Analysis of root-knot nematode and fusarium wilt disease resistance in cotton (Gossypium spp.) using chromosome substitution lines from two alien species.

Chromosome substitution (CS) lines in plants are a powerful genetic resource for analyzing the contribution of chromosome segments to phenotypic variance. In this study, a series of interspecific cotton (Gossypium spp.) CS lines were used to identify a new germplasm resource, and to validate chromosomal regions and favorable alleles associated with nematode or fungal disease resistance traits. The CS lines were developed in the G. hirsutum L. TM-1 background with chromosome or chromosome segment substitutions from G. barbadense L. Pima 3-79 or G. tomentosum. Root-knot nematode (Meloidogyne incognita) and fusarium wilt (Fusarium oxysporum f. sp. vasinfectum) (races 1 and 4) resistance alleles and quantitative trait loci (QTL) previously placed on cotton chromosomes using SSR markers in two interspecific recombinant inbred line populations were chosen for testing. Phenotypic responses of increased resistance or susceptibility in controlled inoculation and infested field assays confirmed the resistance QTLs, based on substitution with the positive or negative allele for resistance. Lines CS-B22Lo, CS-B04, and CS-B18 showed high resistance to nematode root-galling, confirming QTLs on chromosomes 4 and 22 (long arm) with resistance alleles from Pima 3-79. Line CS-B16 had less fusarium race 1-induced vascular root staining and higher percent survival than the TM-1 parent, confirming a major resistance QTL on chromosome 16. Lines CS-B(17-11) and CS-B17 had high fusarium race 4 vascular symptoms and low survival due to susceptible alleles introgressed from Pima 3-79, confirming the localization on chromosome 17 of an identified QTL with resistance alleles from TM1 and other resistant lines. Analyses validated regions on chromosomes 11, 16, and 17 harboring nematode and fusarium wilt resistance genes and demonstrated the value of CS lines as both a germplasm resource for breeding programs and as a powerful genetic analysis tool for determining QTL effects for disease resistance. CS lines carrying small alien chromosome segments with favorable QTL alleles could be used for effective introgression of biotic stress resistance or many other desirable traits by targeting gene interactions and reducing linkage drag effects.

Genome wide linkage disequilibrium in Chinese asparagus bean (Vigna. unguiculata ssp. sesquipedialis) germplasm: implications for domestication history and genome wide association studies.

Association mapping of important traits of crop plants relies on first understanding the extent and patterns of linkage disequilibrium (LD) in the particular germplasm being investigated. We characterize here the genetic diversity, population structure and genome wide LD patterns in a set of asparagus bean (Vigna. unguiculata ssp. sesquipedialis) germplasm from China. A diverse collection of 99 asparagus bean and normal cowpea accessions were genotyped with 1127 expressed sequence tag-derived single nucleotide polymorphism markers (SNPs). The proportion of polymorphic SNPs across the collection was relatively low (39%), with an average number of SNPs per locus of 1.33. Bayesian population structure analysis indicated two subdivisions within the collection sampled that generally represented the 'standard vegetable' type (subgroup SV) and the 'non-standard vegetable' type (subgroup NSV), respectively. Level of LD (r(2)) was higher and extent of LD persisted longer in subgroup SV than in subgroup NSV, whereas LD decayed rapidly (0-2 cM) in both subgroups. LD decay distance varied among chromosomes, with the longest (≈ 5 cM) five times longer than the shortest (≈ 1 cM). Partitioning of LD variance into within- and between-subgroup components coupled with comparative LD decay analysis suggested that linkage group 5, 7 and 10 may have undergone the most intensive epistatic selection toward traits favorable for vegetable use. This work provides a first population genetic insight into domestication history of asparagus bean and demonstrates the feasibility of mapping complex traits by genome wide association study in asparagus bean using a currently available cowpea SNPs marker platform.

Distribution and Prevalence of Parasitic Nematodes of Cowpea (Vigna unguiculata) in Burkina Faso.

