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High temperature stress is a major limiting factor for pepper productivity, which will continue to be a problem under climate change scenarios. Developing heat tolerant cultivars is critical for sustained pepper production, especially in tropical and subtropical regions. In fruiting crops, like pepper, reproductive tissues, especially pollen, are the most sensitive to high temperature stress. Typically, pollen viability and germination are assessed through staining and microscopy, which is tedious and potentially inaccurate. To increase efficiency in assessing pollen traits of pepper, the use of impedance flow cytometry (IFC) has been proposed. We conducted three independent experiments to determine the most effective methodology to use IFC for evaluating pollen traits for heat tolerance in pepper. Seven floral developmental stages were evaluated, and stages 3, 4, and 5 were found to best combine high pollen concentration and activity. Flowers in development stages 3, 4, or 5 were then heat treated at 41, 44, 47, 50, and 55 °C or not heat treated (control). The critical temperature to assess heat tolerance using IFC was found to be 50 °C, with a reduction in pollen activity and concentration occurring at temperatures greater than 47 °C. Twenty-one entries of pepper were then accessed for pollen traits using the staining and IFC methods over 2 months, April (cooler) and June (hotter). Growing environment was found to be the greatest contributor to variability for nearly all pollen traits assessed, with performance during June nearly always being lower. PBC 507 and PBC 831 were identified as being new sources of heat tolerance, based on using IFC for assessing pollen. Pollen viability determined by staining and pollen activity determined using IFC were significantly positively correlated, indicating that IFC is an efficient and accurate method to assess pollen traits in pepper. This work provides a basis for further research in this area and supports more efficient breeding of heat-tolerant cultivars.

Habanero (Capsicum chinense Jacq.) is widely grown and consumed in West and Central African countries, and viral diseases represent an important production challenge. Diagnosis of the viral species affecting habanero productivity in Benin is limited, and understanding this will enable more efficient host resistance breeding. During 2019 and 2020, we characterized the incidence and severity of the viral diseases infecting nine promising habanero breeding lines and one commercial hybrid check under open field conditions in Benin. The horticultural performance, including yield and yield component traits of the entries, was determined during the 2 years of the experiment. A randomized complete block design was used with three replications, each with 24 plants. Data were recorded on days to 50% flowering and 50% fruit maturity, yield and on the yield components of fruit weight (g), fruit length (cm), and fruit width (mm), as well as disease incidence and severity. In total, 35 leaf samples were collected for viral diagnosis among habanero breeding lines. We found that Pepper veinal mottle virus (PVMV; Potyvirus) was the overwhelmingly predominant virus in our trials, with an 80% incidence; however, we found frequent coinfection of PVMV with Cucumber mosaic virus (CMV, Cucumovirus), Polerovirus, and, to a lesser extent, Chili veinal mottle virus (ChiVMV; Potyvirus). The mean disease incidence across all entries was 60%. AVPP1932 and PBC 2010 had the lowest disease incidence (35% and 43%, respectively), whereas AVPP1929 had the highest (86%) disease incidence. The F1 hybrid check Afadja had the overall highest yield, with 30 t⋅ha−1, followed by AVPP1932, with 19 t⋅ha−1, both in 2019. There was a negative correlation between disease incidence and total yield (r = −0.44; P < 0.001), supporting previous studies indicating that viral diseases are major production constraints for habanero in West Africa. This study provides insight regarding the need to improve habanero for resistance to aphid-transmitted viruses and develop integrated pest management strategies to limit losses in Benin.

