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.     

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 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.     

陆地棉品种和骨干品系黄萎病抗性鉴定

选育和推广抗病品种是防治陆地棉黄萎病的主要措施,为了早日实现多类型、多区域大面积抗病品种的应用,本研究选取107份遗传背景差异较大的种质,利用河北省农林科学院棉花研究所小安舍试验站黄萎病病圃进行了3年黄萎病抗性重复鉴定。鉴定得到抗病品系8个,占7.5%;耐病品种(系)20个,占18.7%。本研究表明,当前被作为育种亲本的抗病品系还太少,需要深入开展抗病遗传机制,以及与其他经济性状协同改良的关系,为陆地棉抗病育种提供理论指导;达到抗病或接近抗病水平的大部分品种(系)来自于海陆野远缘后代,具有外源基因血统,证明了远缘杂交是陆地棉黄萎病抗性改良的有效手段。     

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.     

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).     

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.     

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.     

Morphological and molecular screening of French bean (Phaseolus vulgaris L.) germplasm using SCAR markers for Colletotrichum lindemuthianum (Sacc. and Magn.) Scrib. causing anthracnose resistance

This study was carried out to screen Phaseolus vulgaris L. germplasm accessions for anthracnose resistance genes to the fungus Colletotrichum lindemuthianum. (Sacc. and Magn.) Scrib. This fungus is made up of many pathogenic races which poses a challenge in developing resistant plant varieties; however, screening for and selection of resistant plant sources plays an important role in developing resistant plant lines. This screening work involved examining 69 accessions consisting of two resistant lines (D-line, L-line) and two susceptible varieties (Kanchana and Jwala). Fourteen SCAR primers specific to anthracnose disease resistance in the French bean were used. Of these 14 SCAR primers, 5 of them, SAS13, SF10, SC08, SZ04 and SBB14, produced amplification with good monomorphic bands.     

Evaluation of Water Yam (Dioscorea alata L.) Germplasm for Reaction to Yam Anthracnose and Virus Diseases and their Effect on Yield

Use of genetic resistance is the most practical and economic way to manage major diseases of yams. In a search for sources of resistance, 40 water yam (Dioscorea alata L.) accessions from Benin, Ghana, Nigeria and Puerto Rico were screened under natural disease infection conditions in Ibadan, Nigeria. The accessions were evaluated at 1, 3 and 6 months after planting (MAP) for severity of yam anthracnose and viral diseases. The effect of the pathogens on yield was also evaluated at harvest 9 MAP. There were significant differences (P < 0.001) between accessions for severities of anthracnose and viral diseases. Eight (20%) of them had lower anthracnose area under disease progress curves (AUDPC) values than the resistant check while 10 (25%) had AUDPC values below the trial mean. There were significant variations (P < 0.001) in yield components among the accessions. There was significant negative correlation of anthracnose severity with fresh tuber yield (r = −0.51) and with number of tubers per plot (r = −0.40). Similarly, significant negative correlations were observed of virus disease severity with fresh tuber yield (r = −0.78) and number of tubers per plot (r = −0.65). Linear regression models also showed that the fresh yield had significant negative relationships with anthracnose (R² = 0.26) and viral (R² = 0.62) diseases. The accessions identified as resistant constitute a valuable resource for breeding of resistant germplasm.     

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.     

Herbivore-induced resistance in different groundnut germplasm lines to Asian armyworm, Spodoptera litura (Fab.) (Lepidoptera: Noctuidae)

Three groundnut germplasm lines, ICGV86699 (resistant), NCAC 343 (resistant) and TMV 2 (susceptible), were examined for Spodoptera litura (Fab.) resistance. Biochemical parameters such as oxidative enzyme activities, peroxidase (POD) and polyphenol oxidase (PPO), other defensive components such as total phenols, hydrogen peroxide (H₂O₂), malondialdehyde (MDA) and protein contents were evaluated in these germplasm lines after 24, 48, 72 and 96 h following S. litura infestation to characterize the mechanism of resistance. Enzyme activities and total phenols, H₂O₂, MDA and protein contents were increased following infestation; however, significance varied at different time intervals and among germplasm lines depending upon the induced level of resistance. The three germplasm lines differed in resistance mechanisms to S. litura and the resistance may be partly due to higher enzyme activities, and other components studied. Among the three germplasms tested, ICGV86699 showed greater elevation in POD and PPO activities and in phenolic and H₂O₂ contents at different time intervals as compared to NCAC 343 and TMV 2.     

