Association Mapping for Yield Attributing Traits and Yellow Mosaic Disease Resistance in Mung Bean [Vigna radiata (L.) Wilczek]

Mung bean [Vigna radiata (L.) Wilczek] is an important short-duration grain legume widely known for its nutritional, soil ameliorative, and cropping system intensification properties. This study aims at evaluating genetic diversity among mung bean genotypes and detecting genomic regions associated with various yield attributing traits and yellow mosaic disease (YMD) resistance by association mapping. A panel of 80 cultivars and advanced breeding lines was evaluated for 10 yield-related and YMD resistance traits during kharif (monsoon) and summer seasons of 2018–2019 and 2019–2020. A total of 164 genome-wide simple sequence repeat (SSR) markers were initially screened, out of which 89 were found polymorphic which generated 317 polymorphic alleles with an average of 3.56 alleles per SSR locus. The number of alleles at each locus varied from 2 to 7. The population genetic structure analysis grouped different genotypes in three major clusters and three genetically distinct subpopulations (SPs) (i.e., SP-1, SP-2, and SP-3) with one admixture subpopulation (SP-4). Both cluster and population genetic structure analysis categorized the advanced mung bean genotypes in a single group/SP and the released varieties in other groups/SPs, suggesting that the studied genotypes may have common ancestral history at some level. The population genetic structure was also in agreement with the genetic diversity analysis. The estimate of the average degree of linkage disequilibrium (LD) present at the genome level in 80 mung bean genotypes unveiled significant LD blocks. Over the four seasons, 10 marker-trait associations were observed significant for YMD and four seed yield (SY)-related traits viz., days to flowering, days to maturity, plant height, and number of pods per plant using the mixed linear model (MLM) method. These associations may be useful for marker-assisted mung bean yield improvement programs and YMD resistance.

​Standardization of Grafting in Bittergourd (Momordica charantia L.)

Background: Commercial cultivation of bitter gourd is affected by biotic stresses like mosaic disease, fusarium wilt and root-knot nematode as well as abiotic stress like drought. Grafting with resistant rootstocks can be a tool to control these problems. In vegetable production, grafting is exploited commercially in many parts of the world. The cultivated area of grafted solanaceae and cucubitaceae plants has increased tremendously in recent years because of the advantages of grafted plants. Commercial use of vegetable grafting is a relatively recent innovation in India and scientific information on grafting in bittergourd is meager. In this context, identification of suitable rootstocks and standardization of grafting techniques that do not have adverse effect on yield and fruit quality not only lay foundation for further evaluation on tolerance to different biotic and abiotic stresses but also enhance the area and production of bitter gourd especially in sustainable production systems. Methods: Grafting in bitter gourd was carried out with three grafting methods such as hole insertion grafting, one cotyledon grafting and cleft grafting using growth regulators viz., alar and CCC to control height of rootstocks in order to identify suitable method, growth regulator and its concentration. The grafting experiment was done independently to four cucurbitaceous rootstocks viz., sponge gourd, pumpkin, bottle gourd and bitter gourd using bittergourd var. Preethi as scion. Height and diameter of the rootstocks before grafting were recorded and then the growth regulators alar and CCC each at 10 mgL-1 and 50 mgL-1 along with distilled water as control were sprayed on rootstocks in order to prevent the lodging of the root stocks and then grafted using different methods. Days taken for graft union and percentage success were also evaluated after grafting. Result: Our study of grafting bitter gourd scion into four cucurbitaceous rootstocks utilizing three methods and two growth regulators at two different concentration along with control exhibited significant difference in graft success among the methods as well as concentration of growth regulator in all four experiments. This work can be further utilized for imparting resistance against abiotic and biotic stresses in bitter gourd by selecting suitable rootstocks.

Effect of Cultivars and Temperature on Synergistic Interaction Between Panicum Mosaic Virus and Satellite Panicum Mosaic Virus in Switchgrass

Panicum mosaic virus (PMV), the type species of the genus Panicovirus in the family Tombusviridae, naturally infects switchgrass (Panicum virgatum L.). PMV and its molecular partner, satellite panicum mosaic virus (SPMV), interact synergistically in co-infected millets with exacerbated disease phenotype and increased accumulation of PMV, compared to plants infected only by PMV. In this study, we examined the reaction of switchgrass cvs. Summer and Kanlow to PMV and PMV+SPMV infections at 24°C and 32°C. Switchgrass cv. Summer was susceptible to PMV at both temperatures. In contrast, cv. Kanlow was tolerant to PMV at 24°C but not at 32°C, suggesting that Kanlow harbors temperature-sensitive resistance against PMV. At 24°C, PMV was readily detected in inoculated leaves but not in upper non-inoculated leaves of Kanlow, suggesting that resistance to PMV was likely mediated by abrogation of long-distance virus transport. Co-infection by PMV and SPMV at 24°C and 32°C in cv. Summer but not in Kanlow caused increased symptomatic systemic infection and mild disease synergism with slightly increased PMV accumulation compared to plants infected only by PMV. These data suggest that the interaction between PMV and SPMV in switchgrass is cultivar dependent, manifested in Summer but not in Kanlow. However, co-inoculation of cv. Kanlow by PMV+SPMV caused an enhanced asymptomatic infection, suggesting a role for SPMV in enhancing symptomless infection in a tolerant cultivar. These data suggest that enhanced asymptomatic infections in virus-tolerant switchgrass cultivar could serve as a source for virus spread and play an important role in panicum mosaic disease epidemiology under field conditions. Our data revealed that cultivars, co-infection with SPMV, and temperature influenced the severity of symptoms elicited by PMV in switchgrass.

