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.     

国外花生种质资源引种鉴定

从法国国际农艺发展研究中心(CIRAD)引入33份花生种质资源,对它们的主要农艺性状、品质、抗性等进行了较为全面的观察鉴定,筛选出高产种质(单株生产力大于60g)17份,蛋白质含量高种质(大于30%)5份,粗脂肪含量高种质(大于50%)15份。     

Resistance to groundnut rosette disease in wild Arachis species

One hundred and sixteen accessions representing 28 species in the genus Arachis were evaluated for resistance to groundnut rosette disease using an infector row technique during the 1996/97, 1997/98, 1998/99 and 1999/2000 growing seasons at Chitedze, Malawi. Of these, a total of 25 accessions belonging to Arachis diogoi (1 accession), A. hoehnei (2), A. kretschmeri (2), A. cardenasii (2), A. villosa (1), A. pintoi (5), A. kuhlmannii (2), A. appressipila (3), A. stenosperma (5), A. decora (1), and A. triseminata (1) showed resistance to the groundnut rosette disease. No visible disease symptoms were observed in several accessions belonging to A. appressipila, A. cardenasii, A. hoehnei, A. kretschmeri, A. villosa, A. pintoi, A. kuhlmannii, and A. stenosperma. Some accessions in A. appressipila, A. diogoi, A. stenosperma, A. decora, A. triseminata, A. kretschmeri, A. kuhlmannii, and A. pintoi were resistant to all three components of rosette, Groundnut rosette ass is tor virus (GRAV), Groundnut rosette virus (GRV) and its satellite RNA (sat. RNA). Two accessions in A. stenosperma and one accession in A. kuhlmannii showed the presence of all three components of the rosette disease. Several wild Arachis accessions were resistant to GRAV. All the accessions of A. batizocoi (4), A. benensis (2), A. duranensis (46), A. dardani (1), A. ipaensis (1), A. magna (1), A. monticola (3), A. oteroi (1), A. pusilla (4), and A. valida (2) were susceptible to rosette disease. In all these accessions, infected plants were chlorotic and severely stunted. The value of exploitation of the resistance in wild Arachis species in rosette resistance breeding programmes is discussed.     

The pattern of genetic diversity of Guinea-race Sorghum bicolor (L.) Moench landraces as revealed with SSR markers

The Guinea-race of sorghum [Sorghum bicolor (L.) Moench] is a predominantly inbreeding, diploid cereal crop. It originated from West Africa and appears to have spread throughout Africa and South Asia, where it is now the dominant sorghum race, via ancient trade routes. To elucidate the genetic diversity and differentiation among Guinea-race sorghum landraces, we selected 100 accessions from the ICRISAT sorghum Guinea-race Core Collection and genotyped these using 21 simple sequence repeat (SSR) markers. The 21 SSR markers revealed a total of 123 alleles with an average Dice similarity coefficient of 0.37 across 4,950 pairs of accessions, with nearly 50% of the alleles being rare among the accessions analysed. Stratification of the accessions into 11 countries and five eco-regional groups confirmed earlier reports on the spread of Guinea-race sorghum across Africa and South Asia: most of the variation was found among the accessions from semi-arid and Sahelian Africa and the least among accessions from South Asia. In addition, accessions from South Asia most closely resembled those from southern and eastern Africa, supporting earlier suggestions that sorghum germplasm might have reached South Asia via ancient trade routes along the Arabian Sea coasts of eastern Africa, Arabia and South Asia. Stratification of the accessions according to their Snowden classification indicated clear genetic variation between margeritiferum, conspicuum and Roxburghii accessions, whereas the gambicum and guineënse accessions were genetically similar. The implications of these findings for sorghum Guinea-race plant breeding activities are discussed.     

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.     

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.     

Genetic diversity in Bambara groundnut (Vigna subterranea L.) germplasm revealed by RAPD markers.