A comprehensive survey of the plant parasitic nematodes associated with cowpea (Vigna unguiculata) production fields was carried out in the three primary agro-climatic zones of Burkina Faso in West Africa. Across the three zones, a total of 109 samples were collected from the farms of 32 villages to provide a representative coverage of the cowpea production areas. Samples of rhizosphere soil and samples of roots from actively growing cowpea plants were collected during mid- to late-season. Twelve plant-parasitic nematode genera were identified, of which six appeared to have significant parasitic potential on cowpea based on their frequency and abundance. These included Helicotylenchus, Meloidogyne, Pratylenchus, Scutellonema, Telotylenchus, and Tylenchorhynchus. Criconemella and Rotylenchulus also had significant levels of abundance and frequency, respectively. Of the primary genera, Meloidogyne, Pratylenchus, and Scutellonema contained species which are known or suspected to cause losses of cowpea yield in other parts of the world. According to the prevalence and distribution of these genera in Burkina Faso, their potential for damage to cowpea increased from the dry Sahelian semi-desert zone in the north (annual rainfall < 600 mm/year), through the north-central Soudanian zone (annual rainfall of 600-800 mm/year), to the wet Soudanian zone (annual rainfall ≥ 1000 mm) in the more humid south-western region of the country. This distribution trend was particularly apparent for the endoparasitic nematode Meloidogyne and the migratory endoparasite Pratylenchus.

The future of nematode management in cotton.

The importance of plant-parasitic nematodes as yield-limiting pathogens of cotton has received increased recognition and attention in the United States in the recent past. This paper summarizes the remarks made during a symposium of the same title that was held in July 2007 at the joint meeting of the Society of Nematologists and the American Phytopathological Society in San Diego, California. Although several cultural practices, including crop rotation, can be effective in suppressing the populations of the important nematode pathogens of cotton, the economic realities of cotton production limit their use. The use of nematicides is also limited by issues of efficacy and economics. There is a need for development of chemistries that will address these limitations. Also needed are systems that would enable precise nematicide application in terms of rate and placement only in areas where nematode population densities warrant application. Substantial progress is being made in the identification, characterization and mapping of loci for resistance to Meloidogyne incognita and Rotylenchulus reniformis. These data will lead to efficient marker-assisted selection systems that will likely result in development and release of nematode-resistant cotton cultivars with superior yield potential and high fiber quality.

A Fusarium Wilt Resistance Gene in Gossypium barbadense and Its Effect on Root-Knot Nematode-Wilt Disease Complex.

ABSTRACT Fusarium wilt, caused by the soilborne pathogen Fusarium oxysporum f. sp. vasinfectum race 1, is a vascular disease in cotton (Gossypium spp.), and is a component of a disease complex with root-knot nematodes (Meloidogyne incognita). Genetic analysis of two interspecific crosses (G. barbadense Pima S-7 x G. hirsutum Acala NemX and Pima S-7 x Acala SJ-2) showed that one major gene (designated Fov1) with allele dosage effect conferred resistance to F. oxysporum f. sp. vasinfectum race 1 in Pima S-7. Two amplified fragment length polymorphism (AFLP) markers were linked to Fov1 in Pima S-7, with genetic distance from the gene of 9.3 and 14.6 centimorgans. Less severe wilt symptoms in Acala NemX than Acala SJ-2 indicated that Acala NemX possesses one or more minor genes contributing to delay of wilt symptoms. Highly resistant plants in F(2) and F(3) (Pima S-7 x NemX) families indicated transgressive segregation effects of minor genes in Acala NemX combined with Fov1 from Pima S-7. The effects of wilt and nematode resistance on the nematode-wilt disease complex were assayed with two inoculation methods. In the presence of both pathogens, wilt damage measured as shoot and root weight reductions was greatest on wilt- and nematode-susceptible Acala SJ-2 and least in root-knot nematode-resistant and wilt-susceptible Acala NemX. Intermediate damage occurred in wilt-resistant and root-knot nematode-susceptible Pima S-7. The results indicated that nematode resistance was more effective than wilt resistance in suppressing wilt symptoms when either resistance was present alone. Nematode resistance combined with intermediate wilt resistance, as in the F(1) (Pima S-7 x NemX), was highly effective in protecting plants from root-knot nematodes and race 1 of Fusarium wilt as a disease complex.

Dynamics of Meloidogyne incognita Virulence to Resistance Genes Rk and Rk in Cowpea.

