Sources of resistance to groundnut rosette disease in global groundnut germplasm*

About 6800 groundnut germplasm accessions originating from South America, Africa, and Asia were evaluated for resistance to rosette disease using an infector row technique between the 1990/91 and 1996/97 growing seasons. Of these, 116 germplasm accessions, including 15 short-duration Spanish types, have shown high levels of resistance to rosette disease. A high percentage of these resistant accessions were from West Africa and a few were from Asia and southern Africa. Only one out of 1400 accessions from South America showed resistance to rosette disease. All disease-resistant accessions were susceptible to groundnut rosette assistor virus. This is the first report to identify sources of resistance to rosette disease in groundnut germplasm from Asia and South America. These additional sources of resistance provide an opportunity to broaden the genetic base of resistance to rosette disease. The origins of rosette resistance in groundnut are discussed.     

Additional sources of resistance to groundnut rosette disease in groundnut germplasm and breeding lines

Groundnut rosette, a virus disease of groundnut (Arachis hypogaea) transmitted by the aphid, Aphis craccivora Koch, reduces yield in susceptible cultivars by 30–100%. Additional sources were sought in germplasm accessions involving 2301 lines from different sources and from 252 advanced breeding lines derived from crosses involving earlier identified sources of resistance to rosette. The lines were evaluated in field screening trials using an infector row technique during 1996 and 1997 growing seasons. Among the germplasm lines, 65 accessions showed high levels of resistance while 134 breeding lines were resistant. All rosette disease resistant lines were susceptible to groundnut rosette assistor virus. This work identified germplasm and breeding lines that will contribute to an integrated management of groundnut rosette disease. These new sources also provide an opportunity to eliminate yield losses due to the rosette disease.     

The nature of the resistance in groundnut to rosette disease

Groundnut rosette disease is caused by a complex of three agents, groundnut rosette virus (GRV) and its satellite RNA, and groundnut rosette assistor virus (GRAV); the satellite RNA is mainly responsible for the disease symptoms. Groundnut genotypes possessing resistance to rosette disease were shown to be highly resistant (though not immune) to GRV and therefore to its satellite RNA, but were fully susceptible to GRAV.     

Different variants of the satellite RNA of groundnut rosette virus are responsible for the chlorotic and green forms of groundnut rosette disease*

Groundnut plants with symptoms of rosette disease contain groundnut rosette virus (GRV), but GRV is transmitted by Aphis craccivora only from plants that also contain groundnut rosette assistor virus (GRAV). Two main forms of rosette disease are recognised, ‘chlorotic rosette’ and ‘green rosette’. GRV cultures invariably possess a satellite RNA and this is the major cause of rosette symptoms: satellite-free isolates derived from GRV cultures from Nigerian plants with chlorotic or green rosette, or from Malawian plants with chlorotic rosette, induced no symptoms, or only transient mild mottle or interveinal yellowing, in groundnut. When the satellite RNA species from GRV cultures from Nigerian green or Malawian chlorotic rosette were reintroduced into the three satellite-free isolates in homologous and heterologous combinations, the ability to induce rosette symptoms was restored and the type of rosette induced was that of the cultures from which the satellite RNA was derived. Thus different forms of the satellite are responsible for the different forms of rosette disease. Other forms of the satellite induce only mild chlorosis or mottle symptoms in groundnut. Individual plants may contain more than one form of the satellite, and variations in their relative predominance are suggested to account for the variable symptoms (ranging from overall yellowing to mosaic) seen in some plants graft-inoculated with chlorotic rosette.     

