Screening for resistance in wild Lycopersicon species to Fusarium oxysporum f.sp. lycopersici race 1 and race 2

Wild Lycopersicon accessions were screened for resistance to the Fusarium wilt disease caused by Fusarium oxysporum f.sp. lycopersici (Fol) race 1 and race 2. In total, four isolates of each race were used. Among 17 accessions of six Lycopersicon species tested, a wide genetic variation for wilt resistance was observed. Most accessions were highly susceptible, some showed intermediate resistance, but one accession of L. cheesmanii (G1.1615 = PI 266375) and two accessions of L. chilense (G1.1556 and G1.1558) were highly resistant to Fol races 1 and 2. The resistance in the latter three accessions equalled or was higher than the resistance determined by the known I-genes, that have been widely used in breeding programmes. These newly found resistant accessions provide breeders with more opportunities for Fusarium disease resistance and may contribute to our understanding of Fusarium disease resistance gene organisation and evolution.     

Inheritance of resistance to Fusarium wilt in Indian lentil germplasm (Lens culinaris Medik.)

Summary Three lentil genotypes resistant to Fusarium oxysporum f.sp. lentis viz. Pant L 234, JL 446 and LP 286 were crossed with two susceptible ones. The hybrid plants were all resistant in the eight crosses evaluated. Segregation pattern for wilt reaction in F₂, BC(P₁), BC(P₂) and F₃ generations in field and glasshouse conditions indicated that resistance to Fusarium wilt is under the control of two dominant duplicate genes in Pant L 234 and two independent dominant genes with complementary effects in JL 446 and LP 286. A third dominant gene complementary to the dominant genes in JL 446 and LP 286 is present in two susceptible lines. Allelic tests suggest the presence of five independently segregating genes for resistance. Duplicate dominant genes in Pant L 234 are non-allelic to two dominant genes with complementary effects in LP 286 and JL 446 and the third gene complementary to the two genes in JL 446 and LP 286 in susceptible lines JL 641 and L 9–12. Gene symbols among parental genotypes have been designated.     

Cytogenetics of the 'glandless-seed and glanded-plant' trait from Gossypium sturtianum Willis introgressed into upland cotton (Gossypium hirsutum L.)

In order to introgress the ‘glandless-seed and glanded-plant’ trait from Gossypium sturtianum Willis (2n= 2x= 26, C₁ genome) into the cultivated upland cotton Gossypium hirsutum L. (2n= Ax= 52 (AD), genome), two trispecific hybrids have been created using either Gossypium thurberi Torado (2n= 2x= 26, D₁ genome) or Gossypium raimondii Ulbrich (2n= 2x= 26, D₅ genome) as bridge species. The cross of both trispecific hybrids by G. hirsutum produced the first backcross progenies (BCl). Cytogenetic analysis showed that the trispecific hybrids had 52 chromosomes, their chromosome configurations at metaphase I (Ml) being 15.071 + 15.3411 + 0.93III + 0.69IV + 0.26VI in G. thurberi×G. sturtianum×G. hirsutum (TSH) and 14.421 + 17.0311 + 0.82III + 0.15IV + 0.07VI in G. hirsutum × G. raimondii ×. G. sturtianum (HRS), respectively. Among six BCl plants analysed, the only plant expressing the ‘glandless-seed and glanded-plant’ trait had 52 chromosomes and a meiotic configuration of 5.261 + 20.61II + 0.69III + 0.77IV at MI. Pollen fertility was 2.90% in TSH, 8.97% in HRS, and ranged from 0% to 40.28% in the BCl progenies. The introgressed BCl plant is perennial in growth habit. It can be used in breeding programmes aiming at the introgression of the ‘glandless-seed and glanded-plant’ trait into a cultivar of upland cotton.     

