Inferences on pod wall and seed defences against the brown cowpea coreid bug, Clavigralla tomentosicollis Stäl. (Hem., Coreidae) in wild and cultivated Vigna species

Studies were conducted in the laboratory to investigate the different components of the resistance of Vigna vexillata A. Richards, TVnu 72, and several cultivars of cultivated Vigna spp. to infestation and damage by the brown cowpea coreid, Clavigralla tomentosicollis Stäl. The main objective was to determine the different roles of the pod wall and seeds, and the relationship between seed size, number, and damage by this insect. Results showed that both the pod wall and seed clearly contribute different components to the resistance of TVnu 72 to C. tomentosicollis . Analysis of the results suggests that the seed defences and the pod wall pericarp may be more important than are trichomes in this resistance. Seed number was found to influence the extent of damage in a more predictable manner than seed size. However, because both of these traits are generally inversely related and mutually exclusive, their individual effects cannot be completely separated. The implications of these traits in a breeding programme targeted at a specific consumer group are discussed.     

Pod hairs as a factor in Vigna vexillata resistance to the pod-sucking bug, Clavigralla tomentosicollis

Two varieties of Vigna vexillata Benth proved more highly resistant, both under field (free-choice) and screenhouse (no-choice) conditions, to infestation and damage by the pod-sucking bug Clavigralla tomentosicollis (Stål.) (Hemiptera: Coreidae) than cowpea (Vigna unguiculata Walp.) varieties. Trichomes on the pod of V. vexillata partly accounted for the resistance. However, other factors in the pod or seeds are involved in the resistance. Because the pod wall of V. vexillata is not tougher than that of cowpeas, biochemical characteristics of pods and seeds of V. vexillata will be further investigated.

---

Résumé Deux variétés de Vigna vexillata se sont avérées plus résistantes aux infestations et dégâts causés par la punaise des cosses Clavigralla tomentosicollis (Stål.) (Hemiptera: Coreidae) que les variétés de Vigna unguiculata, tant en champ (condition de libre choix) qu'en serre (sans choix possible). Les trichomes des cosses de V. vexillata sont en partie responsables de cette résistance. Cependant d'autres facteurs de la cosses ou des fèves sont impliqués dan la résistance. Comme la paroi de la cosse de V. vexillata n'offre pas plus de résistance mécanique que celle de V. unguiculata, les études futures se porteront sur les caractéristiques biochimiques des cosses et des fèves de V. vexillata.

     

Resistance in accessions of cowpea to the coreid pod-bug Clavigralla tomentosicollis (Hemiptera: Coreidae)

Laboratory and screenhouse experiments were conducted to identify antibiosis and tolerance in four wild accessions of cowpea, Vigna unguiculata subsp. dekindtiana, to Clavigralla tomentosicollis St?l, the most damaging pod-sucking bug on cultivated Vigna genotypes in West Africa. The wild accession TVnu 151 showed antibiosis resistance, causing >50% mortality of the nymphs within 3 d of placing them on pods. Nymphs died more quickly on TVnu 151 than on TVnu 72, the wild and resistant control of the V. vexillata species which affected the weights of surviving insects to a much greater degree than TVnu 151. The three other accessions of the subspecies dekindtiana (TVnu 369, TVnu 517, and TVnu 707) did not cause significant mortality to the bugs, but rather extended their developmental time, with surviving adults showing lower weights and slower oviposition rates than those on the susceptible control IT84S-2246. Wild accessions affected male and female bugs differently, resulting in differential survival. This was reflected by the sex ratio which was male biased on the wild accessions (1:0.3-1:0.9), and female biased on IT84S-2246 (1:1.5). No evidence of tolerance was found in the four accessions of V. unguiculata subsp. dekindtiana. Overall, seed traits seemed to be the major resistance component in these wild accessions.     