Impact of groundnut rosette disease on nutritive value and elemental composition of four varieties of peanut (Arachis hypogaea)

Proximate and elemental composition of four peanut genotypes infected with groundnut rosette disease (GRD) was examined. Moisture and ash content generally decreased while fat and energy content increased in seeds from diseased plants. Protein and carbohydrate varied between seeds of diseased and healthy plants of the different varieties with no consistent pattern. Instrumental neutron activation analysis of 10 elements within leaves, stems and seeds showed elevated levels of K, Al and Cl in leaves, stems and seeds in at least three of the four varieties infected with GRD while Na was decreased in stems but increased in seeds. While significant differences were found, Mg, Mn, Ca and Zn did not show any consistent change with respect to plant part or genotype, between diseased and healthy plants. V and Fe were found at low levels in leaves and stems and not detected in seeds. This represents the first report on the effect of GRD on the nutritive quality of peanuts.     

Inheritance of resistance to groundnut rosette virus in groundnut (Arachis hypogaea L.)

A method of field screening groundnut seedlings for resistance to groundnut rosette virus (GRV), by means of which over 97% incidence was induced in rows of susceptible test plants, was developed at Chitedze Research Station in Malawi. Two GRV-resistant Virginia cultivars (RG 1 and RMP 40) were crossed with three susceptible cultivars, one from each of the Spanish (JL 24), Valencia (ICGM 48) and Virginia (Mani Pintar) botanical groups. Twelve F₁ reciprocal crosses and their F₂ and backcross generations were produced and the material screened in nurseries in 1985/86 and 1986/87. Seedlings raised from plants which did not become infected in the field were inoculated in the glasshouse in order to eliminate susceptible escapees. The numbers of diseased and healthy individuals in each population were subjected to χ² tests. In the majority of the F₂ populations a good fit was obtained for a ratio of one resistant to 15 susceptible plants, a ratio to be expected if resistance to GRV were determined by a pair of independent complementary recessive genes. This was further supported by data from backcross generations.     

A variant of the satellite RNA of groundnut rosette virus that induces brilliant yellow blotch mosaic symptoms in Nicotiana benthamiana

Some Malawian cultures of groundnut rosette virus (GRV) give rise to variants that, although still causing symptoms of the chlorotic type of rosette in groundnut, induce brilliant yellow blotch mosaic symptoms, instead of the usual veinal chlorosis and mild mottle, in Nicotiana benthamiana. One such isolate (YB) induced the formation in infected plants of a 0.9 kbp dsRNA having extensive sequence homology with molecules of similar size in other naturally occurring isolates of GRV. These dsRNA molecules were shown to be double-stranded forms of single-stranded satellite RNA molecules. Experiments in which the satellite was removed from and restored to isolate YB, or exchanged with those from other GRV isolates, showed that it carries the determinant for yellow blotch mosaic symptoms. Plants inoculated with the 0.9 kbp dsRNA (denatured or undenatured) developed yellow blotch mosaic even when the satellite-free GRV helper was not inoculated until 11 days later. The satellite RNA is therefore a very stable molecule. Prior infection of N. benthamiana with a GRV isolate containing a normal form of the satellite protected against expression of yellow blotch mosaic symptoms when the plants were later inoculated with isolate YB, whereas prior infection with satellite-free isolates did not. This provides a simple method of determining whether a GRV isolate has an associated satellite RNA. The YB satellite seems to be a newly recognised variant additional to those known to cause the chlorotic, green and other forms of groundnut rosette disease.     

The Effect of Groundnut rosette assistor virus on the Agronomic Performance of Four Groundnut (Arachis hypogaea L.) Genotypes

The effect of Groundnut rosette assistor virus (GRAV), in the absence of the other two agents (Groundnut rosette virus and its satellite RNA) of the groundnut rosette disease virus complex, was evaluated on the agronomic performance of four groundnut (=peanut) genotypes (CG‐7, ICGV‐SM‐90704, JL‐24 and ICG‐12991) with different botanical characteristics. All genotypes infected with GRAV showed mild yellowing/chlorosis of leaves and the symptoms persisted throughout their growth period. ELISA absorbance values indicated lower amounts of GRAV antigen in ICGV‐SM‐90704 than in the other genotypes. The reduction in leaf area due to GRAV infection varied between 15.5% and 21.7%, whereas the plant height was decreased between 11.3% and 13.4% among the four genotypes. GRAV infection caused 28.4%, 16.9%, 21.7% and 25.5% reduction in the dry weight of haulms in CG‐7, ICGV‐SM‐90704, JL‐24 and ICG‐12991 respectively. Plants infected with GRAV showed greater reduction in seed weight in CG‐7 (52.2%), followed by JL‐24 (46.1%), ICG‐12991 (40.7%) and ICGV‐SM‐90704 (25.7%). These results provide evidence for the first time that GRAV infection, without GRV and sat RNA, affect plant growth and contribute to yield losses in groundnut.     