Chile pepper (Capsicum annuum) is an increasingly important crop worldwide, and Vietnam and Myanmar are major producing countries. The chile pepper markets in Myanmar and Vietnam are different, with production primarily for domestic consumption in Myanmar and for the export market in Vietnam. However, there is an overall lack of domestically developed cultivars in both countries. The objective of this study was to identify high-performing chile pepper entries, adapted to local conditions, for use in domestic breeding programs or direct release. Fruit length, width, weight, and yield were measured during two seasons (2016–17 and 2018–19), and the same entries were evaluated in Hanoi, Vietnam, and Nay Pyi Taw, Myanmar. However, different entries were tested in each season. During the 2016–17 season, AVPP1324 grown in Hanoi had the overall highest yield (15.3 t·ha–1), followed by AVPP1330 (15.0 t·ha–1 in Hanoi) and AVPP1111 (14.4 and 14.9 t·ha–1 in Hanoi and Nay Pyi Taw, respectively). AVPP0303 had the greatest fruit length, fruit width, and fruit weight in both Hanoi and Nay Pyi Taw during the 2016–17 season. During the 2018–19 season, AVPP1345 (24.8 t·ha–1) followed by AVPP9905 (22.5 t·ha–1) in Nay Pyi Taw, and AVPP1245 (17.4 t·ha–1) in Hanoi had the highest yield. AVPP9905 had the greatest fruit weight and width in both locations. AVPP1345 and AVPP9905 had the greatest fruit length during the 2018–19 season. There is an obvious need for domestically produced cultivars in Myanmar and Vietnam that meet local farmer and consumer preferences and that are adapted to the pests, diseases, and stress in each country. Several high-performing lines were identified that can be used as direct release or incorporated in local breeding programs for the development of inbred or F1 hybrid cultivars. This research also provides a basis for future studies on stability of yield and yield components in Southeast Asia.

Wild relatives of domesticated Capsicum represent substantial genetic diversity and thus sources of traits of potential interest. Furthermore, the hybridization compatibility between members of Capsicum species complexes remains unresolved. Improving our understanding of the relationship between Capsicum species relatedness and their ability to form hybrids is a highly pertinent issue. Through the development of novel interspecific hybrids in this study, we demonstrate interspecies compatibility is not necessarily reflected in relatedness according to established Capsicum genepool complexes. Based on a phylogeny constructed by genotyping using simple sequence repeat (SSR) markers and with a portion of the waxy locus, and through principal component analysis (PCA) of phenotypic data, we clarify the relationships among wild and domesticated Capsicum species. Together, the phylogeny and hybridization studies provide evidence for the misidentification of a number of species from the World Vegetable Center genebank included in this study. The World Vegetable Center holds the largest collection of Capsicum genetic material globally, therefore this may reflect a wider issue in the misidentification of Capsicum wild relatives. The findings presented here provide insight into an apparent disconnect between compatibility and relatedness in the Capsicum genus, which will be valuable in identifying candidates for future breeding programs.

Male sterility is of high importance in hybrid seed production of hot and sweet peppers. Genic (or nuclear) male sterility (GMS) is a simply inherited (usually monogenic recessive) and highly stable trait. However, one major disadvantage of using GMS is 1:1 segregation of male sterile to male fertile plants in every subsequent generation. Molecular markers tightly linked to genic male sterility (ms) genes would facilitate an efficient and rapid transfer of ms genes into different genetic backgrounds through marker-assisted backcrossing. The two non-allelic genic male sterility genes ms3 and msw in hot and sweet pepper backgrounds, respectively, are monogenic recessive. Genotyping by sequencing (GBS) in an F2 population segregating for ms3 gene in hot pepper and in an F6 inbred near-isogenic line (NIL) population segregating for msw gene in sweet pepper yielded 9,713 and 7,453 single nucleotide polymorphism markers, respectively. Four candidate SNPs co-segregating with ms3 gene and one co-segregating with msw gene were identified by bulk segregant analysis and physically mapped to chromosomes 1 and 5, respectively. In hot pepper, two markers [HPGMS2 (CAPS) and HPGMS3 (dCAPS)] located 3.83 cM away from the ms3 gene and in sweet pepper the dCAPS marker SPGMS1 co-segregated (completely linked) with the msw gene were developed. These markers will increase the efficacy of the male sterility genes for pepper breeding, as they can be useful in developing the genic male sterile lines in parental inbred lines of commercial hybrids through marker-assisted backcrossing, hybrid seed production, and genetic purity testing of hybrid seeds.