The virulence index of three Meloidogyne incognita field isolates to the resistance gene Rk in cowpea was 0%, 75%, and 120%, with the index measured as reproduction on resistant plants as a percentage of the reproduction on susceptible plants. Continuous culture of the 75% virulent isolate on susceptible tomato for more than 5 years (about 25 generations) resulted in virulence decline to about 4%. The rate of the decline in virulence was described by exponential decay, indicating the progressive loss of virulence on a susceptible host. The 120% virulent isolate declined to 90% virulence during five generations on susceptible cowpea. Following virulence decline, the two isolates were compared over 5 years in inoculated field microplots both separately and as a mixture on susceptible, gene Rk, and gene Rk(2) cowpea plants. At infestation of the plots, the two isolates were 1.2% and 92.0% virulent, respectively, to gene Rk and 0.2% and 8.1% virulent, respectively, to gene Rk(2). Virulence to gene Rk in the two isolates and in mixture increased under 5 years of continuous Rk cowpea plants to 129% to 172% and under Rk(2) cowpea plants to 113% to 139 % by year 5. Virulence to gene Rk(2) increased during continuous cropping with Rk cowpea plants to 42% to 47% and with Rk(2) cowpea plants to 22% to 48% by year 5. Selection of Rk(2)-virulence was slower in the isolate with low itt-virulence. The virulence to both genes Rk and Rk(2) in the mixed population was not different from that in the highly virulent isolate by year 5 of all cropping combinations. Selection of Rk(2)-virulence on plants with Rk, and vice versa, indicated at least partial overlap of gene specificity between Rk and Rk(2) with respect to selection of nematode virulence. This observation should be considered when resistance is used in cowpea rotations.

Phenotypic Expression of rkn1-Mediated Meloidogyne incognita Resistance in Gossypium hirsutum Populations.

The root-knot nematode Meloidogyne incognita is a damaging pest of cotton (Gossypium hirsutum) worldwide. A major gene (rkn1) conferring resistance to M. incognita was previously identified on linkage group A03 in G. hirsutum cv. Acala NemX. To determine the patterns of segregation and phenotypic expression of rkn1, F(1), F(2), F(2:3), BC(1)F(1) and F(2:7) recombinant inbred lines (RIL) from intraspecific crosses between Acala NemX and a closely related susceptible cultivar Acala SJ-2 were inoculated in greenhouse tests with M. incognita race 3. The resistance phenotype was determined by the extent of nematode-induced root galling and nematode egg production on roots. Suppression of root galling and egg production was highly correlated among individuals in all tests. Root galling and egg production on heterozygous plants did not differ from the susceptible parent phenotype 125 d or more after inoculation, but were slightly suppressed with shorter screening (60 d), indicating that rkn1 behaved as a recessive gene or an incompletely recessive gene, depending on the screening condition. In the RIL, rkn1 segregated in an expected 1 resistant: 1 susceptible ratio for a major resistance gene. However, within the resistant class, 21 out of 34 RIL were more resistant than the resistant parent Acala NemX, indicating transgressive segregation. These results suggest that rkn1-based resistance in G. hirsutum can be enhanced in progenies of crosses with susceptible genotypes. Allelism tests and molecular genetic analysis are needed to determine the relationship of rkn1 to other M. incognita resistance sources in cotton.

In support of the telomere concept.

The frequency of recovered X-ray-induced (4000R) rearrangements that, in all probability mimic terminal deletions of the X chromosome was only one of, roughly, 10-5 X chromosomes screened for tip deficiencies. Although the single exception looks terminally deleted, it is probably capped by a very short or nonpolytene telomeric segment. It is apparent from these data that the probability of "healing" or stabilization of a terminally deleted X in the zygotic nucleus or developing embryo of Drosophila melanogaster is vanishingly small. The telomeric caps in two obviously interstitial deficiencies that were recovered represent, roughly, 1/500 of the length of a mitotic chromosome. These findings give some indication of the extreme difficulty of detecting short telomeric segments capping either deleted polytene chromosomes or deleted metaphase chromosomes of, for example, humans.

Rapid change of chromomeric and pairing patterns of polytene chromosome tips in D. melanogaster: migration of polytene-nonpolytene transition zone?