A field screening method to evaluate rice breeding lines for resistance to the hoja blanca virus

A method is presented for rearing large colonies of viruliferous Sogatodes oryzicola, vector of the rice hoja blanca virus (RHBV). These colonies were used for field screening up to 10 000 rice breeding lines per season for resistance to RHBV. Uniform infection of check varieties in the field indicated that the method was adequate. Field release of vectors when plants were 14 days old resulted in satisfactory disease incidence, after 21 days, to distinguish lines segregating for resistance from lines uniformly resistant or susceptible. Various sources of resistance identified earlier continued to be resistant under the screening conditions. Progeny of lines identified as non-segregating resistant continued as non-segregating resistant. Resistant plants from lines segregating for resistance produced progeny lines that were segregating and non-segregating. Ratios of resistant to susceptible plants in F₁ progeny of three-way crosses were consistent with earlier observations that RHBV resistance is a dominant character. The susceptibility of the commercial checks indicates that rice production in RHBV areas of tropical Latin America continues to be at risk from the virus. Virus-resistant commercial cultivars resulting from this method should be available in 2 years.     

Identification and mapping of AFLP markers linked to peanut (<Emphasis Type="Italic">Arachis hypogaea</Emphasis> L.) resistance to the aphid vector of groundnut rosette disease

Groundnut rosette disease is the most destructive viral disease of peanut in Africa and can cause serious yield losses under favourable conditions. The development of disease-resistant cultivars is the most effective control strategy. Resistance to the aphid vector, Aphis craccivora, was identified in the breeding line ICG 12991 and is controlled by a single recessive gene. Bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) analysis were employed to identify DNA markers linked to aphid resistance and for the development of a partial genetic linkage map. A F_2:3 population was developed from a cross using the aphid-resistant parent ICG 12991. Genotyping was carried out in the F₂ generation and phenotyping in the F₃ generation. Results were used to assign individual F₂ lines as homozygous-resistant, homozygous-susceptible or segregating. A total of 308 AFLP (20 EcoRI+3/MseI+3, 144 MluI+3/MseI+3 and 144 PstI+3/MseI+3) primer combinations were used to identify markers associated with aphid resistance in the F_2:3 population. Twenty putative markers were identified, of which 12 mapped to five linkage groups covering a map distance of 139.4 cM. A single recessive gene was mapped on linkage group 1, 3.9 cM from a marker originating from the susceptible parent, that explained 76.1% of the phenotypic variation for aphid resistance. This study represents the first report on the identification of molecular markers closely linked to aphid resistance to groundnut rosette disease and the construction of the first partial genetic linkage map for cultivated peanut.     

Identification of resistance source in wheat germplasm against spot blotch disease caused by Bipolaris sorokiniana

Four hundred and twenty-two spring wheat germplasm (Triticum aestivum L.) lines belonging to Indian, CIMMYT and Chinese wheat programme were evaluated for their tolerance against natural epiphytotic conditions of spot blotch caused by Bipolaris sorokiniana at the hot spot location, Pusa, Bihar, India. Of the 422 entries screened, none of the genotype showed immunity to the disease, whereas 52 were resistant, 180 moderately susceptible, 171 susceptible and 19 highly susceptible. Indian germplasm lines tended to be more susceptible than lines originated from CIMMYT and China. Chirya 3, Chirya 7 and Mayoor from CIMMYT showed high degree of resistance to the disease both under field and polyhouse conditions. On the basis of the disease severity under field conditions, 20 promising resistant genotypes and 10 highly susceptible lines were isolated for further testing under artificial epiphytotic conditions in polyhouse for genetic analysis and their potential for spot blotch resistance breeding.     

Haplotype diversity and population structure in cultivated and wild barley evaluated for Fusarium head blight responses

Fusarium head blight (FHB) is a threat to barley (Hordeum vulgare L.) production in many parts of the world. A number of barley accessions with partial resistance have been reported and used in mapping experiments to identify quantitative trait loci (QTL) associated with FHB resistance. Here, we present a set of barley germplasm that exhibits FHB resistance identified through screening a global collection of 23,255 wild (Hordeum vulgare ssp. spontaneum) and cultivated (Hordeum vulgare ssp. vulgare) accessions. Seventy-eight accessions were classified as resistant or moderately resistant. The collection of FHB resistant accessions consists of 5, 27, 46 of winter, wild and spring barley, respectively. The population structure and genetic relationships of the germplasm were investigated with 1,727 Diversity Array Technology (DArT) markers. Multiple clustering analyses suggest the presence of four subpopulations. Within cultivated barley, substructure is largely centered on spike morphology and growth habit. Analysis of molecular variance indicated highly significant genetic variance among clusters and within clusters, suggesting that the FHB resistant sources have broad genetic diversity. The haplotype diversity was characterized with DArT markers associated with the four FHB QTLs on chromosome 2H bin8, 10 and 13 and 6H bin7. In general, the wild barley accessions had distinct haplotypes from those of cultivated barley. The haplotype of the resistant source Chevron was the most prevalent in all four QTL regions, followed by those of the resistant sources Fredrickson and CIho4196. These resistant QTL haplotypes were rare in the susceptible cultivars and accessions grown in the upper Midwest USA. Some two- and six-rowed accessions were identified with high FHB resistance, but contained distinct haplotypes at FHB QTLs from known resistance sources. These germplasm warrant further genetic studies and possible incorporation into barley breeding programs.     