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.     

Resistance in groundnut (Arachis hypogaea L.) to Aphis craccivora (Koch).

The behaviour, development and reproductive capacity of Aphis craccivora, vector of a number of groundnut viruses, are compared on a range of susceptible and resistant genotypes. Field trials demonstrated no significant difference between genotypes in the rate of arrival of alates, but population development was slower, and subsequent population decline faster, on the genotype EC 36892 (ICG 5240). Behavioural studies in the screenhouse, likewise showed no inhibition to alighting onto EC 36892 though choice tests demonstrated a significant redistribution of the population in favour of the susceptible genotype TMV 2 (ICG 221) over the following 10 h. In clip cage experiments, development was faster and nymphal numbers were higher on the genotype TMV 2 compared to EC 36892.     

Detection of groundnut rosette umbravirus infections with radioactive and non-radioactive probes to its satellite RNA

A cloned cDNA copy of the satellite RNA of groundnut rosette virus (GRV), labelled with either ³²P or digoxigenin, was used as a probe to detect the satellite RNA in infected leaves. The test was successfully applied to N. benthamiana and to groundnuts, infected with isolates of GRV from East and West Africa and with isolates which cause different types of symptom in groundnuts, including one which is almost symptomless. Although the probe did not react with extracts from plants infected with GRV isolates from which the satellite RNA had been artificially eliminated, all naturally occurring GRV isolates contain the satellite RNA. The test therefore provides a reliable indicator of infection by GRV.     

Epidemiology of groundnut rosette virus disease: current status and future research needs@

Rosette is the most destructive virus disease of groundnut in sub-Saharan Africa. It is caused by a complex of three agents, namely groundnut rosette assistor virus, groundnut rosette virus and its satellite RNA. The disease appears to be indigenous to Africa as it has not been recorded elsewhere. Thus rosette represents a new-encounter situation as the disease is thought to have spread to the introduced groundnut from indigenous host plants. Rosette has been known since 1907 and much information has been obtained on the main features of the disease, viz. its biology, transmission, viral aetiology and diagnosis, and the impact of chemical control of the aphid vector, cultural practices and virus-resistant varieties on disease management. However, there are still many gaps in the available knowledge, especially the reasons for the large and unpredictable fluctuations in the incidence and severity of rosette disease throughout sub-Saharan Africa. Three unresolved issues of particular importance concern the nature of the primary source(s) of inoculum, the means of survival of virus and vector during unfavourable periods, and the distances over which the aphid vector can disperse and disseminate virus. Now that the aetiology of the disease is understood and diagnostic tools have been developed, the time is opportune for new initiatives in understanding the ecology and epidemiology of rosette. Substantial progress can be made by developing a co-ordinated multi-disciplinary research programme and making full use of the latest techniques, approaches and experience gained elsewhere with other insect-borne viruses. This information would help to explain the sporadic disease epidemics that cause serious crop losses and sometimes total crop failure, and would also facilitate the development of disease forecasting methods and sustainable integrated disease management strategies.     

Diallel analysis in groundnut (Arachis hypogaea L.)