Targeted transfer of trait for Verticillium wilt resistance from Gossypium barbadense into G. hirsutum using SSR markers

Verticillium wilt (VW) is a soil‐borne disease of cotton that is destructive worldwide. Transferring desired traits from Gossypium barbadense is challenging through traditional interspecific introgression. We previously demonstrated that a molecular marker, BNL3255‐208, is associated with VW resistance in G. barbadense. This breakthrough opens the way for marker‐assisted selection (MAS) breeding. Here, the highly resistant G. barbadense cv. ‘Pima90‐53’ and the severe diseased Gossypium hirsutum cv. ‘CCRI8’ were used as donor parent and recipient parent, respectively. Our goal was to transfer the disease resistance from donor to recipient via MAS. Among 71 MAS obtained lines, as many as 19 lines had enhanced resistance. Among those lines, 11 lines showed high resistance and four lines displayed resistance to VW. Moreover, seven lines displayed improved fibre quality. After combining the markedly improved resistance and fibre properties, we identified two elite innovated introgression lines – ZY2 and ZY31 – that did not seem to differ in other agronomic traits from the recipient parent. This study first successfully transferred of G. barbadense resistance into G. hirsutum by MAS.     

Screening wild Lycopersicon species for resistance to powdery mildew (Oidium lycoperiscum)

Since the late 1980s powdery mildew, designated Oidium lycopersicum, frequently invaded the tomato crop in Western Europe. All commercial cultivars are susceptible. To screen for resistance in wild species a reliable and efficient disease test was developed. Young plants with two to three true leaves are inoculated at high relative humidity by spraying with a freshly prepared suspension of 2×10⁴ conidia, ml^–1. Symptoms are periodically evaluated according to a scale based on the percentage of leaf area with mycelium.One hundred and twenty seven accessions, representing eight wild Lycopersicon species, were screened for resistance to O. lycopersicum. A large variation in resistance was found between species. L. hirsutum was the most resistant species; L. pennellii was moderately resistant; species of the subgeneric group of L. esculentum and of the peruvianum-complex were all susceptible. L. parviflorum was classified separately due to a large variation between accessions. Except for this species, a low variation was found between accessions within species. High levels of resistance were observed in four accessions of L. hirsutum, in one of L. parviflorum and in one of L. peruvianum. This resistance is characterized by a very low disease incidence and a strongly restricted mycelium growth and lack of sporulation.     

Genetic characterization of Gossypium arboreum accession PI 529740 for reniform nematode resistance

Breeding for reniform nematode (Rotylenchulus reniformis) resistance is hindered by the lack of resistance in upland cotton (Gossypium hirsutum) cultivars. Resistance has been frequently identified in accessions from the Gossypium arboreum germplasm collection with accession PI 529740 rated as highly resistant. Accession PI 529740 was crossed with the susceptible G. arboreum accession PI 529729 to develop an F₂ population for genetic characterization. The population showed quantitative variation suggesting multiple genes conferred the resistant phenotype. Thirteen of the 216 F₂ plants showed resistance similar to the resistant parent and these data supported a two recessive gene model. Sixty plants were classified as resistant or moderately resistant, indicating a single recessive gene conferred the moderately resistant phenotype. The classification of 24 F_2:3 families for nematode resistance generally supported the classification of the corresponding F₂ plants; however, most families were highly variable for infection with no families rated as resistant. This information will aid in the introgression of resistance into upland cotton as larger populations will be required to successfully recover resistance conferred by multiple recessive genes.     

Screening of melon (Cucumis melo L.) germplasm for multiple disease resistance

Summary Melon germplasm was screened for cucumber green mottle mosaic virus (CGMMV), powdery mildew (Sphaerotheca fuliginea), downy mildew (Pseudoperonospora cubensis) and Fusarium wilt (Fusarium oxysporum f. sp. melonis) resistance under artificial conditions except downy mildew for which screening was done under natural epiphytotic conditions. High level resistance to all the four diseases was not recorded in any of the collections tested. Nevertheless, ertheless, resistance to three diseases was located in three germplasm. Wild Cucumis species C. figarei exhibited absolute resistance to CGMMV and Fusarium wilt and high level resistance to downy mildew. Phoot or snapmelon (Cucumis melo var. momordica) — a non-dessert from of Indian origin—was highly resistant to downy mildew and resistant to CGMMV and medium resistant to Fusarium wilt. Iroquois was resistant to powdery mildew and medium resistant to downy mildew and CGMMV.     