Development and Evaluation of Stable Sugarcane Mosaic Virus Mild Mutants for Cross-Protection Against Infection by Severe Strain

Sugarcane mosaic virus (SCMV; genus Potyvirus) induces maize dwarf mosaic disease that has caused serious yield losses of maize in China. Cross-protection is one of the efficient strategies to fight against severe virus strains. Although many mild strains have been identified, the spontaneous mutation is one of the challenging problems affecting their application in cross-protection. In this study, we found that the substitution of cysteine (C) at positions 57 or 60 in the zinc finger-like motif of HC-Pro with alanine (A; C57A or C60A) significantly reduced its RNA silencing suppression activity and SCMV virulence. To reduce the risk of mild strains mutating to virulent ones by reverse or complementary mutations, we obtained attenuated SCMV mutants with double-mutations in the zinc finger-like and FRNK motifs of HC-Pro and evaluated their potential application in cross-protection. The results showed that the maize plants infected with FKNK/C60A double-mutant showed symptomless until 95 days post-inoculation and FKNK/C60A cross-protected plants displayed high resistance to severe SCMV strain. This study provides theoretical and material bases for the control of SCMV through cross-protection.

A population based expression atlas provides insights into disease resistance and other physiological traits in cassava (Manihot esculenta Crantz)

Cassava, a food security crop in Africa, is grown throughout the tropics and subtropics. Although cassava can provide high productivity in suboptimal conditions, the yield in Africa is substantially lower than in other geographies. The yield gap is attributable to many challenges faced by cassava in Africa, including susceptibility to diseases and poor soil conditions. In this study, we carried out 3’RNA sequencing on 150 accessions from the National Crops Resources Research Institute, Uganda for 5 tissue types, providing population-based transcriptomics resources to the research community in a web-based queryable cassava expression atlas. Differential expression and weighted gene co-expression network analysis were performed to detect 8820 significantly differentially expressed genes (DEGs), revealing similarity in expression patterns between tissue types and the clustering of detected DEGs into 18 gene modules. As a confirmation of data quality, differential expression and pathway analysis targeting cassava mosaic disease (CMD) identified 27 genes observed in the plant–pathogen interaction pathway, several previously identified CMD resistance genes, and two peroxidase family proteins different from the CMD2 gene. Present research work represents a novel resource towards understanding complex traits at expression and molecular levels for the development of resistant and high-yielding cassava varieties, as exemplified with CMD.

Roles of two distinct alphasatellites modulating geminivirus pathogenesis

Background Alphasatellites are small coding DNA satellites frequently associated with a begomovirus/betasatellite complex, where they are known to modulate virulence and symptom development. Two distinct alphasatellites, namely, Cotton leaf curl Multan alphasatellite (CLCuMuA), and Gossypium darwinii symptomless alphasatellite (GDarSLA) associated with Cotton leaf curl Multan virus-India (CLCuMuV-IN) and Ludwigia leaf distortion betasatellite (LuLDB) were found to be associated with yellow mosaic disease of hollyhock (Alcea rosea) plants. In this study, we show that alphasatellites CLCuMuA and GDarSLA attenuate and delay symptom development in Nicotiana benthamiana. The presence of either alphasatellites reduce the accumulation of the helper virus CLCuMuV-IN. However, the levels of the associated betasatellite, LuLDB, remains unchanged. These results suggest that the alphasatellites could contribute to the host defence and understanding their role in disease development is important for developing resistance strategies. Methods Tandem repeat constructs of two distinct alphasatellites, namely, CLCuMuA and GDarSLA associated with CLCuMuV-IN and LuLDB were generated. N. benthamiana plants were co-agroinoculated with CLCuMuV and its associated alphasatellites and betasatellite molecules and samples were collected at 7, 14 and 21 days post inoculation (dpi). The viral DNA molecules were quantified in N. benthamiana plants by qPCR. The sequences were analysed using the MEGA-X tool, and a phylogenetic tree was generated. Genetic diversity among the CLCuMuA and GDarSLA was analysed using the DnaSP tool. Results We observed a reduction in symptom severity and accumulation of helper virus in the presence of two alphasatellites isolated from naturally infected hollyhock plants. However, no reduction in the accumulation of betasatellite was observed. The phylogenetic and genetic variability study revealed the evolutionary dynamics of these distinct alphasatellites , which could explain the role of hollyhock-associated alphasatellites in plants. Conclusions This study provides evidence that alphasatellites have a role in symptom modulation and suppress helper virus replication without any discernible effect on the replication of the associated betasatellite.