Summary An 8 × 8 full diallel experiment based on 4 bunch plus 4 spreading types of groundnut (Arachis hypogaea L.) was conducted over three environments. For both number of pods and pod yield, additive, nonadditive and reciprocal cross effects were detected and these were also influenced by changes in environments. For number of pods additive genetic variance was predominant whereas it was approximately equal to non-additive genetic variance for pod yield. Graphical analysis revealed the presence of strong non-allelic interaction for number of pods whereas for pod yield absence of dominance and/or presence of non-allelic interaction was evident.Part of Ph.D. Thesis of the first author     

Deep sequencing reveals a novel closterovirus associated with wild rose leaf rosette disease

A bizarre virus‐like symptom of a leaf rosette formed by dense small leaves on branches of wild roses (Rosa multiflora Thunb.), designated as ‘wild rose leaf rosette disease’ (WRLRD), was observed in China. To investigate the presumed causal virus, a wild rose sample affected by WRLRD was subjected to deep sequencing of small interfering RNAs (siRNAs) for a complete survey of the infecting viruses and viroids. The assembly of siRNAs led to the reconstruction of the complete genomes of three known viruses, namely Apple stem grooving virus (ASGV), Blackberry chlorotic ringspot virus (BCRV) and Prunus necrotic ringspot virus (PNRSV), and of a novel virus provisionally named ‘rose leaf rosette‐associated virus’ (RLRaV). Phylogenetic analysis clearly placed RLRaV alongside members of the genus Closterovirus, family Closteroviridae. Genome organization of RLRaV RNA (17 653 nucleotides) showed 13 open reading frames (ORFs), except ORF1 and the quintuple gene block, most of which showed no significant similarities with known viral proteins, but, instead, had detectable identities to fungal or bacterial proteins. Additional novel molecular features indicated that RLRaV seems to be the most complex virus among the known genus members. To our knowledge, this is the first report of WRLRD and its associated closterovirus, as well as two ilarviruses and one capilovirus, infecting wild roses. Our findings present novel information about the closterovirus and the aetiology of this rose disease which should facilitate its control. More importantly, the novel features of RLRaV help to clarify the molecular and evolutionary features of the closterovirus.     

Physiological responses of groundnut (Arachis hypogea L.) to drought stress and its amelioration: a critical review

Groundnut (Arachis hypogea L.), is an important legume cash crop for the tropical farmers and its seeds contain high amounts of edible oil (43–55%) and protein (25–28%). Even though it is a fairly drought-tolerant, production fluctuates considerably as a result of rainfall variability. To develop a water stress response function in groundnut, research works have been done to improve the performance under varying degrees of stress at various physiological stages of crop growth. This review summarizes recent information on drought resistance characteristics of groundnut with a view toward developing appropriate genetic enhancement strategies for water-limited environments. It is suggested that there are considerable gains to be made in increasing yield and stabilizing the yield in environments characterized by terminal drought stress and by shortening crop duration. Many traits conferring dehydration avoidance and dehydration tolerance are available, but integrated traits, expressing at a high level of organization are suggested to be more useful in crop improvement programs. Possible genetic improvement strategies are outlined, ranging from empirical selection for yield in drought environments to a physiological–genetic approach. It was also suggested that in view of recent advances in understanding drought resistance mechanisms, the later strategy is becoming more feasible. It is summarized that application of knowledge into practice in a systematic manner can lead to significant gains in yield and yield stability of the worlds groundnuts production. Research is needed to develop transferable technology to help farmers of arid and semi-arid regions. Increasing soil moisture storage by soil profile management and nutrient management for quick recovery from drought are some of the areas that need to be explored further.     

Managament of peanut bud necrosis disease in groundnut by botanical pesticides

In the management of PBND in groundnut neem seed extract (10%), we recorded significantly lower PBND incidence, AUDPC, apparent infection rate (20.41, 19.5% PBND; 956, 756 AUDPC; 0.0026, 0.002 apparent infection rate) followed by neem cake extract 10% spray (2747, 23.36% PBND; 1275, 1316 AUDPC; 0.0034, 0.0025 apparent rate of infection in 2000–2001 and 2001–2002, respectively). Weekly spray of monocrotophos reduced the PBND (26.77%) and recorded significantly higher pod yields (2547, 2465 kg/ha) and lower AUDPC (861, 1373), and apparent infection rate (0.0021, 0.0019 in 2000–2001 and 2001–2002, respectively) compared to other treatments.     