Morphological and molecular characterization of melon accessions resistant to Fusarium wilts

Fusarium wilt incited by Fusarium oxysporum f. sp. melonis (F.o.m) is one of the most widespread and devastating melon diseases. While resistance to physiological races 0, 1, and 2 is relatively frequent in different botanical varieties, sources of resistance to race 1,2 are restricted to a few Far-Eastern accessions. In this work, the results of a screening for resistance to F.o.m. race 1,2 among 32 accessions are presented. Three Japanese accessions (‘Kogane Nashi Makuwa’, ‘C-211’, and ‘C-40’) showed the highest resistance levels, but useful levels of resistance were also detected in one Russian ‘C-160’ and two Spanish (‘C-300’ and ‘Mollerusa-7’) accessions. These resistant materials, together with other accessions previously described as resistant to F.o.m. races 0, 1, and/or 2 have been morphologically and molecularly characterized. Based on cluster analysis, these accessions have been grouped according to the botanical subspecies they belong to. Assessment of genetic diversity indicated that the resistant accessions to races 0, 1 and 2, are scattered along the established clusters. On the other hand, high levels of resistance to the race 1,2 could be found only among accessions belonging to Cucumis melo subsp. agrestis, nevertheless, a certain degree of resistance to this race could also be found within some accessions belonging to subsp. melo. As far as we know, this is the first report of resistance to F.o.m race 1,2 found out from the Far-Eastern melon material. Based on fruits characteristics, it appears that several inodurus and cantalupensis accessions could be exploited in breeding programs as resistance sources to F.o.m races 0, 1 and/or 2 for the improvement of these melon types. The accessions with the highest levels of resistance to the race 1,2 appeared to be very distant both molecularly and morphologically from the commercial types. Nevertheless ‘C-160’, ‘C-300’, and ‘Mollerusa-7’ classified as var. inodorus are morphologically very similar to the Spanish commercial types and might be used as resistant sources in breeding these melon types.     

海岛棉枯萎病抗性与类黄酮代谢相关基因表达量的相关

枯萎病是危害海岛棉生产的重要因素之一, 研究枯萎病抗性分子机制将为培育抗病海岛棉品种、解决枯萎病对海岛棉的危害问题提供坚实的基础。本研究在前期转录组测序的基础上, 对海岛棉枯萎病抗性差异表达基因进行分析(DEG, Differentially Expressed Gene); 以7个抗病性表现不同的海岛棉品种为材料, 利用qRT-PCR的方法研究抗病差异表达基因在不同接菌时间点的表达量差异, 并分析基因表达量与病情指数的相关性。结果表明, DEG分析得出类黄酮代谢通路相关基因与海岛棉枯萎病抗性有关。qRT-PCR分析显示抗病材料中类黄酮代谢通路关键基因的表达量显著高于感病材料。在接菌后多个时间点, 类黄酮代谢通路中的关键基因TT7、CHI和DFR在抗病材料中的表达量显著或极显著高于感病材料, 其中CHI和DFR基因的表达量与病情指数呈显著负相关。综上所述, 类黄酮代谢通路相关基因对海岛棉枯萎病抗性均有影响, 且CHI、TT7和DFR基因是关键基因。     

Evaluation of resistance to verticillium wilt in diploid,wild potato interspecific hybrids

Summary Verticillium wilt (V. albo-atrum Reinke & Berthold or V. dahliae Kleb) threatens potato (Solanum tuberosum L.) production in most growing areas of the world. Genetic resistance offers the most cost-effective and environmentally-sound control measure. However, there is a dearth of genetic and breeding information on resistance to verticillium wilt in potato, because of obscure parentage of some standard cultivars and the complex segregation at the tetraploid level. The wide range of genetic variability in wild relatives of potatoes can be potentially useful as a source of disease resistance, such as verticillium wilt resistance. Six diploid, wild, interspecific Solanum hybrids involving resistant x resistant and susceptible x resistant crosses, were assayed for verticillium wilt resistance under greenhouse conditions to evaluate their potential as sources of verticillium wilt resistance. The cross between S. gourlayi Oka. and S. chacoense Bitt. and its reciprocal had the most resistant progenies based on mean colony counts. No simple mode of inheritance can be proposed based on the observed segregation ratios. However, the continuous distributions observed on verticillium wilt disease response among hybrid families indicate that inheritance of resistance may be polygenic and complex. In addition, skewness of colony count distributions toward the resistance parents were characteristic of all resistant x susceptible crosses suggesting that resistance may be dominant. By contrast, the susceptible x susceptible cross showed a more normal distribution. Overall, the cross between S. gourlayi and S. chacoense showed the most potential as a source of verticillium wilt resistance.     