Effect of application of different concentrations and appropriate schedules of aqueous garlic (Allium sativum L.) bulb extracts against Maruca vitrata and Clavigralla tomentosicollis on cowpea,Vigna unguiculata (L.) Walp

Abstract

Three concentrations (5, 10 and 20% w/v) and spraying schedules (2×, 4× and 6× weekly applications) of garlic, Allium sativum L. bulb extracts were assayed for efficacy against Maruca vitrata Fab. and Clavigralla tomentosicollis Stal. on cowpea under rain fed conditions in two seasons. The results showed that application of aqueous garlic bulb extracts at all the treatment levels significantly (p < 0.05) reduced the populations of the target pests compared with the untreated check. Application of 10 and 20% garlic bulb extracts with 4 or 6 weekly applications gave better control of these pests, protected the pods from damage and increased grain yields on treated cowpea compared with 5% extract at all three application schedules and were comparable to the synthetic insecticide treatment. This material could play a role in the integrated management of M. vitrata and C. tomentosicollis on cowpea fields managed by limited resource farmers in developing countries. The plant material is readily available, cheap, safe, non phytotoxic and technologically simple to use.     

Isolation of insecticidal lectin-enriched extracts from African yam bean (Sphenostylis stenocarpa) and other legume species

Insecticidal lectins were isolated from 20 resistant Vigna and non-Vigna legumes and tested againstn 3 pests of cowpea namely: Maruca vitrata, Callosobruchus maculatus and Clavigralla tomentosicollis. Crude lectins were separated from seeds using sodium chloride extraction, ammonium sulfate fractionation, and dialysis. SDS-PAGE indicated the molecular size of ca. 30 kDa for the most intense (and presumably active) band. Haemagglutination assays using trypsin-treated rabbit erythrocytes suggested that lectins were among the extracted proteins. Extracts from Lablab purpureus and Sphenostylis stenocarpa both non-Vigna spp., caused greater agglutination than those from the wild Vigna species. Bioassays on all three insect species using the lectin extracts incorporated in either artificial cowpea seeds (5% w/w) or in modified Vanderzant legume pod borer diet (1% w/v) indicated that the non-Vigna extracts were highly toxic to the insects. Mortality after 10 days was >80% in the most toxic extracts. The extract from one of the accessions of Sphenostylis stenocarpa, an edible legume, was singled out for lectin purification and future gene cloning with the view of using it for engineering resistance to cowpea pests.     

Molecular cytogenetic characterisation and phylogenetic analysis of the seven cultivated Vigna species (Fabaceae)

The genomic organisation of the seven cultivated Vigna species, V. unguiculata, V. subterranea, V. angularis, V. umbellata, V. radiata, V. mungo and V. aconitifolia, was determined using sequential combined PI and DAPI (CPD) staining and dual‐colour fluorescence in situ hybridisation (FISH) with 5S and 45S rDNA probes. For phylogenetic analyses, comparative genomic in situ hybridisation (cGISH) onto somatic chromosomes and sequence analysis of the internal transcribed spacer (ITS) of 45S rDNA were used. Quantitative karyotypes were established using chromosome measurements, fluorochrome bands and rDNA FISH signals. All species had symmetrical karyotypes composed of only metacentric or metacentric and submetacentric chromosomes. Distinct heterochromatin differentiation was revealed by CPD staining and DAPI counterstaining after FISH. The rDNA sites among all species differed in their number, location and size. cGISH of V. umbellata genomic DNA to the chromosomes of all species produced strong signals in all centromeric regions of V. umbellata and V. angularis, weak signals in all pericentromeric regions of V. aconitifolia, and CPD‐banded proximal regions of V. mungo var. mungo. Molecular phylogenetic trees showed that V. angularis and V. umbellata were the closest relatives, and V. mungo and V. aconitifolia were relatively closely related; these species formed a group that was separated from another group comprising V. radiata, V. unguiculata ssp. sesquipedalis and V. subterranea. This result was consistent with the phylogenetic relationships inferred from the heterochromatin and cGISH patterns; thus, fluorochrome banding and cGISH are efficient tools for the phylogenetic analysis of Vigna species.     