Effect of soil sodicity on the growth,yield and chemical composition of groundnut (Arachis hypogaea Linn.)

Summary A replicated field experiment was conducted to study the effect of exchangeable sodium percentage (ESP) on the yield, chemical composition, protein and oil content and uptake of nutrients by groundnut (Arachis hypogaea Linn.) variety M-13. ESP over 15 delayed germination and emergence of flowers. There was continuous decrease in dry matter yield at 30 and 60 days of growth, grain and straw yield after harvest and protein, oil and kernel percent with increase in soil ESP. A 50 per cent reduction in groundnut yield was observed at an ESP of 20. Increasing soil ESP, increased Na and decreased K, Ca and N contents, but had no effect on the Mg, P, S, Fe, Mn, Zn and Cu contents of the plant. Sodium content of the plant increased, while potassium and nitrogen decreased with age of the plant. The uptake of all the nutrients decreased with increase in soil ESP. The results showed that groundnut is a relatively sensitive crop to soil sodicity.     

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.     

Probable mechanical transmission of a virus-like agent from rose rosette disease-infected multiflora rose to Nicotiana species

As part of efforts to identify the causal agent of the rose rosette disease (RRD) of multiflora rose (Rosa multiflora Thunb.), root tip extracts from both symptomatic and nonsymptomatic roses were used to mechanically inoculate leaves of Nicotiana glutinosa. Pale green spots were observed along the margins of the major leaf veins only on leaves inoculated with extracts prepared from symptomatic rose plants. Light microscopy revealed abnormal development of the palisade and spongy mesophyll cells in the symptomatic tissue, although no virus‐like particles (VLPs) were observed by electron microscopy. However, VLPs were observed in cells from tissue adjacent to the leaf veins and bordered by the pale green spots. Inoculation of N. benthamiana with extracts from symptomatic N. glutinosa initially did not result in visible symptoms on N. benthamiana inoculated leaves. However, approximately 4 wk post inoculation, splitting of leaf tissue across and along major leaf veins in expanding leaves occurred. In later stages of leaf expansion some leaves split in regions not associated with veins. Light microscopy of thick sections revealed separation between palisade cells and groups of small dead cells in the mesophyll tissue of expanding systemically infected leaf blades. Electron microscopy revealed crystalline arrays in the cytoplasm of mesophyll cells. No abnormal cellular changes were observed in plants inoculated with extracts prepared from nonsymptomatic rose plants.     

In vitro plant regeneration from seed explants of wild groundnut species (Genus Arachis, Section Extranervosae)

In vitro regeneration of wild groundnut species from Section Extranervosae (Arachis villosulicarpa, A. macedoi, A. retusa, A. burchellii, A. pietrarellii, A. prostrata, A. aff. prostrata and a new species) was examined for the purpose of germplasm renewal and conservation. Seeds of different ages, stored at the seed bank of CENARGEN/EMBRAPA were either inoculated on culture medium or used as a source of embryo axis and cotyledon explants. Whole seeds failed to germinate on MS either without growth regulators (MS0) or supplemented with 10 M TDZ. Embryo axes cultured on MS0 produced only single plants. In the presence of 8.8 M BAP these explants showed multi-shoot formation. Cotyledons cultured on MS supplemented with 110 M BAP developed adventitious shoots through direct organogenesis. Plant regeneration was obtained from A. villosulicarpa, A. macedoi, A. retusa, A. burchellii and A. pietrarellii both from embryo axes and cotyledons. Explants from A. prostrata and A. aff. prostrata did not produce regenerants. Rooting of shoots was induced in the presence of 5.4 M NAA. Primary plants derived from these explants were further multiplied by culturing nodal segments on MS medium plus 2.7 M NAA.