Development of AFLP and derived CAPS markers for root-knot nematode resistance in cotton

Resistance to root-knot nematode (Meloidogyne incognita) is determined by a single major gene rkn1 in Gossypium hirsutum Acala NemX cotton. Bulked segregant analysis (BSA) combined with amplified fragment length polymorphism (AFLP) was used to identify molecular markers linked to rkn1. DNA pools from homozygous susceptible (S) and resistant (R) bulks of an F_2:3 originating from the intraspecific cross NemX × SJ-2 were screened with 128 EcoR1/Mse1 primer combinations. Putative AFLP markers were then screened with 60 F_2:7 RIL plants and four AFLP markers were found linked to rkn1. The linkage of AFLP markers to rkn1 was also confirmed in a F₂ population. The closest AFLP marker was converted to a cleaved amplified polymorphic sequence (CAPS) marker (designated GHACC1) by aligning the sequences from both susceptible and resistant parents. GHACC1 linkage to rkn1 was confirmed in the F₂ (1R:3S), F_2:7 RIL (1R:1S) and the backcross population SJ-2 × F₁ (NemX × SJ-2) (1 heterozygous: 1 homozygous). The four AFLP markers, GHACC1 plus two SSR markers (CIR316 and BNL1231) linked to rkn1 from previous work were mapped to intervals of 2.6–14.2 cM from the rkn1 locus, and the genomic region around rkn1 was spanned to about 28.2 cM in the F_2:7 population. The PCR-based GHACC1 and CIR316 markers were tested on 21 nematode resistant and susceptible cotton breeding lines and cultivars. GHACC1 was suitable for nematode resistance screening within G.␣hirsutum, but not G. barbadense, whereas CIR316 was useful in both species, indicating their␣potential for utilization in marker-assisted selection.     

Screening and identification of resistance to bacterial soft rot in Brassica rapa

Mist-chamber, field, and detached leaf inoculation procedures identified plants resistant to bacterial soft rot [Erwinia carotovora subsp. carotovora, (Ecc)] in Brassica rapa and related species. The mist-chamber seedling inoculation provided the best correlation of mean disease severity ratings with the field plant inoculation (r = 0.67^**) and was used to identify resistant materials. The optimum mist-chamber incubation conditions to distinguish the resistance of accessions were 23 ^°C and 100% RH and were used for primary screening. A total of 752 accessions of B. rapa and related Cruciferae were screened. In general, accessions of B. oleracea were more resistant than accessions of B. rapa. Within B. rapa, subspecies pekinensis and chinensis were more susceptible than other subspecies. No completely resistant material was found. In species of B. rapa, only 7% of accessions showed some degree of resistance with plant-to-plant variation within the accessions. G30444, G30449, and AVRDC2837 were identified as the most resistant materials in B. rapa by both mist-chamber and field inoculations. The resistance was correlated between an USA isolate (Geneva-1) and two Chinese isolates (RL4-1 and RL-19) of Ecc. This revised version was published online in July 2006 with corrections to the Cover Date.     

PCR-RFLP analysis of the whole chloroplast DNA from three cultivated species of cotton (Gossypium L.)

Variations of the chloroplast DNA (cpDNA) from three of the four cultivated species of cotton (Malvaceae); Gossypium barbadense L., Gossypium hirsutum L., Gossypium arboreum L. and its synonym Gossypium nanking Meyen., were analyzed. Using specific set of primers, the whole circular cpDNAs from the four test species were amplified. These were subsequently digested with the use of seven restriction enzymes. The amplified fragments of the whole cpDNAs of the diploid cultivated cotton G. arboreum and its synonym G. nanking did not show any differences. However, the allotetraploid cultivated cottons G. barbadense and G. hirsutum, showed some fragment length differences directly visible after amplification and two types of restriction fragment length polymorphism (RFLP), the first appeared as slightly lengthened bands and the other as gain or loss of a restriction site. The results also showed that the chloroplast genomes of the allotetraploid cultivated cottons are highly similar to the diploid cultivated cottons tested in terms of length and digestion patterns. The detected amplified length differences, RFLPs and the restriction sites can be considered as species specific markers for the allotetraploid cultivated cottons, which could be a useful tool for future studies of the cpDNA of the genus Gossypium L.     