Resistance to silverleaf whitefly, Bemisia argentifolii (Hem., Aleyrodidae), in Gossypium thurberi, a wild cotton species

Abstract  Gossypium thurberi Todaro is a wild cotton species native to Mexico and parts of the southwestern USA. Four years of field studies in California's Imperial Valley revealed consistent very high levels of resistance in G. thurberi against silverleaf whitefly, Bemisia argentifolii Bellows and Perring, an important pest of cotton in many regions of the world. Naturally developing field infestations in plots of G. thurberi were significantly lower than in plots of the commercial cotton cultivars DP 5415, Siokra L23, and Stoneville 474. Gossypium thurberi has two morphological traits that, in past research, have been associated with lower levels of whitefly susceptibility: smooth- and okra-leaf; however, the levels of resistance observed in G. thurberi were significantly greater than in the cotton cultivar DP 5415, which is a smooth-leaf cotton, and Siokra L23 which, like G. thurberi , has both smooth- and okra-leaf traits. Therefore, the high level of resistance in G. thurberi seems to be due to factors above and beyond smooth- and okra-leaf. Siokra L23, which is among the least whitefly-susceptible cotton cultivars, developed whitefly populations over 30 times those on G. thurberi in all 3 years that Siokra L23 was tested. The difference in whitefly population development between G. thurberi and the other two cotton cultivars was even more striking, up to a 475-fold difference. In contrast to the clear results on naturally developing field infestations, experiments comparing nymphal survival among G. thurberi and commercial cotton cultivars did not detect antibiosis, and both choice and no-choice oviposition experiments did not detect antixenosis. Thus, the mechanisms of resistance in G. thurberi remain unknown.     

Characterization of resistance to three bruchid species (Callosobruchus spp., Coleoptera,Bruchidae) in cultivated rice bean (Vigna umbellata)

Resistance of wild and cultivated rice bean (Vigna umbellata [Thunberg] Ohwi and Ohashi) to three bruchid species, Callosobruchus chinensis L., Callosobruchus maculatus F., and Callosobruchus analis F., was evaluated. All but three accessions of cultivated, and all wild rice bean accessions tested, exhibited complete resistance to all three bruchid species. Rice bean seeds with seed coat removed also showed complete resistance to the three bruchid species. Results indicate that physical attributes and/or chemical(s) present in the seed coat of rice bean are not the main factors responsible for resistance. Feeding tests were performed by using artificial beans prepared with varying proportions of rice bean (resistant) and azuki bean (susceptible) flour. Number of bruchid adults that emerged decreased, and larval developmental period (days) was extended, when artificial beans with an increasing proportion of rice bean flour were used. These tests revealed that a chemical compound(s) contained in the cotyledon of rice bean has an inhibitory effect on the growth of these bruchid species. The results also indicate that the chemical(s) in rice bean cotyledon is most effective against C. maculatus.     

Genome dissection of traits related to domestication in azuki bean (Vigna angularis) and comparison with other warm-season legumes

BACKGROUND: The objective of this study was to dissect into quantitative trait loci (QTLs) the large morphological and physiological differences between cultivated azuki bean (Vigna angularis) and a wild relative and to infer the commonalities of the QTLs for domestication-related traits across the Asian Vigna and with other warm-season legumes. METHODS: Two linkage maps, for the BC(1)F(1) and F(2) populations, respectively, from the same cross between azuki bean and V. nepalensis were developed. Using these linkage maps QTLs for 33 domestication-related traits were analysed and mapped. The location of mapped QTLs was compared with locations of similar QTLs in other warm-season legumes. KEY RESULTS: QTLs were detected for seed-, pod-, stem- and leaf-related traits. Most traits were controlled by between two and nine QTLs but several traits, such as pod dehiscence, were controlled by single genes. QTLs for domestication-related traits were restricted to particular regions of the azuki bean genome, especially linkage groups 1, 2, 4, 7 and 9. Linkage groups 1 and 2 had QTLs for a suite of traits including pod size, germination, seed size and lower stem length. QTLs on linkage groups 7 and 9 were associated with upper stem length, maximum leaf size and pod and seed size. Pleiotropy or close linkage of genes for domestication-related traits is suggested in these regions. While some QTLs are common to azuki bean and other warm-season legumes, many are recorded for the first time in azuki bean. CONCLUSIONS: QTLs for a large number of domestication-related traits have been mapped for the first time in azuki bean. QTLs with unexpected effect and new QTLs for traits such as seed size have been found. The results provide a foundation that will be useful for improvement of azuki bean and related legumes.     