The high variability of chromomeric patterns in near-terminal regions of polytene chromosome arms has been explored in a number of races, strains and hybrids of Drosophila melanogaster. Traditional explanations for tip differences between strains (differential compaction of chromatin, somatic or germinal deletion) are examined and, in the light of the reported observations, rejected. The range of polytene tip variability and rates of change in wild races are greater than has been supposed: strains formerly considered to be terminally deleted appear to gain terminal bands; others, formerly considered normal, appear to have lost them. Strains with high cell-to-cell tip variability are also described. Cell-to-cell variations, as well as much of the observed rapid changes in tip appearance, are probably due to heritable differences in the location of an abrupt transition zone between polytene and nonpolytene chromatin. A quantitative relationship between the amount of certain subterminal bands present and the frequency of tip association of nonhomologous chromosomes is shown and its possible significance for chromosome is shown and its possible for chromosome pairing discussed.

A possible case of position effect of DNA replication in Drosophila melanogaster.

The behavior of T(3;4)10 provides evidence that the centromere of 4 is not in the doublet 101D3,4, and that most of what has been called the left arm of 4 should be called basal 4R. Basal 4R usually does not replicate as much as the tip of 4R when 4 is in its normal position in the chromocenter. Translocated to the tip of 3R, however, basal 4R attains a width equal to that of the rest of the arm-a possible position effect on DNA replication.

Localization of minutes to specific polytene chromosome bands by means of overlapping duplications.

Overlapping duplications recovered as suppressors of Minute loci have been used to localize M(2)z and M(3)w(124) to specific polytene bands 25A1(2) and 95A1(2). The surprising efficiency of M localization by duplication may result from the tendency of M suppressors to be at least a visible fraction of a polytene band in length.

Properties and evolutionary potential of newly induced tandem duplications in Drosophila melanogaster.

Most of some 33 X-ray-induced duplications recovered as Suppressors of Minute loci proved to be direct tandem duplications. When heterozygous, most duplications were crossover suppressors, and duplications of short to moderate size did not reduce the fitness of their bearers. Crossover suppression by tandem duplication may be attributed to intrastrand foldbacks of the type regularly seen in somatic polytene chromosomes. As a consequence, linkage disequilibrium between duplicated elements and normal chromosomes should be more profound than has been supposed. Tandem duplications appear to be predisposed by reason of frequency of generation, crossover suppression and fitness effects to serve as the primary source of new genes.

An unusual Y chromosome of Drosophila simulans carrying amplified rDNA spacer without rRNA genes.

The X and Y chromosomes of Drosophila melanogaster each contain a cluster of several hundred ribosomal RNA genes (rDNA). A nontranscribed spacer region separates adjacent rRNA genes and contains tandem copies of 240 bp repeats that include the initiation site for RNA polymerase I transcription. We show here that Drosophila simulans, a sibling species of D. melanogaster, contains few, if any, rRNA genes on its Y chromosome but carries instead a large block (3,000 kb or 12,500 copies) of 240 bp nontranscribed spacer repeats. The repeats are located at the tip of the long arm of the simulans Y chromosome, in contrast to their location among rRNA genes on the short arm of the Y chromosome of D. melanogaster. The bobbed mutation in homozygous females of D. melanogaster shortens and thins the bristles, owing to a partial deletion of rRNA genes on the X chromosome. The bristles of bobbed/Y males are normal owing to the presence of a full complement of rRNA genes on the Y chromosome. Peculiarly, in bobbed/Y males of D. simulans the short bristle phenotype does not return to normal but is enhanced by the presence of the Y chromosome. We propose that the 12,500 nontranscribed spacer repeats on the Y chromosome are responsible for this biological effect by competition for a protein factor(s) essential for normal levels of rDNA transcription at the X-linked locus.

Mapping simple repeated DNA sequences in heterochromatin of Drosophila melanogaster.