Resistance in Lycopersicon species to black leaf mold caused by Pseudocercospora Fuligena

Over 540 accessions of wild Lycopersicon species or their crosses with L. esculentum were screened for resistance in a series of trials. Forty-six accessions were selected for the final screening trial based on lower disease ratings in previous trials. Of these, L. hirsutum had the greatest number of resistant accessions, followed by L. esculentum and L. peruvianum. Twenty accessions were quantified for their levels of resistance based on leaf area infected, area under disease progress curve (AUDPC), and the degree of sporulation. There was a significant positive correlation between the AUDPC calculated from 20 accessions evaluated under growth room and field conditions. Five L. hirsutum accessions had no sporulation associated with leaf lesions, whereas L. esculentum accessions had an average of 1.6×10⁴ conidia/cm² of leaf tissue. There was significant positive correlation between the AUDPC values and the number of conidia per cm² of leaf tissue.     

Quantitative trait locus mapping for Verticillium wilt resistance in a backcross inbred line population of cotton (Gossypium hirsutum × Gossypium barbadense) based on RGA-AFLP analysis

Verticillium wilt (VW), caused by Verticillium dahliae Kleb., is one of the most important diseases in cotton. The objective of this study was to map quantitative trait loci (QTLs) conferring VW resistance using resistance gene analog (RGA)-targeted amplified fragment length polymorphism (RGA-AFLP) markers in an interspecific backcross inbred line mapping population, consisting of 146 lines from a susceptible Sure-Grow 747 (Gossypium hirsutum L.) × resistant Pima S-7 (G. barbadense L.) cross. VW resistance was evaluated in replicated tests based on disease incidence in the field, and disease incidence and severity in the greenhouse. Of 160 polymorphic RGA-AFLP markers, 42 were significantly correlated with one or more VW traits and 41 were placed on a linkage map which covered 1,226 cM of the cotton genome and contained 251 other molecular markers. Three QTLs for VW resistance were detected, each of which explained 12.0–18.6 % of the phenotypic variation. Two of these QTLs for disease incidence and severity detected in the greenhouse inoculation tests using root wounding are located on chromosome c4. Both are closely linked to four RGA-AFLP markers and therefore considered as the same QTL for VW resistance. The other QTL detected in the field test was located on c19 and flanked by several RGA-AFLP markers. The desirable QTL allele on c4 for VW resistance detected in the greenhouse was from the VW susceptible Upland parent and absent from the resistant Pima parent which was more VW susceptible due to the disarmament of the first line of defense mechanism due to root wounding during inoculation. The other desirable VW resistance QTL allele, on c19, was from the resistant parent Pima S-7, consistent with the fact that Pima cotton was more resistant to VW when naturally infected in the field. The results should facilitate the development of more sequence specific markers and the transfer of VW resistance from Pima to Upland cotton through marker-assisted selection.     

The exploitation of genetic resources of Brassica oleracea in breeding for resistance to clubroot (Plasmodiophora brassicae)

Summary About 1000 Brassica oleracea accessions were evaluated in glasshouse tests for response to Plasmodiophora brassicae (clubroot). Resistance was confirmed in some north and west European kales and cabbage. A new source of resistance in cabbage, from Eire, is reported. Most other accessions were highly susceptible but lower levels of susceptibility were observed in open pollinated Brussels sprouts and forms of south European cabbage, cauliflower and broccoli. Modern breeding (as in the production of hybrid cultivars) appears to have resulted in increased susceptibility in several crop types. The implications of these results for the exploitation of germplasm are discussed.     