Resistance to the cowpea weevil (Callosobruchus maculatus) larva in pods of cowpea (Vigna unguiculata)

Thirty cowpea, Vigna unguiculata (L.) Walpers, cultivars were evaluated as intact pods to determine if any possessed resistance to the cowpea weevil, Callosobruchus maculatus (Fabr.). Pod resistance was measured as pre-establishment larval mortality (PreM); those larvae dying after egg hatch but before penetrating into the seeds, and as post-establishment within-seed mortality (PostM); those larvae dying after penetrating into the seeds. Among the 30 varieties examined, PreM ranged from 57.9% to 99.4% and PostM ranged from 6.7% to 82.6%. Ten varieties exhibited total intact pod mortality (mortality from egg hatch to adult emergence from the seed) greater than 95%.Physical measurements were made of several pod and seed characteristics to ascertain whether the observed pod resistance was due to seed factors, pod-wall factors, or to interactions between the pod and seeds. Although resistance to breakage (from handling plus pod dehiscence) is a necessary characteristic for overall pod protection against C. maculatus; other factors are also important. Among the other pod and seed characteristics measured to identify major resistance factors, seed coat thickness was the one most highly correlated with pod resistance. Our results suggest that interactions between pod-wall and seed coat characteristics play a large role in pod resistance of cowpeas to C. maculatus.     

The distribution of cultivated species of Porophyllum (Asteraceae) and their wild relatives under climate change

The geographic distribution of plant species is already being affected by climate change. Cropping patterns of edible plant species and their wild relatives will also be affected, making it important to predict possible changes to their distributions in the future. Currently, species distribution models are valuable tools that allow the estimation of species’ potential distributions, in the recent past as well as during other time spans for which climate data have been obtained. With the aim of evaluating how species distributions respond to current and future climate changes, in this work species distribution models were generated for two cultivated species of the Porophyllum genus (Asteraceae), known commonly as ‘pápalos' or ‘pápaloquelites', as well as their Mexican wild relatives, at five points in time (21,000 years ago, present, 2020, 2050, and 2080). Using a database of 1442 entries for 16 species of Porophyllum and 19 environmental variables, species distribution models were constructed for each time period using the Maxent modelling algorithm; those constructed for the future used a severe climate change scenario. The results demonstrate contrasting effects between the two cultivated species; for P. linaria, the future scenario suggests a decrease in distribution area, while for P. macrocephalum distribution is predicted to increase. Similar trends are observed in their wild relatives, where 11 species will tend to decrease in distribution area, while three are predicted to increase. It is concluded that the most important agricultural areas where the cultivated species are grown will not be greatly affected, while the areas inhabited by the wild species will. However, while the results suggest that climate change will affect the distribution of the cultivated species in contrasting ways, evaluations at finer scales are recommended to clarify the impact within cultivation zones.     

Isolation and diversity analysis of resistance gene analogues (RGAs) from cultivated and wild strawberries

Degenerate oligonucleotide primers, designed based on conserved regions of Nucleotide Binding Site (NBS) domains from previously cloned plant resistance genes, were used to isolate Resistance Gene Analogues (RGAs) from wild and cultivated strawberries. Seven distinct families of RGAs of the NBS-LRR type were identified from two related wild species, Fragaria vesca and F. chiloensis, and six different Fragaria × ananassa cultivars. With one exception (GAV-3), the deduced amino acid sequences of strawberry RGAs showed strong similarity to TIR (Toll Interleukin I Receptor)-type R genes from Arabidopsis, tobacco and flax, suggesting the existence of common ancestors. GAV-3 seemed to be more closely related to the non-TIR type. Further studies showed that the recombination level and the ratio of non-synonymous to synonymous substitutions within families were low. These data suggest that NBS-encoding sequences of RGAs in strawberry are subject to a gradual accumulation of mutations leading to purifying selection, rather than to a diversifying process. The present paper is the first report on RGAs in strawberry.Communicated by M.-A. Grandbastien     

Identification of resistance to Peanut bud necrosis virus (PBNV) in wild Arachis germplasm

