Establishment of dual reverse transcriptase-polymerase chain reaction for detection system for Areca palm velarivirus 1.

Areca palm velarivirus 1 (APV1) is one of the main pathogen causing yellow leaf disease, and leading to considerable losses in the Areca palm industry. The detection methods for APV1 are primarily based on phenotype determination and molecular techniques, such as polymerase chain reaction (PCR). However, a single PCR has limitations in accuracy and sensitivity. Therefore, in the present study, we established a dual RT-PCR APV1-detection system with enhanced accuracy and sensitivity using two pairs of specific primers, YLDV2-F/YLDV2-R and YLDV4-F/YLDV4-R. Moreover, two cDNA fragments covering different regions of the viral genome were simultaneously amplified, with PCR amplicon of 311 and 499 bp, respectively. The dual RT-PCR detection system successfully amplified the two target regions of the APV1, demonstrating high specificity and sensitivity and compensating for the limitations of single-primer detection methods. We tested 60 Areca palm samples from different geographical regions, highlighting its advantages in that the dual RT-PCR system efficiently and accurately detected APV1 in samples across diverse areas. The dual RT-PCR APV1 detection system provides a rapid, accurate, and sensitive method for detecting the virus and offers valuable technical support for research in preventing and managing yellow leaf diseases caused by APV1 in Areca palms. Moreover, the findings of this study can serve as a reference for establishing similar plants viral detection systems in the future.

Characteristics of NAC transcription factors in Solanaceae crops and their roles in responding to abiotic and biotic stresses.

As one of the largest transcription factor (TF) families in plants, the NAC (NAM, ATAF1/2, and CUC2) family plays important roles in response pathways to various abiotic and biotic stresses, such as drought, high salinity, low temperature, and pathogen infection. Although, there are a number of reviews on the involvement of NAC TF in plant responses to biotic and abiotic stresses, most of them are focused on the model plants Arabidopsis thaliana and Oryza sativa, and there is a lack of systematic evaluation of specific species. Solanaceae, the world's third most significant cash crop, has been seriously affected by environmental disturbances in recent years in terms of yield and quality, posing a severe threat to global food security. This review focuses on the functional roles of NAC transcription factors in response to external stresses involved in five important Solanaceae crops: tomato, potato, pepper, eggplant and tobacco, and analyzes the affinities between them. It will provide resources for stress-resistant breeding of Solanaceae crops using transgenic technology.

Characterization of anthocyanin accumulation, nutritional properties, and postharvest attributes of transgenic purple tomato.

Anthocyanins are natural pigments with diverse physiological roles and protective effects, but most tomatoes produce little. In this study, the anthocyanin characteristics, nutritional properties, and postharvest attributes of purple tomato (SlMYB75-OE) obtained by overexpression of SlMYB75 gene were first analyzed. Compared to wild-type (WT), eight monomeric anthocyanins were newly produced by overexpression of SlMYB75, and further study demonstrated the expression of dihydroflavonol-4-reductase (SlDFR) and two UDP-glycosyltransferase (SlUGTs) genes was activated by SlMYB75. The contents of sugars (sucrose, glucose, and fructose) and citric acid content in SlMYB75-OE were higher and lower, respectively, than in WT. In addition, FRAP and DPPH assays indicated SlMYB75-OE had higher antioxidant capacity, when compared to WT. Moreover, SlMYB75-OE exhibited a longer shelf life and stronger resistance to Botrytis cinerea than WT, and this characteristic was positively correlated with anthocyanin content. These results help to clarify the function of SlMYB75 and provide a reference for tomato breeding.

Candidate gene discovery for salt tolerance in rice (Oryza sativa L.) at the germination stage based on genome-wide association study.

Salt stress affects rice seed germination and seedling formation, seriously restricting rice production. Screening salt-tolerant rice varieties and analyzing the genetic mechanisms underlying salt tolerance are therefore very important to ensure rice production. In this study, 313 Oryza sativa ssp. japonica germplasm were used to conduct a genome-wide association study (GWAS) using 1% NaCl as a salt stress treatment during germination stage. The germination potential (GP) on different days and the germination index (GI) under salt stress were used as salt tolerance indicators. The results of population structure analysis showed that the 313 germplasm studied could be divided into two subpopulations, consistent with the geographical origins of the materials. There were 52 loci significantly related to salt tolerance during germination, and the phenotypic contribution rate of 29 loci was > 10%. A region on chromosome 11 (17049672-17249672 bp) was repeatedly located, and the candidate gene LOC_Os11g29490, which encodes a plasma membrane ATPase, was identified in this locus. Further haplotype analysis showed the GP of germplasm with different haplotypes at that locus significantly differed under salt stress (p < 0.05), and germplasm carrying Hap2 displayed strong salt tolerance during the germination stage. Two other promising candidate genes for salt tolerance were identified: LOC_Os01g27170 (OsHAK3), which encodes a potassium transporter, and LOC_Os10g42550 (OsITPK5), which encodes an inositol 1, 3, 4-trisphosphate 5/6-kinase. The results of this study provide a theoretical basis for salt-tolerant gene cloning and molecular design breeding in rice.

Mapping QTLs for yield component traits using overwintering cultivated rice.