Background: Mungbean (Vigna radiata [L.] R. Wilczek) is an important legume crop with high nutritional value in South and Southeast Asia. The crop plant is susceptible to a storage pest caused by bruchids (Callosobruchus spp.). Some wild and cultivated mungbean accessions show resistance to bruchids. Genomic and transcriptomic comparison of bruchid-resistant and -susceptible mungbean could reveal bruchid-resistant genes (Br) for this pest and give insights into the bruchid resistance of mungbean. Results: Flow cytometry showed that the genome size varied by 61 Mb (mega base pairs) among the tested mungbean accessions. Next generation sequencing followed by de novo assembly of the genome of the bruchid-resistant recombinant inbred line 59 (RIL59) revealed more than 42,000 genes. Transcriptomic comparison of bruchid-resistant and -susceptible parental lines and their offspring identified 91 differentially expressed genes (DEGs) classified into 17 major and 74 minor bruchid-resistance-associated genes. We found 408 nucleotide variations (NVs) between bruchid-resistant and -susceptible lines in regions spanning 2 kb (kilo base pairs) of the promoters of 68 DEGs. Furthermore, 282 NVs were identified on exons of 148 sequence-changed-protein genes (SCPs). DEGs and SCPs comprised genes involved in resistant-related, transposable elements (TEs) and conserved metabolic pathways. A large number of these genes were mapped to a region on chromosome 5. Molecular markers designed for variants of putative bruchid-resistance-associated genes were highly diagnostic for the bruchid-resistant genotype. Conclusions: In addition to identifying bruchid-resistance-associated genes, we found that conserved metabolism and TEs may be modifier factors for bruchid resistance of mungbean. The genome sequence of a bruchid-resistant inbred line, candidate genes and sequence variations in promoter regions and exons putatively conditioning resistance as well as markers detecting these variants could be used for development of bruchid-resistant mungbean varieties. [ABSTRACT FROM AUTHOR]

Background: Large ex situ germplasm collections generally harbor a wide range of crop diversity. AVRDC - The World Vegetable Center is holding in trust the world's second largest mungbean (Vigna radiata) germplasm collection with more than 6,700 accessions. Screening large collections for traits of interest is laborious and expensive. To enhance the access of breeders to the diversity of the crop, mungbean core and mini core collections have been established. Results: The core collection of 1,481 entries has been built by random selection of 20% of the accessions after geographical stratification and subsequent cluster analysis of eight phenotypic descriptors in the whole collection. Summary statistics, especially the low differences of means, equal variance of the traits in both the whole and core collection and the visual inspection of quantile-quantile plots comparing the variation of phenotypic traits present in both collections indicated that the core collection well represented the pattern of diversity of the whole collection. The core collection was genotyped with 20 simple sequence repeat markers and a mini core set of 289 accessions was selected, which depicted the allele and genotype diversity of the core collection. Conclusions: The mungbean core and mini core collections plus their phenotypic and genotypic data are available for distribution to breeders. It is expected that these collections will enhance the access to biodiverse mungbean germplasm for breeding. [ABSTRACT FROM AUTHOR]

Phytophthora capsici is one of the most devastating pathogens facing pepper (Capsicum annuum) producers worldwide. Numerous factors, such as the race of the pathogen, the growing environment, and the source of resistance, have resulted in an overall lack of widely applicable molecular markers associated with resistance. Our objective was to determine the effect of the rating system on quantitative trait locus (QTL) detection and understand inheritance patterns of host resistance that can influence selection and molecular marker accuracy. We evaluated an F2:1| recombinant inbred line population screened against the highly virulent strain (Pc 134) and scored using two widely used methods, developed by Bosland and Lindsey and by Black. The rating system developed by Bosland and Lindsey resulted in slightly higher logarithm of odds for the QTL on chromosome 5, and we detected a QTL on chromosome 12 uniquely using this rating system. A QTL on chromosome 10 was detected using both rating systems, but Black resulted in considerably higher logarithm of odds for this QTL compared with the Bosland and Lindsey system. Molecular markers developed were nominally better at accurately predicting the phenotype than previously published molecular markers but did not completely explain resistance in our validation populations. The inheritance pattern of resistance in one of our F2 populations did not significantly deviate from a 7:9 segregation ratio, indicating duplicative recessive epistasis. However, these results could be confounded by the presence of incomplete gene action, which was found through the improved selection accuracy when the phenotypes of heterozygous individuals were grouped with those with susceptible alleles. [ABSTRACT FROM AUTHOR]