Random amplified polymorphic DNA (RAPD) markers were used to assess genetic diversity in Bambara groundnut (Vigna subterranea L.) germplasm using 25 African accessions from the collection in the International Institute for Tropical Agriculture, Ibadan, Nigeria. Fifty random decamer primers were screened to assess their ability to detect polymorphism in bambara; 17 of them were selected for this study. Considerable genetic diversity was found among the V. subterranea accessions studied. The relationships among the 25 accessions were studied by cluster analysis. The dendrograms showed two main groups of accessions mainly along the lines of their geographic origin. It is concluded that RAPD can be used for germplasm classification in bambara groundnut and hence for improving this crop.     

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.     

Use of Simple Sequence Repeat Markers for Estimation of Genetic Diversity in Coconut (Cocos nucifera L) Germplasm Accessions

Simple Sequence Repeat (SSR) markers were used to determine the genetic relationship among 33 coconut germplasm accessions collected from different geographical regions. Wide range of Jaccard’s similarity coefficient (0.00–0.86) reflects the informativeness of the SSR markers. Dwarf and intermediate accessions showed highest similarity among them. The tall accessions belonging to South East Asia, South Asia and South Pacific were clustered based on their geographical regions, but dwarf and intermediate accessions were clustered separately. Clustering of accessions belonging to Atlantic and America revealed the spread of coconut from Far East to South Pacific. Principal coordinate plot and UPGMA analyses formed similar groups of the accessions.     

Genetic diversity and structure of the zombi pea (<Emphasis Type="Italic">Vigna vexillata</Emphasis> (L.) A. Rich) gene pool based on SSR marker analysis

Zombi pea (Vigna vexillata (L.) A. Rich) is an underutilized legume species and a useful gene source for resistance to biotic and abiotic stresses, although there is little understanding on its genetic diversity and structure. In this study, 422 (408 wild and 14 cultivated) accessions of zombi pea from diverse origins (201 from Africa, 126 from America, 85 from Australia, 5 from Asia and 5 from unknown origin) were analyzed with 20 simple sequence repeat (SSR) markers to determine its genetic diversity and genetic structure. The SSR markers detected 273 alleles in total with a mean of 13.6 alleles per locus. Polymorphism information content values of the markers varied from 0.58 to 0.90 with an average of 0.76. Overall gene diversity was 0.715. Gene diversity and average allelic richness was highest in Africa (0.749 and 8.08, respectively) and lowest in America (0.435 and 4.10, respectively). Nei’s genetic distance analysis revealed that the highest distance was between wild Australia and cultivated Africa (0.559), followed by wild West Africa and wild Australia (0.415). STRUCTURE, neighbor-joining (NJ), and principal coordinate analyses consistently showed that these zombi pea accessions were clustered into three major groups, viz. America, Africa and Asia, and Australia. NJ tree also suggested that American and Australian accessions are originated from East African zombi peas, and that the cultivated accessions from Africa and Asia were genetically distinct, while those from America were clustered with some cultivated accessions from Africa. These results suggest that Africa is the center of origin and diversity of zombi pea, and that domestication of this pea took place more than once in different regions.     

Mechanisms and diversity of resistance to insect pests in wild relatives of groundnut