Eighty three wild Arachis germplasm accessions, belonging to 24 species of five sections and one natural hybrid derivative of a cross between the cultivated and a wild Arachis species, were evaluated along with a susceptible groundnut cultivar for resistance to Peanut bud necrosis virus (PBNV) in a replicated field trial at ICRISAT, Patancheru, India. Thirty days after sowing, the percentage of infected plants were recorded for all the accessions and subsequently young leaflets from all these accessions were tested for the presence of the virus by enzyme linked immunosorbent assay (ELISA). One accession each of A. benensis and A. cardenasii, and two accessions of A. villosa, in the section Arachis, two accessions of A. appressipila in the section Procumbentes, and one accession of A. triseminata under section Triseminatae were not infected by PBNV. These seven field‐resistant accessions were tested under glasshouse conditions for virus resistance by mechanical sap inoculations. One accession of A. cardenasii and two accessions of A. villosa did not show systemic infection. Similarly, in another glasshouse test, where 13 A. cardenasii accessions of section Arachis were evaluated, two accessions did not show systemic infection. In all these resistant accessions, the inoculated leaves showed infection, but the systemic leaves did not show the presence of virus in spite of repeated mechanical sap inoculations. So, the resistance in these accessions appears to be due to a block in systemic movement of the virus. To our knowledge this is the first report on the identification of resistance to PBNV in wild Arachis species. Since both A. cardenasii and A. villosa are the progenitors of cultivated groundnut and can be hybridised with the latter, the resistant accessions are being utilised in conventional breeding programmes to transfer PBNV resistance to widely adapted groundnut cultivars.     

Broad-based resistance to pigeonpea sterility mosaic disease in wild relatives of pigeonpea (Cajanus: Phaseoleae)

Sterility mosaic disease (SMD), an important biotic constraint on pigeonpea (Cajanus cajan) in the Indian subcontinent, is caused by Pigeonpea sterility mosaic virus (PPSMV) transmitted by the eriophyid mite, Aceria cajani. Distinct PPSMV isolates occur in different geographical regions and broad‐based resistance to all these isolates is scarce in cultivated pigeonpea germplasm. Wild relatives of pigeonpea, which are known to possess resistance to several pests and diseases, were evaluated for broad‐based SMD resistance. One hundred and fifteen wild Cajanus accessions from six species (C. albicans, C. platycarpus, C. cajanifolius, C. lineatus, C. scarabaeoides and C. sericeus) were evaluated against three PPSMV isolates prevailing in peninsular India. Evaluations were done under greenhouse conditions in endemic locations of each isolate through mite‐mediated virus inoculation. Fifteen accessions showed resistance to all three isolates: ICP 15614, 15615, 15626, 15684, 15688, 15700, 15701, 15725, 15734, 15736, 15737, 15740, 15924, 15925 and 15926. Most of the wild accessions did not support mite multiplication. The majority of the accessions resistant to PPSMV following inoculations with viruliferous mites were susceptible by graft inoculation, suggesting that vector resistance is conferring resistance to infection with PPSMV. The 15 accessions identified as being resistant to infection to all three virus isolates tested are cross compatible with pigeonpea by traditional breeding. They are therefore useful for exploitation in breeding programmes to increase both the level of SMD resistance and to diversify its genetic base in the cultivated pigeonpea gene pool.     

Mapping of quantitative trait loci for a new source of resistance to bruchids in the wild species Vigna nepalensis Tateishi & Maxted (Vigna subgenus Ceratotropis)

Azuki bean breeders have long been interested in producing azuki bean [Vigna angularis (Willd.) Ohwi & Ohashi] varieties with bruchid resistance. A new bruchid (Callosobruchus spp.) resistance source was found in V. nepalensis Tateishi & Maxted, a species that is cross compatible with azuki bean. Quantitative trait loci (QTLs) analysis for resistance to C. chinensis (L.) and C. maculatus (F.) was conducted using F(2) (V. nepalensis x V. angularis) and BC(1)F(1) [(V. nepalensis x V. angularis) x V. angularis] populations derived from crosses between the bruchid resistant species V. nepalensis and bruchid susceptible species V. angularis. Resistance was measured using two traits, percentage of seeds damaged by bruchids and the time taken for adult bruchids to emerge from seeds. Based on the results from both populations seven QTLs were detected for bruchid resistance; five QTLs for resistance to C. chinensis and two QTLs for resistance to C. maculatus. The different locations found for some resistance QTL to the two bruchid species suggests different resistance mechanisms. QTLs on linkage group (LG) 1 and LG2 for bruchid resistance to C. chinensis co-localized with seed size QTLs suggesting that incremental increase in seed size accompanied susceptibility to C. chinensis. Based on linked markers the QTL on these two linkage groups appear to be the same as previously reported in other Asian Vigna. However, several other QTLs were newly detected including one on LG4 that appears unrelated to seed size. Transfer of these new sources of bruchid resistance from V. nepalensis to azuki bean will be aided by the progress being made in azuki genome mapping.     