Heterochromatin in Drosophila has unusual genetic, cytological and molecular properties. Highly repeated DNA sequences (satellites) are the principal component of heterochromatin. Using probes from cloned satellites, we have constructed a chromosome map of 10 highly repeated, simple DNA sequences in heterochromatin of mitotic chromosomes of Drosophila melanogaster. Despite extensive sequence homology among some satellites, chromosomal locations could be distinguished by stringent in situ hybridizations for each satellite. Only two of the localizations previously determined using gradient-purified bulk satellite probes are correct. Eight new satellite localizations are presented, providing a megabase-level chromosome map of one-quarter of the genome. Five major satellites each exhibit a multi-chromosome distribution, and five minor satellites hybridize to single sites on the Y chromosome. Satellites closely related in sequence are often located near one another on the same chromosome. About 80% of Y chromosome DNA is composed of nine simple repeated sequences, in particular (AAGAC)n (8 Mb), (AAGAG)n (7 Mb) and (AATAT)n (6 Mb). Similarly, more than 70% of the DNA in chromosome 2 heterochromatin is composed of five simple repeated sequences. We have also generated a high resolution map of satellites in chromosome 2 heterochromatin, using a series of translocation chromosomes whose breakpoints in heterochromatin were ordered by N-banding. Finally, staining and banding patterns of heterochromatic regions are correlated with the locations of specific repeated DNA sequences. The basis for the cytochemical heterogeneity in banding appears to depend exclusively on the different satellite DNAs present in heterochromatin.

Isofemale Line Analysis of Meloidogyne incognita Virulence to Cowpea Resistance Gene Rk.

Isofemale lines (IFL) from single egg masses were studied for genetic variation in Meloidogyne incognita isolates avirulent and virulent to the resistance gene Rk in cowpea (Vigna unguiculata). In parental isolates cultured on susceptible and resistant cowpea, the virulent isolate contained 100% and the avirulent isolate 7% virulent lineages. Virulence was selected from the avirulent isolate within eight generations on resistant cowpea (lineage selection). In addition, virulence was selected from avirulent females (individual selection). Virulence differed (P

Fitness of Virulent Meloidogyne incognita Isolates on Susceptible and Resistant Cowpea.

A study of life-history traits was made to determine factors associated with the fitness of Meloidogyne incognita isolates virulent to resistance gene Rk in cowpea. Egg hatch, root penetration, egg mass production, and fecundity (eggs per egg mass) of avirulent and virulent phenotypes were compared among M. incognita isolates, isofemale lines, and single descent lines over multiple generations on resistant and susceptible cowpea. Variation (P

Can mutagenesis reveal major genes affecting senescence?

The Drosophila melanogaster genome was screened for sex-linked recessive and autosomal dominant mutations affecting the senescent decline in motor abilities. No persistent identifiable genes delaying the onset of senescence were recovered among 18,089 F1 male offspring of males exposed to radiation or the chemical mutagen, EMS.

The consequences of fat body transplantation into young and old Drosophila.

Despite some age-related changes in cells, we observed no massive fat body involution during senescence of two species of Drosophila. Transplants of larval fat failed to prolong the life of old D. gibberosa. Transplanted larval fat persisted for weeks and underwent similar changes (except for glycogen stores) in old as in young hosts, suggesting that only minor changes in hemolymph composition occur with age.

Cardiovascular effects of microinjection of adenosine into the nucleus tractus solitarius.

Rats were anesthetized with urethane and limited occipital craniotomy was conducted to expose the caudal medulla in the region of the obex. Microinjections of adenosine were made into the nucleus tractus solitarius and heart rate and blood pressure responses recorded. Adenosine produced dose-related decreases in blood pressure and heart rate. The data indicate that adenosine may play a neuromodulatory role in central cardiovascular control areas.

Blood-brain barrier: a definition of normal and altered function.

The anatomical component of the blood-brain barrier (BBB) has been shown to be the cerebral capillary. These capillary endothelial cells are connected by continuous tight intercellular junctions and under normal conditions do not demonstrate transendothelial channels or pinocytotic vesicles. The rate that substances penetrate the BBB is related to molecular size, lipid solubility, and the presence of a specific carrier-mediated transport system. This latter mechanism for transendothelial passage is utilized for the movement of a wide variety of biologically important compounds such as sugars, amino acids, and organic acids. In certain pathological conditions, the permeability of the BBB is altered so that normally excluded plasma proteins and fluid enter the brain extracellular space, with the subsequent development of cerebral edema. In other abnormal conditions, alterations in the specialized transport systems operating across the cerebral capillary result in adverse changes in cerebral and neurotransmitter metabolism. An understanding of the unique properties of the BBB and of the changes that occur in pathological conditions has allowed the development of rational therapeutic strategies for a wide variety of diseases of the central nervous system.