The levels of resistance to insect pests in cultivated groundnut (Arachis hypogaea) germplasm are quite low, and therefore, we screened 30 accessions of Arachis spp. and 12 derived lines for resistance to insect pests under field and greenhouse conditions. Accessions belonging to Arachis cardenasii, Arachis duranensis, Arachis kempff-mercadoi, Arachis monticola, Arachis stenosperma, Arachis paraguariensis, Arachis pusilla, and Arachis triseminata showed multiple resistance to the leaf miner Aproaerema modicella, Helicoverpa armigera, Empoasca kerri, and to rust, Puccnia arachidis Speg., and late leaf spot, Cercosporidium personatum (Berk. et Curt.). Arachis cardenasii (ICG 8216), Arachis ipaensis (ICG 8206), A. paraguariensis (ICG 8130), and Arachis appressipila (ICG 8946) showed resistance to leaf feeding and antibiosis to Spodoptera litura under no-choice conditions. Six lines, derived from wild relatives, showed resistance to H. armigera and S. litura, and/or leaf miner. Plant morphological characteristics such as main stem thickness, hypanthium length, leaflet shape and length, leaf hairiness, standard petal length and petal markings, basal leaflet width, main stem thickness and hairiness, stipule adnation length and width, and peg length showed significant correlation and/or regression coefficients with damage by H. armigera, S. litura, and leafhoppers, and these traits can possibly be used as markers to select for resistance to these insect pests. Principal component analysis placed the Arachis spp. accessions into five groups, and these differences can be exploited to diversify resistance to the target insect pests in groundnut.     

Simple sequence repeat diversity in diploid and tetraploid Coffea species.

Thirty-four fluorescently labeled microsatellite markers were used to assess genetic diversity in a set of 30 Coffea accessions from the CENICAFE germplasm bank in Colombia. The plant material included one sample per accession of seven East African accessions representing five diploid species and 23 wild and cultivated tetraploid accessions of Coffea arabica from Africa, Indonesia, and South America. More allelic diversity was detected among the five diploid species than among the 23 tetraploid genotypes. The diploid species averaged 3.6 alleles/locus and had an average polymorphism information content (PIC) value of 0.6, whereas the wild tetraploids averaged 2.5 alleles/locus and had an average PIC value of 0.3 and the cultivated tetraploids (C. arabica cultivars) averaged 1.9 alleles/locus and had an average PIC value of 0.22. Fifty-five percent of the alleles found in the wild tetraploids were not shared with cultivated C. arabica genotypes, supporting the idea that the wild tetraploid ancestors from Ethiopia could be used productively as a source of novel genetic variation to expand the gene pool of elite C. arabica germplasm.     

Genetic Relationships among Tall Coconut Palm (Cocos nucifera L.) Accessions of the International Coconut Genebank for Latin America and the Caribbean (ICG-LAC), Evaluated Using Microsatellite Markers (SSRs)

The diversity and genetic relationships among two accessions of tall coconut palms collected in Brazil and seven accessions introduced from different geographic regions of the world were analyzed using 25 microsatellite primers, 19 of which were polymorphic and detected between 4 and 10 alleles per locus, with an average of 6.57. The observed and expected heterozygosity ranged from 0.25 and 0.40 in the Rennell Islands Tall (RIT) accession to 0.54 and 0.62 in the Polynesian Tall (PYT) accession. The analysis of genetic structure resulted in the formation of five distinct groups. The first group was formed by the accessions Brazilian Tall—Praia do Forte (BRTPF), Brazilian Tall—Merepe (BRTMe) and West African Tall (WAT); the second group consisted of Malaysian Tall (MLT); the third group of RIT; the fourth group of Vanuatu Tall (VTT); and the fifth group of Rotuman Tall (RTMT), Tonga Tall (TONT) and PYT. The dendrogram based on the nearest-neighbor method detected the formation of two main groups and five subgroups, indicating that the genetic relationships of the accessions are based on their geographic regions of origin. The analyses revealed genetic relationships between the accessions collected in Brazil and the accession from Africa, and among palms from South East Asia and the South Pacific, confirming the common origin of these accessions. The information obtained in this study can guide decisions on germplasm conservation activities and the efficient selection of genetically divergent parents for use in coconut breeding programs in Brazil, which are attempting to select for disease resistance, mainly to lethal yellowing, among other characteristics.     