Zingiberene-mediated resistance to the South American tomato pinworm derived from Lycopersicon hirsutum var. hirsutum

The Lycopersicon hirsutum var. hirsutum accession PI 127826 is recognized as a good source of resistance to arthropod pests due to the action of the allelochemical zimgiberene, a sesquiterpene present in its glandular trichomes. Five genotypes were selected from the F₂ generation of the interspecific cross Lycopersicon esculentum ‘TOM-556’ × Lycopersicon hirsutum var. hirsutum ‘PI 127826’, based on their low levels (BPX-368-clone#56) or high levels(BPX-368-clone#92, BPX-368-clone#105,BPX-368-clone#179, BPX-368-clone#250) of zingiberene. The five F₂ genotypes were tested for resistance to the South American tomato pinworm Tuta absolutaalong with accession L. esculentum ‘TOM-556’ (pinworm susceptible), and the accessions L. hirsutum var. hirsutum ‘PI 127826’ and L. pennellii ‘LA716’ (resistant). The F₂ clones selected for high foliar zingiberene levels showed lower scores for leaflet lesion type(LLT), percent leaflets attacked (PLA) and overall plant damage (OPD) than the low zingiberene genotypes. The results indicated that zingiberene mediates resistance to the South American pinworm, based on feeding and on ovipositing deterrence, in populations derived from the interspecific cross between Lycopersicon. esculentum and Lycopersicon hirsutum var. hirsutum. Indirect selection for high foliar zingiberene content is suggested as an efficient technique for breeding tomatoes for resistance to the South American tomato pinworm. This revised version was published online in July 2006 with corrections to the Cover Date.     

Sources of resistance to Verticillium wilt in Capsicum

Summary A total of 125 novel accessions of Capsicum annuum L. and Capsicum baccatum L. were screened for sources of resistance to Verticillium wilt (Verticillium dahliae Kleb). A soil infestation method with 2000 microsclerotia of V. dahliae per gram of planting medium, and a soil temperature of 25±3°C, identified 27 Capsicum accessions with resistance to Verticillium wilt. P.I. 215699 (a mixture of Capsicum baccatum var. microcarpum and Capsicum annuum), P.I. 535616 (Capsicum annuum), and P.I. 555614 (Capsicum annuum), had the lowest disease severity and the highest percentage of resistant plants.     

Germplasm evaluation and transfer of Verticillium wilt resistance from Pima (<Emphasis Type="Italic">Gossypium barbadense</Emphasis>) to Upland cotton (<Emphasis Type="Italic">G. hirsutum</Emphasis>)

Verticillium wilt (VW, Verticillium dahliae) is a worldwide destructive soil-borne fungal disease and employment of VW resistant cultivars is the most economic and efficient method in sustainable cotton production. However, information concerning VW resistance in current commercial cotton cultivars and transfer of VW resistance from Pima (Gossypium barbadense) to Upland (Gossypium hirsutum) cotton is lacking. The objective of the current study was to report findings in evaluating commercial cotton cultivars and germplasm lines for VW resistance in field and greenhouse (GH) experiments conducted in 2003, 2006, and 2007. In the study, 267 cultivars and germplasm lines were screened in the GH, while 357 genotypes were screened in the field. The results indicated that (1) VW significantly reduced cotton yield, lint percentage, 50% span length and micronaire, but not 2.5% span length and fiber strength, when healthy and diseased plants in 23 cultivars were compared; (2) some commercial cotton cultivars developed by major cotton seed companies in the US displayed good VW resistance; (3) many Acala cotton cultivars released in the past also had good VW resistance, but not all Acala cotton germplasm are resistant; (4) Pima cotton possessed higher levels of VW resistance than Upland cotton, but the performance was reversed when the root system was wounded after inoculation; (5) VW resistance in some conventional cultivars was transferred into their transgenic version through backcrossing; and (6) some advanced backcross inbred lines developed from a cross between Upland and Pima cotton showed good VW resistance. The successful development of VW resistant transgenic cultivars and transfer of VW resistance from Pima to Upland cotton implies that VW resistance is associated with a few genes if not a major one.     

半野生棉抗黄萎病筛选及根系分泌物对黄萎病菌的影响

 对陆地棉7个野生种系的220份材料进行黄萎病菌的抗性筛选,用8份不同抗性材料进行了根系分泌物分析。从220份材料中筛选出18份抗黄萎病材料。根系分泌物研究表明,抗病品种根系分泌物对黄萎病菌菌丝生长及孢子萌发均具有抑制作用,而感病品种根系分泌物对其则有促进作用。另外通过对根系分泌物中氨基酸和可溶性糖的分析发现,抗病品种根系分泌物中氨基酸种类少于感病品种,并且抗病品种根系分泌物中氨基酸总量和可溶性糖含量显著少于感病品种。同时发现,精氨酸为抗病品种特有氨基酸。