Comparative RFLP mapping of an allotetraploid wild rice species (Oryza latifolia) and cultivated rice (O. sativa)

The purpose of this study was to construct a comparative RFLP map of an allotetraploid wild rice species, Oryza latifolia, and to study the relationship between the CCDD genome of O. latifolia and the AA genome of O. sativa. A set of RFLP markers, which had been previously mapped to the AA genome of cultivated rice, were used to construct the comparative map. Fifty-eight F₂ progeny, which were derived from a single F₁ plant, were used for segregation analysis. The comparative RFLP map contains 149 DNA markers, including 145 genomic DNA markers from cultivated rice, 3 cDNA markers from oat, and one known gene (waxy, from maize). Segregation patterns reflected the allotetraploid ancestry of O. latifolia, and the CC and DD genomes were readily distinguished by most probes tested. There is a high degree of conservation between the CCDD genome of O. latifolia and the AA genome of O. sativa based on our data, but some inversions and translocations were noted.     

Inheritance of resistance to yellow mosaic virus in blackgram (Vigna mungo (L.) Hepper)

Summary The inheritance of resistance to mungbean yellow mosaic virus (MYMV) was studied in blackgram (Vigna mungo (L.) Hepper). The highly resistant donors Pant U-84 and UPU-2 and a highly susceptible line, UL-2, their F₁'s, F₂'s and backcrosses were grown with spreader located every 5 to 6 rows. The resistance was found to be digenic and recessive in all the crosses and free from cytoplasmic effect.     

Research Note: Simple molecular discrimination of cultivated Porphyra species (Porphyra yezoensis and Porphyra tenera) and related wild species (Bangiales,Rhodophyta)

To discriminate between cultivated Porphyra species (Porphyra yezoensis and Porphyra tenera) and closely related wild Porphyra species, we developed a polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) analysis of the rbcL gene using five restriction enzymes. Although our previous PCR‐RFLP analyses of internal transcribed spacer (ITS) rDNA and plastid RuBisCO spacer regions could not always discriminate wild P. yezoensis, wild P. tenera, and closely related wild species, the PCR‐RFLP profiles of the rbcL gene were useful in discriminating samples collected from natural habitats. Therefore, PCR‐RFLP analysis of the rbcL gene will help in the simple identification of a large number of samples, not only for the establishment of reliable cultures as breeding material, but also for the taxonomic investigations of species that are closely related to cultivated Porphyra.     

An ancient R gene from the wild potato species Solanum bulbocastanum confers broad-spectrum resistance to Phytophthora infestans in cultivated potato and tomato

Late blight, caused by the oomycete pathogen Phytophthora infestans, is the most devastating disease for potato cultivation. Here, we describe the positional cloning of the Rpi-blb1 gene from the wild potato species Solanum bulbocastanum known for its high levels of resistance to late blight. The Rpi-blb1 locus, which confers full resistance to complex isolates of P. infestans and for which race specificity has not yet been demonstrated, was mapped in an intraspecific S. bulbocastanum population on chromosome 8, 0.3 cM from marker CT88. Molecular analysis of a bacterial artificial chromosome (BAC) clone spanning the Rpi-blb1 locus identified a cluster of four candidate resistance gene analogues of the coiled coil, nucleotide-binding site, leucine-rich repeat (CC-NBS-LRR) class of plant resistance (R) genes. One of these candidate genes, designated the Rpi-blb1 gene, was able to complement the susceptible phenotype in a S. tuberosum and tomato background, demonstrating the potential of interspecific transfer of broad-spectrum late blight resistance to cultivated Solanaceae from sexually incompatible host species. Paired comparisons of synonymous and non-synonymous nucleotide substitutions between different regions of Rpi-blb1 paralogues revealed high levels of synonymous divergence, also in the LRR region. Although amino acid diversity between Rpi-blb1 homologues is centred on the putative solvent exposed residues of the LRRs, the majority of nucleotide differences in this region have not resulted in an amino acid change, suggesting conservation of function. These data suggest that Rpi-blb1 is relatively old and may be subject to balancing selection.