Identification of Rotylenchulus reniformis Resistant Glycine Lines

Identification of resistance to reniform nematode (Rotylenchulus reniformis) is the first step in developing resistant soybean (Glycine max) cultivars that will benefit growers in the mid-South region of the United States. This study was conducted to identify soybean (G. max and G. soja) lines with resistance to this pathogen. Sixty-one wild and domestic soybean lines were evaluated in replicated growth chamber tests. Six previously untested soybean lines with useful levels of resistance to reniform nematode were identified in both initial screening and subsequent confirmation tests: released germplasm lines DS4-SCN05 (PI 656647) and DS-880 (PI 659348); accession PI 567516 C; and breeding lines DS97-84-1, 02011-126-1-1-2-1 and 02011-126-1-1-5-1. Eleven previously untested moderately susceptible or susceptible lines were also identified: released germplasm lines D68-0099 (PI 573285) and LG01-5087-5; accessions PI 200538, PI 416937, PI 423941, PI 437697, PI 467312, PI 468916, PI 594692, and PI 603751 A; and cultivar Stafford (PI 508269). Results of previously tested lines evaluated in the current study agreed with published reports 69.6% of the time for resistant lines and 87.5% of the time for susceptible lines. Soybean breeders may benefit from incorporating the newly identified resistant lines into their breeding programs.     

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。     

Variation for Resistance to <i>Alternaria tenuissima</i> and Potential Structural Mechanism among Different Cultivars of <i>Chrysanthemum morifolium</i>

Black spot disease, caused by the necrotrophic fungus Alternaria tenuissima (Fr.) Wiltsh (A. tenuissima), is considered a highly destructive disease of Chrysanthemum (Chrysanthemum morifolium Ramat.). A set of 17 accessions of commercial chrysanthemum cultivars were evaluated for resistance to A. tenuissima by seedling artificial inoculation. It was found that the reaction of the accessions to artificial inoculation ranged from resistant to highly susceptible. Five varieties of chrysanthemum (‘Zhongshan Taogui’, ‘Jinba’, ‘Zhongshan Jinguan’, ‘Jinling Wanhuang’ and ‘Jinling Yangguang’) were resistant; two varieties of chrysanthemum (‘Zhongshan Xinggui’ and ‘Zhongshan Jinkui’) were moderately resistant; and others were susceptible to various degrees, four varieties of chrysanthemum (‘Zhongshan Zihe’, ‘Zhongshan Jiuhong’, ‘Zaoyihong’ and ‘Jinling Jiaohuang’) were highly susceptible, especially. Some leaf morphological features of two resistant and two highly susceptible cultivars were further researched. Trichome density, length, height, gland size and stomata density were found to be associated with plant passive resistance. Resistant varieties that were identified in present study will be promising germplasm for exploitation of breeding programmes aimed at developing A. tenuissima-resistant cultivars and increasing genetic diversity.     

Divergence and allelomorphic relationship of a soybean virus resistance gene based on tightly linked DNA microsatellite and RFLP markers

The use of genetically diverse resistance sources is important in breeding for durable disease resistance. Detection and evaluation of resistance genes by conventional inheritance experiments, however, often require laborious screening and genetic testing. In the present study, a marker-assisted screening for resistance sources was initiated in soybean [Glycine max (L.) Merr] using one DNA microsatellite and two RFLP markers tightly linked to a soybean mosaic virus (SMV) resistance gene (Rsv1). The three marker loci were used to screen 67 diverse soybean cultivars, breeding lines, and plant introductions. Five variants were found at the microsatellite locus (HSP176L), and the two RFLP loci (pA186 and pK644a) near Rsv1 show a remarkably higher level of restriction polymorphism than Rsv1-independent RFLP loci. Several specific variants at the three marker loci were found to be correlated with virus resistance, among which HSP176L-2 can be detected by PCR, thus may be useful for germplasm screening. The grouping of the 67 accessions according to their multilocus marker variants agrees with the available pedigree information. When all, or most, of the cultivars within a given group with the same Rsv1-linked marker variant are resistant, their SMV resistance is most likely conferred by Rsv1. These putatively Rsv1-carrying groups contain a total of 38 SMV-resistant lines including six differential cultivars that are known to carry Rsv1. The remaining seven resistant accessions (Columbia, Holladay, Peking, Virginia, FFR-471, PI 507403, and PI 556949) do not carry resistance marker variants, and at least some of them could be sources of resistance genes independent of Rsv1.