post inoculation, the inoculated fungus was re-isolated from root and stem sections of these plants. Plants were smaller in size and displayed the classical symptoms of dry rot. The fungal colonies were confirmed to be F. solani based on fungal morphology and PCR. This is the first report of F. solani infecting sour orange rootstock plants in Texas

Transcriptome analysis of Huanglongbing-infected sweet orange leaves using RNA sequencing and quantitative PCR

RNA sequencing (RNA-seq) methods provide a complete description of RNA transcripts including alternative splicing and small RNA characterization. RNA-seq performed in our laboratory on healthy and Huanglongbing (HLB)-infected young leaves revealed that 4,044 transcripts were up-regulated and 2,562 were down-regulated in the diseased trees. Moreover, a number of genes showed alternative splicing events including exon skipping, intron retention, and 5' and 3' alternative splicing. Furthermore, quantitative PCR (qPCR) performed on 20 randomly chosen genes with high differential expression (10 up and 10 down regulated) showed that all were consistent with RNA-seq data. Additionally, variation in levels of gene expression was observed between young and mature leaves. These early host plant response genes due to HLB-infection might be useful in the development of early HLB-detection methods before manifestation of disease symptoms in the infected plants

Candidatus Liberibacter asisticus detection in the leaves, roots from infected trees and leaves of new shoots from the stumps of the infected sweet orange trees in Texas

A total of 108 root and corresponding symptomatic leaf samples from four different quadrants were collected from 27 6 year old sweet orange trees in which the presence of Candidatus Liberibacter asiaticus (CLas) was previously confirmed by quantitative polymerase chain reaction (qPCR). There was no significant difference (p>0.05) in the level of infection as determined by threshold cycle (Ct) values between different types of tissue tested. Analysis of variance (ANOVA) showed that there is no significant difference (p>0.05) in test results among different distances from the trunk or quandrants where the root samples were collected. The stumps of the infected trees were covered in a psyllid-proof cage and leaves from the new shoots emerged from these stumps did not show the presence of CLas where as the roots showed the presence of CLas.  Moreover, there was no significant difference between roots from infected trees and roots from infected stumps.  Additionally, there was a significant difference with an average Ct value difference of 2.97 cycles between the DNA samples extracted from roots using two different commercially available kits

Quantifying Citrus Tree Health Using True Color UAV Images

Huanglongbing (HLB) and Phytophthora foot and root rot are diseases that affect citrus production and profitability. The symptoms and physiological changes associated with these diseases are diagnosed through expensive and time-consuming field measurements. Unmanned aerial vehicles (UAVs) using red/green/blue (RGB, true color) imaging, may be an economic alternative to diagnose diseases. A methodology using a UAV with a RGB camera was developed to assess citrus health. The UAV was flown in April 2018 on a grapefruit field infected with HLB and foot rot. Ten trees were selected for each of the following disease classifications: (HLB-, foot rot–), (HLB+, foot rot–), (HLB-, foot rot+) (HLB+, foot rot+). Triangular greenness index (TGI) images were correlated with field measurements such as tree nutritional status, leaf area, SPAD (leaf greenness), foot rot disease severity and HLB. It was found that 61% of the TGI differences could be explained by Na, Fe, foot rot, Ca, and K. This study shows that diseased citrus trees can be monitored using UAVs equipped with RGB cameras, and that TGI can be used to explain subtle differences in tree health caused by multiple diseases

Candidatus Liberibacter asisticus detection in the leaves, roots from infected trees and leaves of new shoots from the stumps of the infected sweet orange trees in Texas

A total of 108 root and corresponding symptomatic leaf samples from four different quadrants were collected from 27 6 year old sweet orange trees in which the presence of Candidatus Liberibacter asiaticus (CLas) was previously confirmed by quantitative polymerase chain reaction (qPCR). There was no significant difference (p>0.05) in the level of infection as determined by threshold cycle (Ct) values between different types of tissue tested. Analysis of variance (ANOVA) showed that there is no significant difference (p>0.05) in test results among different distances from the trunk or quandrants where the root samples were collected. The stumps of the infected trees were covered in a psyllid-proof cage and leaves from the new shoots emerged from these stumps did not show the presence of CLas where as the roots showed the presence of CLas.  Moreover, there was no significant difference between roots from infected trees and roots from infected stumps.  Additionally, there was a significant difference with an average Ct value difference of 2.97 cycles between the DNA samples extracted from roots using two different commercially available kits

Screening Potential Citrus Rootstocks for Phytophthora nicotianae Tolerance

Seeds from four citrus rootstocks including sour orange, Bitters-C22 citrandarin, Sarawak pummelo 3 Rio Red grapefruit, and Sarawak pummelo 3Bower mandarin were exposed to high inoculum levels of Phytophthora nicotianae to screen for tolerance. Inoculation of pregerminated seeds (PGIS) and non-PGIS was carried out. The average P. nicotianae propagule counts from the soil samples where these seedlings were raised ranged from 424 to 1361 colony forming units/cm3. The proportion of live to dead plants was recorded at 11months postinoculation, which showed that Sarawak3Bower performed significantly better than other rootstocks. Evaluation of the rootstocks 18 months postinoculation resulted in only one surviving sour orange plant, which suggests potential rootstock resistance