Habanero (Capsicum chinense Jacq.) is widely grown and consumed in West and Central African countries, and viral diseases represent an important production challenge. Diagnosis of the viral species affecting habanero productivity in Benin is limited, and understanding this will enable more efficient host resistance breeding. During 2019 and 2020, we characterized the incidence and severity of the viral diseases infecting nine promising habanero breeding lines and one commercial hybrid check under open field conditions in Benin. The horticultural performance, including yield and yield component traits of the entries, was determined during the 2 years of the experiment. A randomized complete block design was used with three replications, each with 24 plants. Data were recorded on days to 50% flowering and 50% fruit maturity, yield and on the yield components of fruit weight (g), fruit length (cm), and fruit width (mm), as well as disease incidence and severity. In total, 35 leaf samples were collected for viral diagnosis among habanero breeding lines. We found that Pepper veinal mottle virus (PVMV; Potyvirus) was the overwhelmingly predominant virus in our trials, with an 80% incidence; however, we found frequent coinfection of PVMV with Cucumber mosaic virus (CMV, Cucumovirus), Polerovirus, and, to a lesser extent, Chili veinal mottle virus (ChiVMV; Potyvirus). The mean disease incidence across all entries was 60%. AVPP1932 and PBC 2010 had the lowest disease incidence (35% and 43%, respectively), whereas AVPP1929 had the highest (86%) disease incidence. The F1 hybrid check Afadja had the overall highest yield, with 30 t·ha-1, followed by AVPP1932, with 19 t·ha-1, both in 2019. There was a negative correlation between disease incidence and total yield (r 5 20.44; P < 0.001), supporting previous studies indicating that viral diseases are major production constraints for habanero in West Africa. This study provides insight regarding the need to improve habanero for resistance to aphid-transmitted viruses and develop integrated pest management strategies to limit losses in Benin. [ABSTRACT FROM AUTHOR]

Chile pepper (Capsicum annuum L.) is an increasingly important vegetable and spice crop. Among the most devastating chile pepper-infecting viruses, especially in tropical and subtropical regions, are members of the whitefly transmitted Begomovirus, which cause pepper yellow leaf curl (PYLC). An effective PYLC management strategy is the development of resistant cultivars. However, genetic recombination, acquisition of extra DNA components, and synergistic interactions among different begomoviruses have resulted in the rapid emergence of new viruses that can infect new hosts, cause new disease symptoms, and overcome host resistance. In this project, 98 Capsicum entries comprising breeding lines, open pollinated varieties, genebank accessions, and wild species were screened for resistance to strains of Pepper yellow leaf curl Thailand virus (PepYLCThV). We used a randomized complete block design with three replications and 10 plants per replication in field net-houses at two locations (Khon Kaen and Kamphaeng Saen, Thailand) using augmented inoculation by viruliferous whiteflies. Scoring was done at '60, 90, and 120 days after inoculation using a standardized 6-point scale (1 = no symptoms to 6 = very severe symptoms), and the average of the scores of 10 plants within each replication was used for analysis. Although no entry was immune to the disease, the breeding line 9852-123 was highly resistant. Several accessions and lines were moderately resistant at both locations, although a high level of variability within these entries was observed. Overall, the disease severity at the Khon Kaen location was greater compared with Kamphaeng Saen, highlighting the importance of multilocation testing for disease resistance. The resistant entry identified here can be used to study gene action and to move resistance genes into well-adapted germplasm. [ABSTRACT FROM AUTHOR]

Heat tolerance is important for the sustainable production of many crops, including chile pepper. Tolerance to high temperature is complex and involves various component traits, with pollen viability being among the most important. in vitro pollen assays for heat tolerance have been widely used in chile pepper; however, associations between the pollen treatment and pollination have not been widely explored. The objectives of this study were to validate the utility of in vitro heat stress pollen characterization through in vivo pollination during summer and winter seasons and to evaluate the cross-compatibility among wild and domesticated species to initiate introgression population development. Seven entries of wild and domestic Capsicum species grown during the summer and winter seasons were used to evaluate pollination success rate. Pollen was either used directly or treated at 38 °C for four hours before making reciprocal self- and cross-pollination among all the entries. Significant associations between in vitro pollen treatment and pollination success rate during summer and winter seasons were identified. Heat treatment was a greater contributor to variability than the growing environment, which validates previous reports on the usefulness of studying pollen in vitro in selection for heat tolerance. Accessions of the wild progenitor C. annuum var glabriusculum, PBC 1969 and PBC 1970, were identified as a potential heat-tolerant source for use in breeding and future research. This work provides a basis for future research in exploring additional heat tolerance components as well as for the development of phenotyping assays for pollen or other floral traits.