Host range and some properties of groundnut rosette virus

Two isolates of groundnut rosette virus from East Africa (GRVE₁ and GRVE₂) and from West Africa (GRVW₁ and GRVW₂) were transmitted by Aphis craccivora obtained from West Africa. A third isolate from West Africa (GRVW₃) was not transmitted by A. craccivora from three widely separated sources. GRVW₁, GRVW₂ and GRVW₃ caused leaf-symptoms in groundnut of a mosaic pattern in light and dark green. GRVE₁ and GRVE₂ caused chlorosis or chlorosis and leaf distortion as well as mosaic symptoms. Groundnut plants with GRVW₁ could not be infected by means of aphids with GRVE₁, and GRVE₁ gave similar protection against GRVW₁, which suggests that they are strains of the same virus. All isolates were transmissible manually from groundnut to groundnut (Arachis hypogea), Trifolium incarnatum and T. repens, and caused systemic infection. Inoculated Nicotiana clevelandii and N. rustica developed symptoms but virus could not be recovered from them. Chenopodium amaranticolor, C. hybridum and C. quinoa showed local lesions on inoculated leaves. Virus could be acquired by aphids from groundnut or Trifolium repens infected by means of aphids, but not from those infected by manual inoculation. Virus could not be recovered from T. incarnatum manually or by aphids, but was transmitted by cleft-grafting from clover to groundnut. Saps extracted in borax buffer plus zinc sulphate at pH 9 from plants infected with GRVW₁ and GRVE₁ remained infective at 18° C. for 1 week, and at — 20° C. for up to 4 weeks. Virus could be recovered from frozen leaves. Buffered saps lost infectivity when heated above 50° C. for 10 min.; most were still infective when diluted 1/10 and some at 1/100. Electron micrographs of partially purified preparations contained spherical particles 25–28 mμ in diameter. There were usually only about five per microscope field and they resembled those of some other viruses.     

Soybean germplasm pools in Asia revealed by nuclear SSRs

Soybean was domesticated in East Asia, where various kinds of landraces have been established as a result of adaptation to different environments and the diversification of food cultures. Asia is thus an important germplasm pool of soybean. In order to evaluate the genetic structure of the Asian soybean population, we analyzed allelic profiles at 20 simple-sequence repeat (SSR) loci of 131 accessions introduced from 14 Asian countries. The SSR loci produced an average of 11.9 alleles and a mean gene diversity of 0.782 in the accessions tested. Quantification theory III analysis and cluster analysis with the UPGMA method clearly separated the Japanese from the Chinese accessions, suggesting that the Japanese and Chinese populations formed different germplasm pools. The Korean accessions were involved in both germplasm pools, whereas most of the accessions from southeast and south/central Asia were derived from the Chinese pool. Relatively high genetic diversity and the absence of region-specific clusters in the southeast and south/central Asian populations suggest that soybean in these areas has been introduced repeatedly and independently from the diverse Chinese germplasm pool. The present study indicates that the two germplasm pools can be used as exotic genetic resources to enlarge the genetic bases of the respective Asian soybean populations.     

Studies on the transmission of groundnut rosette virus by Aphis craccivora Koch

Four strains of groundnut rosette virus were transmitted by a race of Aphis craccivora (Koch) from groundnut in Nigeria. Two of these strains, both from East Africa, were transmitted only by A. craccivora from Kenya. A fifth isolate, from Nigeria, was not transmissible by either race. The two races of aphids have been shown elsewhere to be distinct biotypes. Most A. craccivora needed longer than 24 h feeding on infected groundnuts to acquire virus, and many needed 2–3 days of feeding on healthy plants to cause infection, even after several days on infected plants. The delays partly reflect the slow uptake of virus and possibly a period needed for virus multiplication in aphid tissue but some is lost through resistance of the test plants to infection. In consecutive feeding experiments Natal Common variety could be infected soon after aphids had left the source of virus, but a more resistant Nigerian variety sometimes needed several more days. The frequency of inoculation by aphids, or the concentration of virus in the inocula or both, increased with time, but the times at which aphids were able to infect plants was also dependent on variety.     

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.     