The present study conducted QTL (quantitative trait locus) mapping using F2 and F2:3 generations derived from a cross between an overwintering cultivated rice Nuodao89-1 and Shuhui527, to identify potential yield component related QTLs. A total of 37 QTLs were detected across all chromosome except chromosome 7, with LOD values ranging from 3.10 to 12.67. Three QTLs including .qTGW3.1, qTGW6.1 and qNEG9.1 were repeatedly detected in both generations. qTGW3.1 was from Nuodao89-1, a total of 46 functional genes involved in 229 gene ontology terms were identified within its locus. Six QTL clusters were founded and corresponding agronomic traits of those QTLs showed highly significant correlation. Three types of epistatic interaction including 47 epistatic QTL pairs for eight yield component traits were detected, but the epistatic QTLs was not so important in controlling the genetic expression of the yield-related Nuodao89-1 as the additive QTLs. Overall, this research provides a theoretical basis for the mining of yield-related genes from overwintering cultivated rice.

Chemical composition analysis and antioxidant activity of black rice pigment.

The purpose of this research was conducted to determine anthocyanin constituent from the grains of Purple black rice No. 6. Moreover, the in vitro antioxidant activity of black rice pigment (BRP) was evaluated. The crude extract of black rice was isolated and purified by silica gel thin layer chromatography. Anthocyanins were elucidated using ultraviolet visible spectroscopy, infrared spectroscopy, nuclear magnetic resonance spectroscopy, and high-performance liquid chromatography. Meanwhile, salicylic acid Fenton system, superoxide radical system, H2 O2 scavenging system, DPPH-free radical scavenging system, and Prussian blue method were used to determine the free radical scavenging ability and total reducing ability of BRP. Overall results suggest that the main component of BRP is cyanidin-3-glucoside, which is a kind of anthocyanins. It also had significant scavenging capacity to OH, O2- , H2 O2 , DPPH, and its scavenging rate and reducing ability increased with the increase in pigment concentration.

Locating QTLs controlling overwintering seedling rate in perennial glutinous rice 89-1 (Oryza sativa L.).

A new cold tolerant germplasm resource named glutinous rice 89-1 (Gr89-1, Oryza sativa L.) can overwinter using axillary buds, with these buds being ratooned the following year. The overwintering seedling rate (OSR) is an important factor for evaluating cold tolerance. Many quantitative trait loci (QTLs) controlling cold tolerance at different growth stages in rice have been identified, with some of these QTLs being successfully cloned. However, no QTLs conferring to the OSR trait have been located in the perennial O. sativa L. To identify QTLs associated with OSR and to evaluate cold tolerance. 286 F12 recombinant inbred lines (RILs) derived from a cross between the cold tolerant variety Gr89-1 and cold sensitive variety Shuhui527 (SH527) were used. A total of 198 polymorphic simple sequence repeat (SSR) markers that were distributed uniformly on 12 chromosomes were used to construct the linkage map. The gene ontology (GO) annotation of the major QTL was performed through the rice genome annotation project system. Three main-effect QTLs (qOSR2, qOSR3, and qOSR8) were detected and mapped on chromosomes 2, 3, and 8, respectively. These QTLs were located in the interval of RM14208 (35,160,202 base pairs (bp))-RM208 (35,520,147 bp), RM218 (8,375,236 bp)-RM232 (9,755,778 bp), and RM5891 (24,626,930 bp)-RM23608 (25,355,519 bp), and explained 19.6%, 9.3%, and 11.8% of the phenotypic variations, respectively. The qOSR2 QTL displayed the largest effect, with a logarithm of odds score (LOD) of 5.5. A total of 47 candidate genes on the qOSR2 locus were associated with 219 GO terms. Among these candidate genes, 11 were related to cell membrane, 7 were associated with cold stress, and 3 were involved in response to stress and biotic stimulus. OsPIP1;3 was the only one candidate gene related to stress, biotic stimulus, cold stress, and encoding a cell membrane protein. After QTL mapping, a total of three main-effect QTLs-qOSR2, qOSR3, and qOSR8-were detected on chromosomes 2, 3, and 8, respectively. Among these, qOSR2 explained the highest phenotypic variance. All the QTLs elite traits come from the cold resistance parent Gr89-1. OsPIP1;3 might be a candidate gene of qOSR2.

Development of a highly sensitive and specific monoclonal antibody-based enzyme-linked immunosorbent assay for determination of doxycycline in chicken muscle, liver and egg.

A modified indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) method was developed using a highly sensitive and specific monoclonal antibody (McAb) to determine doxycycline (DC) residues in chicken tissues and egg. The McAb against DC was produced by hybridoma technique and a modified ic-ELISA was characterised in terms of sensitivity, specificity, precision and accuracy. At optimal experimental conditions, the standard curve was constructed at concentrations ranged from 0.01 to 100 ng/ml. The IC(50) value was 1.32±0.18 ng/ml. The limit of detection was 0.14±0.02 ng/g. The recoveries of DC from spiked chicken liver, muscle, and egg at levels of 50-600 ng/g were 84.6-85.5%, 88.2-89.1%, and 84.4-89.3%, respectively. The coefficient variations (CVs) were 5.1-9.3%, 3.7-11.3%, and 4.7-9.8%, respectively. Linear regression analysis showed good correlation, with r(2) values 0.9909 for chicken liver and 0.9916 for chicken muscle.