Viruses associated with chlorotic rosette and green rosette diseases of groundnut in Nigeria*

Groundnut (Arachis hypogaea) plants from Nigeria with chlorotic rosette disease contained a manually transmissible virus, considered to be a strain of groundnut rosette virus (GRV(C)). GRV(C) infected nine out of 32 species in three out of nine families. It caused local lesions without systemic infection in Chenopodium amaranticolor, C. murale and C. quinoa, and systemic symptoms in Glycine max, Nicotiana benthamiana, N. clevelandii and Phaseolus vulgaris as well as in groundnut. Some ‘rosette-resistant’ groundnut lines were also infected. GRV(C) was transmitted by Aphis craccivora, but only from groundnut plants that were also infected with an aphid-transmissible second virus, which was not manually transmissible and was considered to be groundnut rosette assistor virus (GRAV). Plants infected with GRAV contained isometric particles c. 25 nm in diameter which were detectable by immunosorbent electron microscopy on grids coated with antisera to several luteoviruses, especially with antisera to bean leaf roll, potato leafroll and beet western yellows viruses. No virus-like particles were observed in extracts from plants infected with GRV(C) alone. A single groundnut plant obtained from Nigeria with symptoms of green rosette contained luteovirus particles, presumed to be of GRAV, and yielded a manually transmissible virus that induced symptoms similar to those of GRV(C) in C. amaranticolor but gave only mild or symptomless infection of N. benthamiana and N. clevelandii. It was considered to be a strain of GRV and designated GRV(G).     

Genetic diversity of the Bambara groundnut (Vigna subterranea (L.) Verdc.) as assessed by SSR markers

Bambara groundnut ( Vigna subterranea (L.) Verdc.) is an important African legume crop. In this study, a collection consisting of 240 accessions was analyzed using 22 simple sequence repeat (SSR) markers. In total, 166 alleles were detected, with a mean of 7.59 alleles per locus. Allelic and gene diversities were higher in the west African and Cameroon/Nigeria regions with 6.68 and 6.18 alleles per locus, and 0.601 and 0.571, respectively. The genetic distance showed high similarity between west African and Cameroon/Nigeria accessions. Principal coordinate analyses and neighbor-joining analysis consistently revealed that the majority of west African accessions were grouped with Cameroon/Nigeria accessions, but they were differentiated from east African, central African, and southeast Asian accessions. Population structure analysis showed that two subpopulations existed, and most of the east African accessions were restricted to one subpopulation with some Cameroon/Nigeria accessions, whereas most of the west African accessions were associated with most of the Cameroon/Nigeria accessions in the other subpopulation. Comparison with SSR analysis of other Vigna cultigens, i.e., cultivated azuki bean ( Vigna angularis ) and mungbean ( Vigna radiata ), reveals that the mean gene diversity of Bambara groundnut was lower than azuki bean but higher than mungbean.     

Genetic diversity analysis of North Africa’s barley using SSR markers

It was demonstrated that some North Africa barley accessions have diverse tolerance sources for abiotic stresses and a good nutritional quality, but the studies done were incomplete since they were realized separately in each country apart.To implement a more complete analysis, 31 barley accessions originated from North Africa (Algeria, Tunisia and Egypt) were analyzed using 11 SSR markers selected from the seven barley linkage groups for studying the genetic diversity among these chosen barley accessions.Over the 11 SSR markers, a total of 478 reproducible bands were scored with an average of 2.13 alleles/primer and the average polymorphism information content of 0.5.Genetic distance analysis of the 31 accessions showed a large genetic diversity and high number of different groups. The most accessions are clustered according to their eco-geographical origin, according to their pedigree and agronomic characters or according to the caryopsis character (hulled or naked caryopsis). This high number of obtained groups is an invaluable aid in crop improvement strategies and confirms the opinion suggesting that North Africa could be a secondary center of origin of barley. The various growing conditions and the multiple uses of barley in each country may be the cause of the large variability of the barley germplasm in each region.